PMID- 10984461 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Opportunities at the intersection of bioinformatics and health informatics: a case study. PG - 431-8 AB - This paper provides a "viewpoint discussion" based on a presentation made to the 2000 Symposium of the American College of Medical Informatics. It discusses potential opportunities for researchers in health informatics to become involved in the rapidly growing field of bioinformatics, using the activities of the Yale Center for Medical Informatics as a case study. One set of opportunities occurs where bioinformatics research itself intersects with the clinical world. Examples include the correlations between individual genetic variation with clinical risk factors, disease presentation, and differential response to treatment; and the implications of including genetic test results in the patient record, which raises clinical decision support issues as well as legal and ethical issues. A second set of opportunities occurs where bioinformatics research can benefit from the technologic expertise and approaches that informaticians have used extensively in the clinical arena. Examples include database organization and knowledge representation, data mining, and modeling and simulation. Microarray technology is discussed as a specific potential area for collaboration. Related questions concern how best to establish collaborations with bioscientists so that the interests and needs of both sets of researchers can be met in a synergistic fashion, and the most appropriate home for bioinformatics in an academic medical center. AD - Yale University, New Haven, Connecticut, USA. FAU - Miller, P L AU - Miller PL LA - eng GR - G08-LM-05583/LM/NLM PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM CIN - J Am Med Inform Assoc. 2000 Sep-Oct;7(5):512-6. PMID: 10984470 MH - *Computational Biology/organization & administration MH - Cooperative Behavior MH - Decision Support Systems, Clinical MH - Gene Expression MH - Genome MH - *Medical Informatics/organization & administration MH - Neurosciences MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):431-8. PMID- 10984462 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - The interactions between clinical informatics and bioinformatics: a case study. PG - 439-43 AB - For the past decade, Stanford Medical Informatics has combined clinical informatics and bioinformatics research and training in an explicit way. The interest in applying informatics techniques to both clinical problems and problems in basic science can be traced to the Dendral project in the 1960s. Having bioinformatics and clinical informatics in the same academic unit is still somewhat unusual and can lead to clashes of clinical and basic science cultures. Nevertheless, the benefits of this organization have recently become clear, as the landscape of academic medicine in the next decades has begun to emerge. The author provides examples of technology transfer between clinical informatics and bioinformatics that illustrate how they complement each other. AD - Stanford University, Stanford, California 94305-5479, USA. russ.altman@stanford.edu FAU - Altman, R B AU - Altman RB LA - eng GR - GM-61374/GM/NIGMS GR - LM-05652/LM/NLM GR - LM-06422/LM/NLM PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM CIN - J Am Med Inform Assoc. 2000 Sep-Oct;7(5):512-6. PMID: 10984470 MH - Academic Medical Centers/organization & administration MH - California MH - *Computational Biology/education/methods/organization & administration MH - Cooperative Behavior MH - *Medical Informatics/organization & administration MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Research Support, U.S. Gov't, P.H.S. MH - *Technology Transfer EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):439-43. PMID- 10984463 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20001218 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Electronic health record meets digital library: a new environment for achieving an old goal. PG - 444-52 AB - Linking the electronic health record to the digital library is a Web-era reformulation of the long-standing informatics goal of seamless integration of automated clinical data and relevant knowledge-based information to support informed decisions. The spread of the Internet, the development of the World Wide Web, and converging format standards for electronic health data and digital publications make effective linking increasingly feasible. Some existing systems link electronic health data and knowledge-based information in limited settings or limited ways. Yet many challenging informatics research problems remain to be solved before flexible and seamless linking becomes a reality and before systems become capable of delivering the specific piece of information needed at the time and place a decision must be made. Connecting the electronic health record to the digital library also requires positive resolution of important policy issues, including health data privacy, government encouragement of high-speed communications, electronic intellectual property rights, and standards for health data and for digital libraries. Both the research problems and the policy issues should be important priorities for the field of medical informatics. AD - Library of Medicine, Bethesda, Maryland 20894, USA. blh@nlm.nih.gov FAU - Humphreys, B L AU - Humphreys BL LA - eng PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - *Internet/standards MH - Libraries/*organization & administration/standards MH - Medical Records Systems, Computerized/*organization & administration MH - Publishing/*organization & administration/standards MH - Systems Integration EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):444-52. PMID- 10984464 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20001218 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Improving clinical communication: a view from psychology. PG - 453-61 AB - Recent research has studied the communication behaviors of clinical hospital workers and observed a tendency for these workers to use communication behaviors that were often inefficient. Workers were observed to favor synchronous forms of communication, such as telephone calls and chance face-to-face meetings with colleagues, even when these channels were not effective. Synchronous communication also contributes to a highly interruptive working environment, increasing the potential for clinical errors to be made. This paper reviews these findings from a cognitive psychological perspective, focusing on current understandings of how human memory functions and on the potential consequences of interruptions on the ability to work effectively. It concludes by discussing possible communication technology interventions that could be introduced to improve the clinical communication environment and suggests directions for future research. AD - Hewlett-Packard Research Laboratories, Bristol, England (JSP). jsp@hplb.hpl.hp.com FAU - Parker, J AU - Parker J FAU - Coiera, E AU - Coiera E LA - eng PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - Attitude of Health Personnel MH - Cognitive Science MH - *Communication MH - Memory MH - Personnel, Hospital/*psychology EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):453-61. PMID- 10984465 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Comparative evaluation of three continuous speech recognition software packages in the generation of medical reports. PG - 462-8 AB - OBJECTIVE: To compare out-of-box performance of three commercially available continuous speech recognition software packages: IBM ViaVoice 98 with General Medicine Vocabulary; Dragon Systems NaturallySpeaking Medical Suite, version 3.0; and L&H Voice Xpress for Medicine, General Medicine Edition, version 1.2. DESIGN: Twelve physicians completed minimal training with each software package and then dictated a medical progress note and discharge summary drawn from actual records. MEASUREMENTS: Errors in recognition of medical vocabulary, medical abbreviations, and general English vocabulary were compared across packages using a rigorous, standardized approach to scoring. RESULTS: The IBM software was found to have the lowest mean error rate for vocabulary recognition (7.0 to 9.1 percent) followed by the L&H software (13.4 to 15.1 percent) and then Dragon software (14.1 to 15.2 percent). The IBM software was found to perform better than both the Dragon and the L&H software in the recognition of general English vocabulary and medical abbreviations. CONCLUSION: This study is one of a few attempts at a robust evaluation of the performance of continuous speech recognition software. Results of this study suggest that with minimal training, the IBM software outperforms the other products in the domain of general medicine; however, results may vary with domain. Additional training is likely to improve the out-of-box performance of all three products. Although the IBM software was found to have the lowest overall error rate, successive generations of speech recognition software are likely to surpass the accuracy rates found in this investigation. AD - Boston Veterans Administration Medical Center, Boston, Massachusetts 02130, USA. devine.eric@boston.va.gov FAU - Devine, E G AU - Devine EG FAU - Gaehde, S A AU - Gaehde SA FAU - Curtis, A C AU - Curtis AC LA - eng PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - Comparative Study MH - Evaluation Studies MH - Humans MH - *Medical Records Systems, Computerized MH - Research Support, U.S. Gov't, Non-P.H.S. MH - *Software MH - *Speech EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):462-8. PMID- 10984466 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Variables that may enhance medical students' perceived preparedness for computer-based testing. PG - 469-74 AB - OBJECTIVE: To identify variables that may enhance medical student's preparedness for computer-based administration of the United States Medical Licensing Examination (USMLE). DESIGN: A cross-sectional survey of 301 medical students who completed a self-administered questionnaire. MEASUREMENTS: The questionnaire was designed to obtain information about students' computer resources, personal experience with computers, computer expertise, opinions about computers, experience with computer-based testing, perceived preparedness for the computer-based USMLE, and demographic variables. Variables related to students' perceived preparedness for the computer-based USMLE were identified by ordinal logistic regression. RESULTS: A significant regression model yielded four significant predictors: perceived preparedness for USMLE content (P: < 0.0001), opinions about computers (P: < 0.0012), gender (P: < 0.0001), and a gender by computer-based testing experience interaction (P: < 0. 0004). Computer resources, personal experience with computers, computer expertise, age, race, and year of medical school were not significant predictors. CONCLUSION: Students' perceived preparedness for computer-based administration of high-stakes examinations may be facilitated by preparing them for examination content, by enhancing their opinions about computers, and by increasing their computer-based testing experiences. AD - East Carolina University, Greenville, North Carolina, USA. dlynch@med-scape.com FAU - Lynch, D C AU - Lynch DC FAU - Whitley, T W AU - Whitley TW FAU - Emmerling, D A AU - Emmerling DA FAU - Brinn, J E AU - Brinn JE LA - eng PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - Attitude to Computers MH - *Computer Literacy MH - Computer User Training MH - Continental Population Groups MH - Cross-Sectional Studies MH - Educational Measurement/*methods MH - Female MH - Humans MH - Licensure, Medical MH - Logistic Models MH - Male MH - Questionnaires MH - Sex Factors MH - *Students, Medical/psychology MH - United States EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):469-74. PMID- 10984467 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Exploring performance issues for a clinical database organized using an entity-attribute-value representation. PG - 475-87 AB - BACKGROUND: The entity-attribute-value representation with classes and relationships (EAV/CR) provides a flexible and simple database schema to store heterogeneous biomedical data. In certain circumstances, however, the EAV/CR model is known to retrieve data less efficiently than conventionally based database schemas. OBJECTIVE: To perform a pilot study that systematically quantifies performance differences for database queries directed at real-world microbiology data modeled with EAV/CR and conventional representations, and to explore the relative merits of different EAV/CR query implementation strategies. METHODS: Clinical microbiology data obtained over a ten-year period were stored using both database models. Query execution times were compared for four clinically oriented attribute-centered and entity-centered queries operating under varying conditions of database size and system memory. The performance characteristics of three different EAV/CR query strategies were also examined. RESULTS: Performance was similar for entity-centered queries in the two database models. Performance in the EAV/CR model was approximately three to five times less efficient than its conventional counterpart for attribute-centered queries. The differences in query efficiency became slightly greater as database size increased, although they were reduced with the addition of system memory. The authors found that EAV/CR queries formulated using multiple, simple SQL statements executed in batch were more efficient than single, large SQL statements. CONCLUSION: This paper describes a pilot project to explore issues in and compare query performance for EAV/CR and conventional database representations. Although attribute-centered queries were less efficient in the EAV/CR model, these inefficiencies may be addressable, at least in part, by the use of more powerful hardware or more memory, or both. AD - Yale University, New Haven, Connecticut, USA. FAU - Chen, R S AU - Chen RS FAU - Nadkarni, P AU - Nadkarni P FAU - Marenco, L AU - Marenco L FAU - Levin, F AU - Levin F FAU - Erdos, J AU - Erdos J FAU - Miller, P L AU - Miller PL LA - eng GR - G08-LM05583/LM/NLM GR - T15-LM07056/LM/NLM GR - U01-CA78266/CA/NCI PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - Comparative Study MH - *Database Management Systems MH - Databases/organization & administration MH - *Databases, Factual MH - Information Storage and Retrieval/*methods MH - Microbiology MH - Pilot Projects MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):475-87. PMID- 10984468 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - GEM: a proposal for a more comprehensive guideline document model using XML. PG - 488-98 AB - OBJECTIVE: To develop a guideline document model that includes a sufficiently broad set of concepts to be useful throughout the guideline life cycle. DESIGN: Current guideline document models are limited in that they reflect the specific orientation of the stakeholder who created them; thus, developers and disseminators often provide few constructs for conceptualizing recommendations, while implementers de-emphasize concepts related to establishing guideline validity. The authors developed the Guideline Elements Model (GEM) using XML to better represent the heterogeneous knowledge contained in practice guidelines. Core constructs were derived from the Institute of Medicine's Guideline Appraisal Instrument, the National Guideline Clearinghouse, and the augmented decision table guideline representation. These were supplemented by additional concepts from a literature review. RESULTS: The GEM hierarchy includes more than 100 elements. Major concepts relate to a guideline's identity, developer, purpose, intended audience, method of development, target population, knowledge components, testing, and review plan. Knowledge components in guideline documents include recommendations (which in turn comprise conditionals and imperatives), definitions, and algorithms. CONCLUSION: GEM is more comprehensive than existing models and is expressively adequate to represent the heterogeneous information contained in guidelines. Use of XML contributes to a flexible, comprehensible, shareable, and reusable knowledge representation that is both readable by human beings and processible by computers. AD - Yale University, New Haven, Connecticut, USA. richard.shiffman@yale.edu FAU - Shiffman, R N AU - Shiffman RN FAU - Karras, B T AU - Karras BT FAU - Agrawal, A AU - Agrawal A FAU - Chen, R AU - Chen R FAU - Marenco, L AU - Marenco L FAU - Nath, S AU - Nath S LA - eng GR - 1-R29-LM-05552/LM/NLM GR - T15-LM07056/LM/NLM PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - Algorithms MH - *Hypermedia MH - Models, Theoretical MH - *Practice Guidelines MH - *Programming Languages MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):488-98. PMID- 10984469 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20001218 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Corpus-based statistical screening for phrase identification. PG - 499-511 AB - PURPOSE: The authors study the extraction of useful phrases from a natural language database by statistical methods. The aim is to leverage human effort by providing preprocessed phrase lists with a high percentage of useful material. METHOD: The approach is to develop six different scoring methods that are based on different aspects of phrase occurrence. The emphasis here is not on lexical information or syntactic structure but rather on the statistical properties of word pairs and triples that can be obtained from a large database. MEASUREMENTS: The Unified Medical Language System (UMLS) incorporates a large list of humanly acceptable phrases in the medical field as a part of its structure. The authors use this list of phrases as a gold standard for validating their methods. A good method is one that ranks the UMLS phrases high among all phrases studied. Measurements are 11-point average precision values and precision-recall curves based on the rankings. RESULT: The authors find of six different scoring methods that each proves effective in identifying UMLS quality phrases in a large subset of MEDLINE. These methods are applicable both to word pairs and word triples. All six methods are optimally combined to produce composite scoring methods that are more effective than any single method. The quality of the composite methods appears sufficient to support the automatic placement of hyperlinks in text at the site of highly ranked phrases. CONCLUSION: Statistical scoring methods provide a promising approach to the extraction of useful phrases from a natural language database for the purpose of indexing or providing hyperlinks in text. AD - National Library of Medicine, Bethesda, Maryland 20894, USA. wonkim@ncbi.nlm.nih.gov FAU - Kim, W AU - Kim W FAU - Wilbur, W J AU - Wilbur WJ LA - eng PT - Journal Article PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM MH - *Abstracting and Indexing MH - *Hypermedia MH - Information Storage and Retrieval/*methods MH - MEDLINE MH - *Natural Language Processing MH - Statistics MH - *Unified Medical Language System MH - Vocabulary, Controlled EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):499-511. PMID- 10984470 OWN - NLM STAT- MEDLINE DA - 20001016 DCOM- 20001016 LR - 20041117 PUBM- Print IS - 1067-5027 VI - 7 IP - 5 DP - 2000 Sep-Oct TI - Bioinformatics and clinical informatics: the imperative to collaborate. PG - 512-6 FAU - Kohane, I S AU - Kohane IS LA - eng PT - Comment PT - Editorial PL - UNITED STATES TA - J Am Med Inform Assoc JID - 9430800 SB - IM CON - J Am Med Inform Assoc. 2000 Sep-Oct;7(5):431-8. PMID: 10984461 CON - J Am Med Inform Assoc. 2000 Sep-Oct;7(5):439-43. PMID: 10984462 MH - *Computational Biology/organization & administration MH - Confidentiality MH - Cooperative Behavior MH - Costs and Cost Analysis MH - False Positive Reactions MH - Genome, Human MH - Humans MH - *Medical Informatics/organization & administration MH - Vocabulary, Controlled EDAT- 2000/09/14 11:00 MHDA- 2000/10/21 11:01 PST - ppublish SO - J Am Med Inform Assoc 2000 Sep-Oct;7(5):512-6. PMID- 11818032 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021125 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 TI - Kinetic comparison of tissue non-specific and placental human alkaline phosphatases expressed in baculovirus infected cells: application to screening for Down's syndrome. PG - 2 AB - BACKGROUND: In humans, there are four alkaline phosphatases, and each form exhibits a characteristic pattern of tissue distribution. The availability of an easy method to reveal their activity has resulted in large amount of data reporting correlations between variations in activity and illnesses. For example, alkaline phosphatase from neutrophils of mothers pregnant with a trisomy 21 fetus (Down's syndrome) displays significant differences both in its biochemical and immunological properties, and in its affinity for some specific inhibitors. RESULTS: To analyse these differences, the biochemical characteristics of two isozymes (non specific and placental alkaline phosphatases) were expressed in baculovirus infected cells. Comparative analysis of the two proteins allowed us to estimate the kinetic constants of denaturation and sensitivity to two inhibitors (L-p-bromotetramisole and thiophosphate), allowing better discrimination between the two enzymes. These parameters were then used to estimate the ratio of the two isoenzymes in neutrophils of pregnant mothers with or without a trisomy 21 fetus. It appeared that the placental isozyme represented 13% of the total activity of neutrophils of non pregnant women. This proportion did not significantly increase with normal pregnancy. By contrast, in pregnancies with trisomy 21 fetus, the proportion reached 60-80% of activity. CONCLUSION: Over-expression of the placental isozyme compared with the tissue-nonspecific form in neutrophils of mother with a trisomy 21 fetus may explain why the characteristics of the alkaline phosphatase in these cells is different from normal. Application of this knowledge could improve the potential of using alkaline phosphatase measurements to screen for Down's syndrome. AD - Laboratoire de Synthese et Physicochimie de Molecules d'Interet Biologique, Universite Paul Sabatier (UMR 5068), 118 route de Narbonne, 31 062, Toulouse, France. denier@cict.fr FAU - Denier, Colette C AU - Denier CC FAU - Brisson-Lougarre, Andree A AU - Brisson-Lougarre AA FAU - Biasini, Ghislaine G AU - Biasini GG FAU - Grozdea, Jean J AU - Grozdea JJ FAU - Fournier, Didier D AU - Fournier DD LA - eng PT - Journal Article DEP - 20020115 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Enzyme Inhibitors) RN - 0 (Isoenzymes) RN - 0 (Nitrophenols) RN - 0 (Organophosphorus Compounds) RN - 0 (Phosphates) RN - 0 (carcinoplacental isoenzymes) RN - 13598-51-1 (thiophosphoric acid) RN - 330-13-2 (nitrophenylphosphate) RN - 5036-02-2 (Tetramisole) RN - 6646-46-4 (4-bromotetramisole) RN - EC 3.1.3.1 (Alkaline Phosphatase) SB - IM MH - Alkaline Phosphatase/*analysis/genetics/metabolism MH - Baculoviridae/genetics MH - Comparative Study MH - Down Syndrome/diagnosis MH - Enzyme Inhibitors/chemistry/pharmacology MH - Enzyme Stability MH - Enzyme Tests MH - Female MH - Humans MH - Isoenzymes/*analysis/genetics/metabolism MH - Kinetics MH - Neutrophils/enzymology MH - Nitrophenols/chemistry/metabolism MH - Organophosphorus Compounds/chemistry/metabolism MH - Phosphates/chemistry/pharmacology MH - Placenta/*enzymology MH - Pregnancy MH - Protein Denaturation MH - Research Support, Non-U.S. Gov't MH - Tetramisole/*analogs & derivatives/chemistry/pharmacology EDAT- 2002/01/31 10:00 MHDA- 2002/11/26 04:00 PHST- 2001/08/23 [received] PHST- 2002/01/15 [accepted] PHST- 2002/01/15 [aheadofprint] PST - ppublish SO - BMC Biochem 2002;3(1):2. Epub 2002 Jan 15. PMID- 11825341 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021125 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 TI - Decreased insulin binding to mononuclear leucocytes and erythrocytes from dogs after S-nitroso-N-acetypenicillamine administration. PG - 1 AB - BACKGROUND: Nitric oxide (NO) and oxygen free-radicals play an important part in the destruction of beta-cells in auto- immune diabetes although the precise mechanism of interaction is still not known. This study was designed to examine any possible diabetogenic effect of NO by investigating any differences in cellular binding of insulin to its receptor on the cell membranes of erythrocytes and mononuclear leucocytes of dogs treated with the NO donor, S-nitroso-N-acetylpenicillamine (SNAP) and controls treated with captopril. RESULTS: The result obtained showed decreased binding of insulin to its receptor on the cell membranes of erythrocytes and mononuclear leucocytes. Mononuclear leucocytes from SNAP-treated dogs had decreased ability to bind insulin (16.30 +/- 1.24 %) when compared to mononuclear leucocytes from captopril-treated controls (20.30 +/- 1.93 %). Similar results were obtained for erythrocytes from dogs treated with SNAP (27.20 +/- 1.33 %) compared with dogs treated with captopril (34.70 +/- 3.58 %). Scatchard analysis demonstrated that this decrease in insulin binding was accounted for by a decrease in insulin receptor sites per cell, with mononuclear leucocytes of SNAP-treated dogs having 55 % less insulin receptor sites per cell compared with those of captopril-treated controls (P < 0.05). Average affinity and kinetic analysis revealed a 35 % decrease in the average receptor affinity, with mononuclear leucocytes from captopril-treated controls having an empty site affinity of 12.36 +/- 1.12 x 10(-8) M(-1) compared with 9.64 +/- 0.11 x 10(-8) M(-1) in SNAP-treated dogs (P < 0.05). CONCLUSION: These results suggest that acute alteration of the insulin receptor on the membranes of mononuclear leucocytes and erythrocytes occurred in dogs treated with S-nitroso-N-acetylpenicillamine. These findings suggest the first evidence of the novel role of NO as a modulator of insulin binding and the involvement of NO in the aetiology of diabetes mellitus. AD - Department of Basic Medical Sciences, University of the West Indies, Kingston, Jamaica. dmcgrowd@yahoo.com FAU - McGrowder, Donovan AU - McGrowder D FAU - Ragoobirsingh, Dalip AU - Ragoobirsingh D FAU - Dasgupta, Tara AU - Dasgupta T LA - eng PT - Journal Article DEP - 20020102 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Nitric Oxide Donors) RN - 11061-68-0 (Insulin) RN - 62571-86-2 (Captopril) RN - 79032-48-7 (S-Nitroso-N-Acetylpenicillamine) RN - EC 2.7.1.112 (Receptor, Insulin) SB - IM MH - Animals MH - Binding, Competitive MH - Captopril/pharmacology MH - Cell Membrane/metabolism MH - Dogs MH - Erythrocytes/drug effects/metabolism MH - Female MH - Insulin/*metabolism MH - Kinetics MH - Leukocytes, Mononuclear/drug effects/metabolism MH - Male MH - Nitric Oxide Donors/*pharmacology MH - Receptor, Insulin/*metabolism MH - Research Support, Non-U.S. Gov't MH - S-Nitroso-N-Acetylpenicillamine/*pharmacology EDAT- 2002/02/05 10:00 MHDA- 2002/11/26 04:00 PHST- 2001/08/06 [received] PHST- 2002/01/02 [accepted] PHST- 2002/01/02 [aheadofprint] PST - ppublish SO - BMC Biochem 2002;3(1):1. Epub 2002 Jan 2. PMID- 11825346 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030424 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 TI - Quality and methods of developing practice guidelines. PG - 1 AB - BACKGROUND: It is not known whether there are differences in the quality and recommendations between evidence-based (EB) and consensus-based (CB) guidelines. We used breast cancer guidelines as a case study to assess for these differences. METHODS: Five different instruments to evaluate the quality of guidelines were identified by a literature search. We also searched MEDLINE and the Internet to locate 8 breast cancer guidelines. These guidelines were classified in three categories: evidence based, consensus based and consensus based with no explicit consideration of evidence (CB-EB). Each guideline was evaluated by three of the authors using each of the instruments. For each guideline we assessed the agreement among 14 decision points which were selected from the NCCN (National Cancer Comprehensive Network) guidelines algorithm. For each decision point we recorded the level of the quality of the information used to support it. A regression analysis was performed to assess if the percentage of high quality evidence used in the guidelines development was related to the overall quality of the guidelines. RESULTS: Three guidelines were classified as EB, three as CB-EB and two as CB. The EB guidelines scored better than CB, with the CB-EB scoring in the middle among all instruments for guidelines quality assessment. No major disagreement in recommendations was detected among the guidelines regardless of the method used for development, but the EB guidelines had a better agreement with the benchmark guideline for any decision point. When the source of evidence used to support decision were of high quality, we found a higher level of full agreement among the guidelines' recommendations. Up to 94% of variation in the quality score among guidelines could be explained by the quality of evidence used for guidelines development. CONCLUSION: EB guidelines have a better quality than CB guidelines and CB-EB guidelines. Explicit use of high quality evidence can lead to a better agreement among recommendations. However, no major disagreement among guidelines was noted regardless of the method for their development. AD - H Lee Moffitt Cancer Center and Research institute, the University of South Florida, Tampa, FL, USA. crusehh@moffitt.usf.edu FAU - Cruse, Hugh AU - Cruse H FAU - Winiarek, Magdalena AU - Winiarek M FAU - Marshburn, Jan AU - Marshburn J FAU - Clark, Otavio AU - Clark O FAU - Djulbegovic, Benjamin AU - Djulbegovic B LA - eng PT - Evaluation Studies PT - Journal Article DEP - 20020111 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Algorithms MH - Benchmarking MH - Breast Neoplasms/*therapy MH - Comparative Study MH - *Consensus MH - *Evidence-Based Medicine MH - Female MH - Humans MH - Observer Variation MH - Peer Review, Research MH - Practice Guidelines/*standards MH - Quality Control MH - Regional Medical Programs/standards MH - Reproducibility of Results MH - United States EDAT- 2002/02/05 10:00 MHDA- 2003/04/25 05:00 PHST- 2001/10/20 [received] PHST- 2002/01/11 [accepted] PHST- 2002/01/11 [aheadofprint] PST - ppublish SO - BMC Health Serv Res 2002;2(1):1. Epub 2002 Jan 11. PMID- 11825347 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030424 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 TI - Resource use data by patient report or hospital records: do they agree? PG - 2 AB - BACKGROUND: Economic evaluations alongside clinical trials are becoming increasingly common. Cost data are often collected through the use of postal questionnaires; however, the accuracy of this method is uncertain. We compared postal questionnaires with hospital records for collecting data on physiotherapy service use. METHODS: As part of a randomised trial of orthopaedic medicine compared with orthopaedic surgery we collected physiotherapy use data on a group of patients from retrospective postal questionnaires and from hospital records. RESULTS: 315 patients were referred for physiotherapy. Hospital data on attendances was available for 30% (n = 96), compared with 48% (n = 150) of patients completing questionnaire data (95% Cl for difference = 10% to 24%); 19% (n = 59) had data available from both sources. The two methods produced an intraclass correlation coefficient of 0.54 (95% Cl 0.31 to 0.70). However, the two methods produced significantly different estimates of resource use with patient self report recalling a mean of 1.3 extra visits (95% Cl 0.4 to 2.2) compared with hospital records. CONCLUSIONS: Using questionnaires in this study produced data on a greater number of patients compared with examination of hospital records. However, the two data sources did differ in the quantity of physiotherapy used and this should be taken into account in any analysis. AD - Health Economics Research Group, Brunel University, Uxbridge, Middlesex, UK. hesradk@brunel.ac.uk FAU - Kennedy, Andrew D M AU - Kennedy AD FAU - Leigh-Brown, Anne P AU - Leigh-Brown AP FAU - Torgerson, David J AU - Torgerson DJ FAU - Campbell, James AU - Campbell J FAU - Grant, Adrian AU - Grant A LA - eng PT - Clinical Trial PT - Journal Article PT - Randomized Controlled Trial DEP - 20020117 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Adult MH - Comparative Study MH - Female MH - Great Britain MH - Health Resources/*utilization MH - *Hospital Records MH - Humans MH - Male MH - Middle Aged MH - Musculoskeletal Diseases/*therapy MH - Orthopedics/statistics & numerical data MH - Patient Compliance/*statistics & numerical data MH - Physical Therapy Department, Hospital/*utilization MH - Questionnaires MH - Referral and Consultation MH - Reproducibility of Results MH - *Self Disclosure EDAT- 2002/02/05 10:00 MHDA- 2003/04/25 05:00 PHST- 2001/11/26 [received] PHST- 2002/01/17 [accepted] PHST- 2002/01/17 [aheadofprint] PST - ppublish SO - BMC Health Serv Res 2002;2(1):2. Epub 2002 Jan 17. PMID- 11835696 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021125 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 TI - Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches' broom disease. PG - 3 AB - BACKGROUND: Several Trichoderma strains have been reported to be effective in controlling plant diseases, and the action of fungal hydrolytic enzymes has been considered as the main mechanism involved in the antagonistic process. However, although Trichoderma strains were found to impair development of Crinipellis perniciosa, the causal agent of cocoa plant witches' broom disease, no fungal strain is available for effective control of this disease. We have then undertaken a program of construction of hydrolytic enzyme-overproducing Trichoderma strains aiming improvement of the fungal antagonistic capacity. The protease of an indian Trichoderma isolate showing antagonistic activity against C. perniciosa was purified to homogeneity and characterized for its kinetic properties and action on the phytopathogen cell wall. RESULTS: A protease produced by the Trichoderma harzianum isolate 1051 was purified to homogeneity by precipitation with ammonium sulfate followed by hydrophobic chromatography. The molecular mass of this protease as determined by SDS-polyacrylamide gel electrophoresis was about 18.8 kDa. Its N-terminal amino acid sequence shares no homology with any other protease. The purified enzyme substantially affected the cell wall of the phytopathogen C. perniciosa. Western-blotting analysis showed that the enzyme was present in the culture supernatant 24 h after the Trichoderma started to grow in casein-containing liquid medium. CONCLUSIONS: The capacity of the Trichoderma harzianum protease to hydrolyze the cell wall of C. perniciosa indicates that this enzyme may be actually involved in the antagonistic process between the two fungi. This fact strongly suggest that hydrolytic enzyme over-producing transgenic fungi may show superior biocontrol capacity. AD - Departamento de Biologia Celular, Universidade de Brasilia, 70910-900, Brasilia, Brazil. jdemarco@uol.com.br FAU - De Marco, Janice L AU - De Marco JL FAU - Felix, Carlos Roberto AU - Felix CR LA - eng PT - Journal Article DEP - 20020122 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Antifungal Agents) RN - EC 3.4.- (Endopeptidases) SB - IM MH - Agaricales/*drug effects/ultrastructure MH - Antifungal Agents/*analysis/isolation & purification/pharmacology MH - Cacao/microbiology MH - Cell Wall/drug effects MH - Endopeptidases/*analysis/isolation & purification/pharmacology MH - Hydrogen-Ion Concentration MH - Microscopy, Electron, Scanning MH - Pest Control, Biological MH - Plant Diseases/microbiology MH - Research Support, Non-U.S. Gov't MH - Temperature MH - Trichoderma/*enzymology/isolation & purification EDAT- 2002/02/12 10:00 MHDA- 2002/11/26 04:00 PHST- 2001/09/17 [received] PHST- 2002/01/22 [accepted] PHST- 2002/01/22 [aheadofprint] PST - ppublish SO - BMC Biochem 2002;3(1):3. Epub 2002 Jan 22. PMID- 11876827 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021125 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 TI - Probing substrate binding to metallo-beta-lactamase L1 from Stenotrophomonas maltophilia by using site-directed mutagenesis. PG - 4 AB - BACKGROUND: The metallo-beta-lactamases are Zn(II)-containing enzymes that hydrolyze the beta-lactam bond in penicillins, cephalosporins, and carbapenems and are involved in bacterial antibiotic resistance. There are at least 20 distinct organisms that produce a metallo-beta-lactamase, and these enzymes have been extensively studied using X-ray crystallographic, computational, kinetic, and inhibition studies; however, much is still unknown about how substrates bind and the catalytic mechanism. In an effort to probe substrate binding to metallo-beta-lactamase L1 from Stenotrophomonas maltophilia, nine site-directed mutants of L1 were prepared and characterized using metal analyses, CD spectroscopy, and pre-steady state and steady state kinetics. RESULTS: Site-directed mutations were generated of amino acids previously predicted to be important in substrate binding. Steady-state kinetic studies using the mutant enzymes and 9 different substrates demonstrated varying Km and kcat values for the different enzymes and substrates and that no direct correlation between Km and the effect of the mutation on substrate binding could be drawn. Stopped-flow fluorescence studies using nitrocefin as the substrate showed that only the S224D and Y228A mutants exhibited weaker nitrocefin binding. CONCLUSIONS: The data presented herein indicate that Ser224, Ile164, Phe158, Tyr228, and Asn233 are not essential for tight binding of substrate to metallo-beta-lactamase L1. The results in this work also show that Km values are not reliable for showing substrate binding, and there is no correlation between substrate binding and the amount of reaction intermediate formed during the reaction. This work represents the first experimental testing of one of the computational models of the metallo-beta-lactamases. AD - Department of Chemistry and Biochemistry, Miami University, Oxford, OH, USA. sutsong@yahoo.com FAU - Carenbauer, Anne L AU - Carenbauer AL FAU - Garrity, James D AU - Garrity JD FAU - Periyannan, Gopal AU - Periyannan G FAU - Yates, Robert B AU - Yates RB FAU - Crowder, Michael W AU - Crowder MW LA - eng GR - R29 AI40052/AI/NIAID PT - Journal Article DEP - 20020213 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Carbapenems) RN - 0 (Cephalosporins) RN - 0 (Metals) RN - 0 (Penicillins) RN - 41906-86-9 (nitrocefin) RN - 55520-40-6 (Tyrosine) RN - 56-45-1 (Serine) RN - 63-91-2 (Phenylalanine) RN - 7006-34-0 (Asparagine) RN - 73-32-5 (Isoleucine) RN - EC 3.5.2.- (beta-lactamase L1) RN - EC 3.5.2.6 (beta-Lactamases) SB - IM MH - Asparagine/genetics MH - Binding Sites MH - Carbapenems/metabolism MH - Cephalosporins/metabolism MH - Computational Biology MH - Isoleucine/genetics MH - Kinetics MH - Metals/analysis MH - Models, Molecular MH - Mutagenesis, Site-Directed MH - Penicillins/metabolism MH - Phenylalanine/genetics MH - Protein Binding MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Research Support, U.S. Gov't, P.H.S. MH - Serine/genetics MH - Stenotrophomonas maltophilia/*enzymology MH - Tyrosine/genetics MH - beta-Lactamases/chemistry/*genetics/*metabolism EDAT- 2002/03/06 10:00 MHDA- 2002/11/26 04:00 PHST- 2001/11/06 [received] PHST- 2002/02/13 [accepted] PHST- 2002/02/13 [aheadofprint] PST - ppublish SO - BMC Biochem 2002;3(1):4. Epub 2002 Feb 13. PMID- 11882257 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030424 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 TI - What kind of evidence is it that Evidence-Based Medicine advocates want health care providers and consumers to pay attention to? PG - 3 AB - BACKGROUND: In 1992, Evidence-Based Medicine advocates proclaimed a "new paradigm", in which evidence from health care research is the best basis for decisions for individual patients and health systems. Hailed in New York Times Magazine in 2001 as one of the most influential ideas of the year, this approach was initially and provocatively pitted against the traditional teaching of medicine, in which the key elements of knowing for clinical purposes are understanding of basic pathophysiologic mechanisms of disease coupled with clinical experience. This paper reviews the origins, aspirations, philosophical limitations, and practical challenges of evidence-based medicine. DISCUSSION: EBM has long since evolved beyond its initial (mis)conception, that EBM might replace traditional medicine. EBM is now attempting to augment rather than replace individual clinical experience and understanding of basic disease mechanisms. EBM must continue to evolve, however, to address a number of issues including scientific underpinnings, moral stance and consequences, and practical matters of dissemination and application. For example, accelerating the transfer of research findings into clinical practice is often based on incomplete evidence from selected groups of people, who experience a marginal benefit from an expensive technology, raising issues of the generalizability of the findings, and increasing problems with how many and who can afford the new innovations in care. SUMMARY: Advocates of evidence-based medicine want clinicians and consumers to pay attention to the best findings from health care research that are both valid and ready for clinical application. Much remains to be done to reach this goal. AD - Department of Clinical Epidemiology and Biostatistics Faculty of Health Sciences, McMaster University, Hamilton, Ontario, Canada. bhaynes@mcmaster.ca FAU - Haynes, R Brian AU - Haynes RB LA - eng PT - Journal Article DEP - 20020306 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - *Decision Making MH - Diffusion of Innovation MH - *Evidence-Based Medicine/ethics/trends MH - *Health Services Research MH - Humans MH - Knowledge MH - Patient Participation MH - Physician's Practice Patterns MH - Practice Guidelines MH - Probability MH - Randomized Controlled Trials MH - Resource Allocation/ethics MH - Uncertainty EDAT- 2002/03/08 10:00 MHDA- 2003/04/25 05:00 PHST- 2001/12/24 [received] PHST- 2002/03/06 [accepted] PHST- 2002/03/06 [aheadofprint] PST - ppublish SO - BMC Health Serv Res 2002;2(1):3. Epub 2002 Mar 6. PMID- 11884248 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030424 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 TI - Reporting of measures of accuracy in systematic reviews of diagnostic literature. PG - 4 AB - BACKGROUND: There are a variety of ways in which accuracy of clinical tests can be summarised in systematic reviews. Variation in reporting of summary measures has only been assessed in a small survey restricted to meta-analyses of screening studies found in a single database. Therefore, we performed this study to assess the measures of accuracy used for reporting results of primary studies as well as their meta-analysis in systematic reviews of test accuracy studies. METHODS: Relevant reviews on test accuracy were selected from the Database of Abstracts of Reviews of Effectiveness (1994-2000), which electronically searches seven bibliographic databases and manually searches key resources. The structured abstracts of these reviews were screened and information on accuracy measures was extracted from the full texts of 90 relevant reviews, 60 of which used meta-analysis. RESULTS: Sensitivity or specificity was used for reporting the results of primary studies in 65/90 (72%) reviews, predictive values in 26/90 (28%), and likelihood ratios in 20/90 (22%). For meta-analysis, pooled sensitivity or specificity was used in 35/60 (58%) reviews, pooled predictive values in 11/60 (18%), pooled likelihood ratios in 13/60 (22%), and pooled diagnostic odds ratio in 5/60 (8%). Summary ROC was used in 44/60 (73%) of the meta-analyses. There were no significant differences in measures of test accuracy among reviews published earlier (1994-97) and those published later (1998-2000). CONCLUSIONS: There is considerable variation in ways of reporting and summarising results of test accuracy studies in systematic reviews. There is a need for consensus about the best ways of reporting results of test accuracy studies in reviews. AD - Academic Dept, of Obstetrics & Gynaecology, Birmingham Women's Hospital, Birmingham B15 2TG, United Kingdom. h.honest@bham.ac.uk FAU - Honest, Honest AU - Honest H FAU - Khan, Khalid S AU - Khan KS LA - eng PT - Journal Article DEP - 20020307 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Bias (Epidemiology) MH - *Data Interpretation, Statistical MH - Databases, Bibliographic MH - Humans MH - Laboratory Techniques and Procedures/*standards MH - *Meta-Analysis MH - Pathology, Clinical/*standards MH - Quality Control MH - Reproducibility of Results MH - Research Design/*standards MH - Research Support, Non-U.S. Gov't MH - *Review Literature MH - Sensitivity and Specificity EDAT- 2002/03/09 10:00 MHDA- 2003/04/25 05:00 PHST- 2001/09/05 [received] PHST- 2002/03/07 [accepted] PHST- 2002/03/07 [aheadofprint] PST - ppublish SO - BMC Health Serv Res 2002;2(1):4. Epub 2002 Mar 7. PMID- 11914161 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030422 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Mar 21 TI - A randomised controlled trial of a patient based Diabetes Recall and Management System: the DREAM trial: a study protocol [ISRCTN32042030]. PG - 5 AB - BACKGROUND: Whilst there is broad agreement on what constitutes high quality health care for people with diabetes, there is little consensus on the most efficient way of delivering it. Structured recall systems can improve the quality of care but the systems evaluated to date have been of limited sophistication and the evaluations have been carried out in small numbers of relatively unrepresentative settings. Hartlepool, Easington and Stockton currently operate a computerised diabetes register which has to date produced improvements in the quality of care but performance has now plateaued leaving substantial scope for further improvement. This study will evaluate the effectiveness and efficiency of an area wide 'extended' system incorporating a full structured recall and management system, actively involving patients and including clinical management prompts to primary care clinicians based on locally-adapted evidence based guidelines. METHODS: The study design is a two-armed cluster randomised controlled trial of 61 practices incorporating evaluations of the effectiveness of the system, its economic impact and its impact on patient wellbeing and functioning. AD - Centre for Health Services Research, University of Newcastle, Newcastle upon Tyne, UK. martin.eccles@ncl.ac.uk FAU - Eccles, Martin AU - Eccles M FAU - Hawthorne, Gillian AU - Hawthorne G FAU - Whitty, Paula AU - Whitty P FAU - Steen, Nick AU - Steen N FAU - Vanoli, Alessandra AU - Vanoli A FAU - Grimshaw, Jeremy AU - Grimshaw J FAU - Wood, Linda AU - Wood L LA - eng PT - Clinical Trial PT - Journal Article PT - Randomized Controlled Trial DEP - 20020321 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - *Database Management Systems MH - Diabetes Mellitus/diagnosis/*prevention & control MH - Efficiency, Organizational MH - England MH - *Evidence-Based Medicine MH - Family Practice/organization & administration/*standards MH - Guideline Adherence MH - Humans MH - Medical Audit MH - Patient Compliance MH - Preventive Health Services/standards/utilization MH - Primary Health Care/organization & administration/standards MH - Quality Assurance, Health Care/*organization & administration MH - Registries MH - *Reminder Systems EDAT- 2002/03/27 10:00 MHDA- 2003/04/23 05:00 PHST- 2001/12/14 [received] PHST- 2002/03/21 [accepted] PHST- 2002/03/21 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Mar 21;2(1):5. PMID- 11914162 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030423 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Mar 25 TI - Organization specific predictors of job satisfaction: findings from a Canadian multi-site quality of work life cross-sectional survey. PG - 6 AB - BACKGROUND: Organizational features can affect how staff view their quality of work life. Determining staff perceptions about quality of work life is an important consideration for employers interested in improving employee job satisfaction. The purpose of this study was to identify organization specific predictors of job satisfaction within a health care system that consisted of six independent health care organizations. METHODS: 5,486 full, part and causal time (non-physician) staff on active payroll within six organizations (2 community hospitals, 1 community hospital/long-term care facility, 1 long-term care facility, 1 tertiary care/community health centre, and 1 visiting nursing agency) located in five communities in Central West Ontario, Canada were asked to complete a 65-item quality of work life survey. The self-administered questionnaires collected staff perceptions of: co-worker and supervisor support; teamwork and communication; job demands and decision authority; organization characteristics; patient/resident care; compensation and benefits; staff training and development; and impressions of the organization. Socio-demographic data were also collected. RESULTS: Depending on the organization, between 15 and 30 (of the 40 potential predictor) variables were found to be statistically associated with job satisfaction (univariate analyses). Logistic regression analyses identified the best predictors of job satisfaction and these are presented for each of the six organizations and for all organizations combined. CONCLUSIONS: The findings indicate that job satisfaction is a multidimensional construct and although there appear to be some commonalities across organizations, some predictors of job satisfaction appear to be organization and context specific. AD - St, Joseph's Health System Research Network, Father Sean O'Sullivan Research Centre, Hamilton, Ontario. kruegerp@mcmaster.ca FAU - Krueger, Paul AU - Krueger P FAU - Brazil, Kevin AU - Brazil K FAU - Lohfeld, Lynne AU - Lohfeld L FAU - Edward, H Gayle AU - Edward HG FAU - Lewis, David AU - Lewis D FAU - Tjam, Erin AU - Tjam E LA - eng PT - Journal Article DEP - 20020325 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Adult MH - *Attitude of Health Personnel MH - Communication MH - Community Health Centers/organization & administration MH - Community Health Nursing/organization & administration MH - Decision Making, Organizational MH - Delivery of Health Care, Integrated/*organization & administration MH - Female MH - Forecasting MH - Hospitals, Community/organization & administration MH - Humans MH - *Job Satisfaction MH - Logistic Models MH - Male MH - Middle Aged MH - Ontario MH - Patient Care Team MH - Personnel Management/*methods MH - Questionnaires MH - Research Support, Non-U.S. Gov't MH - Residential Facilities/organization & administration EDAT- 2002/03/27 10:00 MHDA- 2003/04/24 05:00 PHST- 2001/10/01 [received] PHST- 2002/03/25 [accepted] PHST- 2002/03/25 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Mar 25;2(1):6. PMID- 11914163 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030423 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Mar 25 TI - Outcomes research in the development and evaluation of practice guidelines. PG - 7 AB - BACKGROUND: Practice guidelines have been developed in response to the observation that variations exist in clinical medicine that are not related to variations in the clinical presentation and severity of the disease. Despite their widespread use, however, practice guideline evaluation lacks a rigorous scientific methodology to support its development and application. DISCUSSION: Firstly, we review the major epidemiological foundations of practice guideline development. Secondly, we propose a chronic disease epidemiological model in which practice patterns are viewed as the exposure and outcomes of interest such as quality or cost are viewed as the disease. Sources of selection, information, confounding and temporal trend bias are identified and discussed. SUMMARY: The proposed methodological framework for outcomes research to evaluate practice guidelines reflects the selection, information and confounding biases inherent in its observational nature which must be accounted for in both the design and the analysis phases of any outcomes research study. AD - Division of Clinical Epidemiology, Research Institute of the McGill University Health Center, Montreal, Canada. louise.pilote@mcgill.ca FAU - Pilote, Louise AU - Pilote L FAU - Tager, Ira B AU - Tager IB LA - eng PT - Journal Article DEP - 20020325 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Biomedical Research MH - Chronic Disease/*epidemiology/therapy MH - Confounding Factors (Epidemiology) MH - Data Collection MH - Databases MH - Epidemiologic Methods MH - Evaluation Studies MH - Humans MH - Outcome Assessment (Health Care)/*methods MH - Physician's Practice Patterns/*statistics & numerical data MH - Practice Guidelines/*standards MH - Selection Bias MH - Treatment Outcome MH - Uncertainty EDAT- 2002/03/27 10:00 MHDA- 2003/04/24 05:00 PHST- 2001/10/01 [received] PHST- 2002/03/25 [accepted] PHST- 2002/03/25 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Mar 25;2(1):7. PMID- 11914164 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030423 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Mar 26 TI - Inter-rater agreement in the scoring of abstracts submitted to a primary care research conference. PG - 8 AB - BACKGROUND: Checklists for peer review aim to guide referees when assessing the quality of papers, but little evidence exists on the extent to which referees agree when evaluating the same paper. The aim of this study was to investigate agreement on dimensions of a checklist between two referees when evaluating abstracts submitted for a primary care conference. METHODS: Anonymised abstracts were scored using a structured assessment comprising seven categories. Between one (poor) and four (excellent) marks were awarded for each category, giving a maximum possible score of 28 marks. Every abstract was assessed independently by two referees and agreement measured using intraclass correlation coefficients. Mean total scores of abstracts accepted and rejected for the meeting were compared using an unpaired t test. RESULTS: Of 52 abstracts, agreement between reviewers was greater for three components relating to study design (adjusted intraclass correlation coefficients 0.40 to 0.45) compared to four components relating to more subjective elements such as the importance of the study and likelihood of provoking discussion (0.01 to 0.25). Mean score for accepted abstracts was significantly greater than those that were rejected (17.4 versus 14.6, 95% CI for difference 1.3 to 4.1, p = 0.0003). CONCLUSIONS: The findings suggest that inclusion of subjective components in a review checklist may result in greater disagreement between reviewers. However in terms of overall quality scores, abstracts accepted for the meeting were rated significantly higher than those that were rejected. AD - Division of Primary Health Care, University of Bristol, Cotham House, Cotham Hill, Bristol BS8 2PR, UK. alan.a.montgomery@bristol.ac.uk FAU - Montgomery, Alan A AU - Montgomery AA FAU - Graham, Anna AU - Graham A FAU - Evans, Philip H AU - Evans PH FAU - Fahey, Tom AU - Fahey T LA - eng PT - Journal Article DEP - 20020326 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Abstracting and Indexing/classification/*standards MH - Congresses MH - *Consensus MH - Data Interpretation, Statistical MH - Great Britain MH - Humans MH - Judgment MH - Manuscripts, Medical MH - Observer Variation MH - Peer Review, Research/methods/*standards MH - Primary Health Care MH - Research Support, Non-U.S. Gov't EDAT- 2002/03/27 10:00 MHDA- 2003/04/24 05:00 PHST- 2001/12/03 [received] PHST- 2002/03/26 [accepted] PHST- 2002/03/26 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Mar 26;2(1):8. PMID- 11943069 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Mar 27 TI - The Caenorhabditis elegans Y87G2A.14 Nudix hydrolase is a peroxisomal coenzyme A diphosphatase. PG - 5 AB - BACKGROUND: The number of Nudix hydrolase family members varies widely among different organisms. In order to understand the reasons for the particular spectrum possessed by a given organism, the substrate specificity and function of different family members must be established. RESULTS: The Y87G2A.14 Nudix hydrolase gene product of Caenorhabditis elegans has been expressed as a thioredoxin fusion protein in Escherichia coli and shown to be a CoA diphosphatase with catalytic activity towards CoA and its derivatives. The products of CoA hydrolysis were 3',5'-ADP and 4'-phosphopantetheine with Km and kcat values of 220 microM and 13.8 s(-1) respectively. CoA esters yielded 3',5'-ADP and the corresponding acyl-phosphopantetheine. Activity was optimal at pH 9.5 with 5 mM Mg2+ and fluoride was inhibitory with a Ki of 3 microM. The Y87G2A.14 gene product has a potential C-terminal tripeptide PTS1 peroxisomal targeting signal - SKI. By fusing a Y87G2A.14 cDNA to the C-terminus of yeast-enhanced green fluorescent protein, the enzyme appeared to be targeted to peroxisomes by the SKI signal when transfected into yeast cells. Deletion of SKI abolished specific targeting. CONCLUSIONS: The presence of related sequences with potential PTS1 or PTS2 peroxisomal targeting signals in other organisms suggests a conserved peroxisomal function for the CoA diphosphatase members of this group of Nudix hydrolases. AD - School of Biological Sciences, Life Sciences Building, University of Liverpool, P,O, Box 147, Liverpool L69 7ZB, UK. salamara@liv.ac.uk FAU - AbdelRaheim, Salama R AU - AbdelRaheim SR FAU - McLennan, Alexander G AU - McLennan AG LA - eng PT - Journal Article DEP - 20020327 PL - England TA - BMC Biochem JID - 101084098 RN - 85-61-0 (Coenzyme A) RN - EC 3.6.1.- (Pyrophosphatases) RN - EC 3.6.1.- (nudix hydrolase) SB - IM MH - Amino Acid Sequence MH - Animals MH - Caenorhabditis elegans/*enzymology MH - Cloning, Molecular MH - Coenzyme A/*metabolism MH - Kinetics MH - Molecular Sequence Data MH - Peroxisomes/*metabolism MH - Pyrophosphatases/genetics/isolation & purification/*metabolism MH - Research Support, Non-U.S. Gov't MH - Sequence Alignment MH - Substrate Specificity EDAT- 2002/04/12 10:00 MHDA- 2002/11/26 04:00 PHST- 2002/02/12 [received] PHST- 2002/03/27 [accepted] PHST- 2002/03/27 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Mar 27;3(1):5. PMID- 11972899 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20041215 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Apr 24 TI - Interconversion of Anthozoa GFP-like fluorescent and non-fluorescent proteins by mutagenesis. PG - 7 AB - BACKGROUND: Within the family of green fluorescent protein (GFP) homologs, one can mark two main groups, specifically, fluorescent proteins (FPs) and non-fluorescent or chromoproteins (CPs). Structural background of differences between FPs and CPs are poorly understood to date. RESULTS: Here, we applied site-directed and random mutagenesis in order to to transform CP into FP and vice versa. A purple chromoprotein asCP (asFP595) from Anemonia sulcata and a red fluorescent protein DsRed from Discosoma sp. were selected as representatives of CPs and FPs, respectively. For asCP, some substitutions at positions 148 and 165 (numbering in accordance to GFP) were found to dramatically increase quantum yield of red fluorescence. For DsRed, substitutions at positions 148, 165, 167, and 203 significantly decreased fluorescence intensity, so that the spectral characteristics of these mutants became more close to those of CPs. Finally, a practically non-fluorescent mutant DsRed-NF was generated. This mutant carried four amino acid substitutions, specifically, S148C, I165N, K167M, and S203A. DsRed-NF possessed a high extinction coefficient and an extremely low quantum yield (< 0.001). These spectral characteristics allow one to regard DsRed-NF as a true chromoprotein. CONCLUSIONS: We located a novel point in asCP sequence (position 165) mutations at which can result in red fluorescence appearance. Probably, this finding could be applied onto other CPs to generate red and far-red fluorescent mutants. A possibility to transform an FP into CP was demonstrated. Key role of residues adjacent to chromophore's phenolic ring in fluorescent/non-fluorescent states determination was revealed. AD - Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117997 Moscow, Russia. biomasha@mail.ru FAU - Bulina, Maria E AU - Bulina ME FAU - Chudakov, Dmitry M AU - Chudakov DM FAU - Mudrik, Nikolay N AU - Mudrik NN FAU - Lukyanov, Konstantin A AU - Lukyanov KA LA - eng PT - Journal Article DEP - 20020424 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (FP595 protein, Anemonia sulcata) RN - 0 (Luminescent Proteins) RN - 0 (fluorescent protein 583) RN - 147336-22-9 (Green Fluorescent Proteins) SB - IM MH - Amino Acid Sequence MH - *Anthozoa MH - Fluorescence MH - Green Fluorescent Proteins MH - Luminescent Proteins/*chemistry/*genetics MH - Models, Molecular MH - Molecular Sequence Data MH - Mutagenesis MH - Mutagenesis, Site-Directed MH - Research Support, Non-U.S. Gov't MH - Sequence Alignment MH - Spectrometry, Fluorescence EDAT- 2002/04/26 10:00 MHDA- 2002/11/26 04:00 PHST- 2001/12/14 [received] PHST- 2002/04/24 [accepted] PHST- 2002/04/24 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Apr 24;3(1):7. PMID- 11996675 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021218 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 May 7 TI - Hisactophilin is involved in osmoprotection in Dictyostelium. PG - 10 AB - BACKGROUND: Dictyostelium cells exhibit an unusual stress response as they protect themselves against hyperosmotic stress. Cytoskeletal proteins are recruited from the cytosolic pool to the cell cortex, thereby reinforcing it. In order to gain more insight into the osmoprotective mechanisms of this amoeba, we used 1-D and 2-D gel electrophoresis to identify new proteins that are translocated during osmotic shock. RESULTS: We identified hisactophilin as one of the proteins that are enriched in the cytoskeletal fraction during osmotic shock. In mutants lacking hisactophilin, viability is reduced under hyperosmotic stress conditions. In wild type cells, serine phosphorylation of hisactophilin was specifically induced by hypertonicity, but not when other stress conditions were imposed on cells. The phosphorylation kinetics reveals a slow accumulation of phosphorylated hisactophilin from 20-60 min after onset of the hyperosmotic shock condition. CONCLUSION: In the present study, we identified hisactophilin as an essential protein for the osmoprotection of Dictyostelium cells. The observed phosphorylation kinetics suggest that hisactophilin regulation is involved in long-term osmoprotection and that phosphorylation occurs in parallel with inactivation of the dynamic actin cytoskeleton. AD - Max-Planck-Institute for Biochemistry, 82152 Martinsried, Germany. pintsch@switch-biotech.com FAU - Pintsch, Tanja AU - Pintsch T FAU - Zischka, Hans AU - Zischka H FAU - Schuster, Stephan C AU - Schuster SC LA - eng PT - Journal Article DEP - 20020507 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Carrier Proteins) RN - 0 (Hypertonic Solutions) RN - 0 (Microfilament Proteins) RN - 0 (Protozoan Proteins) RN - 122962-20-3 (hisactophilin protein, Protozoan) RN - 50-70-4 (Sorbitol) RN - 56-45-1 (Serine) SB - IM MH - Animals MH - Carrier Proteins/drug effects/genetics/*metabolism MH - Cell Division/drug effects/genetics MH - Cytoskeleton/drug effects/metabolism MH - Dictyostelium/cytology/*drug effects/metabolism MH - Dose-Response Relationship, Drug MH - Electrophoresis, Gel, Two-Dimensional MH - Hypertonic Solutions/pharmacology MH - *Microfilament Proteins MH - Mutation MH - Osmotic Pressure MH - Phosphorylation/drug effects MH - Protozoan Proteins/drug effects/genetics/*metabolism MH - Research Support, Non-U.S. Gov't MH - Serine/metabolism MH - Sorbitol/*pharmacology EDAT- 2002/05/09 10:00 MHDA- 2002/12/19 04:00 PHST- 2001/12/19 [received] PHST- 2002/05/07 [accepted] PHST- 2002/05/07 [aheadofprint] PST - epublish SO - BMC Biochem 2002 May 7;3(1):10. PMID- 11997336 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - A complete sequence of the T. tengcongensis genome. PG - 689-700 AB - Thermoanaerobacter tengcongensis is a rod-shaped, gram-negative, anaerobic eubacterium that was isolated from a freshwater hot spring in Tengchong, China. Using a whole-genome-shotgun method, we sequenced its 2,689,445-bp genome from an isolate, MB4(T) (Genbank accession no. AE008691). The genome encodes 2588 predicted coding sequences (CDS). Among them, 1764 (68.2%) are classified according to homology to other documented proteins, and the rest, 824 CDS (31.8%), are functionally unknown. One of the interesting features of the T. tengcongensis genome is that 86.7% of its genes are encoded on the leading strand of DNA replication. Based on protein sequence similarity, the T. tengcongensis genome is most similar to that of Bacillus halodurans, a mesophilic eubacterium, among all fully sequenced prokaryotic genomes up to date. Computational analysis on genes involved in basic metabolic pathways supports the experimental discovery that T. tengcongensis metabolizes sugars as principal energy and carbon source and utilizes thiosulfate and element sulfur, but not sulfate, as electron acceptors. T. tengcongensis, as a gram-negative rod by empirical definitions (such as staining), shares many genes that are characteristics of gram-positive bacteria whereas it is missing molecular components unique to gram-negative bacteria. A strong correlation between the G + C content of tDNA and rDNA genes and the optimal growth temperature is found among the sequenced thermophiles. It is concluded that thermophiles are a biologically and phylogenetically divergent group of prokaryotes that have converged to sustain extreme environmental conditions over evolutionary timescale. AD - Beijing Genomics Institute/Genomics and Bioinformatics Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences (CAS), Beijing 100101, China. FAU - Bao, Qiyu AU - Bao Q FAU - Tian, Yuqing AU - Tian Y FAU - Li, Wei AU - Li W FAU - Xu, Zuyuan AU - Xu Z FAU - Xuan, Zhenyu AU - Xuan Z FAU - Hu, Songnian AU - Hu S FAU - Dong, Wei AU - Dong W FAU - Yang, Jian AU - Yang J FAU - Chen, Yanjiong AU - Chen Y FAU - Xue, Yanfen AU - Xue Y FAU - Xu, Yi AU - Xu Y FAU - Lai, Xiaoqin AU - Lai X FAU - Huang, Li AU - Huang L FAU - Dong, Xiuzhu AU - Dong X FAU - Ma, Yanhe AU - Ma Y FAU - Ling, Lunjiang AU - Ling L FAU - Tan, Huarong AU - Tan H FAU - Chen, Runsheng AU - Chen R FAU - Wang, Jian AU - Wang J FAU - Yu, Jun AU - Yu J FAU - Yang, Huanming AU - Yang H LA - eng SI - GENBANK/AE008691 PT - Journal Article PL - United States TA - Genome Res JID - 9518021 RN - 0 (Codon) RN - 7704-34-9 (Sulfur) SB - IM MH - Bacillaceae/cytology/*genetics/metabolism/physiology MH - Base Composition/genetics MH - Codon/genetics MH - DNA Repair/genetics MH - DNA Replication/genetics MH - GC Rich Sequence/genetics MH - Genes, Structural, Bacterial/genetics MH - *Genome, Bacterial MH - Genomics/methods MH - Heat MH - Ion Transport/genetics MH - Molecular Sequence Data MH - Oxygen Consumption/genetics MH - Protein Biosynthesis/genetics MH - Recombination, Genetic/genetics MH - Repetitive Sequences, Nucleic Acid/genetics MH - Replication Origin/genetics MH - Research Support, Non-U.S. Gov't MH - Sequence Analysis, DNA/methods MH - Sulfur/metabolism MH - Transcription, Genetic EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.219302 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):689-700. PMID- 11997337 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041215 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Identification of a novel cis-regulatory element involved in the heat shock response in Caenorhabditis elegans using microarray gene expression and computational methods. PG - 701-12 AB - We report here the identification of a previously unknown transcription regulatory element for heat shock (HS) genes in Caenorhabditis elegans. We monitored the expression pattern of 11,917 genes from C. elegans to determine the genes that were up-regulated on HS. Twenty eight genes were observed to be consistently up-regulated in several different repetitions of the experiments. We analyzed the upstream regions of these genes using computational DNA pattern recognition methods. Two potential cis-regulatory motifs were identified in this way. One of these motifs (TTCTAGAA) was the DNA binding motif for the heat shock factor (HSF), whereas the other (GGGTGTC) was previously unreported in the literature. We determined the significance of these motifs for the HS genes using different statistical tests and parameters. Comparative sequence analysis of orthologous HS genes from C. elegans and Caenorhabditis briggsae indicated that the identified DNA regulatory motifs are conserved across related species. The role of the identified DNA sites in regulation of HS genes was tested by in vitro mutagenesis of a green fluorescent protein (GFP) reporter transgene driven by the C. elegans hsp-16-2 promoter. DNA sites corresponding to both motifs are shown to play a significant role in up-regulation of the hsp-16-2 gene on HS. This is one of the rare instances in which a novel regulatory element, identified using computational methods, is shown to be biologically active. The contributions of individual sites toward induction of transcription on HS are nonadditive, which indicates interaction and cross-talk between the sites, possibly through the transcription factors (TFs) binding to these sites. AD - Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63114, USA. FAU - GuhaThakurta, Debraj AU - GuhaThakurta D FAU - Palomar, Lisanne AU - Palomar L FAU - Stormo, Gary D AU - Stormo GD FAU - Tedesco, Pat AU - Tedesco P FAU - Johnson, Thomas E AU - Johnson TE FAU - Walker, David W AU - Walker DW FAU - Lithgow, Gordon AU - Lithgow G FAU - Kim, Stuart AU - Kim S FAU - Link, Christopher D AU - Link CD LA - eng GR - AG12423/AG/NIA GR - HG00249/HG/NHGRI GR - R25 GM62495-01/GM/NIGMS PT - Journal Article PL - United States TA - Genome Res JID - 9518021 RN - 0 (Luminescent Proteins) RN - 147336-22-9 (Green Fluorescent Proteins) RN - 9007-49-2 (DNA) SB - IM EIN - Genome Res 2002 Aug;12(8):1301 MH - Animals MH - Base Sequence MH - Binding Sites/genetics MH - Caenorhabditis elegans/*genetics MH - Comparative Study MH - Computational Biology/*methods MH - Conserved Sequence/genetics MH - DNA/metabolism MH - Gene Expression Profiling/*methods MH - Genes, Structural, Helminth/genetics MH - Green Fluorescent Proteins MH - Heat-Shock Response/*genetics MH - Luminescent Proteins/biosynthesis MH - Molecular Sequence Data MH - Mutagenesis, Site-Directed/genetics MH - Oligonucleotide Array Sequence Analysis/*methods MH - Probability MH - Promoter Regions (Genetics)/genetics MH - Regulatory Sequences, Nucleic Acid/*physiology MH - Research Support, U.S. Gov't, P.H.S. MH - Species Specificity MH - Statistics, Nonparametric MH - Up-Regulation/genetics EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.228902 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):701-12. PMID- 11997338 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041203 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Genes in a refined Smith-Magenis syndrome critical deletion interval on chromosome 17p11.2 and the syntenic region of the mouse. PG - 713-28 AB - Smith-Magenis syndrome (SMS) is a multiple congenital anomaly/mental retardation syndrome associated with behavioral abnormalities and sleep disturbance. Most patients have the same approximately 4 Mb interstitial genomic deletion within chromosome 17p11.2. To investigate the molecular bases of the SMS phenotype, we constructed BAC/PAC contigs covering the SMS common deletion interval and its syntenic region on mouse chromosome 11. Comparative genome analysis reveals the absence of all three approximately 200-kb SMS-REP low-copy repeats in the mouse and indicates that the evolution of SMS-REPs was accompanied by transposition of adjacent genes. Physical and genetic map comparisons in humans reveal reduced recombination in both sexes. Moreover, by examining the deleted regions in SMS patients with unusual-sized deletions, we refined the minimal Smith-Magenis critical region (SMCR) to an approximately 1.1-Mb genomic interval that is syntenic to an approxiamtely 1.0-Mb region in the mouse. Genes within the SMCR and its mouse syntenic region were identified by homology searches and by gene prediction programs, and their gene structures and expression profiles were characterized. In addition to 12 genes previously mapped, we identified 8 new genes and 10 predicted genes in the SMCR. In the mouse syntenic region of the human SMCR, 16 genes and 6 predicted genes were identified. The SMCR is highly conserved between humans and mice, including 19 genes with the same gene order and orientation. Our findings will facilitate both the identification of gene(s) responsible for the SMS phenotype and the engineering of an SMS mouse model. AD - Department of Molecular & Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA. FAU - Bi, Weimin AU - Bi W FAU - Yan, Jiong AU - Yan J FAU - Stankiewicz, Pawe AU - Stankiewicz P FAU - Park, Sung-Sup AU - Park SS FAU - Walz, Katherina AU - Walz K FAU - Boerkoel, Cornelius F AU - Boerkoel CF FAU - Potocki, Lorraine AU - Potocki L FAU - Shaffer, Lisa G AU - Shaffer LG FAU - Devriendt, Koen AU - Devriendt K FAU - Nowaczyk, Magorzata J M AU - Nowaczyk MJ FAU - Inoue, Ken AU - Inoue K FAU - Lupski, James R AU - Lupski JR LA - eng GR - P01 CA75719/CA/NCI GR - P01 HD38420/HD/NICHD GR - R01 NS27042/NS/NINDS PT - Journal Article PL - United States TA - Genome Res JID - 9518021 RN - 0 (Chaperonins) RN - 0 (Chromosomes, Artificial, Bacterial) RN - 0 (Chromosomes, Artificial, P1 Bacteriophage) RN - 0 (Fungal Proteins) RN - 0 (Saccharomyces cerevisiae Proteins) RN - EC 3.6.3.14 (ATPAF2 protein, human) RN - EC 3.6.3.14 (Proton-Translocating ATPases) SB - IM MH - Abnormalities, Multiple/*genetics MH - Animals MH - *Chaperonins MH - *Chromosome Deletion MH - Chromosomes, Artificial, Bacterial/genetics MH - Chromosomes, Artificial, P1 Bacteriophage/genetics MH - Chromosomes, Human, Pair 17/*genetics MH - Comparative Study MH - Contig Mapping/methods MH - Female MH - Fungal Proteins/genetics MH - Gene Order/genetics MH - Humans MH - Male MH - Mental Retardation/*genetics MH - Mice MH - Physical Chromosome Mapping/methods MH - *Proton-Translocating ATPases MH - Pseudogenes/genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - *Saccharomyces cerevisiae Proteins MH - Syndrome MH - Synteny/*genetics EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.73702 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):713-28. PMID- 11997339 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Structure and evolution of the Smith-Magenis syndrome repeat gene clusters, SMS-REPs. PG - 729-38 AB - An approximately 4-Mb genomic segment on chromosome 17p11.2, commonly deleted in patients with the Smith-Magenis syndrome (SMS) and duplicated in patients with dup(17)(p11.2p11.2) syndrome, is flanked by large, complex low-copy repeats (LCRs), termed proximal and distal SMS-REP. A third copy, the middle SMS-REP, is located between them. SMS-REPs are believed to mediate nonallelic homologous recombination, resulting in both SMS deletions and reciprocal duplications. To delineate the genomic structure and evolutionary origin of SMS-REPs, we constructed a bacterial artificial chromosome/P1 artificial chromosome contig spanning the entire SMS region, including the SMS-REPs, determined its genomic sequence, and used fluorescence in situ hybridization to study the evolution of SMS-REP in several primate species. Our analysis shows that both the proximal SMS-REP (approximately 256 kb) and the distal copy (approximately 176 kb) are located in the same orientation and derived from a progenitor copy, whereas the middle SMS-REP (approximately 241 kb) is inverted and appears to have been derived from the proximal copy. The SMS-REP LCRs are highly homologous (>98%) and contain at least 14 genes/pseudogenes each. SMS-REPs are not present in mice and were duplicated after the divergence of New World monkeys from pre-monkeys approximately 40-65 million years ago. Our findings potentially explain why the vast majority of SMS deletions and dup(17)(p11.2p11.2) occur at proximal and distal SMS-REPs and further support previous observations that higher-order genomic architecture involving LCRs arose recently during primate speciation and may predispose the human genome to both meiotic and mitotic rearrangements. AD - Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, 77030, USA. FAU - Park, Sung-Sup AU - Park SS FAU - Stankiewicz, Pawel AU - Stankiewicz P FAU - Bi, Weimin AU - Bi W FAU - Shaw, Christine AU - Shaw C FAU - Lehoczky, Jessica AU - Lehoczky J FAU - Dewar, Ken AU - Dewar K FAU - Birren, Bruce AU - Birren B FAU - Lupski, James R AU - Lupski JR LA - eng GR - PO1 HD39420/HD/NICHD GR - R01 NS27042/NS/NINDS GR - U54 HG02045/HG/NHGRI PT - Journal Article PL - United States TA - Genome Res JID - 9518021 SB - IM MH - Abnormalities, Multiple/*genetics MH - Base Composition/genetics MH - Cell Line MH - Cell Line, Transformed MH - Chromosomes, Human, Pair 17/genetics MH - Cloning, Molecular/methods MH - Contig Mapping/methods MH - DNA Fingerprinting/methods MH - *Evolution, Molecular MH - Gene Dosage MH - Gene Duplication MH - Genome, Human MH - Humans MH - Mental Retardation/*genetics MH - Multigene Family/*genetics MH - Repetitive Sequences, Nucleic Acid/*genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Sequence Alignment/methods MH - Sequence Analysis, DNA/methods MH - Sequence Homology, Nucleic Acid MH - Syndrome EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.82802 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):729-38. PMID- 11997340 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Discovery of regulatory elements by a computational method for phylogenetic footprinting. PG - 739-48 AB - Phylogenetic footprinting is a method for the discovery of regulatory elements in a set of orthologous regulatory regions from multiple species. It does so by identifying the best conserved motifs in those orthologous regions. We describe a computer algorithm designed specifically for this purpose, making use of the phylogenetic relationships among the sequences under study to make more accurate predictions. The program is guaranteed to report all sets of motifs with the lowest parsimony scores, calculated with respect to the phylogenetic tree relating the input species. We report the results of this algorithm on several data sets of interest. A large number of known functional binding sites are identified by our method, but we also find several highly conserved motifs for which no function is yet known. AD - Department of Computer Science and Engineering, University of Washington, Seattle, Washington 98195-2350, USA. FAU - Blanchette, Mathieu AU - Blanchette M FAU - Tompa, Martin AU - Tompa M LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Fish Proteins) RN - 0 (Interleukin-3) RN - 11061-68-0 (Insulin) RN - 122784-93-4 (growth hormone type I, Salmo salar) RN - 9002-72-6 (Growth Hormone) RN - 9038-94-2 (Metallothionein) SB - IM MH - Algorithms MH - Animals MH - Comparative Study MH - Computational Biology/*methods MH - DNA Footprinting/*methods MH - *Fish Proteins MH - Genes, fos/genetics MH - Genes, myc/genetics MH - Growth Hormone/genetics MH - Humans MH - Insulin/genetics MH - Interleukin-3/genetics MH - Introns/genetics MH - Metallothionein/genetics MH - Multigene Family/genetics MH - *Phylogeny MH - Promoter Regions (Genetics)/genetics MH - Regulatory Sequences, Nucleic Acid/*genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.6902 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):739-48. PMID- 11997341 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041203 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Transcriptional regulation of the stem cell leukemia gene (SCL)--comparative analysis of five vertebrate SCL loci. PG - 749-59 AB - The stem cell leukemia (SCL) gene encodes a bHLH transcription factor with a pivotal role in hematopoiesis and vasculogenesis and a pattern of expression that is highly conserved between mammals and zebrafish. Here we report the isolation and characterization of the zebrafish SCL locus together with the identification of three neighboring genes, IER5, MAP17, and MUPP1. This region spans 68 kb and comprises the longest zebrafish genomic sequence currently available for comparison with mammalian, chicken, and pufferfish sequences. Our data show conserved synteny between zebrafish and mammalian SCL and MAP17 loci, thus suggesting the likely genomic domain necessary for the conserved pattern of SCL expression. Long-range comparative sequence analysis/phylogenetic footprinting was used to identify noncoding conserved sequences representing candidate transcriptional regulatory elements. The SCL promoter/enhancer, exon 1, and the poly(A) region were highly conserved, but no homology to other known mouse SCL enhancers was detected in the zebrafish sequence. A combined homology/structure analysis of the poly(A) region predicted consistent structural features, suggesting a conserved functional role in mRNA regulation. Analysis of the SCL promoter/enhancer revealed five motifs, which were conserved from zebrafish to mammals, and each of which is essential for the appropriate pattern or level of SCL transcription. AD - Cambridge Institute for Medical Research, Cambridge University, Cambridge, CB2 2XY, United Kingdom. bg200@cam.ac.uk FAU - Gottgens, Berthold AU - Gottgens B FAU - Barton, Linda M AU - Barton LM FAU - Chapman, Michael A AU - Chapman MA FAU - Sinclair, Angus M AU - Sinclair AM FAU - Knudsen, Bjarne AU - Knudsen B FAU - Grafham, Darren AU - Grafham D FAU - Gilbert, James G R AU - Gilbert JG FAU - Rogers, Jane AU - Rogers J FAU - Bentley, David R AU - Bentley DR FAU - Green, Anthony R AU - Green AR LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (5' Untranslated Regions) RN - 0 (Chromosomes, Artificial, P1 Bacteriophage) RN - 0 (DNA-Binding Proteins) RN - 0 (Genetic Markers) RN - 0 (Proto-Oncogene Proteins) RN - 0 (Tal1 protein, mouse) RN - 0 (Transcription Factors) RN - 0 (Zebrafish Proteins) RN - 0 (tal1 protein, zebrafish) RN - 135471-20-4 (TAL1 protein, human) RN - 24937-83-5 (Poly A) SB - IM MH - 5' Untranslated Regions/genetics MH - Amino Acid Sequence MH - Animals MH - Cell Line MH - Chickens MH - Chromosomes, Artificial, P1 Bacteriophage/genetics MH - Cloning, Molecular MH - Comparative Study MH - Conserved Sequence MH - DNA-Binding Proteins/biosynthesis/*genetics/metabolism MH - Exons/genetics MH - Gene Expression Regulation, Neoplastic/*genetics MH - Genetic Markers/genetics/physiology MH - Humans MH - Leukemia, T-Cell, Acute/*genetics MH - Mice MH - Mice, Transgenic MH - Molecular Sequence Data MH - Poly A/metabolism MH - Promoter Regions (Genetics)/genetics MH - *Proto-Oncogene Proteins MH - Rats MH - Research Support, Non-U.S. Gov't MH - Sequence Homology, Nucleic Acid MH - Tetraodontiformes MH - Transcription Factors/biosynthesis/chemistry/*genetics MH - Zebrafish/genetics MH - *Zebrafish Proteins EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.45502 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):749-59. PMID- 11997342 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Determination of redundancy and systems properties of the metabolic network of Helicobacter pylori using genome-scale extreme pathway analysis. PG - 760-9 AB - The capabilities of genome-scale metabolic networks can be described through the determination of a set of systemically independent and unique flux maps called extreme pathways. The first study of genome-scale extreme pathways for the simultaneous formation of all nonessential amino acids or ribonucleotides in Helicobacter pylori is presented. Three key results were obtained. First, the extreme pathways for the production of individual amino acids in H. pylori showed far fewer internal states per external state than previously found in Haemophilus influenzae, indicating a more rigid metabolic network. Second, the degree of pathway redundancy in H. pylori was essentially the same for the production of individual amino acids and linked amino acid sets, but was approximately twice that of the production of the ribonucleotides. Third, the metabolic network of H. pylori was unable to achieve extensive conversion of amino acids consumed to the set of either nonessential amino acids or ribonucleotides and thus diverted a large portion of its nitrogen to ammonia production, a potentially important result for pH regulation in its acidic habitat. Genome-scale extreme pathways elucidate emergent system-wide properties. Extreme pathway analysis is emerging as a potentially important method to analyze the link between the metabolic genotype and its phenotypes. AD - Department of Bioengineering, University of California at San Diego, La Jolla, California 92093, USA. FAU - Price, Nathan D AU - Price ND FAU - Papin, Jason A AU - Papin JA FAU - Palsson, Bernhard O AU - Palsson BO LA - eng GR - GM57089/GM/NIGMS PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Amino Acids) RN - 0 (Nucleotides) RN - 7440-44-0 (Carbon) RN - 7727-37-9 (Nitrogen) SB - IM MH - Amino Acids/biosynthesis/metabolism MH - Biomass MH - Carbon/metabolism MH - Comparative Study MH - Escherichia coli/metabolism MH - *Genome, Bacterial MH - Genotype MH - Helicobacter pylori/*genetics/*metabolism/physiology MH - Nitrogen/metabolism MH - Nucleotides/biosynthesis MH - Phenotype MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.218002. Article published online before print in April 2002 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):760-9. PMID- 11997343 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Evidence suggesting that a fifth of annotated Caenorhabditis elegans genes may be pseudogenes. PG - 770-5 AB - Only a minority of the genes, identified in the Caenorhabditis elegans genome sequence data by computer analysis, have been characterized experimentally. We attempted to determine the expression patterns for a random sample of the annotated genes using reporter gene fusions. A low success rate was obtained for evolutionarily recently duplicated genes. Analysis of the data suggests that this is not due to conditional or low-level expression. The remaining explanation is that most of the annotated genes in the recently duplicated category are pseudogenes, a proportion corresponding to 20% of all of the annotated C. elegans genes. Further support for this surprisingly high figure was sought by comparing sequences for families of recently duplicated C. elegans genes. Although only a preliminary analysis, clear evidence for a gene having been recently inactivated by genetic drift was found for many genes in the recently duplicated category. At least 4% of the annotated C. elegans genes can be recognized as pseudogenes simply from closer inspection of the sequence data. Lessons learned in identifying pseudogenes in C. elegans could be of value in the annotation of the genomes of other species where, although there may be fewer pseudogenes, they may be harder to detect. AD - School of Biology, University of Leeds, Leeds, LS2 9JT, United Kingdom. FAU - Mounsey, Andrew AU - Mounsey A FAU - Bauer, Petra AU - Bauer P FAU - Hope, Ian A AU - Hope IA LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Caenorhabditis elegans Proteins) SB - IM MH - Amino Acid Sequence MH - Animals MH - Caenorhabditis elegans/*genetics MH - Caenorhabditis elegans Proteins/genetics MH - Computational Biology/*methods MH - Gene Expression Regulation MH - Gene Fusion/methods MH - Genes, Reporter/genetics MH - Genes, Structural, Helminth/*genetics MH - Molecular Sequence Data MH - Pseudogenes/*genetics MH - Research Support, Non-U.S. Gov't EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr208802. Article published online before print in April 2002 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):770-5. PMID- 11997344 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Analyses of the extent of shared synteny and conserved gene orders between the genome of Fugu rubripes and human 20q. PG - 776-84 AB - Cosmid and BAC contig maps have been constructed across two Fugu genomic regions containing the orthologs of human genes mapping to human chromosome 20q. Contig gene contents have been assessed by sample sequencing and comparative database analyses. Contigs are centered around two Fugu topoisomerase1 (top1) genes that were initially identified by sequence similarity to human TOP1 (20q12). Two other genes (SNAI1 and KRML) mapping to human chromosome 20 are also duplicated in Fugu. The two contigs have been mapped to separate Fugu chromosomes. Our data indicate that these linkage groups result from the duplication of an ancestral chromosome segment containing at least 40 genes that now map to the long arm of human chromosome 20. Although there is considerable conservation of synteny, gene orders are not well conserved between Fugu and human, with only very short sections of two to three adjacent genes being maintained in both organisms. Comparative analyses have allowed this duplication event to be dated before the separation of Fugu and zebrafish. Our data (which are best explained by regional duplication, followed by substantial gene loss) support the hypothesis that there have been a large number of gene and regional duplications (and corresponding gene loss) in the fish lineage, possibly resulting from a single whole genome duplication event. AD - Fugu Genomics, United Kingdom Human Genome Mapping Project Resource Centre, Wellcome Genome Campus, Hinxton Hall, Hinxton, Cambridgeshire, CB10 1SB, United Kingdom. sfsmith@hgmp.mrc.ac.uk FAU - Smith, Sarah F AU - Smith SF FAU - Snell, Philip AU - Snell P FAU - Gruetzner, Frank AU - Gruetzner F FAU - Bench, Anthony J AU - Bench AJ FAU - Haaf, Thomas AU - Haaf T FAU - Metcalfe, Judith A AU - Metcalfe JA FAU - Green, Anthony R AU - Green AR FAU - Elgar, Greg AU - Elgar G LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Cosmids) SB - IM MH - Animals MH - Cell Line MH - Chromosomes, Human, Pair 20/*genetics MH - Comparative Study MH - Conserved Sequence/*genetics MH - Contig Mapping/methods MH - Cosmids/genetics MH - Gene Duplication MH - Gene Order/*genetics MH - *Genome MH - Genome, Human MH - Humans MH - Microscopy, Fluorescence MH - Sequence Tagged Sites MH - Synteny/*genetics MH - Takifugu/*genetics MH - Zebrafish/genetics EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.221802. Article published online before print in April 2002 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):776-84. PMID- 11997345 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Large-scale protein annotation through gene ontology. PG - 785-94 AB - Recent progress in genomic sequencing, computational biology, and ontology development has presented an opportunity to investigate biological systems from a unique perspective, that is, examining genomes and transcriptomes through the multiple and hierarchical structure of Gene Ontology (GO). We report here our development of GO Engine, a computational platform for GO annotation, and analysis of the resultant GO annotations of human proteins. Protein annotation was centered on sequence homology with GO-annotated proteins and protein domain analysis. Text information analysis and a multiparameter cellular localization predictive tool were also used to increase the annotation accuracy, and to predict novel annotations. The majority of proteins corresponding to full-length mRNA in GenBank, and the majority of proteins in the NR database (nonredundant database of proteins) were annotated with one or more GO nodes in each of the three GO categories. The annotations of GenBank and SWISS-PROT proteins are available to the public at the GO Consortium web site. AD - Compugen Inc., Jamesburg, New Jersey 08831, USA. han@cgen.com FAU - Xie, Hanqing AU - Xie H FAU - Wasserman, Alon AU - Wasserman A FAU - Levine, Zurit AU - Levine Z FAU - Novik, Amit AU - Novik A FAU - Grebinskiy, Vladimir AU - Grebinskiy V FAU - Shoshan, Avi AU - Shoshan A FAU - Mintz, Liat AU - Mintz L LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Proteins) SB - IM MH - Animals MH - Computational Biology/*methods MH - Databases, Genetic MH - Databases, Protein MH - Genome, Human MH - Humans MH - Multigene Family MH - Proteins/*classification/*genetics/physiology MH - Sequence Analysis, Protein MH - Sequence Homology, Amino Acid EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.86902 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):785-94. PMID- 11997346 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Integration of Cot analysis, DNA cloning, and high-throughput sequencing facilitates genome characterization and gene discovery. PG - 795-807 AB - Cot-based sequence discovery represents a powerful means by which both low-copy and repetitive sequences can be selectively and efficiently fractionated, cloned, and characterized. Based upon the results of a Cot analysis, hydroxyapatite chromatography was used to fractionate sorghum (Sorghum bicolor) genomic DNA into highly repetitive (HR), moderately repetitive (MR), and single/low-copy (SL) sequence components that were consequently cloned to produce HRCot, MRCot, and SLCot genomic libraries. Filter hybridization (blotting) and sequence analysis both show that the HRCot library is enriched in sequences traditionally found in high-copy number (e.g., retroelements, rDNA, centromeric repeats), the SLCot library is enriched in low-copy sequences (e.g., genes and "nonrepetitive ESTs"), and the MRCot library contains sequences of moderate redundancy. The Cot analysis suggests that the sorghum genome is approximately 700 Mb (in agreement with previous estimates) and that HR, MR, and SL components comprise 15%, 41%, and 24% of sorghum DNA, respectively. Unlike previously described techniques to sequence the low-copy components of genomes, sequencing of Cot components is independent of expression and methylation patterns that vary widely among DNA elements, developmental stages, and taxa. High-throughput sequencing of Cot clones may be a means of "capturing" the sequence complexity of eukaryotic genomes at unprecedented efficiency. AD - Center for Applied Genetic Technologies and Department of Crop and Soil Sciences, University of Georgia, Athens, Georgia 30602, USA. dgp@arches.uga.edu FAU - Peterson, Daniel G AU - Peterson DG FAU - Schulze, Stefan R AU - Schulze SR FAU - Sciara, Erica B AU - Sciara EB FAU - Lee, Scott A AU - Lee SA FAU - Bowers, John E AU - Bowers JE FAU - Nagel, Alexander AU - Nagel A FAU - Jiang, Ning AU - Jiang N FAU - Tibbitts, Deanne C AU - Tibbitts DC FAU - Wessler, Susan R AU - Wessler SR FAU - Paterson, Andrew H AU - Paterson AH LA - eng SI - GENBANK/AZ921847 SI - GENBANK/AZ921848 SI - GENBANK/AZ921849 SI - GENBANK/AZ921850 SI - GENBANK/AZ921851 SI - GENBANK/AZ921852 SI - GENBANK/AZ921853 SI - GENBANK/AZ921854 SI - GENBANK/AZ921855 SI - GENBANK/AZ921856 SI - GENBANK/AZ921857 SI - GENBANK/AZ921858 SI - GENBANK/AZ921859 SI - GENBANK/AZ921860 SI - GENBANK/AZ921861 SI - GENBANK/AZ921862 SI - GENBANK/AZ921863 SI - GENBANK/AZ921864 SI - GENBANK/AZ921865 SI - GENBANK/AZ921866 SI - GENBANK/AZ921867 SI - 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GENBANK/AZ922834 SI - GENBANK/AZ922835 SI - GENBANK/AZ922836 SI - GENBANK/AZ922837 SI - GENBANK/AZ922838 SI - GENBANK/AZ922839 SI - GENBANK/AZ922840 SI - GENBANK/AZ922841 SI - GENBANK/AZ922842 SI - GENBANK/AZ922843 SI - GENBANK/AZ922844 SI - GENBANK/AZ922845 SI - GENBANK/AZ922846 PT - Journal Article PL - United States TA - Genome Res JID - 9518021 RN - 0 (Chromosomes, Artificial, Bacterial) RN - 0 (Genetic Markers) RN - 0 (Plant Proteins) RN - 0 (kafirin protein, Sorghum bicolor) SB - IM MH - Base Composition/genetics MH - Blotting, Southern/methods MH - Chromosomes, Artificial, Bacterial/genetics MH - Cloning, Molecular/*methods MH - Expressed Sequence Tags MH - GC Rich Sequence/genetics MH - Gene Dosage MH - *Genes, Structural, Plant MH - Genetic Markers/genetics MH - *Genome, Plant MH - Genomic Library MH - Molecular Sequence Data MH - Nucleic Acid Denaturation MH - Nucleic Acid Hybridization MH - Plant Proteins/genetics MH - Poaceae/*genetics MH - Repetitive Sequences, Nucleic Acid/genetics MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Sequence Analysis, DNA/*methods MH - Temperature EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.226102. Article published online before print in April 2002 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):795-807. PMID- 11997347 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - Protein coding palindromes are a unique but recurrent feature in Rickettsia. PG - 808-16 AB - Rickettsia are unique in inserting in-frame a number of palindromic sequences within protein coding regions. In this study, we extensively analyzed repeated sequences in the genome of Rickettsia conorii and examined their locations in regard to coding versus noncoding regions. We identified 656 interspersed repeated sequences classified into 10 distinct families. Of the 10 families, three palindromic sequence families showed clear cases of insertions into open reading frames (ORFs). The location of those in-frame insertions appears to be always compatible with the encoded protein three-dimensional (3-D) fold and function. We provide evidence for a progressive loss of the palindromic property over time after the insertions. This comprehensive study of Rickettsia repeats confirms and extends our previous observations and further indicates a significant role of selfish DNAs in the creation and modification of proteins. AD - Information Genetique & Structurale, CNRS-AVENTIS UMR 1889, 13402 Marseille Cedex 20, France. Hiroyuki.Ogata@igs.cnrs-mrs.fr FAU - Ogata, Hiroyuki AU - Ogata H FAU - Audic, Stephane AU - Audic S FAU - Abergel, Chantal AU - Abergel C FAU - Fournier, Pierre-Edouard AU - Fournier PE FAU - Claverie, Jean-Michel AU - Claverie JM LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Bacterial Proteins) SB - IM MH - Amino Acid Sequence/genetics MH - Bacterial Proteins/chemistry/*genetics MH - Base Composition/genetics MH - Base Sequence MH - Computational Biology/methods MH - *Genome, Bacterial MH - Molecular Sequence Data MH - Protein Structure, Quaternary/genetics MH - Rickettsia conorii/chemistry/*genetics EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.227602. Article published online before print in April 2002 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):808-16. PMID- 11997348 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - A fine physical map of the rice chromosome 4. PG - 817-23 AB - As part of an international effort to completely sequence the rice genome, we have produced a fine bacterial artificial chromosome (BAC)-based physical map of the Oryza sativa japonica Nipponbare chromosome 4 through an integration of 114 sequenced BAC clones from a taxonomically related subspecies O. sativa indica Guangluai 4 and 182 RFLP and 407 expressed sequence tag (EST) markers with the fingerprinted data of the Nipponbare genome. The map consists of 11 contigs with a total length of 34.5 Mb covering 94% of the estimated chromosome size (36.8 Mb). BAC clones corresponding to telomeres, as well as to the centromere position, were determined by BAC-pachytene chromosome fluorescence in situ hybridization (FISH). This gave rise to an estimated length ratio of 5.13 for the long arm and 2.9 for the short arm (on the basis of the physical map), which indicates that the short arm is a highly condensed one. The FISH analysis and physical mapping also showed that the short arm and the pericentromeric region of the long arm are rich in heterochromatin, which occupied 45% of the chromosome, indicating that this chromosome is likely very difficult to sequence. To our knowledge, this map provides the first example of a rapid and reliable physical mapping on the basis of the integration of the data from two taxonomically related subspecies. AD - National Center for Gene Research, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200233, China. FAU - Zhao, Qiang AU - Zhao Q FAU - Zhang, Yu AU - Zhang Y FAU - Cheng, Zhukuan AU - Cheng Z FAU - Chen, Mingsheng AU - Chen M FAU - Wang, Shengyue AU - Wang S FAU - Feng, Qi AU - Feng Q FAU - Huang, Yucheng AU - Huang Y FAU - Li, Ying AU - Li Y FAU - Tang, Yesheng AU - Tang Y FAU - Zhou, Bo AU - Zhou B FAU - Chen, Zhehua AU - Chen Z FAU - Yu, Shuliang AU - Yu S FAU - Zhu, Jingjie AU - Zhu J FAU - Hu, Xin AU - Hu X FAU - Mu, Jie AU - Mu J FAU - Ying, Kai AU - Ying K FAU - Hao, Pei AU - Hao P FAU - Zhang, Lei AU - Zhang L FAU - Lu, Yiqi AU - Lu Y FAU - Zhang, Lei S AU - Zhang LS FAU - Liu, Yilei AU - Liu Y FAU - Yu, Zhen AU - Yu Z FAU - Fan, Danlin AU - Fan D FAU - Weng, Qijun AU - Weng Q FAU - Chen, Ling AU - Chen L FAU - Lu, Tingting AU - Lu T FAU - Liu, Xiaohui AU - Liu X FAU - Jia, Peixin AU - Jia P FAU - Sun, Tongguo AU - Sun T FAU - Wu, Yongrui AU - Wu Y FAU - Zhang, Yujun AU - Zhang Y FAU - Lu, Ying AU - Lu Y FAU - Li, Can AU - Li C FAU - Wang, Rong AU - Wang R FAU - Lei, Haiyan AU - Lei H FAU - Li, Tao AU - Li T FAU - Hu, Hao AU - Hu H FAU - Wu, Mei AU - Wu M FAU - Zhang, Runquan AU - Zhang R FAU - Guan, Jianping AU - Guan J FAU - Zhu, Jia AU - Zhu J FAU - Fu, Gang AU - Fu G FAU - Gu, Minghong AU - Gu M FAU - Hong, Guofan AU - Hong G FAU - Xue, Yongbiao AU - Xue Y FAU - Wing, Rod AU - Wing R FAU - Jiang, Jiming AU - Jiang J FAU - Han, Bin AU - Han B LA - eng PT - Letter PL - United States TA - Genome Res JID - 9518021 RN - 0 (Chromosomes, Artificial, Bacterial) RN - 0 (DNA, Plant) SB - IM MH - Chromosomes/chemistry/*genetics MH - Chromosomes, Artificial, Bacterial MH - Contig Mapping MH - Cytogenetic Analysis/methods MH - DNA, Plant/genetics MH - In Situ Hybridization, Fluorescence/methods MH - Oryza sativa/chemistry/*genetics MH - Physical Chromosome Mapping/*methods MH - Recombination, Genetic/genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Seeds/genetics EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.48902 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):817-23. PMID- 11997349 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - RePS: a sequence assembler that masks exact repeats identified from the shotgun data. PG - 824-31 AB - We describe a sequence assembler, RePS (repeat-masked Phrap with scaffolding), that explicitly identifies exact 20mer repeats from the shotgun data and removes them prior to the assembly. The established software is used to compute meaningful error probabilities for each base. Clone-end-pairing information is used to construct scaffolds that order and orient the contigs. We show with real data for human and rice that reasonable assemblies are possible even at coverages of only 4x to 6x, despite having up to 42.2% in exact repeats. AD - Hangzhou Genomics Institute, Institute of Bioinformatics of Zhejiang University, Key Laboratory of Bioinformatics of Zhejiang Province, Hangzhou 310007, China. wangj@genomics.org.cn FAU - Wang, Jun AU - Wang J FAU - Wong, Gane Ka-Shu AU - Wong GK FAU - Ni, Peixiang AU - Ni P FAU - Han, Yujun AU - Han Y FAU - Huang, Xiangang AU - Huang X FAU - Zhang, Jianguo AU - Zhang J FAU - Ye, Chen AU - Ye C FAU - Zhang, Yong AU - Zhang Y FAU - Hu, Jianfei AU - Hu J FAU - Zhang, Kunlin AU - Zhang K FAU - Xu, Xin AU - Xu X FAU - Cong, Lijuan AU - Cong L FAU - Lu, Hong AU - Lu H FAU - Ren, Xide AU - Ren X FAU - Ren, Xiaoyu AU - Ren X FAU - He, Jun AU - He J FAU - Tao, Lin AU - Tao L FAU - Passey, Douglas A AU - Passey DA FAU - Wang, Jian AU - Wang J FAU - Yang, Huanming AU - Yang H FAU - Yu, Jun AU - Yu J FAU - Li, Songgang AU - Li S LA - eng GR - 1 R01 ES09909/ES/NIEHS PT - Journal Article PL - United States TA - Genome Res JID - 9518021 SB - IM MH - Cloning, Molecular/methods MH - Comparative Study MH - Contig Mapping/*methods MH - Humans MH - Oryza sativa/genetics MH - Repetitive Sequences, Nucleic Acid/*genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Sequence Analysis, DNA/methods MH - *Software EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.165102 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):824-31. PMID- 11997350 OWN - NLM STAT- MEDLINE DA - 20020508 DCOM- 20020605 LR - 20041117 PUBM- Print IS - 1088-9051 VI - 12 IP - 5 DP - 2002 May TI - rVista for comparative sequence-based discovery of functional transcription factor binding sites. PG - 832-9 AB - Identifying transcriptional regulatory elements represents a significant challenge in annotating the genomes of higher vertebrates. We have developed a computational tool, rVista, for high-throughput discovery of cis-regulatory elements that combines clustering of predicted transcription factor binding sites (TFBSs) and the analysis of interspecies sequence conservation to maximize the identification of functional sites. To assess the ability of rVista to discover true positive TFBSs while minimizing the prediction of false positives, we analyzed the distribution of several TFBSs across 1 Mb of the well-annotated cytokine gene cluster (Hs5q31; Mm11). Because a large number of AP-1, NFAT, and GATA-3 sites have been experimentally identified in this interval, we focused our analysis on the distribution of all binding sites specific for these transcription factors. The exploitation of the orthologous human-mouse dataset resulted in the elimination of > 95% of the approximately 58,000 binding sites predicted on analysis of the human sequence alone, whereas it identified 88% of the experimentally verified binding sites in this region. AD - Genome Sciences Department, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA. ggloots@lbl.gov FAU - Loots, Gabriela G AU - Loots GG FAU - Ovcharenko, Ivan AU - Ovcharenko I FAU - Pachter, Lior AU - Pachter L FAU - Dubchak, Inna AU - Dubchak I FAU - Rubin, Edward M AU - Rubin EM LA - eng PT - Journal Article PT - Validation Studies PL - United States TA - Genome Res JID - 9518021 RN - 0 (Cytokines) RN - 0 (Transcription Factors) SB - IM MH - Animals MH - Base Sequence/genetics MH - Binding Sites/genetics MH - Comparative Study MH - Computational Biology/methods MH - Cytokines/chemistry/genetics MH - Humans MH - Mice MH - Multigene Family/genetics MH - Promoter Regions (Genetics)/genetics MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Research Support, U.S. Gov't, P.H.S. MH - *Software MH - Transcription Factors/*chemistry/*genetics EDAT- 2002/05/09 10:00 MHDA- 2002/06/06 10:01 AID - 10.1101/gr.225502. Article published online before print in April 2002 [doi] PST - ppublish SO - Genome Res 2002 May;12(5):832-9. PMID- 12006105 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030423 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 May 13 TI - Management of obstetric anal sphincter injury: a systematic review & national practice survey. PG - 9 AB - BACKGROUND: We aim to establish the evidence base for the recognition and management of obstetric anal sphincter injury (OASI) and to compare this with current practice amongst UK obstetricians and coloproctologists. METHODS: A systematic review of the literature and a postal questionnaire survey of consultant obstetricians, trainee obstetricians and consultant coloproctologists was carried out. RESULTS: We found a wide variation in experience of repairing acute anal sphincter injury. The group with largest experience were consultant obstetricians (46.5% undertaking > or = 5 repairs/year), whilst only 10% of responding colorectal surgeons had similar levels of experience (p < 0.001). There was extensive misunderstanding in terms of the definition of obstetric anal sphincter injuries. Overall, trainees had a greater knowledge of the correct classification (p < 0.01). Observational studies suggest that a new 'overlap' repair using PDS sutures with antibiotic cover gives better functional results. However, our literature search found only one randomised controlled trial (RCT) on the technique of repair of OASI, which showed no difference in incidence of anal incontinence at three months. Despite this, there was a wide variation in practice, with 337(50%) consultants, 82 (55%) trainees and 80 (89%) coloproctologists already using the 'overlap' method for repair of a torn EAS (p < 0.001). Although over 50% of colorectal surgeons would undertake long-term follow-up of their patients, this was the practice of less than 10% of obstetricians (p < 0.001). Whilst over 70% of coloproctologists would recommend an elective caesarean section in a subsequent pregnancy, only 22% of obstetric consultants and 14% of trainees (p < 0.001). CONCLUSION: An agreed classification of OASI, development of national guidelines, formalised training, multidisciplinary management and further definitive research is strongly recommended. AD - Academic Department of Obstetrics & Gynaecology North Staffordshire Hospital Trust/Keele University Stoke on Trent, England. ruwanfernando@hotmail.com FAU - Fernando, Ruwan J AU - Fernando RJ FAU - Sultan, Abdul H AU - Sultan AH FAU - Radley, Simon AU - Radley S FAU - Jones, Peter W AU - Jones PW FAU - Johanson, Richard B AU - Johanson RB LA - eng PT - Journal Article PT - Review DEP - 20020513 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Anus/*injuries MH - Clinical Competence MH - Colorectal Surgery/methods/*standards MH - Continuity of Patient Care MH - Delivery, Obstetric/*adverse effects MH - Fecal Incontinence/etiology/surgery MH - Female MH - Great Britain/epidemiology MH - Humans MH - Labor Complications/diagnosis/epidemiology/*surgery MH - Obstetrics/education/methods/*standards MH - Patient Care Management MH - *Physician's Practice Patterns MH - Pregnancy MH - Randomized Controlled Trials MH - Research Support, Non-U.S. Gov't MH - Rupture/diagnosis/epidemiology/*surgery MH - Treatment Outcome RF - 55 EDAT- 2002/05/15 10:00 MHDA- 2003/04/24 05:00 PHST- 2001/09/27 [received] PHST- 2002/05/13 [accepted] PHST- 2002/05/13 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 May 13;2(1):9. PMID- 12014993 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021216 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Apr 25 TI - Mitochondria from cultured cells derived from normal and thiamine-responsive megaloblastic anemia individuals efficiently import thiamine diphosphate. PG - 8 AB - BACKGROUND: Thiamine diphosphate (ThDP) is the active form of thiamine, and it serves as a cofactor for several enzymes, both cytosolic and mitochondrial. Isolated mitochondria have been shown to take up thiamine yet thiamine diphosphokinase is cytosolic and not present in mitochondria. Previous reports indicate that ThDP can also be taken up by rat mitochondria, but the kinetic constants associated with such uptake seemed not to be physiologically relevant. RESULTS: Here we examine ThDP uptake by mitochondria from several human cell types, including cells from patients with thiamine-responsive megaloblastic anemia (TRMA) that lack a functional thiamine transporter of the plasma membrane. Although mitochondria from normal lymphoblasts took up thiamine in the low micromolar range, surprisingly mitochondria from TRMA lymphoblasts lacked this uptake component. ThDP was taken up efficiently by mitochondria isolated from either normal or TRMA lymphoblasts. Uptake was saturable and biphasic with a high affinity component characterized by a Km of 0.4 to 0.6 microM. Mitochondria from other cell types possessed a similar high affinity uptake component with variation seen in uptake capacity as revealed by differences in Vmax values. CONCLUSIONS: The results suggest a shared thiamine transporter for mitochondria and the plasma membrane. Additionally, a high affinity component of ThDP uptake by mitochondria was identified with the apparent affinity constant less than the estimates of the cytosolic concentration of free ThDP. This finding indicates that the high affinity uptake is physiologically significant and may represent the main mechanism for supplying phosphorylated thiamine for mitochondrial enzymes. AD - Department of Biological Sciences, Vanderbilt University, VU Station B 351634, Nashville TN 37235-1634, USA. tony.song@vanderbilt.edu FAU - Song, Qilin AU - Song Q FAU - Singleton, Charles K AU - Singleton CK LA - eng GR - AA12014-02/AA/NIAAA PT - Journal Article DEP - 20020425 PL - England TA - BMC Biochem JID - 101084098 RN - 154-87-0 (Thiamine Pyrophosphate) RN - 59-43-8 (Thiamine) SB - IM MH - Anemia, Megaloblastic/drug therapy/*metabolism MH - Animals MH - Biological Transport MH - CHO Cells MH - Cell Line MH - Hamsters MH - Humans MH - Kinetics MH - Lymphocytes/cytology/metabolism MH - Mitochondria/*metabolism MH - Research Support, U.S. Gov't, P.H.S. MH - Thiamine/*therapeutic use MH - Thiamine Pyrophosphate/*pharmacokinetics MH - Tumor Cells, Cultured EDAT- 2002/05/17 10:00 MHDA- 2002/12/18 04:00 PHST- 2002/02/06 [received] PHST- 2002/04/25 [accepted] PHST- 2002/04/25 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Apr 25;3(1):8. PMID- 12019031 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021218 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 May 4 TI - Expression of monolysocardiolipin acyltransferase activity is regulated in concert with the level of cardiolipin and cardiolipin biosynthesis in the mammalian heart. PG - 9 AB - BACKGROUND: Monolysocardiolipin acyltransferase (MLCL AT) catalyzes the acylation of monolysocardiolipin to cardiolipin in mammalian tissues. We previously reported that cardiac cardiolipin levels, MLCL AT and cardiolipin synthase activities were all elevated in rats made hyperthyroid by thyroxine treatment. In this study, we examined if cardiac mitochondrial MLCL AT activity was dependent upon the biosynthesis and level of cardiolipin in the heart. Rat heart mitochondrial MLCL AT activity was determined under conditions in which the levels of cardiac cardiolipin and cardiolipin synthase activity were either reduced or unaltered using four different disease models in the rat. In addition, these parameters were examined in a murine model of cardiac cell differentiation. RESULTS: In rats made hypothyroid by treatment with 6-n-propyl-2-thiouracil in the drinking water for 34 days, cardiac cardiolipin content was decreased 29% (p < 0.025) and this was associated with a 32% decrease (p < 0.025) in cardiolipin synthase and a 35% reduction (p < 0.025) in MLCL AT activities. Streptozotocin-induced diabetes or hyperinsulinemia in rats did not affect cardiac cardiolipin content nor MLCL AT and cardiolipin synthase activities. Finally, cardiolipin content, MLCL AT and cardiolipin synthase activities were unaltered during murine P19 teratocarcinoma cell differentiation into cardiac myocytes. In all models, phospholipase A2 activities were unaltered compared with controls. CONCLUSION: We propose a general model in which the expression of MLCL AT activity is regulated in concert with the biosynthesis and level of cardiolipin in the heart. AD - Department of Pharmacology, Centre on Aging, University of Manitoba, Winnipeg, Canada. taylorw@ms.umanitoba.ca FAU - Taylor, William A AU - Taylor WA FAU - Xu, Fred Y AU - Xu FY FAU - Ma, Brian J AU - Ma BJ FAU - Mutter, Thomas C AU - Mutter TC FAU - Dolinsky, Vernon W AU - Dolinsky VW FAU - Hatch, Grant M AU - Hatch GM LA - eng PT - Journal Article DEP - 20020504 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Cardiolipins) RN - 0 (Membrane Proteins) RN - 0 (RNA, Messenger) RN - EC 2.3. (Acyltransferases) RN - EC 2.3.1.- (monolysocardiolipin acyltransferase) RN - EC 2.7.8 (Transferases (Other Substituted Phosphate Groups)) RN - EC 2.7.8.- (cardiolipin synthetase) SB - IM MH - Acyltransferases/genetics/*metabolism MH - Animals MH - Cardiolipins/*metabolism MH - Cell Differentiation MH - Diabetes Mellitus, Experimental/enzymology/metabolism MH - Gene Expression Regulation, Enzymologic MH - Hyperinsulinism/enzymology/metabolism MH - Hypothyroidism/enzymology/metabolism MH - Male MH - *Membrane Proteins MH - Myocardium/cytology/*metabolism MH - RNA, Messenger/genetics/metabolism MH - Rats MH - Rats, Sprague-Dawley MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Transferases (Other Substituted Phosphate Groups)/metabolism MH - Tumor Cells, Cultured EDAT- 2002/05/23 10:00 MHDA- 2002/12/19 04:00 PHST- 2002/02/06 [received] PHST- 2002/05/04 [accepted] PHST- 2002/05/04 [aheadofprint] PST - epublish SO - BMC Biochem 2002 May 4;3(1):9. PMID- 12022922 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021216 LR - 20041027 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Apr 4 TI - Identification of protein tyrosine phosphatase 1B and casein as substrates for 124-v-Mos. PG - 6 AB - BACKGROUND: The mos proto-oncogene encodes a cytoplasmic serine/threonine-specific protein kinase with crucial function during meiotic cell division in vertebrates. Based on oncogenic amino acid substitutions the viral derivative, 124-v-Mos, displays constitutive protein kinase activity and functions independent of unknown upstream effectors of mos protein kinase. We have utilized this property of 124-v-Mos and screened for novel mos substrates in immunocomplex kinase assays in vitro. RESULTS: We generated recombinant 124-v-Mos using the baculovirus expression system in Spodoptera frugiperda cells and demonstrated constitutive kinase activity by the ability of 124-v-Mos to auto-phosphorylate and to phosphorylate vimentin, a known substrate of c-Mos. Using this approach we analyzed a panel of acidic and basic substrates in immunocomplex protein kinase assays and identified novel in vitro substrates for 124-v-Mos, the protein tyrosine phosphatase 1B (PTP1B), alpha-casein and beta-casein. We controlled mos-specific phosphorylation of PTP1B and casein in comparative assays using a synthetic kinase-inactive 124-v-Mos mutant and further, tryptic digests of mos-phosphorylated beta-casein identified a phosphopeptide specifically targeted by wild-type 124-v-Mos. Two-dimensional phosphoamino acid analyses showed that 124-v-mos targets serine and threonine residues for phosphorylation in casein at a 1:1 ratio but auto-phosphorylation occurs predominantly on serine residues. CONCLUSION: The mos substrates identified in this study represent a basis to approach the identification of the mos-consensus phosphorylation motif, important for the development of specific inhibitors of the Mos protein kinase. AD - Temple University, Fels Institute for Cancer Research and Molecular Biology, Philadelphia, PA, USA. alexandropolis@yahoo.com FAU - Proikas-Cezanne, Tassula AU - Proikas-Cezanne T FAU - Stabel, Silvia AU - Stabel S FAU - Riethmacher, Dieter AU - Riethmacher D LA - eng PT - Journal Article DEP - 20020404 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Caseins) RN - 0 (Peptide Fragments) RN - 0 (Recombinant Fusion Proteins) RN - 0 (Tubulin) RN - 0 (Vimentin) RN - EC 2.7.1.37 (Protein-Serine-Threonine Kinases) RN - EC 2.7.1.37 (Proto-Oncogene Proteins c-mos) RN - EC 3.1.3.48 (Protein-Tyrosine-Phosphatase) RN - EC 3.1.3.48 (protein tyrosine phosphatase 1B) SB - IM MH - Animals MH - Binding Sites MH - Caseins/*metabolism MH - Cell Line MH - Peptide Fragments/genetics/metabolism MH - Phosphorylation MH - Protein-Serine-Threonine Kinases/metabolism MH - Protein-Tyrosine-Phosphatase/*metabolism MH - Proto-Oncogene Proteins c-mos/chemistry/genetics/*metabolism MH - Recombinant Fusion Proteins/genetics/metabolism MH - Substrate Specificity MH - Tubulin/metabolism MH - Vimentin/metabolism EDAT- 2002/05/23 10:00 MHDA- 2002/12/18 04:00 PHST- 2002/02/01 [received] PHST- 2002/04/04 [accepted] PHST- 2002/04/04 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Apr 4;3(1):6. PMID- 12028592 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 May 17 TI - The regulation of protein synthesis and translation factors by CD3 and CD28 in human primary T lymphocytes. PG - 11 AB - BACKGROUND: Activation of human resting T lymphocytes results in an immediate increase in protein synthesis. The increase in protein synthesis after 16-24 h has been linked to the increased protein levels of translation initiation factors. However, the regulation of protein synthesis during the early onset of T cell activation has not been studied in great detail. We studied the regulation of protein synthesis after 1 h of activation using alphaCD3 antibody to stimulate the T cell receptor and alphaCD28 antibody to provide the co-stimulus. RESULTS: Activation of the T cells with both antibodies led to a sustained increase in the rate of protein synthesis. The activities and/or phosphorylation states of several translation factors were studied during the first hour of stimulation with alphaCD3 and alphaCD28 to explore the mechanism underlying the activation of protein synthesis. The initial increase in protein synthesis was accompanied by activation of the guanine nucleotide exchange factor, eukaryotic initiation factor (eIF) 2B, and of p70 S6 kinase and by dephosphorylation of eukaryotic elongation factor (eEF) 2. Similar signal transduction pathways, as assessed using signal transduction inhibitors, are involved in the regulation of protein synthesis, eIF2B activity and p70 S6 kinase activity. A new finding was that the p38 MAPK alpha/beta pathway was involved in the regulation of overall protein synthesis in primary T cells. Unexpectedly, no changes were detected in the phosphorylation state of the cap-binding protein eIF4E and the eIF4E-binding protein 4E-BP1, or the formation of the cap-binding complex eIF4F. CONCLUSIONS: Both eIF2B and p70 S6 kinase play important roles in the regulation of protein synthesis during the early onset of T cell activation. AD - Division of Molecular Physiology, School of Life Sciences, University of Dundee, Dundee, MSI/Wellcome Trust Biocentre, DD1 5EH United Kingdom. m.scheperkleijn@dundee.ac.uk FAU - Kleijn, Miranda AU - Kleijn M FAU - Proud, Christopher G AU - Proud CG LA - eng PT - Journal Article DEP - 20020517 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Antibodies) RN - 0 (Antigens, CD28) RN - 0 (Antigens, CD3) RN - 0 (Carrier Proteins) RN - 0 (EIF4EBP1 protein, human) RN - 0 (Eukaryotic Initiation Factor-2B) RN - 0 (Eukaryotic Initiation Factor-4E) RN - 0 (Eukaryotic Initiation Factors) RN - 0 (Phosphoproteins) RN - EC 2.7.1.37 (Ribosomal Protein S6 Kinases, 70-kDa) SB - IM MH - Antibodies/pharmacology MH - Antigens, CD28/immunology/physiology MH - Antigens, CD3/immunology/physiology MH - Carrier Proteins/metabolism MH - Cells, Cultured MH - Eukaryotic Initiation Factor-2B/metabolism MH - Eukaryotic Initiation Factor-4E/metabolism MH - Eukaryotic Initiation Factors/*metabolism MH - Humans MH - Kinetics MH - *Lymphocyte Activation MH - Phosphoproteins/metabolism MH - Phosphorylation MH - *Protein Biosynthesis MH - Research Support, Non-U.S. Gov't MH - Ribosomal Protein S6 Kinases, 70-kDa/metabolism MH - Signal Transduction MH - T-Lymphocytes/immunology/*metabolism EDAT- 2002/05/25 10:00 MHDA- 2002/11/26 04:00 PHST- 2002/03/14 [received] PHST- 2002/05/17 [accepted] PHST- 2002/05/17 [aheadofprint] PST - epublish SO - BMC Biochem 2002 May 17;3(1):11. PMID- 12052258 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030423 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 May 27 TI - Hospital competition, resource allocation and quality of care. PG - 10 AB - BACKGROUND: A variety of approaches have been used to contain escalating hospital costs. One approach is intensifying price competition. The increase in price based competition, which changes the incentives hospitals face, coupled with the fact that consumers can more easily evaluate the quality of hotel services compared with the quality of clinical care, may lead hospitals to allocate more resources into hotel rather than clinical services. METHODS: To test this hypothesis we studied hospitals in California in 1982 and 1989, comparing resource allocations prior to and following selective contracting, a period during which the focus of competition changed from quality to price. We estimated the relationship between clinical outcomes, measured as risk-adjusted-mortality rates, and resources. RESULTS: In 1989, higher competition was associated with lower clinical expenditures levels compared with 1982. The trend was stronger for non-profit hospitals. Lower clinical resource use was associated with worse risk adjusted mortality outcomes. CONCLUSIONS: This study raises concerns that cost reductions may be associated with increased mortality. AD - University of Rochester Medical Center, Rochester, New York 14642, USA. dana_mukamel@urm.rochester.edu FAU - Mukamel, Dana B AU - Mukamel DB FAU - Zwanziger, Jack AU - Zwanziger J FAU - Bamezai, Anil AU - Bamezai A LA - eng GR - HS09545/HS/AHCPR PT - Journal Article DEP - 20020527 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - California/epidemiology MH - Cost Control/methods MH - Decision Making, Organizational MH - *Economic Competition MH - Health Expenditures/*trends MH - Health Services Research MH - Hospital Charges MH - Hospital Departments/classification/*economics/*standards MH - Hospital Mortality MH - Hospital-Patient Relations MH - Hospitals/classification MH - Humans MH - Managed Care Programs/legislation & jurisprudence/organization & administration MH - Quality of Health Care/*trends MH - Research Support, U.S. Gov't, P.H.S. MH - *Resource Allocation MH - Risk Adjustment MH - Treatment Outcome EDAT- 2002/06/08 10:00 MHDA- 2003/04/24 05:00 PHST- 2001/12/21 [received] PHST- 2002/05/27 [accepted] PHST- 2002/05/27 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 May 27;2(1):10. PMID- 12052259 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 May 29 TI - The role of the Zn(II) binding domain in the mechanism of E. coli DNA topoisomerase I. PG - 13 AB - BACKGROUND: Escherichia coli DNA topoisomerase I binds three Zn(II) with three tetracysteine motifs which, together with the 14 kDa C-terminal region, form a 30 kDa DNA binding domain (ZD domain). The 67 kDa N-terminal domain (Top67) has the active site tyrosine for DNA cleavage but cannot relax negatively supercoiled DNA. We analyzed the role of the ZD domain in the enzyme mechanism. RESULTS: Addition of purified ZD domain to Top67 partially restored the relaxation activity, demonstrating that covalent linkage between the two domains is not necessary for removal of negative supercoils from DNA. The two domains had similar affinities to ssDNA. However, only Top67 could bind dsDNA with high affinity. DNA cleavage assays showed that the Top67 had the same sequence and structure selectivity for DNA cleavage as the intact enzyme. DNA rejoining also did not require the presence of the ZD domain. CONCLUSIONS: We propose that during relaxation of negatively supercoiled DNA, Top67 by itself can position the active site tyrosine near the junction of double-stranded and single-stranded DNA for cleavage. However, the interaction of the ZD domain with the passing single-strand of DNA, coupled with enzyme conformational change, is needed for removal of negative supercoils. AD - Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY, USA. a_ahumada@msn.com FAU - Ahumada, Adriana AU - Ahumada A FAU - Tse-Dinh, Yuk-Ching AU - Tse-Dinh YC LA - eng GR - GM17315/GM/NIGMS GR - GM54226/GM/NIGMS PT - Journal Article DEP - 20020529 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (DNA, Circular) RN - 0 (DNA, Single-Stranded) RN - 0 (DNA, Superhelical) RN - 7439-95-4 (Magnesium) RN - 7440-66-6 (Zinc) RN - 7647-14-5 (Sodium Chloride) RN - 9007-49-2 (DNA) RN - EC 5.99.1.- (DNA Topoisomerases, Type I, Bacterial) SB - IM MH - Binding Sites MH - Catalytic Domain MH - DNA/chemistry/metabolism MH - DNA Topoisomerases, Type I, Bacterial/*chemistry/*metabolism MH - DNA, Circular/metabolism MH - DNA, Single-Stranded/chemistry/metabolism MH - DNA, Superhelical/metabolism MH - Escherichia coli/*enzymology MH - Magnesium/pharmacology MH - Models, Genetic MH - Protein Binding MH - Protein Structure, Tertiary MH - Research Support, U.S. Gov't, P.H.S. MH - Sodium Chloride/pharmacology MH - Substrate Specificity MH - Zinc/*metabolism EDAT- 2002/06/08 10:00 MHDA- 2002/11/26 04:00 PHST- 2002/03/11 [received] PHST- 2002/05/29 [accepted] PHST- 2002/05/29 [aheadofprint] PST - epublish SO - BMC Biochem 2002 May 29;3(1):13. PMID- 12052260 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20050201 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 May 28 TI - Mutational analysis of human profilin I reveals a second PI(4,5)-P2 binding site neighbouring the poly(L-proline) binding site. PG - 12 AB - BACKGROUND: Profilin is a small cytoskeletal protein which interacts with actin, proline-rich proteins and phosphatidylinositol 4,5-bisphosphate (PI(4,5)-P2). Crystallography, NMR and mutagenesis of vertebrate profilins have revealed the amino acid residues that are responsible for the interactions with actin and poly(L-proline) peptides. Although Arg88 of human profilin I was shown to be involved in PI(4,5)-P2-binding, it was suggested that carboxy terminal basic residues may be involved as well. RESULTS: Using site directed mutagenesis we have refined the PI(4,5)-P2 binding site of human profilin I. For each mutant we assessed the stability and studied the interactions with actin, a proline-rich peptide and PI(4,5)-P2 micelles. We identified at least two PI(4,5)-P2-binding regions in human profilin I. As expected, one region comprises Arg88 and overlaps with the actin binding site. The second region involves Arg136 in the carboxy terminal helix and neighbours the poly(L-proline) binding site. In addition, we show that adding a small protein tag to the carboxy terminus of profilin strongly reduces binding to poly(L-proline), suggesting local conformational changes of the carboxy terminal alpha-helix may have dramatic effects on ligand binding. CONCLUSIONS: The involvement of the two terminal alpha-helices of profilin in ligand binding imposes important structural constraints upon the functions of this region. Our data suggest a model in which the competitive interactions between PI(4,5)-P2 and actin and PI(4,5)-P2 and poly(L-proline) regulate profilin functions. AD - Department of Medical Protein Research (VIB09), Flanders Interuniversity Institute of Biotechnology, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium. anja.lambrechts@rug.ac.be FAU - Lambrechts, Anja AU - Lambrechts A FAU - Jonckheere, Veronique AU - Jonckheere V FAU - Dewitte, Daisy AU - Dewitte D FAU - Vandekerckhove, Joel AU - Vandekerckhove J FAU - Ampe, Christophe AU - Ampe C LA - eng PT - Journal Article DEP - 20020528 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Actins) RN - 0 (Contractile Proteins) RN - 0 (Ligands) RN - 0 (Microfilament Proteins) RN - 0 (Peptides) RN - 0 (Phosphatidylinositol 4,5-Diphosphate) RN - 0 (profilins) RN - 25191-13-3 (polyproline) RN - 73-22-3 (Tryptophan) SB - IM MH - Actins/metabolism MH - Binding Sites MH - *Contractile Proteins MH - DNA Mutational Analysis MH - Humans MH - Ligands MH - Microfilament Proteins/*chemistry/*genetics/metabolism MH - Models, Molecular MH - Mutagenesis, Site-Directed MH - Peptides/*metabolism MH - Phosphatidylinositol 4,5-Diphosphate/*metabolism MH - Protein Folding MH - Protein Structure, Secondary MH - Research Support, Non-U.S. Gov't MH - Tryptophan/physiology EDAT- 2002/06/08 10:00 MHDA- 2002/11/26 04:00 PHST- 2002/02/13 [received] PHST- 2002/05/28 [accepted] PHST- 2002/05/28 [aheadofprint] PST - epublish SO - BMC Biochem 2002 May 28;3(1):12. PMID- 12057022 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030422 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Jun 2 TI - Training practitioners in preparing systematic reviews: a cross-sectional survey of participants in the Australasian Cochrane Centre training program. PG - 11 AB - BACKGROUND: Although systematic reviews of health care interventions are an invaluable tool for health care providers and researchers, many potential authors never publish reviews. This study attempts to determine why some people with interest in performing systematic reviews do not subsequently publish a review; and what steps could possibly increase review completion. METHODS: Cross-sectional survey by email and facsimile of the 179 participants in Australasian Cochrane Centre training events between 1998 and 2000. RESULTS: Ninety-two participants responded to the survey (51 percent). Response rate of deliverable surveys was 82 percent (92/112). The remainder of the participants had invalid or no contact information on file. More than 75 percent of respondents felt that the current workshops met their needs for training. The most critical barriers to completion of a Cochrane review were: lack of time (80 percent), lack of financial support (36 percent), methodological problems (23 percent) and problems with group dynamics (10 percent). CONCLUSIONS: Strategies to protect reviewer time and increase the efficiency of the review process may increase the numbers of trained reviewers completing a systematic review. AD - Australasian Cochrane Centre, Monash Institute of Health Services Research, Monash Medical Centre, Melbourne, Australia. janet.piehl@med.monash.edu.au FAU - Piehl, Janet H AU - Piehl JH FAU - Green, Sally AU - Green S FAU - Silagy, Chris AU - Silagy C LA - eng PT - Journal Article DEP - 20020602 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - *Attitude of Health Personnel MH - Australia MH - Cross-Sectional Studies MH - Evidence-Based Medicine MH - Humans MH - *Meta-Analysis MH - Publishing/*statistics & numerical data MH - Questionnaires MH - *Randomized Controlled Trials MH - Research Personnel/*education/statistics & numerical data MH - Time EDAT- 2002/06/12 10:00 MHDA- 2003/04/23 05:00 PHST- 2002/02/20 [received] PHST- 2002/06/02 [accepted] PHST- 2002/06/02 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Jun 2;2(1):11. PMID- 12069692 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021220 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jun 10 TI - Gene expression profile of HIV-1 Tat expressing cells: a close interplay between proliferative and differentiation signals. PG - 14 AB - BACKGROUND: Expression profiling holds great promise for rapid host genome functional analysis. It is plausible that host expression profiling in an infection could serve as a universal phenotype in virally infected cells. Here, we describe the effect of one of the most critical viral activators, Tat, in HIV-1 infected and Tat expressing cells. We utilized microarray analysis from uninfected, latently HIV-1 infected cells, as well as cells that express Tat, to decipher some of the cellular changes associated with this viral activator. RESULTS: Utilizing uninfected, HIV-1 latently infected cells, and Tat expressing cells, we observed that most of the cellular host genes in Tat expressing cells were down-regulated. The down-regulation in Tat expressing cells is most apparent on cellular receptors that have intrinsic receptor tyrosine kinase (RTK) activity and signal transduction members that mediate RTK function, including Ras-Raf-MEK pathway. Co-activators of transcription, such as p300/CBP and SRC-1, which mediate gene expression related to hormone receptor genes, were also found to be down-regulated. Down-regulation of receptors may allow latent HIV-1 infected cells to either hide from the immune system or avoid extracellular differentiation signals. Some of the genes that were up-regulated included co-receptors for HIV-1 entry, translation machinery, and cell cycle regulatory proteins. CONCLUSIONS: We have demonstrated, through a microarray approach, that HIV-1 Tat is able to regulate many cellular genes that are involved in cell signaling, translation and ultimately control the host proliferative and differentiation signals. AD - Department of Biochemistry and Molecular Biology George Washington University School of Medicine Washington DC 20037, USA. bcmfxk@gwumc.edu FAU - de la Fuente, Cynthia AU - de la Fuente C FAU - Santiago, Francisco AU - Santiago F FAU - Deng, Longwen AU - Deng L FAU - Eadie, Carolyne AU - Eadie C FAU - Zilberman, Irene AU - Zilberman I FAU - Kehn, Kylene AU - Kehn K FAU - Maddukuri, Anil AU - Maddukuri A FAU - Baylor, Shanese AU - Baylor S FAU - Wu, Kaili AU - Wu K FAU - Lee, Chee Gun AU - Lee CG FAU - Pumfery, Anne AU - Pumfery A FAU - Kashanchi, Fatah AU - Kashanchi F LA - eng GR - 13969/PHS GR - AI43894/AI/NIAID GR - AI44357/AI/NIAID PT - Journal Article DEP - 20020610 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Chromatin) RN - 0 (Gene Products, tat) RN - 0 (Transcription Factors) RN - 61512-21-8 (Thymosin) SB - IM MH - Cell Cycle/genetics MH - Cell Differentiation/genetics MH - Cell Division/genetics MH - Chromatin/genetics MH - Gene Expression Profiling/*methods MH - Gene Expression Regulation/genetics MH - Gene Products, tat/*genetics MH - HIV-1/*genetics MH - Hela Cells/cytology/metabolism/virology MH - Humans MH - Protein Biosynthesis/genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Signal Transduction/*genetics MH - Thymosin/genetics MH - Transcription Factors/genetics MH - Transcription, Genetic/genetics MH - Tumor Cells, Cultured EDAT- 2002/06/19 10:00 MHDA- 2002/12/21 04:00 PHST- 2002/01/29 [received] PHST- 2002/06/10 [accepted] PHST- 2002/06/10 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jun 10;3(1):14. PMID- 12079498 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021218 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jun 17 TI - Low solubility of unconjugated bilirubin in dimethylsulfoxide--water systems: implications for pKa determinations. PG - 17 AB - BACKGROUND: Aqueous pKa values of unconjugated bilirubin are important determinants of its solubility and transport. Published pKa data on an analog, mesobilirubin-XIIIalpha, studied by 13C-NMR in buffered solutions containing 27 and 64 vol% (C2H3)2SO because of low aqueous solubility of mesobilirubin, were extrapolated to obtain pKa values in water of 4.2 and 4.9. Previous chloroform-water partition data on bilirubin diacid led to higher estimates of its pKa, 8.12 and 8.44, and its aqueous solubility. A thermodynamic analysis, using this solubility and a published solubility in DMSO, suggested that the systems used to measure 13C-NMR shifts were highly supersaturated. This expectation was assessed by measuring the residual concentrations of bilirubin in the supernatants of comparable DMSO-buffer systems, after mild centrifugation to remove microprecipitates. RESULTS: Extensive sedimentation was observed from numerous systems, many of which appeared optically clear. The very low supernatant concentrations at the lowest pH values (4.1-5.9) were compatible with the above thermodynamic analysis. Extensive sedimentation and low supernatant concentrations occurred also at pH as high as 7.2. CONCLUSIONS: The present study strongly supports the validity of the aqueous solubility of bilirubin diacid derived from partition data, and, therefore, the corresponding high pKa values. Many of the mesobilirubin systems in the 13C-NMR studies were probably supersaturated, contained microsuspensions, and were not true solutions. This, and previously documented errors in pH determinations that caused serious errors in pKa values of the many soluble reference acids and mesobilirubin, raise doubts regarding the low pKa estimates for mesobilirubin from the 13C-NMR studies. AD - School of Pharmacy, University of Wisconsin, 777 Highland Avenue, Madison, WI 53705-2222, USA. pmukerjee@aol.com FAU - Mukerjee, Pasupati AU - Mukerjee P FAU - Ostrow, J Donald AU - Ostrow JD FAU - Tiribelli, Claudio AU - Tiribelli C LA - eng PT - Journal Article DEP - 20020617 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Serum Albumin) RN - 635-65-4 (Bilirubin) RN - 67-68-5 (Dimethyl Sulfoxide) RN - 7732-18-5 (Water) SB - IM MH - Bilirubin/*chemistry MH - Dimethyl Sulfoxide/*chemistry MH - Humans MH - Hydrogen-Ion Concentration MH - Serum Albumin/chemistry MH - Solubility MH - Thermodynamics MH - Water/*chemistry EDAT- 2002/06/25 10:00 MHDA- 2002/12/19 04:00 PHST- 2002/04/23 [received] PHST- 2002/06/17 [accepted] PHST- 2002/06/17 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jun 17;3(1):17. PMID- 12079499 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021220 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jun 13 TI - Functional group interactions of a 5-HT3R antagonist. PG - 16 AB - BACKGROUND: Lerisetron, a competitive serotonin type 3 receptor (5-HT3R) antagonist, contains five functional groups capable of interacting with amino acids in the 5-HT3R binding site. Site directed mutagenesis studies of the 5-HT3AR have revealed several amino acids that are thought to form part of the binding domain of this receptor. The specific functional groups on the ligand that interact with these amino acids are, however, unknown. Using synthetic analogs of lerisetron as molecular probes in combination with site directed mutagenesis, we have identified some of these interactions and have proposed a model of the lerisetron binding site. RESULTS: Two analogs of lerisetron were synthesized to probe 5-HT3R functional group interactions with this compound. Analog 1 lacks the N1 benzyl group of lerisetron and analog 2 contains oxygen in place of the distal piperazine nitrogen. Both analogs show significantly decreased binding affinity to wildtype 5-HT3ASRs. Mutations at W89, R91, Y142 and Y152 produced significant decreases in binding compared to wildtype receptors. Binding affinities of analogs 1 and 2 were altered only by mutations at W89, and Y152. CONCLUSIONS: Based on the data obtained for lerisetron and analogs 1 and 2, we have proposed a tentative model of the lerisetron binding pocket of the 5-HT3ASR. According to this model, The N-benzyl group interacts in a weak interaction with R91 while the benzimidazole group interacts with W89. Our data support an interaction of the distal amino nitrogen with Y142 and Y152. AD - Department of Neurobiology and Physiology, Northwestern University, Evanston, IL 60208-3520, USA. padma964@hotmail.com FAU - Venkataraman, Padmavati AU - Venkataraman P FAU - Joshi, Prasad AU - Joshi P FAU - Venkatachalan, Srinivasan P AU - Venkatachalan SP FAU - Muthalagi, Mani AU - Muthalagi M FAU - Parihar, Harish S AU - Parihar HS FAU - Kirschbaum, Karen S AU - Kirschbaum KS FAU - Schulte, Marvin K AU - Schulte MK LA - eng PT - Journal Article DEP - 20020613 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Benzimidazoles) RN - 0 (Benzyl Compounds) RN - 0 (Piperazines) RN - 0 (Piperidines) RN - 0 (Receptors, Serotonin) RN - 0 (Receptors, Serotonin, 5-HT3) RN - 0 (Serotonin Antagonists) RN - 110-85-0 (piperazine) RN - 143257-98-1 (lerisetron) RN - 55520-40-6 (Tyrosine) RN - 73-22-3 (Tryptophan) RN - 74-79-3 (Arginine) RN - 7727-37-9 (Nitrogen) SB - IM MH - Animals MH - Arginine/physiology MH - Benzimidazoles/metabolism MH - Benzyl Compounds/metabolism MH - Cell Line MH - Humans MH - Kidney/cytology/embryology MH - Mice MH - Mutagenesis, Site-Directed/genetics MH - Nitrogen/metabolism MH - Patch-Clamp Techniques MH - Piperazines/chemistry/metabolism MH - Piperidines/metabolism MH - Protein Binding/physiology MH - Protein Interaction Mapping/*methods MH - Receptors, Serotonin/genetics/*metabolism MH - Receptors, Serotonin, 5-HT3 MH - Research Support, Non-U.S. Gov't MH - Serotonin Antagonists/*metabolism MH - Structure-Activity Relationship MH - Tryptophan/physiology MH - Tyrosine/physiology EDAT- 2002/06/25 10:00 MHDA- 2002/12/21 04:00 PHST- 2002/02/05 [received] PHST- 2002/06/13 [accepted] PHST- 2002/06/13 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jun 13;3(1):16. PMID- 12079500 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021220 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jun 13 TI - Identification of critical residues in loop E in the 5-HT3ASR binding site. PG - 15 AB - BACKGROUND: The serotonin type 3 receptor (5-HT3R) is a member of a superfamily of ligand gated ion channels. All members of this family share a large degree of sequence homology and presumably significant structural similarity. A large number of studies have explored the structure-function relationships of members of this family, particularly the nicotinic and GABA receptors. This information can be utilized to gain additional insights into specific structural and functional features of other receptors in this family. RESULTS: Thirteen amino acids in the mouse 5-HT3ASR that correspond to the putative E binding loop of the nicotinic alpha7 receptor were chosen for mutagenesis. Due to the presence of a highly conserved glycine in this region, it has been suggested that this binding loop is comprised of a hairpin turn and may form a portion of the ligand-binding site in this ion channel family. Mutation of the conserved glycine (G147) to alanine eliminated binding of the 5-HT3R antagonist [3H]granisetron. Three tyrosine residues (Y140, Y142 and Y152) also significantly altered the binding of 5-HT3R ligands. Mutations in neighboring residues had little or no effect on binding of these ligands to the 5-HT3ASR. CONCLUSION: Our data supports a role for the putative E-loop region of the 5-HT3R in the binding of 5-HT, mCPBG, d-tc and lerisetron. 5-HT and mCPBG interact with Y142, d-tc with Y140 and lerisetron with both Y142 and Y152. Our data also provides support for the hypothesis that this region of the receptor is present in a loop structure. AD - Department of Neurobiology, Northwestern University, Evanston, IL 60201. USA. padma964@hotmail.com FAU - Venkataraman, Padmavati AU - Venkataraman P FAU - Venkatachalan, Srinivasan P AU - Venkatachalan SP FAU - Joshi, Prasad R AU - Joshi PR FAU - Muthalagi, Mani AU - Muthalagi M FAU - Schulte, Marvin K AU - Schulte MK LA - eng PT - Journal Article DEP - 20020613 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Amino Acids) RN - 0 (Receptors, Serotonin) RN - 0 (Receptors, Serotonin, 5-HT3) RN - 55520-40-6 (Tyrosine) RN - 56-87-1 (Lysine) SB - IM MH - Amino Acid Sequence MH - Amino Acids/*analysis/genetics MH - Animals MH - Binding Sites/genetics MH - Cell Line MH - Humans MH - Kidney/chemistry/cytology/embryology/metabolism MH - Lysine/analysis/genetics MH - Mice MH - Molecular Sequence Data MH - Mutagenesis, Site-Directed/genetics MH - Protein Structure, Tertiary/genetics MH - Receptors, Serotonin/*chemistry/genetics/*physiology MH - Receptors, Serotonin, 5-HT3 MH - Research Support, Non-U.S. Gov't MH - Sequence Alignment/methods MH - Sequence Homology, Amino Acid MH - Transfection MH - Tyrosine/analysis/genetics EDAT- 2002/06/25 10:00 MHDA- 2002/12/21 04:00 PHST- 2002/01/30 [received] PHST- 2002/06/13 [accepted] PHST- 2002/06/13 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jun 13;3(1):15. PMID- 12084180 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030422 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Jun 25 TI - A comparison of hospital readmission rates between two general physicians with different outpatient review practices. PG - 12 AB - BACKGROUND: There has been a relentless increase in emergency medical admissions in the UK over recent years. Many of these patients suffer with chronic conditions requiring continuing medical attention. We wished to determine whether conventional outpatient clinic follow up after discharge has any impact on the rate of readmission to hospital. METHODS: Two consultant general physicians with the same patient case-mix but markedly different outpatient follow-up practice were chosen. Of 1203 patients discharged, one consultant saw twice as many patients in the follow-up clinic than the other (Dr A 9.8% v Dr B 19.6%). The readmission rate in the twelve months following discharge was compared in a retrospective analysis of hospital activity data. Due to the specialisation of the admitting system, patients mainly had cardiovascular or cerebrovascular disease or had taken an overdose. Few had respiratory or infectious diseases. Outpatient follow-up was focussed on patients with cardiac disease. RESULTS: Risk of readmission increased significantly with age and length of stay of the original episode and was less for digestive system and musculo-skeletal disorders. 28.7% of patients discharged by Dr A and 31.5 % of those discharged by Dr B were readmitted at least once. Relative readmission risk was not significantly different between the consultants and there was no difference in the length of stay of readmissions. CONCLUSIONS: Increasing the proportion of patients with this age- and case-mix who are followed up in a hospital general medical outpatient clinic is unlikely to reduce the demand for acute hospital beds. AD - Birmingham Heartlands and Solihull NHS Trust (Teaching), Birmingham, UK. Raynerh@heartsol.wmids.nhs.uk FAU - Rayner, Hugh C AU - Rayner HC FAU - Temple, R Mark AU - Temple RM FAU - Marshall, Tim AU - Marshall T FAU - Clarke, Dianne AU - Clarke D LA - eng PT - Journal Article DEP - 20020625 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Adolescent MH - Adult MH - Age Factors MH - Aged MH - Chronic Disease MH - Cohort Studies MH - Comparative Study MH - *Continuity of Patient Care MH - *Diagnosis-Related Groups MH - Episode of Care MH - Family Practice/*organization & administration/standards/statistics & numerical data MH - Great Britain/epidemiology MH - Humans MH - International Classification of Diseases MH - Length of Stay/statistics & numerical data MH - *Medical Audit MH - Middle Aged MH - Outpatient Clinics, Hospital/*utilization MH - Patient Readmission/*statistics & numerical data MH - Risk Factors MH - *Utilization Review EDAT- 2002/06/27 10:00 MHDA- 2003/04/23 05:00 PHST- 2001/10/22 [received] PHST- 2002/06/25 [accepted] PHST- 2002/06/25 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Jun 25;2(1):12. PMID- 12086585 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021218 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jun 18 TI - Insertion of a small peptide of six amino acids into the beta7-beta8 loop of the p51 subunit of HIV-1 reverse transcriptase perturbs the heterodimer and affects its activities. PG - 18 AB - BACKGROUND: HIV-1 RT is a heterodimeric enzyme, comprising of the p66 and p51 subunits. Earlier, we have shown that the beta7-beta8 loop of p51 is a key structural element for RT dimerization (Pandey et al., Biochemistry 40: 9505, 2001). Deletion or alanine substitution of four amino acid residues of this loop in the p51 subunit severely impaired DNA binding and catalytic activities of the enzyme. To further examine the role of this loop in HIV-1 RT, we have increased its size such that the six amino acids loop sequences are repeated in tandem and examined its impact on the dimerization process and catalytic function of the enzyme. RESULTS: The polymerase and the RNase H activities of HIV-1 RT carrying insertion in the beta7-beta8 loop of both the subunits (p66INS/p51INS) were severely impaired with substantial loss of DNA binding ability. These enzymatic activities were restored when the mutant p66INS subunit was dimerized with the wild type p51. Glycerol gradient sedimentation analysis revealed that the mutant p51INS subunit was unable to form stable dimer either with the wild type p66 or mutant p66INS. Furthermore, the p66INS/p66INS mutant sedimented as a monomeric species, suggesting its inability to form stable homodimer. CONCLUSION: The data presented herein indicates that any perturbation in the beta7-beta8 loop of the p51 subunit of HIV-1 RT affects the dimerization process resulting in substantial loss of DNA binding ability and catalytic function of the enzyme. AD - Department of Biochemistry and Molecular Biology, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, 185 South Orange Avenue, Newark, NJ 07103, USA. pandeypk@umdnj.edu FAU - Pandey, Pradeep K AU - Pandey PK FAU - Kaushik, Neerja AU - Kaushik N FAU - Singh, Kamalendra AU - Singh K FAU - Sharma, Bechan AU - Sharma B FAU - Upadhyay, Alok K AU - Upadhyay AK FAU - Kumar, Suriender AU - Kumar S FAU - Harris, Dylan AU - Harris D FAU - Pandey, Virendra N AU - Pandey VN LA - eng GR - CA72821/CA/NCI PT - Journal Article DEP - 20020618 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Amino Acids) RN - 0 (DNA-Binding Proteins) RN - 0 (Protein Subunits) RN - 9007-49-2 (DNA) RN - EC 2.7.7.- (HIV-1 Reverse Transcriptase) RN - EC 2.7.7.49 (RNA-Directed DNA Polymerase) RN - EC 2.7.7.7 (DNA-Directed DNA Polymerase) RN - EC 3.1.26.4 (Ribonuclease H, Calf Thymus) SB - IM MH - Amino Acids/genetics MH - DNA/metabolism MH - DNA-Binding Proteins/chemistry/genetics/metabolism MH - DNA-Directed DNA Polymerase/*metabolism MH - Dimerization MH - HIV-1 Reverse Transcriptase/chemistry/genetics/*metabolism MH - Models, Molecular MH - Mutagenesis, Insertional MH - Mutation MH - Protein Binding MH - *Protein Conformation MH - Protein Structure, Tertiary MH - Protein Subunits MH - RNA-Directed DNA Polymerase/metabolism MH - Research Support, U.S. Gov't, P.H.S. MH - Ribonuclease H, Calf Thymus/*metabolism MH - Ultracentrifugation/methods EDAT- 2002/06/28 10:00 MHDA- 2002/12/19 04:00 PHST- 2002/02/26 [received] PHST- 2002/06/18 [accepted] PHST- 2002/06/18 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jun 18;3(1):18. PMID- 12097150 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021218 LR - 20040924 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jun 25 TI - Molecular cloning and tissue distribution of mammalian L-threonine 3-dehydrogenases. PG - 19 AB - BACKGROUND: In mammals, L-threonine is an indispensable amino acid. The conversion of L-threonine to glycine occurs through a two-step biochemical pathway involving the enzymes L-threonine 3-dehydrogenase and 2-amino-3-ketobutyrate coenzyme A ligase. The L-threonine 3-dehydrogenase enzyme has been purified and characterised, but the L-threonine 3-dehydrogenase gene has not previously been identified in mammals. RESULTS: Transcripts for L-threonine 3-dehydrogenase from both the mouse and pig are reported. The ORFs of both L-threonine dehydrogenase cDNAs encode proteins of 373 residues (41.5 kDa) and they share 80% identity. The mouse gene is located on chromosome 14, band C. The amino-terminal regions of these proteins have characteristics of a mitochondrial targeting sequence and are related to the UDP-galactose 4-epimerases, with both enzyme families having an amino-terminal NAD+ binding domain. That these cDNAs encode threonine dehydrogenases was shown, previously, by tiling 13 tryptic peptide sequences, obtained from purified L-threonine dehydrogenase isolated from porcine liver mitochondria, on to the pig ORF. These eukaryotic L-threonine dehydrogenases also have significant similarity with the prokaryote L-threonine dehydrogenase amino-terminus peptide sequence of the bacterium, Clostridium sticklandii. In murine tissues, the expression of both L-threonine dehydrogenase and 2-amino-3-ketobutyrate coenzyme A ligase mRNAs were highest in the liver and were also present in brain, heart, kidney, liver, lung, skeletal muscle, spleen and testis. CONCLUSIONS: The first cloning of transcripts for L-threonine dehydrogenase from eukaryotic organisms are reported. However, they do not have any significant sequence homology to the well-characterised Escherichia coli L-threonine dehydrogenase. AD - Tissue Engineering and Regenerative Medicine Centre, Division of Investigative Science, Faculty of Medicine, Imperial College of Science, Technology and Medicine, Chelsea & Westminster Hospital, London, United Kingdom. alasdair.edgar@ic.ac.uk FAU - Edgar, Alasdair J AU - Edgar AJ LA - eng SI - GENBANK/AY095535 SI - GENBANK/AY116662 PT - Journal Article DEP - 20020625 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (DNA, Complementary) RN - 0 (RNA, Messenger) RN - 53-84-9 (NAD) RN - EC 1.1 (Alcohol Oxidoreductases) RN - EC 1.1.1.103 (L-threonine 3-dehydrogenase) SB - IM MH - Alcohol Oxidoreductases/*genetics/metabolism MH - Amino Acid Sequence MH - Animals MH - Base Sequence MH - Binding Sites MH - Biological Transport MH - DNA, Complementary/chemistry/genetics MH - Gene Expression MH - Mice MH - Mitochondria/metabolism MH - Molecular Sequence Data MH - NAD/metabolism MH - RNA, Messenger/genetics/*metabolism MH - Sequence Alignment MH - Sequence Analysis, DNA MH - Sequence Homology, Amino Acid MH - Swine EDAT- 2002/07/05 10:00 MHDA- 2002/12/19 04:00 PHST- 2002/06/17 [received] PHST- 2002/06/25 [accepted] PHST- 2002/06/25 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jun 25;3(1):19. PMID- 12110157 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030422 LR - 20041117 PUBM- Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Jul 11 TI - Application of the development stages of a cluster randomized trial to a framework for valuating complex health interventions. PG - 13 AB - INTRODUCTION: Trials of complex health interventions often pose difficult methodologic challenges. The objective of this paper is to assess the extent to which the various development steps of a cluster randomized trial to optimize antibiotic use in nursing homes are represented in a recently published framework for the design and evaluation of complex health interventions. In so doing, the utility of the framework for health services researchers is evaluated. METHODS: Using the five phases of the framework (theoretical, identification of components of the intervention, definition of trial and intervention design, methodological issues for main trial, promoting effective implementation), corresponding stages in the development of the cluster randomized trial using diagnostic and treatment algorithms to optimize the use of antibiotics in nursing homes are identified and described. RESULTS: Synthesis of evidence needed to construct the algorithms, survey and qualitative research used to define components of the algorithms, a pilot study to assess the feasibility of delivering the algorithms, methodological issues in the main trial including choice of design, allocation concealment, outcomes, sample size calculation, and analysis are adequately represented using the stages of the framework. CONCLUSIONS: The framework is a useful resource for researchers planning a randomized clinical trial of a complex intervention. AD - Department of Pathology, McMaster University, Hamilton, Ontario, Canada. loebm@mcmaster.ca FAU - Loeb, Mark B AU - Loeb MB LA - eng PT - Journal Article DEP - 20020711 PL - England TA - BMC Health Serv Res JID - 101088677 RN - 0 (Anti-Bacterial Agents) SB - IM MH - Aged MH - Algorithms MH - Anti-Bacterial Agents/*therapeutic use MH - Cluster Analysis MH - Critical Pathways MH - Drug Resistance, Bacterial MH - Female MH - Health Services Research/*methods MH - Humans MH - *Intervention Studies MH - Male MH - Nursing Homes/standards/*statistics & numerical data MH - Nursing Staff/education MH - Ontario MH - Qualitative Research MH - Randomized Controlled Trials MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Urinary Tract Infections/*drug therapy/nursing/urine EDAT- 2002/07/12 10:00 MHDA- 2003/04/23 05:00 PHST- 2002/03/13 [received] PHST- 2002/07/11 [accepted] PHST- 2002/07/11 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Jul 11;2(1):13. PMID- 12121577 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021220 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jul 16 TI - Cloning and characterisation of hAps1 and hAps2, human diadenosine polyphosphate-metabolising Nudix hydrolases. PG - 20 AB - BACKGROUND: The human genome contains at least 18 genes for Nudix hydrolase enzymes. Many have similar functions to one another. In order to understand their roles in cell physiology, these proteins must be characterised. RESULTS: We have characterised two novel human gene products, hAps1, encoded by the NUDT11 gene, and hAps2, encoded by the NUDT10 gene. These cytoplasmic proteins are members of the DIPP subfamily of Nudix hydrolases, and differ from each other by a single amino acid. Both metabolise diadenosine-polyphosphates and, weakly, diphosphoinositol polyphosphates. An apparent polymorphism of hAps1 has also been identified, which leads to the point mutation S39N. This has also been characterised. The favoured nucleotides were diadenosine 5',5"'-pentaphosphate (kcat/Km = 11, 8 and 16 x 10(3) M(-1) x s(-1) respectively for hAps1, hAps1-39N and hAps2) and diadenosine 5',5"'-hexaphosphate (kcat/Km = 13, 14 and 11 x 10(3) M(-1) x s(-1) respectively for hAps1, hAps1-39N and hAps2). Both hAps1 and hAps2 had pH optima of 8.5 and an absolute requirement for divalent cations, with manganese (II) being favoured. Magnesium was not able to activate the enzymes. Therefore, these enzymes could be acutely regulated by manganese fluxes within the cell. CONCLUSIONS: Recent gene duplication has generated the two Nudix genes, NUDT11 and NUDT10. We have characterised their gene products as the closely related Nudix hydrolases, hAps1 and hAps2. These two gene products complement the activity of previously described members of the DIPP family, and reinforce the concept that Ap5A and Ap6A act as intracellular messengers. AD - Division of Cell Signalling, School of Life Sciences, The University of Dundee, Dundee, DD1 5EH, UK. n.r.leslie@dundee.ac.uk FAU - Leslie, Nick R AU - Leslie NR FAU - McLennan, Alexander G AU - McLennan AG FAU - Safrany, Stephen T AU - Safrany ST LA - eng PT - Journal Article DEP - 20020716 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (RNA, Messenger) RN - EC 3.6 (Acid Anhydride Hydrolases) RN - EC 3.6.1.- (Pyrophosphatases) RN - EC 3.6.1.- (diadenosine polyphosphate hydrolase) RN - EC 3.6.1.- (nudix hydrolase) RN - EC 3.6.1.- (nudix hydrolase Aps1, human) RN - EC 3.6.1.- (nudix hydrolase Aps2, human) SB - IM MH - *Acid Anhydride Hydrolases/analysis/genetics/metabolism MH - Amino Acid Sequence MH - Animals MH - Cattle MH - Cloning, Molecular MH - Cytosol/enzymology MH - Humans MH - Mice MH - Molecular Sequence Data MH - *Pyrophosphatases/analysis/genetics/metabolism MH - RNA, Messenger/biosynthesis MH - Research Support, Non-U.S. Gov't MH - Sequence Alignment MH - Substrate Specificity MH - Tissue Distribution MH - X Chromosome EDAT- 2002/07/18 10:00 MHDA- 2002/12/21 04:00 PHST- 2002/05/17 [received] PHST- 2002/07/16 [accepted] PHST- 2002/07/16 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jul 16;3(1):20. PMID- 12126482 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030422 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Jul 18 TI - Priority setting for new technologies in medicine: a transdisciplinary study. PG - 14 AB - BACKGROUND: Decision makers in health care organizations struggle with how to set priorities for new technologies in medicine. Traditional approaches to priority setting for new technologies in medicine are insufficient and there is no widely accepted model that can guide decision makers. DISCUSSION: Daniels and Sabin have developed an ethically based account about how priority setting decisions should be made. We have developed an empirically based account of how priority setting decisions are made. In this paper, we integrate these two accounts into a transdisciplinary model of priority setting for new technologies in medicine that is both ethically and empirically based. SUMMARY: We have developed a transdisciplinary model of priority setting that provides guidance to decision makers that they can operationalize to help address priority setting problems in their institution. AD - University of Toronto Joint Centre for Bioethics, 88 College St, Toronto, Canada M5G-1L4. jegibson@chass.utoronto.ca FAU - Gibson, Jennifer L AU - Gibson JL FAU - Martin, Douglas K AU - Martin DK FAU - Singer, Peter A AU - Singer PA LA - eng PT - Journal Article DEP - 20020718 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Consensus MH - *Decision Making, Organizational MH - *Ethics, Institutional MH - Health Priorities/*ethics MH - Humans MH - Managed Care Programs/ethics MH - Models, Organizational MH - Research Support, Non-U.S. Gov't MH - Resource Allocation MH - Social Justice MH - Social Responsibility MH - Social Values MH - Technology Assessment, Biomedical/*ethics MH - United States EDAT- 2002/07/20 10:00 MHDA- 2003/04/23 05:00 PHST- 2001/12/21 [received] PHST- 2002/07/18 [accepted] PHST- 2002/07/18 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Jul 18;2(1):14. PMID- 12149129 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20041117 PUBM- Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Jul 30 TI - Improvement of Drosophila acetylcholinesterase stability by elimination of a free cysteine. PG - 21 AB - BACKGROUND: Acetylcholinesterase is irreversibly inhibited by organophosphate and carbamate insecticides allowing its use for residue detection with biosensors. Drosophila acetylcholinesterase is the most sensitive enzyme known and has been improved by in vitro mutagenesis. However, it is not sufficiently stable for extensive utilization. It is a homodimer in which both subunits contain 8 cysteine residues. Six are involved in conserved intramolecular disulfide bridges and one is involved in an interchain disulfide bridge. The 8th cysteine is not conserved and is present at position 290 as a free thiol pointing toward the center of the protein. RESULTS: The free cysteine has been mutated to valine and the resulting protein has been assayed for stability using various denaturing agents: temperature, urea, acetonitrile, freezing, proteases and spontaneous-denaturation at room temperature. It was found that the C290V mutation rendered the protein 1.1 to 2.7 fold more stable depending on the denaturing agent. CONCLUSION: It seems that stabilization resulting from the cysteine to valine mutation originates from a decrease of thiol-disulfide interchanges and from an increase in the hydrophobicity of the buried side chain. AD - Laboratoire de Synthese et Physicochimie des Molecules d'Interet Biologique, UMR 5068, Universite Paul Sabatier, 31062, Toulouse, France. isa10yann@yahoo.fr FAU - Fremaux, Isabelle AU - Fremaux I FAU - Mazeres, Serge AU - Mazeres S FAU - Brisson-Lougarre, Andree AU - Brisson-Lougarre A FAU - Arnaud, Muriel AU - Arnaud M FAU - Ladurantie, Caroline AU - Ladurantie C FAU - Fournier, Didier AU - Fournier D LA - eng PT - Journal Article DEP - 20020730 PL - England TA - BMC Biochem JID - 101084098 RN - 52-90-4 (Cysteine) RN - 7004-03-7 (Valine) RN - EC 3.1.1.7 (Acetylcholinesterase) SB - IM MH - Acetylcholinesterase/*chemistry/genetics/*metabolism MH - Animals MH - Cysteine/*chemistry/genetics MH - Drosophila melanogaster/*enzymology MH - Enzyme Stability MH - Hydrophobicity MH - Models, Molecular MH - Mutagenesis MH - Protein Denaturation MH - Research Support, Non-U.S. Gov't MH - Valine/genetics EDAT- 2002/08/01 10:00 MHDA- 2002/11/26 04:00 PHST- 2002/05/18 [received] PHST- 2002/07/30 [accepted] PHST- 2002/07/30 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Jul 30;3(1):21. PMID- 12153701 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030422 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Aug 1 TI - Accuracy of responses from postal surveys about continuing medical education and information behavior: experiences from a survey among German diabetologists. PG - 15 AB - BACKGROUND: Postal surveys are a popular instrument for studies about continuing medical education habits. But little is known about the accuracy of responses in such surveys. The objective of this study was to quantify the magnitude of inaccurate responses in a postal survey among physicians. METHODS: A sub-analysis of a questionnaire about continuing medical education habits and information management was performed. The five variables used for the quantitative analysis are based on a question about the knowledge of a fictitious technical term and on inconsistencies in contingency tables of answers to logically connected questions. RESULTS: Response rate was 52%. Non-response bias is possible but seems not very likely since an association between demographic variables and inconsistent responses could not be found. About 10% of responses were inaccurate according to the definition. CONCLUSION: It was shown that a sub-analysis of a questionnaire makes a quantification of inaccurate responses in postal surveys possible. This sub-analysis revealed that a notable portion of responses in a postal survey about continuing medical education habits and information management was inaccurate. AD - Research Group Medicine/Research Unit Biotechnology, Society, and Environment, University of Hamburg, Germany. trelle@uni-hamburg.de FAU - Trelle, Sven AU - Trelle S LA - eng PT - Journal Article PT - Validation Studies DEP - 20020801 PL - England TA - BMC Health Serv Res JID - 101088677 SB - IM MH - Clinical Competence MH - Decision Making MH - Diabetes Mellitus/*therapy MH - Education, Medical, Continuing/*statistics & numerical data MH - Evidence-Based Medicine/*statistics & numerical data MH - Family Practice/education/standards MH - Female MH - Germany MH - Humans MH - Internal Medicine/education/standards MH - Male MH - Meta-Analysis MH - Pediatrics/education/standards MH - Physician's Practice Patterns/*statistics & numerical data MH - Postal Service MH - Primary Health Care/*standards MH - Problem Solving MH - *Questionnaires MH - Reproducibility of Results MH - Risk Reduction Behavior EDAT- 2002/08/03 10:00 MHDA- 2003/04/23 05:00 PHST- 2002/04/12 [received] PHST- 2002/08/01 [accepted] PHST- 2002/08/01 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Aug 1;2(1):15. PMID- 12154231 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041215 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Restriction of lentivirus in monkeys. PG - 11920-5 AB - Retroviruses are able to cross species barriers and have done so many times throughout evolution. Perhaps as a consequence, dominant mechanisms have arisen to block infection by murine retroviruses in mice (restriction factor Fv1) and humans (restriction factor Ref1), as well as in other mammals. Here we describe a block to HIV and simian immunodeficiency virus in monkeys. Like previously described restrictions the block is saturable and gives rise to multiple-hit infection kinetics. Furthermore, like restriction of murine leukemia virus in humans, the block is before reverse transcription. Intriguingly, African green monkey cells are able to block both HIV and simian immunodeficiency virus, and each virus is able to saturate and abrogate the restriction of the other, suggesting that a common factor is responsible. AD - Wohl Virion Centre, Department of Immunology and Molecular Pathology, University College London, 46 Cleveland Street, London W1T 4JF, United Kingdom. FAU - Besnier, Caroline AU - Besnier C FAU - Takeuchi, Yasuhiro AU - Takeuchi Y FAU - Towers, Greg AU - Towers G LA - eng PT - Journal Article DEP - 20020801 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (DNA Primers) RN - 0 (Luminescent Proteins) RN - 0 (Virus Inhibitors) RN - 147336-22-9 (Green Fluorescent Proteins) SB - IM CIN - Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11549-51. PMID: 12195025 MH - Animals MH - Base Sequence MH - Cell Line MH - DNA Primers MH - DNA Replication MH - Green Fluorescent Proteins MH - HIV-1/genetics/*physiology MH - Haplorhini MH - Luminescent Proteins/genetics MH - Rabbits MH - Research Support, Non-U.S. Gov't MH - SIV/genetics/*physiology MH - Transcription, Genetic MH - *Virus Inhibitors EDAT- 2002/08/03 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/01 [aheadofprint] AID - 10.1073/pnas.172384599 [doi] AID - 172384599 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11920-5. Epub 2002 Aug 1. PMID- 12167173 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021122 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Aug 7 TI - Combinatorial diversity of fission yeast SCF ubiquitin ligases by homo- and heterooligomeric assemblies of the F-box proteins Pop1p and Pop2p. PG - 22 AB - BACKGROUND: SCF ubiquitin ligases share the core subunits cullin 1, SKP1, and HRT1/RBX1/ROC1, which associate with different F-box proteins. F-box proteins bind substrates following their phosphorylation upon stimulation of various signaling pathways. Ubiquitin-mediated destruction of the fission yeast cyclin-dependent kinase inhibitor Rum1p depends on two heterooligomerizing F-box proteins, Pop1p and Pop2p. Both proteins interact with the cullin Pcu1p when overexpressed, but it is unknown whether this reflects their co-assembly into bona fide SCF complexes. RESULTS: We have identified Psh1p and Pip1p, the fission yeast homologues of human SKP1 and HRT1/RBX1/ROC1, and show that both associate with Pop1p, Pop2p, and Pcu1p into a ~500 kDa SCFPop1p-Pop2p complex, which supports polyubiquitylation of Rum1p. Only the F-box of Pop1p is required for SCFPop1p-Pop2p function, while Pop2p seems to be attracted into the complex through binding to Pop1p. Since all SCFPop1p-Pop2p subunits, except for Pop1p, which is exclusively nuclear, localize to both the nucleus and the cytoplasm, the F-box of Pop2p may be critical for the assembly of cytoplasmic SCFPop2p complexes. In support of this notion, we demonstrate individual SCFPop1p and SCFPop2p complexes bearing ubiquitin ligase activity. CONCLUSION: Our data suggest that distinct homo- and heterooligomeric assemblies of Pop1p and Pop2p generate combinatorial diversity of SCFPop function in fission yeast. Whereas a heterooligomeric SCFPop1p-Pop2p complex mediates polyubiquitylation of Rum1p, homooligomeric SCFPop1p and SCFPop2p complexes may target unknown nuclear and cytoplasmic substrates. AD - Department of Cancer Cell Biology, Harvard School of Public Health, Boston, MA, USA. vseibert@europroteome.com FAU - Seibert, Volker AU - Seibert V FAU - Prohl, Corinna AU - Prohl C FAU - Schoultz, Ida AU - Schoultz I FAU - Rhee, Edward AU - Rhee E FAU - Lopez, Rebecca AU - Lopez R FAU - Abderazzaq, Kareem AU - Abderazzaq K FAU - Zhou, Chunshui AU - Zhou C FAU - Wolf, Dieter A AU - Wolf DA LA - eng GR - ES-00002/ES/NIEHS GR - GM50780/GM/NIGMS PT - Journal Article DEP - 20020807 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Macromolecular Substances) RN - 0 (Protein Subunits) RN - 0 (Rum1 protein, S pombe) RN - 0 (Schizosaccharomyces pombe Proteins) RN - 0 (Ubiquitins) RN - EC 6.3.2. (Peptide Synthases) RN - EC 6.3.2.19 (SKP Cullin F-Box Protein Ligases) SB - IM MH - Binding Sites MH - Cell Compartmentation MH - Macromolecular Substances MH - Mutation MH - Peptide Synthases/chemistry/genetics/*physiology MH - Protein Structure, Tertiary MH - Protein Subunits MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - SKP Cullin F-Box Protein Ligases MH - Schizosaccharomyces/*enzymology/metabolism MH - Schizosaccharomyces pombe Proteins/chemistry/genetics/metabolism/*physiology MH - Ubiquitins/metabolism EDAT- 2002/08/09 10:00 MHDA- 2002/11/26 04:00 PHST- 2002/06/07 [received] PHST- 2002/08/07 [accepted] PHST- 2002/08/07 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Aug 7;3(1):22. PMID- 12183222 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20031114 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Resistance to macrolides and related antibiotics in Streptococcus pneumoniae. PG - 2727-34 AD - Laboratoire de Microbiologie, CHRU de Caen, Caen, France. leclercq-r@chu-caen.fr FAU - Leclercq, Roland AU - Leclercq R FAU - Courvalin, Patrice AU - Courvalin P LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (RNA, Bacterial) RN - 114-07-8 (Erythromycin) RN - EC 2.1.1. (Methyltransferases) RN - EC 2.1.1.66 (rRNA (adenosine-O-2'-)methyltransferase) SB - IM MH - Anti-Bacterial Agents/metabolism/*pharmacology MH - Base Sequence MH - Binding Sites/drug effects MH - Drug Resistance MH - Erythromycin/metabolism/pharmacology MH - Methylation MH - Methyltransferases/genetics MH - Nucleic Acid Conformation MH - Phenotype MH - RNA, Bacterial/chemistry/genetics MH - Ribosomes/drug effects/genetics/metabolism MH - Streptococcus pneumoniae/*drug effects/genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2727-34. PMID- 12183223 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Importance of the fourth alpha-helix within the CAP homology domain of type II topoisomerase for DNA cleavage site recognition and quinolone action. PG - 2735-46 AB - We report that point mutations causing alteration of the fourth alpha-helix (alpha4-helix) of the CAP homology domain of eukaryotic (Saccharomyces cerevisiae) type II topoisomerases (Ser(740)Trp, Gln(743)Pro, and Thr(744)Pro) change the selection of type II topoisomerase-mediated DNA cleavage sites promoted by Ca(2+) or produced by etoposide, the fluoroquinolone CP-115,953, or mitoxantrone. By contrast, Thr(744)Ala substitution had minimal effect on Ca(2+)- and drug-stimulated DNA cleavage sites, indicating the selectivity of single amino acid substitutions within the alpha4-helix on type II topoisomerase-mediated DNA cleavage. The equivalent mutation in the gene for Escherichia coli gyrase causing Ser(83)Trp also changed the DNA cleavage pattern generated by Ca(2+) or quinolones. Finally, Thr(744)Pro substitution in the yeast type II topoisomerase rendered the enzyme sensitive to antibacterial quinolones. This study shows that the alpha4-helix within the conserved CAP homology domain of type II topoisomerases is critical for selecting the sites of DNA cleavage. It also demonstrates that selective amino acid residues in the alpha4-helix are important in determining the activity and possibly the binding of quinolones to the topoisomerase II-DNA complexes. AD - Department of Internal Medicine and Medical Oncology, West German Cancer Center, University Medical School of Essen, 45122 Essen, Germany. dirk.strumberg@uni-essen.de FAU - Strumberg, Dirk AU - Strumberg D FAU - Nitiss, John L AU - Nitiss JL FAU - Dong, Jiaowang AU - Dong J FAU - Walker, Jerrylaine AU - Walker J FAU - Nicklaus, Marc C AU - Nicklaus MC FAU - Kohn, Kurt W AU - Kohn KW FAU - Heddle, Jonathan G AU - Heddle JG FAU - Maxwell, Anthony AU - Maxwell A FAU - Seeber, Siegfried AU - Seeber S FAU - Pommier, Yves AU - Pommier Y LA - eng GR - CA21765/CA/NCI GR - CA52814/CA/NCI PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (4-Quinolones) RN - 0 (Anti-Infective Agents) RN - 0 (DNA, Bacterial) RN - 0 (Fluoroquinolones) RN - 0 (RNA Cap-Binding Proteins) RN - 136440-70-5 (CP 115953) RN - 7440-70-2 (Calcium) RN - EC 5.99.1.- (DNA Gyrase) RN - EC 5.99.1.- (DNA Topoisomerases, Type II, Bacterial) SB - IM MH - 4-Quinolones MH - Amino Acid Sequence MH - Anti-Infective Agents/*pharmacology MH - Calcium/pharmacology MH - DNA Fragmentation MH - DNA Gyrase/antagonists & inhibitors/metabolism MH - DNA Topoisomerases, Type II, Bacterial/biosynthesis/*metabolism MH - DNA, Bacterial/chemistry/*metabolism MH - Electrophoresis, Polyacrylamide Gel MH - Escherichia coli/enzymology MH - *Fluoroquinolones MH - Protein Conformation MH - RNA Cap-Binding Proteins/*metabolism MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Reverse Transcriptase Polymerase Chain Reaction MH - Saccharomyces cerevisiae/enzymology MH - Sequence Homology, Nucleic Acid EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2735-46. PMID- 12183224 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20031114 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Evaluation of antibiotic susceptibilities of three rickettsial species including Rickettsia felis by a quantitative PCR DNA assay. PG - 2747-51 AB - Rickettsiae grow only intracellularly, and the antibiotic susceptibilities of these bacteria have been assessed by either plaque, dye uptake, or immunofluorescence assays, which are time-consuming. We used a quantitative PCR (with the LightCycler instrument) to assess the levels of inhibition of Rickettisa felis, R. conorii, and R. typhi DNA synthesis in the presence of various antibiotics. We established the kinetics of rickettsial DNA during growth and showed that R. conorii grows more quickly than R. typhi in cell culture, with maximum replication occurring after 5 and 7 days, respectively. The MICs of the antibiotics tested for R. conorii and R. typhi by the quantitative PCR assay were similar to those previously obtained by plaque and dye uptake assays. We found that R. felis is susceptible to doxycycline, rifampin, thiamphenicol, and fluoroquinolones but not to gentamicin, erythromycin, amoxicillin, or trimethoprim-sulfamethoxazole. The resistance of this new species to erythromycin is consistent with its current taxonomic position within the spotted fever group. We believe that quantitative PCR could be used in the future to simplify and shorten antibiotic susceptibility assays of other rickettsiae and other strict intracellular pathogens. AD - Unite des Rickettsies CNRS UPRES-A 6020, Faculte de Medecine, Universite de la Mediterranee, 13385 Marseille Cedex 05, France. FAU - Rolain, Jean-Marc AU - Rolain JM FAU - Stuhl, Laetitia AU - Stuhl L FAU - Maurin, Max AU - Maurin M FAU - Raoult, Didier AU - Raoult D LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (DNA, Bacterial) RN - 0 (Dyes) SB - IM MH - Anti-Bacterial Agents/*pharmacology MH - Calibration MH - DNA, Bacterial/*drug effects/genetics/isolation & purification MH - Dyes MH - Kinetics MH - Microbial Sensitivity Tests/instrumentation/*methods MH - Reverse Transcriptase Polymerase Chain Reaction/instrumentation/*methods MH - Rickettsia/*drug effects MH - Rickettsia conorii/drug effects MH - Rickettsia felis/drug effects MH - Rickettsia typhi/drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2747-51. PMID- 12183225 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - N-alkyl urea hydroxamic acids as a new class of peptide deformylase inhibitors with antibacterial activity. PG - 2752-64 AB - Peptide deformylase (PDF) is a prokaryotic metalloenzyme that is essential for bacterial growth and is a new target for the development of antibacterial agents. All previously reported PDF inhibitors with sufficient antibacterial activity share the structural feature of a 2-substituted alkanoyl at the P(1)' site. Using a combination of iterative parallel synthesis and traditional medicinal chemistry, we have identified a new class of PDF inhibitors with N-alkyl urea at the P(1)' site. Compounds with MICs of 200 micro M for matrilysin and other mammalian metalloproteases. Structure-activity relationship analysis identified preferred substitutions resulting in improved potency and decreased cytotoxity. One of the compounds (VRC4307) was cocrystallized with PDF, and the enzyme-inhibitor structure was determined at a resolution of 1.7 A. This structural information indicated that the urea compounds adopt a binding position similar to that previously determined for succinate hydroxamates. Two compounds, VRC4232 and VRC4307, displayed in vivo efficacy in a mouse protection assay, with 50% protective doses of 30.8 and 17.9 mg/kg of body weight, respectively. These N-alkyl urea hydroxamic acids provide a starting point for identifying new PDF inhibitors that can serve as antimicrobial agents. AD - Versicor, Inc., Fremont, California 94555, USA. FAU - Hackbarth, Corinne J AU - Hackbarth CJ FAU - Chen, Dawn Z AU - Chen DZ FAU - Lewis, Jason G AU - Lewis JG FAU - Clark, Kirk AU - Clark K FAU - Mangold, James B AU - Mangold JB FAU - Cramer, Jeffrey A AU - Cramer JA FAU - Margolis, Peter S AU - Margolis PS FAU - Wang, Wen AU - Wang W FAU - Koehn, Jim AU - Koehn J FAU - Wu, Charlotte AU - Wu C FAU - Lopez, S AU - Lopez S FAU - Withers, George 3rd AU - Withers G 3rd FAU - Gu, Helen AU - Gu H FAU - Dunn, Elina AU - Dunn E FAU - Kulathila, R AU - Kulathila R FAU - Pan, Shi-Hao AU - Pan SH FAU - Porter, Wilma L AU - Porter WL FAU - Jacobs, Jeff AU - Jacobs J FAU - Trias, Joaquim AU - Trias J FAU - Patel, Dinesh V AU - Patel DV FAU - Weidmann, Beat AU - Weidmann B FAU - White, Richard J AU - White RJ FAU - Yuan, Zhengyu AU - Yuan Z LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (DNA Primers) RN - 0 (Hydroxamic Acids) RN - 0 (Protease Inhibitors) RN - 57-13-6 (Urea) RN - EC 3.4.11 (Aminopeptidases) RN - EC 3.5. (Amidohydrolases) RN - EC 3.5.1.88 (peptide deformylase) SB - IM MH - *Amidohydrolases MH - Aminopeptidases/*antagonists & inhibitors MH - Animals MH - Bacteria/drug effects MH - Biotransformation MH - Crystallography, X-Ray MH - DNA Primers MH - Drug Resistance MH - Drug Screening Assays, Antitumor MH - Escherichia coli/metabolism MH - Female MH - Haemophilus influenzae/drug effects/genetics MH - Humans MH - Hydroxamic Acids/*chemical synthesis/pharmacokinetics/*pharmacology MH - In Vitro MH - Male MH - Mice MH - Microbial Sensitivity Tests MH - Microsomes, Liver/metabolism MH - Molecular Conformation MH - Protease Inhibitors/*chemical synthesis/pharmacokinetics/*pharmacology MH - Rats MH - Rats, Sprague-Dawley MH - Septicemia/drug therapy/microbiology MH - Streptococcus pneumoniae/drug effects/genetics MH - Structure-Activity Relationship MH - Tumor Cells, Cultured MH - Urea/*analogs & derivatives/chemical synthesis/pharmacokinetics/pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2752-64. PMID- 12183226 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Oxidative stress increases susceptibility of Mycobacterium tuberculosis to isoniazid. PG - 2765-71 AB - Isoniazid is a first-line antibiotic used in the treatment of infections caused by Mycobacterium tuberculosis. Isoniazid is a prodrug requiring oxidative activation by the catalase-peroxidase hemoprotein, KatG. Resistance to isoniazid can be obtained by point mutations in the katG gene, with one of the most common being a threonine-for-serine substitution at position 315 (S315T). The S315T mutation is found in more than 50% of isoniazid-resistant clinical isolates and results in an approximately 200-fold increase in the MIC of isoniazid compared to that for M. tuberculosis H37Rv. In the present study we investigated the hypothesis that superoxide plays a role in KatG-mediated isoniazid activation. Plumbagin and clofazimine, compounds capable of generating superoxide anion, resulted in a lower MIC of isoniazid for M. tuberculosis H37Rv and a strain carrying the S315T mutation. These agents did not cause as great of an increase in isoniazid susceptibility in the mutant strain when the susceptibilities were assessed by using the inhibitory concentration that causes a 50% decrease in growth. These results provide evidence that superoxide can play a role in isoniazid activation. Since clofazimine alone has antitubercular activity, the observation of synergism between clofazimine and isoniazid raises the interesting possibility of using both drugs in combination to treat M. tuberculosis infections. AD - Section of Hematology Research and Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA. FAU - Bulatovic, Vanja M AU - Bulatovic VM FAU - Wengenack, Nancy L AU - Wengenack NL FAU - Uhl, James R AU - Uhl JR FAU - Hall, Leslie AU - Hall L FAU - Roberts, Glenn D AU - Roberts GD FAU - Cockerill, Franklin R 3rd AU - Cockerill FR 3rd FAU - Rusnak, Frank AU - Rusnak F LA - eng GR - AI47142/AI/NIAID PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antitubercular Agents) RN - 0 (Bacterial Proteins) RN - 0 (DNA, Bacterial) RN - 0 (Naphthoquinones) RN - 0 (Oxidants) RN - 11062-77-4 (Superoxides) RN - 2030-63-9 (Clofazimine) RN - 481-42-5 (plumbagin) RN - 54-85-3 (Isoniazid) RN - EC 1.11.1. (Peroxidases) RN - EC 1.11.1.6 (catalase HPI) SB - IM MH - Antitubercular Agents/*pharmacology MH - *Bacterial Proteins MH - Base Sequence MH - Clofazimine/pharmacology MH - DNA, Bacterial/chemistry MH - Drug Resistance, Microbial MH - Isoniazid/*pharmacology MH - Microbial Sensitivity Tests MH - Molecular Sequence Data MH - Mycobacterium tuberculosis/*drug effects/genetics MH - Naphthoquinones/pharmacology MH - Oxidants/metabolism MH - Oxidative Stress/*physiology MH - Peroxidases/genetics MH - Research Support, U.S. Gov't, P.H.S. MH - Superoxides/metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2765-71. PMID- 12183227 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Heterologous expression of epothilone biosynthetic genes in Myxococcus xanthus. PG - 2772-8 AB - Epothilones are potential anticancer drugs that stabilize microtubules in a manner similar to paclitaxel (Taxol). Epothilones are produced from the myxobacterium Sorangium cellulosum, which has a 16-h doubling time and produces only milligram-per-liter amounts of epothilone A and epothilone B. Furthermore, genetic manipulation of S. cellulosum is difficult. To produce epothilones in a more genetically amenable and rapidly growing host, we chose the closely related and best-characterized myxobacteria Myxococcus xanthus. We inserted 65.4 kb of S. cellulosum DNA that encompassed the entire epothilone gene cluster into the chromosome of M. xanthus by a series of homologous recombination events. The resulting strain produced epothilones A and B. Construction of a strain that contained a mutation in epoK, the P450 epoxidase, resulted in production of epothilones C and D. AD - Kosan Biosciences, Inc., Hayward, California 94545, USA. julien@kosan.com FAU - Julien, Bryan AU - Julien B FAU - Shah, Sanjay AU - Shah S LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Bacterial Proteins) RN - 0 (Culture Media) RN - 0 (DNA, Bacterial) RN - 0 (Epothilones) RN - 0 (Plasmids) RN - 0 (epothilone C) RN - 0 (epothilone D) RN - 9035-51-2 (Cytochrome P-450 Enzyme System) RN - EC 1. (Oxidoreductases) RN - EC 1.- (EpoK protein, Polyangium cellulosum) SB - IM MH - Bacterial Proteins/genetics MH - Chromosomes, Bacterial/genetics MH - Culture Media MH - Cytochrome P-450 Enzyme System/genetics MH - DNA, Bacterial/genetics MH - Epothilones/*biosynthesis/*genetics MH - Escherichia coli/genetics MH - Gene Expression Regulation, Bacterial/*genetics MH - Multigene Family/genetics MH - Mutation/genetics MH - Myxococcus xanthus/*genetics/*metabolism MH - Oxidoreductases/genetics MH - Plasmids/genetics MH - Protein Engineering EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2772-8. PMID- 12183228 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Different levels of genetic homogeneity in vancomycin-resistant and -susceptible Enterococcus faecium isolates from different human and animal sources analyzed by amplified-fragment length polymorphism. PG - 2779-83 AB - The genetic relationship among fecal vancomycin-resistant Enterococcus faecium (VREF) and vancomycin-susceptible E. faecium (VSEF) isolates (n = 178) from the same populations of pigs, human healthy volunteers, and hospitalized patients (from The Netherlands) and chickens (from The Netherlands and Greece) was studied by amplified-fragment length polymorphism (AFLP). The majority of VREF isolates from pigs, healthy volunteers, and hospitalized patients grouped together (genetic similarity, >or=65%). In a previous AFLP study by our group the VREF isolates from hospitalized patients grouped separately, most likely because these were clinical and not fecal isolates as in the present study. Furthermore, VSEF isolates from humans and pigs were found much more genetically diverse than VREF isolates, whereas VREF and VSEF isolates from chickens clustered together in a separate genogroup (genetic similarity, >or=65%), a pattern clearly distinct from the patterns for human and pig isolates. The present study suggests that pigs are a more important source of VREF for humans than chickens and that human- and pig-derived VSEF isolates seem much more heterogeneous than VREF isolates. AD - Department of Medical Microbiology, University Hospital Maastricht, The Netherlands. FAU - Bruinsma, Nienke AU - Bruinsma N FAU - Willems, Rob J L AU - Willems RJ FAU - van den Bogaard, Anthony E AU - van den Bogaard AE FAU - van Santen-Verheuvel, Marga AU - van Santen-Verheuvel M FAU - London, Nancy AU - London N FAU - Driessen, Christel AU - Driessen C FAU - Stobberingh, Ellen E AU - Stobberingh EE LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Glycopeptide) RN - 1404-90-6 (Vancomycin) SB - IM MH - Algorithms MH - Animals MH - Antibiotics, Glycopeptide/pharmacology MH - Enterococcus faecium/*drug effects/*genetics MH - Feces/microbiology MH - Genotype MH - Gram-Positive Bacterial Infections/microbiology/transmission MH - Humans MH - Phenotype MH - Polymorphism, Restriction Fragment Length MH - Poultry MH - Research Support, Non-U.S. Gov't MH - Swine MH - Vancomycin/pharmacology MH - Vancomycin Resistance/*genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2779-83. PMID- 12183229 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - The LisRK signal transduction system determines the sensitivity of Listeria monocytogenes to nisin and cephalosporins. PG - 2784-90 AB - The Listeria monocytogenes two-component signal transduction system, LisRK, initially identified in strain LO28, plays a significant role in the virulence potential of this important food-borne pathogen. Here, it is shown that, in addition to its major contribution in responding to ethanol, pH, and hydrogen peroxide stresses, LisRK is involved in the ability of the cell to tolerate important antimicrobials used in food and in medicine, e.g., the lantibiotic nisin and the cephalosporin family of antibiotics. A (Delta)lisK mutant (lacking the LisK histidine kinase sensor component) displays significantly enhanced resistance to the lantibiotic nisin, a greatly enhanced sensitivity to the cephalosporins, and a large reduction in the expression of three genes thought to encode a penicillin-binding protein, another histidine kinase (other than LisK), and a protein of unknown function. Confirmation of the role of LisRK was obtained when the response regulator, LisR, was overexpressed using both constitutive and inducible (nisin-controlled expression) systems. Under these conditions we observed a reversion of the (Delta)lisK mutant to wild-type growth kinetics in the presence of nisin. It was also found that overexpression of LisR complemented the reduced expression of two of the aforementioned genes. These results demonstrate the important role of LisRK in the response of L. monocytogenes to a number of antimicrobial agents. AD - Department of Microbiology and National Food Biotechnology Centre, University College Cork, Cork, Ireland. FAU - Cotter, Paul D AU - Cotter PD FAU - Guinane, Caitriona M AU - Guinane CM FAU - Hill, Colin AU - Hill C LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Peptide) RN - 0 (Cephalosporins) RN - 0 (Culture Media) RN - 0 (Plasmids) RN - 0 (Transcription Factors) RN - 1414-45-5 (Nisin) RN - EC 2.7.1.37 (Protein Kinases) RN - EC 2.7.3.- (protein-histidine kinase) SB - IM MH - Antibiotics, Peptide/*pharmacology MH - Cephalosporins/*pharmacology MH - Culture Media MH - Gene Expression Regulation, Bacterial/drug effects/genetics MH - Listeria monocytogenes/*drug effects/genetics MH - Microbial Sensitivity Tests MH - Nisin/*pharmacology MH - Plasmids MH - Protein Kinases/*genetics/physiology MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Signal Transduction/*drug effects/genetics MH - Transcription Factors/*genetics/physiology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2784-90. PMID- 12183230 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Genetic and biochemical characterization of CGB-1, an Ambler class B carbapenem-hydrolyzing beta-lactamase from Chryseobacterium gleum. PG - 2791-6 AB - Chryseobacterium gleum (previously included in the Flavobacterium IIb species) is a gram-negative aerobe that is a source of nosocomial infections. An Ambler class B beta-lactamase gene was cloned and expressed in Escherichia coli from reference strain C. gleum CIP 103039 that had reduced susceptibility to expanded-spectrum cephalosporins and carbapenems. The purified beta-lactamase, CGB-1, with a pI value of 8.6 and a determined relative molecular mass of ca. 26 kDa, hydrolyzed penicillins; narrow- and expanded-spectrum cephalosporins; and carbapenems. CGB-1 was a novel member of the molecular subclass B1 of metallo-enzymes. It had 83 and 42% amino acid identity with IND-1 from Chryseobacterium indologenes and BlaB from C. meningosepticum, respectively. Thus, in addition to the previously characterized clavulanic acid-inhibited extended-spectrum beta-lactamase CGA-1 of Ambler class A, C. gleum produces a very likely chromosome-borne class B beta-lactamase. AD - Service de Bacteriologie-Virologie, Hopital de Bicetre, Assistance Publique/Hopitaux de Paris, Faculte de Medecine Paris-Sud, 94275 Le Kremlin-Bicetre Cedex, France. FAU - Bellais, Samuel AU - Bellais S FAU - Naas, Thierry AU - Naas T FAU - Nordmann, Patrice AU - Nordmann P LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Carbapenems) RN - 0 (DNA, Bacterial) RN - 0 (DNA, Recombinant) RN - 0 (Enzyme Inhibitors) RN - 0 (Plasmids) RN - 58001-44-8 (Clavulanic Acid) RN - EC 3.5.2.6 (beta-Lactamases) RN - EC 3.5.2.6 (beta-lactamase CGB-1, Chryseobacterium gleum) SB - IM MH - Amino Acid Sequence MH - Bacteria/drug effects/enzymology/*genetics MH - Base Sequence MH - Carbapenems/*pharmacology MH - Clavulanic Acid/pharmacology MH - Cloning, Molecular MH - Conjugation, Genetic MH - DNA, Bacterial/analysis MH - DNA, Recombinant/genetics MH - Drug Resistance MH - Enzyme Induction/drug effects MH - Enzyme Inhibitors/pharmacology MH - In Situ Hybridization MH - Isoelectric Focusing MH - Kinetics MH - Microbial Sensitivity Tests MH - Molecular Sequence Data MH - Plasmids/genetics MH - Research Support, Non-U.S. Gov't MH - beta-Lactamases/*genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2791-6. PMID- 12183231 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Frequency of disinfectant resistance genes and genetic linkage with beta-lactamase transposon Tn552 among clinical staphylococci. PG - 2797-803 AB - A total of 61 strains of Staphylococcus aureus and 177 coagulase-negative staphylococcal strains were isolated from the blood of patients with bloodstream infections and from the skin of both children under cancer treatment and human immunodeficiency virus-positive patients. The MIC analyses revealed that 118 isolates (50%) were resistant to quaternary ammonium compound-based disinfectant benzalkonium chloride (BC). The frequencies of resistance to a range of antibiotics were significantly higher among BC-resistant staphylococci than among BC-sensitive staphylococci. Of 78 BC-resistant staphylococcal isolates, plasmid DNA from 65 (83%), 2 (3%), 43 (55%), and 15 (19%) isolates hybridized to qacA or -B (qacA/B), qacC, blaZ, and tetK probes, respectively. The qacA/B and blaZ probes hybridized to the same plasmid in 19 (24%) staphylococcal strains. The plasmids harboring both qacA/B and blaZ genes varied from approximately 20 to 40 kb. The Staphylococcus epidermidis Fol62 isolate, harboring multiresistance plasmid pMS62, contained qacA/B and blaZ together with tetK. Molecular and genetic studies indicated different structural arrangements of blaZ and qacA/B, including variable intergenic distances and transcriptional directions of the two genes on the same plasmid within the strains. The different organizations may be due to the presence of various genetic elements involved in cointegration, recombination, and rearrangements. These results indicate that qac resistance genes are common and that linkage between resistance to disinfectants and penicillin resistance occurs frequently in clinical isolates in Norway. Moreover, the higher frequency of antibiotic resistance among BC-resistant strains indicates that the presence of either resistance determinant selects for the other during antimicrobial therapy and disinfection in hospitals. AD - MATFORSK, Norwegian Food Research Institute, N-1430 As, Norway. FAU - Sidhu, Maan Singh AU - Sidhu MS FAU - Heir, Even AU - Heir E FAU - Leegaard, Truls AU - Leegaard T FAU - Wiger, Karianne AU - Wiger K FAU - Holck, Askild AU - Holck A LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Culture Media) RN - 0 (DNA Transposable Elements) RN - 0 (DNA, Bacterial) RN - 0 (Disinfectants) RN - 0 (Plasmids) SB - IM MH - Culture Media MH - DNA Transposable Elements/*genetics MH - DNA, Bacterial/biosynthesis/genetics MH - Disinfectants/*pharmacology MH - Drug Resistance, Microbial MH - Genes, Bacterial/genetics MH - Linkage (Genetics)/*genetics MH - Microbial Sensitivity Tests MH - Nucleic Acid Hybridization MH - Plasmids/genetics MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Staphylococcal Infections/microbiology MH - Staphylococcus/*drug effects/*genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2797-803. PMID- 12183232 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Isoniazid-induced transient high-level resistance in Mycobacterium tuberculosis. PG - 2804-10 AB - An American Type Culture Collection reference strain and eight clinical strains of Mycobacterium tuberculosis, all of which were susceptible to isoniazid (INH) (mean MIC, 0.06 mg/liter) and negative for the Ser315Thr katG mutation, were left in their BACTEC 12B vials (for use with the BACTEC 460-TB method) containing 0.1 mg of INH per liter for periods of up to 28 days after the completion of the antibiotic susceptibility test. Each eventually grew to levels compatible with those of INH-resistant strains. Successive passages in INH-containing BACTEC 12B vials and onto solid media showed that the resistance noted above was maintained. Successive passages of these M. tuberculosis strains in which INH resistance had been induced into BACTEC 12B vials or solid media containing stepwise increases in INH concentrations eventually yielded organisms resistant to 20 mg of INH per liter. Transfer of cells in which INH resistance had been induced to drug-free medium followed by repeated passages in that medium eventually yielded organisms whose susceptibility to INH was identical to that of the original parent strains. The cycle of induced INH resistance could be repeated with these now INH-susceptible cells. The use of M. tuberculosis identification probes and IS6110-based restriction fragment length polymorphism analyses of cultures throughout the induction of INH resistance and the reversal of resistance in drug-free medium eliminated the possibility that the culture was contaminated or that the initial specimen had a mixed type of infection. Induced high-level resistance to INH (20 mg/liter) could be reduced 100-fold with a subinhibitory concentration of reserpine but not with verapamil. These results collectively suggest that high-level resistance to INH can be induced in INH-susceptible M. tuberculosis strains by the induction of a reserpine-sensitive efflux mechanism. AD - Unit of Mycobacteriology, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, P-1349-008 Lisbon, Portugal. FAU - Viveiros, Miguel AU - Viveiros M FAU - Portugal, Isabel AU - Portugal I FAU - Bettencourt, Rosario AU - Bettencourt R FAU - Victor, Thomas C AU - Victor TC FAU - Jordaan, Annemarie M AU - Jordaan AM FAU - Leandro, Clara AU - Leandro C FAU - Ordway, Diane AU - Ordway D FAU - Amaral, Leonard AU - Amaral L LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antitubercular Agents) RN - 0 (Bacterial Proteins) RN - 0 (Calcium Channel Blockers) RN - 50-55-5 (Reserpine) RN - 52-53-9 (Verapamil) RN - 54-85-3 (Isoniazid) RN - EC 1.11.1. (Peroxidases) RN - EC 1.11.1.6 (catalase HPI) SB - IM MH - Antitubercular Agents/metabolism/*pharmacology MH - *Bacterial Proteins MH - Calcium Channel Blockers/pharmacology MH - Drug Resistance, Bacterial MH - Isoniazid/metabolism/*pharmacology MH - Microbial Sensitivity Tests MH - Mycobacterium tuberculosis/*drug effects/metabolism MH - Peroxidases/genetics MH - Polymorphism, Restriction Fragment Length MH - Research Support, Non-U.S. Gov't MH - Reserpine/pharmacology MH - Reverse Transcriptase Polymerase Chain Reaction MH - Verapamil/pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2804-10. PMID- 12183233 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Identification and analysis of amino acid mutations in porin IB that mediate intermediate-level resistance to penicillin and tetracycline in Neisseria gonorrhoeae. PG - 2811-20 AB - PenB is the third resistance determinant in the stepwise acquisition of multiple resistance genes in chromosomally mediated resistant Neisseria gonorrhoeae (CMRNG). Alterations in por(IB), one of two alleles at the por locus that encodes the outer membrane protein porin IB (PIB), were recently reported to be responsible for the increased resistance to penicillin and tetracycline conferred by penB, but the specific mutations conferring antibiotic resistance were not identified experimentally. To determine which amino acids in PIB confer increased resistance, we transformed a recipient strain with chimeras of the por(IB) genes from strains FA1090 and FA140 (penB2). These studies revealed that two amino acid changes, G120D and A121D, were both necessary and sufficient to confer increased resistance to penicillin and tetracycline. Site-saturation and site-directed mutagenesis of Gly-120 and Ala-121 revealed that both a single mutation, G120K, and the double mutations G120R A121H and G120P A121P also conferred antibiotic resistance to the recipient strain. The identical mutations in PIA increased penicillin and tetracycline resistance either moderately or not at all. Analysis of por(IB) genes present in the GenBank database from 51 clinical isolates demonstrated that lysine and aspartate mutations at positions 120 and/or 121 also occur in nature. These studies demonstrate that charged amino acids at positions 120 and 121 in PIB are highly preferential for conferring resistance to penicillin and tetracycline in N. gonorrhoeae. AD - Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7365, USA. FAU - Olesky, Melanie AU - Olesky M FAU - Hobbs, Marcia AU - Hobbs M FAU - Nicholas, Robert A AU - Nicholas RA LA - eng GR - AI-36901/AI/NIAID PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Bacterial Outer Membrane Proteins) RN - 0 (Chimeric Proteins) RN - 0 (DNA, Bacterial) RN - 0 (Porins) RN - 0 (porin proteins, Neisseria) SB - IM MH - Amino Acid Sequence MH - Amino Acid Substitution MH - Bacterial Outer Membrane Proteins/drug effects/metabolism MH - Blotting, Western MH - Chimeric Proteins/chemistry/genetics MH - DNA, Bacterial/drug effects/genetics MH - Electrophoresis, Polyacrylamide Gel MH - Microbial Sensitivity Tests MH - Molecular Sequence Data MH - Mutation/*genetics MH - Neisseria gonorrhoeae/drug effects/*genetics/growth & development MH - Penicillin Resistance/*genetics MH - Porins/*genetics MH - Research Support, U.S. Gov't, P.H.S. MH - Reverse Transcriptase Polymerase Chain Reaction MH - Tetracycline Resistance/*genetics MH - Transformation, Bacterial EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2811-20. PMID- 12183234 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Antibiotic resistance genes and Salmonella genomic island 1 in Salmonella enterica serovar Typhimurium isolated in Italy. PG - 2821-8 AB - Fifty-four epidemiologically unrelated multidrug-resistant Salmonella enterica serovar Typhimurium isolates, collected between 1992 and 2000 in Italy, were analyzed for the presence of integrons. Strains were also tested for Salmonella genomic island 1 (SGI1), carrying antibiotic resistance genes in DT104 strains. A complete SGI1 was found in the majority of the DT104 strains. Two DT104 strains, showing resistance to streptomycin-spectinomycin and sulfonamides, carried a partially deleted SGI1 lacking the flo(st), tetR, and tetA genes, conferring chloramphenicol-florfenicol and tetracycline resistance, and the integron harboring the pse-1 gene cassette, conferring ampicillin resistance. The presence of SGI1 was also observed in serovar Typhimurium strains belonging to other phage types, suggesting either the potential mobility of this genomic island or changes in the phage-related phenotype of DT104 strains. AD - Laboratory of Bacteriology and Medical Mycology, Istituto Superiore di Sanita, Rome, Italy. alecara@iss.it FAU - Carattoli, Alessandra AU - Carattoli A FAU - Filetici, Emma AU - Filetici E FAU - Villa, Laura AU - Villa L FAU - Dionisi, Anna Maria AU - Dionisi AM FAU - Ricci, Antonia AU - Ricci A FAU - Luzzi, Ida AU - Luzzi I LA - eng SI - GENBANK/AF261825 PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (DNA, Bacterial) SB - IM MH - Blotting, Southern MH - Cloning, Molecular MH - DNA, Bacterial/genetics MH - Drug Resistance, Microbial MH - Electrophoresis, Gel, Pulsed-Field MH - Genes, Bacterial/*genetics MH - Genome, Bacterial MH - Humans MH - Italy MH - Molecular Sequence Data MH - Research Support, Non-U.S. Gov't MH - Salmonella Infections/microbiology MH - Salmonella enterica/*drug effects/*genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2821-8. PMID- 12183235 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - The Candida dubliniensis CdCDR1 gene is not essential for fluconazole resistance. PG - 2829-41 AB - The present study investigated the role of the Candida dubliniensis CdCDR1 and CdCDR2 genes in the development of fluconazole resistance. The C. dubliniensis CdCDR1 gene was 92% identical at the nucleotide sequence level to the corresponding C. albicans gene. However, 58% (14 of 24) of C. dubliniensis genotype 1 isolates tested harbored a nonsense mutation in the CdCDR1 open reading frame that converted codon 756 (TAT) to a TAG translational stop codon. Analysis of five of these C. dubliniensis isolates by Western immunoblotting showed that they expressed a truncated 85-kDa CdCdr1p compared to the full-length 170-kDa CdCdr1p. Expression of CdCDR1 alleles from six C. dubliniensis isolates in a pdr5 Saccharomyces cerevisiae strain revealed that CdCDR1 alleles from three isolates that encoded truncated proteins were unable to confer resistance to drugs and antifungals. However, reassignment of the TAG sequence at codon 756 to TAT (encoding tyrosine) in an allele from strain CD36 conferred the ability to mediate resistance to multiple drugs. Fluconazole-resistant isolates of C. dubliniensis harboring functional alleles of CdCDR1 were found to exhibit two- to ninefold-higher levels of CdCDR1 mRNA than did matched fluconazole-susceptible isolates. By comparison, levels of CdMDR1 expression ranged from approximately 50- to 100-fold greater in resistant isolates. Fluconazole resistance was also identified in isolates harboring nonfunctional CdCDR1 alleles, but resistance in these isolates was only associated with increased CdMDR1 expression. Targeted disruption of two functional alleles of CdCDR1 in a fluconazole-resistant derivative of C. dubliniensis that overexpressed both CdCDR1 and CdMDR1 revealed that although CdCDR1 was important for mediating reduced susceptibility to itraconazole and ketoconazole, there was no affect on fluconazole susceptibility in the double mutant. Evidence presented in this study reveals that CdCDR1 is not essential for the development of fluconazole resistance in C. dubliniensis. AD - Microbiology Research Unit, Department of Oral Surgery, Oral Medicine and Pathology, School of Dental Science, Trinity College, University of Dublin, Republic of Ireland. FAU - Moran, Gary AU - Moran G FAU - Sullivan, Derek AU - Sullivan D FAU - Morschhauser, Joachim AU - Morschhauser J FAU - Coleman, David AU - Coleman D LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (ATP-Binding Cassette Transporters) RN - 0 (Antifungal Agents) RN - 0 (Azoles) RN - 0 (CDR1 protein, Candida albicans) RN - 0 (CDR2 protein, fungal) RN - 0 (DNA Primers) RN - 0 (DNA, Fungal) RN - 0 (Fungal Proteins) RN - 0 (Membrane Transport Proteins) RN - 0 (RNA, Messenger) RN - 86386-73-4 (Fluconazole) SB - IM MH - ATP-Binding Cassette Transporters/biosynthesis/genetics MH - Alleles MH - Antifungal Agents/*pharmacology MH - Azoles/pharmacology MH - Blotting, Southern MH - Blotting, Western MH - Candida/*drug effects/*genetics MH - DNA Primers/pharmacology MH - DNA, Fungal/drug effects/genetics MH - Drug Resistance, Fungal MH - Fluconazole/*pharmacology MH - Fungal Proteins/biosynthesis/genetics MH - Gene Targeting MH - Membrane Transport Proteins/biosynthesis/*genetics MH - Mutagenesis, Site-Directed MH - RNA, Messenger/biosynthesis MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Saccharomyces cerevisiae/drug effects/genetics MH - Transformation, Genetic/genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2829-41. PMID- 12183236 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Effect of 5-iodo-2'-deoxyuridine on vaccinia virus (orthopoxvirus) infections in mice. PG - 2842-7 AB - There is a concern that there may be unregistered stocks of smallpox that can be used for bioterrorism or biological warfare. According to the WHO Advisory Committee on Variola Research, there is a need to develop strategies to treat smallpox infections should they reappear. It would also be important to have an effective drug at hand for the treatment of monkeypox disease in humans. We show here that 5-iodo-2'-deoxyuridine (IDU) is a potent inhibitor of vaccinia virus (VV) replication and that IDU inhibits VV DNA synthesis in a dose-dependent way. The in vivo protective effect of IDU was assessed in the VV tail lesion model in immunocompetent mice and in a lethal model for VV infection in SCID (severe combined immune deficiency) mice that had been infected either intranasally, intraperitoneally, or intravenously. Subcutaneous treatment with IDU at 150 and 100 mg/kg of body weight markedly reduced the number of tail lesions in immunocompetent NMRI mice. Untreated intranasally VV-infected SCID mice died at 20.8 +/- 3.1 days after infection (mean +/- standard deviation). Treatment with IDU (subcutaneously, 150 mg/kg/day [from day 0 to 4] and 75 mg/kg/day [from day 6 to 11]) delayed-virus induced mortality by 15 days (mean day of death +/- standard deviation, 35.8 +/- 6.7; P < 0.0001). This protective effect was associated with (i) an improvement of lung histology and (ii) a marked reduction in lung viral titers. IDU also delayed VV-induced mortality when mice had either been infected intraperitoneally or intravenously. Even when the start of treatment with IDU (in intraperitoneally VV-infected mice) was postponed until 2 or 4 days after infection, an important delay in virus-induced mortality was noted. AD - Rega Institute for Medical Research, Katholieke Universiteit Leuven, 3000 Leuven, Belgium. johan.neyts@rega.kuleuven.ac.be FAU - Neyts, Johan AU - Neyts J FAU - Verbeken, Erik AU - Verbeken E FAU - De Clercq, Erik AU - De Clercq E LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antiviral Agents) RN - 0 (DNA, Viral) RN - 54-42-2 (Idoxuridine) SB - IM MH - Animals MH - Antiviral Agents/*therapeutic use MH - Body Weight/physiology MH - Cells, Cultured MH - DNA, Viral/biosynthesis/genetics MH - Growth/physiology MH - Humans MH - Idoxuridine/*therapeutic use MH - Immunologic Deficiency Syndromes/immunology MH - Lung/virology MH - Mice MH - Mice, SCID MH - Plaque Assay MH - Research Support, Non-U.S. Gov't MH - Tail/pathology MH - Vaccinia/*drug therapy/pathology/virology MH - *Vaccinia virus EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2842-7. PMID- 12183237 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Penciclovir susceptibilities of herpes simplex virus isolates from patients using penciclovir cream for treatment of recurrent herpes labialis. PG - 2848-53 AB - The antiherpesvirus agent penciclovir (PCV) shares an identical activation pathway and a similar mode of action with acyclovir (ACV). However, since PCV represents a relatively recent treatment option, the clinical resistance profile to PCV is less well known. A susceptibility program was established to assess the resistance profile for serial herpes simplex virus isolates from immunocompetent patients with recurrent herpes labialis obtained throughout a 4-day period of treatment with topical PCV (1% cream) or a placebo. Two isolates (2 of 1,035 [0.19%]), representing 0.34% of the patients (2 of 585), were confirmed to be PCV-resistant (Pcv(r)) herpes simplex virus type 1 by a plaque reduction assay in MRC-5 cells. These two viruses were highly resistant to PCV (50% inhibitory concentrations [IC(50)s], >55 micro g/ml) and were isolated less than 17 h after the start of patient-initiated treatment. However, subsequent isolates on days 2 and 3 from these patients were completely susceptible to PCV (IC(50)s, <2.0 micro g/ml). Thus, it is not clear whether the resistance to PCV for these two early-treatment isolates was directly associated with the 17 h of PCV treatment; several possible explanations are discussed. In an analysis of the distribution of IC(50) differences between the first and last isolates, there were three patients with minor IC(50) increases in the PCV-treated population and one in the placebo-treated group, although statistically, only the latter was an outlier. No patients were found to have Pcv(r) virus at the end of acute treatment, regardless of treatment group. Overall, the prevalence of Pcv(r) was found to be similar to the 0.3% Acv(r) reported for immunocompetent, untreated populations. AD - Department of Host Defense, The Antimicrobial and Host Defense Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, Collegeville, Pennsylvania 19426-0989, USA. robert_t_sarisky@gsk.com FAU - Sarisky, Robert T AU - Sarisky RT FAU - Bacon, Teresa AU - Bacon T FAU - Boon, Ron AU - Boon R FAU - Locke, Leslie AU - Locke L FAU - Nguyen, Tammy T AU - Nguyen TT FAU - Leary, Jeffry AU - Leary J FAU - Esser, Klaus AU - Esser K FAU - Saltzman, Robin AU - Saltzman R LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antiviral Agents) RN - 39809-25-1 (penciclovir) RN - 59277-89-3 (Acyclovir) RN - EC 2.7.1.112 (Protein-Tyrosine Kinase) SB - IM MH - Acyclovir/*analogs & derivatives/*pharmacology MH - Antiviral Agents/*pharmacology MH - Autoradiography MH - Drug Resistance, Viral MH - Frameshift Mutation/genetics MH - Herpes Labialis/*drug therapy/*virology MH - Herpesvirus 1, Human/*drug effects/genetics/isolation & purification MH - Humans MH - Microbial Sensitivity Tests MH - Plaque Assay MH - Protein-Tyrosine Kinase/metabolism MH - Recurrence EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2848-53. PMID- 12183238 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Samarangenin B from Limonium sinense suppresses herpes simplex virus type 1 replication in Vero cells by regulation of viral macromolecular synthesis. PG - 2854-64 AB - Inhibitory effects of ethanolic extracts from 10 Chinese herbs on herpes simplex virus type 1 (HSV-1) replication were investigated. By a bioassay-guided fractionation procedure, samarangenin B (Sam B) was isolated from Limonium sinense; Sam B significantly suppressed HSV-1 multiplication in Vero cells without apparent cytotoxicity. Time-of-addition experiments suggested that the inhibitory action of Sam B on HSV-1 replication was not due to the blocking of virus adsorption. In an attempt to further localize the point in the HSV-1 replication cycle where arrest occurred, a set of key regulatory events leading to viral multiplication was examined, including viral immediate-early (alpha), early (beta), and late (gamma) gene expression and DNA replication. Results indicated that levels of glycoprotein B (gB), gC, gD, gG, and infected-cell protein 5 (ICP5) expression and gB mRNA expression in Vero cells were impeded by Sam B. Data from PCR showed that replication of HSV-1 DNA in Vero cells was arrested by Sam B. Furthermore, Sam B decreased DNA polymerase, ICP0, and ICP4 gene expression in Vero cells. Results of an electrophoretic mobility shift assay demonstrated that Sam B interrupted the formation of an alpha-trans-induction factor/C1/Oct-1/GARAT multiprotein complex. The mechanisms of antiviral action of Sam B seem to be mediated, at least in part, by inhibiting HSV-1 alpha gene expression, including expression of the ICP0 and ICP4 genes, by blocking beta transcripts such as DNA polymerase mRNA, and by arresting HSV-1 DNA synthesis and structural protein expression in Vero cells. These results show that Sam B is an antiviral agent against HSV-1 replication. AD - National Research Institute of Chinese Medicine,Taipei, Taiwan, Republic of China. kuo9111@cma23.nricm.edu.tw FAU - Kuo, Yuh-Chi AU - Kuo YC FAU - Lin, Lie-Chwen AU - Lin LC FAU - Tsai, Wei-Jern AU - Tsai WJ FAU - Chou, Cheng-Jen AU - Chou CJ FAU - Kung, Szu-Hao AU - Kung SH FAU - Ho, Yen-Hui AU - Ho YH LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antiviral Agents) RN - 0 (Benzopyrans) RN - 0 (DNA, Complementary) RN - 0 (DNA, Viral) RN - 0 (Drugs, Chinese Herbal) RN - 0 (RNA, Viral) RN - 0 (samarangenin B) RN - EC 2.7.7.7 (DNA-Directed DNA Polymerase) SB - IM MH - Animals MH - Antiviral Agents/*pharmacology MH - Benzopyrans/isolation & purification/*pharmacology MH - Blotting, Northern MH - Blotting, Western MH - Cell Survival/drug effects MH - Cercopithecus aethiops MH - DNA Replication/drug effects MH - DNA, Complementary/biosynthesis/genetics MH - DNA, Viral/*biosynthesis MH - DNA-Directed DNA Polymerase/biosynthesis/genetics MH - Drugs, Chinese Herbal/pharmacology MH - Electrophoretic Mobility Shift Assay MH - Herpesvirus 1, Human/*drug effects/*metabolism MH - Plaque Assay MH - Plumbaginaceae/*chemistry MH - RNA, Viral/biosynthesis/genetics MH - Research Support, Non-U.S. Gov't MH - Vero Cells MH - Virus Replication/*drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2854-64. PMID- 12183239 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Immunomodulatory effect of zidovudine (ZDV) on cytotoxic T lymphocytes previously exposed to ZDV. PG - 2865-71 AB - In a previous study, zidovudine (ZDV) was shown to cause a concentration-dependent inhibition of antigen-specific cytotoxic T-lymphocyte (CTL) clonal expansion (S. Francke, C. G. Orosz, K. A. Hayes, and L. E. Mathes, Antimicrob. Agents Chemother. 44:1900-1905, 2000). However, this suppressive effect was lost if exposure to ZDV was delayed for 24 to 48 h during the antigen sensitization period, suggesting that antigen-primed CTL may be less susceptible than naive T lymphocytes to the suppressive effects of ZDV. The present study was undertaken to determine if naive T lymphocytes were more sensitive to the suppressive effects of ZDV than T lymphocytes previously exposed to antigen. The 50% inhibitory concentration (IC(50)) values of ZDV were determined on naive and antigen-primed T-cell responses in an alloantigen system. Lymphocyte cultures with continuous antigen exposure (double prime) were more resistant to ZDV suppression (IC(50) = 316 micro M) than were naive lymphocytes (IC(50) = 87.5 micro M). Interestingly, lymphocytes that were antigen primed but deprived of antigen during the final 7 days of culture (prime/hold) were exquisitely sensitive to ZDV suppression (IC(50) = 29.3 micro M). The addition of 80 micro M ZDV during the initial priming of the single-prime (prime/hold) and double-prime cultures did not select for a more drug-resistant cell population. The differences in ZDV sensitivities are likely a reflection of the physiological properties of the lymphocytes related to their activation state. AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio 43210, USA. FAU - Francke, Sabine AU - Francke S FAU - Orosz, Charles G AU - Orosz CG FAU - Hsu, Jason AU - Hsu J FAU - Mathes, Lawrence E AU - Mathes LE LA - eng GR - P30 CA16058/CA/NCI GR - R01 AI40855/AI/NIAID PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Adjuvants, Immunologic) RN - 0 (Anti-HIV Agents) RN - 0 (Antigens, Viral) RN - 0 (Chromium Radioisotopes) RN - 0 (Interleukin-2) RN - 30516-87-1 (Zidovudine) SB - IM MH - Adjuvants, Immunologic/*pharmacology MH - Animals MH - Anti-HIV Agents/*pharmacology MH - Antigens, Viral/immunology MH - Cell Separation MH - Chromium Radioisotopes/diagnostic use MH - Drug Resistance, Viral MH - Female MH - In Vitro MH - Interleukin-2/pharmacology MH - Lymphocyte Culture Test, Mixed MH - Mice MH - Mice, Inbred DBA MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - T-Lymphocytes, Cytotoxic/*drug effects/*immunology MH - Zidovudine/*pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2865-71. PMID- 12183240 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Antiviral activities of MCC-478, a novel and specific inhibitor of hepatitis B virus. PG - 2872-7 AB - MCC-478 is a newly synthesized 2-amino-6-arylthio-9-phosphonomethoxyethylpurine bis(2,2,2-trifluoroethyl) ester derivative. MCC-478 showed a substantially higher (ca. 80-fold) anti-hepatitis B virus (HBV) activity than that of lamivudine, despite no significant anti-human immunodeficiency virus activity. Since the bis(2,2,2-trifluoroethyl) ester group was used to improve the oral bioavailability of the phosphonomethoxyethylpurine derivatives, two monoester derivatives and one phosphonic acid derivative were also evaluated. It was suggested that these hydrolyzed derivatives, which appeared in animals given MCC-478, have enough anti-HBV activity to contribute to efficacy in vivo. Furthermore, no apparent cytotoxic effects or reductions of mitochondrial DNA content by MCC-478 and its derivatives were observed. These results indicated that MCC-478 may be a new promising anti-HBV agent. AD - Research Laboratory IV, Mitsubishi Pharma Corporation, Yokohama, Japan. Kamiya.Naohiro@mh.m-pharma.co.jp FAU - Kamiya, Naohiro AU - Kamiya N FAU - Kubota, Atsushi AU - Kubota A FAU - Iwase, Yumiko AU - Iwase Y FAU - Sekiya, Kouichi AU - Sekiya K FAU - Ubasawa, Masaru AU - Ubasawa M FAU - Yuasa, Satoshi AU - Yuasa S LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (2-amino-6-(4-methoxyphenylthio)-9-(2-(phosphonomethoxy)ethyl)purine bis(2,2,2-trifluoroethyl) ester) RN - 0 (Anti-HIV Agents) RN - 0 (Antiviral Agents) RN - 0 (DNA, Mitochondrial) RN - 0 (Purines) SB - IM MH - Animals MH - Anti-HIV Agents/pharmacology MH - Antiviral Agents/*pharmacology/toxicity MH - Cell Survival/drug effects MH - Cells, Cultured MH - DNA, Mitochondrial/drug effects MH - HIV-1/drug effects MH - Hepatitis B virus/*drug effects MH - Humans MH - Purines/*pharmacology MH - Tumor Cells, Cultured MH - Virus Replication/drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2872-7. PMID- 12183241 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Effects of neutrophils on cefazolin activity and penicillin-binding proteins in Staphylococcus aureus abscesses. PG - 2878-84 AB - Bacteria survive within abscesses despite antimicrobial therapy, usually necessitating drainage. Our previous work showed that bacterial killing is diminished within the neutrophils of animals with abscesses. To further assess the role of neutrophils in Staphylococcus aureus survival and the poor activities of beta-lactams in abscesses, tissue cage abscess-bearing rats were given polymorphonuclear leukocyte (PMN)-depleting antibody prior to and several times following inoculation of the tissue cages with S. aureus. Cefazolin (300 mg/kg of body weight/day) was administered to all animals in appropriately divided doses. After 7 days of antimicrobial therapy, the 17 animals that received anti-PMN serum had significantly fewer abscess neutrophils than the 18 controls and fewer abscess bacteria (5.55 versus 3.79 log(10) CFU/ml [P = 0.04]) than the 18 controls. The data were consistent with the premise that cefazolin is more effective in abscesses depleted of neutrophils. To investigate further, S. aureus was incubated with rat peritoneal neutrophils; and bacterial cell membrane proteins were isolated, labeled with biotinylated ampicillin, separated by electrophoresis, blotted onto nitrocellulose, and stained for biotin reactivity. PBP 2 expression was consistently and significantly decreased after a brief, nonkilling PMN exposure. These experiments showed that PMN depletion enhanced the activity of cefazolin in the abscess milieu. Furthermore, altered bacterial cell wall cefazolin targets may be the mechanism by which the PMN diminishes antimicrobial activity, suggesting the importance of the staphylococcus-PMN interaction in the outcome of established infections. AD - Section of Infectious Diseases, Department of Medicine, University of Missouri--Kansas City, Kansas City, Missouri 64108, USA. bambergerd@umkc.edu FAU - Bamberger, David M AU - Bamberger DM FAU - Herndon, Betty L AU - Herndon BL FAU - Fitch, Jeffrey AU - Fitch J FAU - Florkowski, Aaron AU - Florkowski A FAU - Parkhurst, Vera AU - Parkhurst V LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Bacterial Proteins) RN - 0 (Carrier Proteins) RN - 0 (Fluorescent Dyes) RN - 0 (Penicillin-Binding Proteins) RN - 25953-19-9 (Cefazolin) RN - 58-85-5 (Biotin) RN - 69-53-4 (Ampicillin) RN - EC 2.3.2.12 (Peptidyl Transferases) RN - EC 2.4.1.- (Hexosyltransferases) RN - EC 3.4.17.8 (Muramoylpentapeptide Carboxypeptidase) SB - IM MH - Abscess/immunology/*microbiology MH - Ampicillin/pharmacology MH - Animals MH - Anti-Bacterial Agents/pharmacology MH - *Bacterial Proteins MH - Biotin MH - Carrier Proteins/*metabolism MH - Cefazolin/*pharmacology MH - Fluorescent Dyes MH - *Hexosyltransferases MH - Lymphocyte Count MH - Male MH - Muramoylpentapeptide Carboxypeptidase/*metabolism MH - Neutropenia/microbiology MH - Neutrophils/drug effects/*physiology MH - Penicillin-Binding Proteins MH - *Peptidyl Transferases MH - Phagocytosis/drug effects MH - Rats MH - Research Support, Non-U.S. Gov't MH - Staphylococcal Infections/immunology/*microbiology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2878-84. PMID- 12183242 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20031114 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Efficacy of quinupristin-dalfopristin in preventing vascular graft infection due to Staphylococcus epidermidis with intermediate resistance to glycopeptides. PG - 2885-8 AB - A rat model was used to investigate the efficacy of quinupristin-dalfopristin (Q-D) in the prevention of vascular prosthetic graft infection due to methicillin-resistant Staphylococcus epidermidis with intermediate resistance to glycopeptides. The in vitro activity of the compound was compared to that of vancomycin by MIC determination and time-kill study. Moreover, the efficacy of collagen-sealed Q-D-soaked Dacron was evaluated in a rat model of graft infection. Graft infections were established in the subcutaneous tissue of the backs of 120 adult male Wistar rats. The in vivo study included a control group, one contaminated group that did not receive any antibiotic prophylaxis, two contaminated groups that received grafts soaked with 10 and 100 micro g of Q-D per ml, respectively, and two contaminated groups that received grafts soaked with 10 and 100 micro g of vancomycin per ml, respectively. Rats that received Dacron grafts soaked with 100 micro g of Q-D per ml showed no evidence of infection (<10 CFU/ml). In contrast, for rats that received Dacron grafts soaked with 10 micro g of Q-D per ml and Dacron grafts soaked with 10 or 100 micro g of vancomycin per ml, the quantitative graft cultures demonstrated 2.2 x 10(2) +/- 1.3 x 10(2), 2.2 x 10(6) +/- 1.9 x 10(5), and 5.6 x 10(2) +/- 0.3 x 10(2) CFU/ml, respectively. Taken together the results of the study demonstrate that the use of Dacron grafts soaked with Q-D can result in significant bacterial growth inhibition and show that this compound is potentially valuable for prevention of vascular prosthetic graft infection. AD - Institute of Infectious Diseases and Public Health, University of Ancona, Italy. anconacmi@interfree.it FAU - Giacometti, Andrea AU - Giacometti A FAU - Cirioni, Oscar AU - Cirioni O FAU - Ghiselli, Roberto AU - Ghiselli R FAU - Orlando, Fiorenza AU - Orlando F FAU - Mocchegiani, Federico AU - Mocchegiani F FAU - Riva, Alessandra AU - Riva A FAU - Del Prete, Maria Simona AU - Del Prete MS FAU - Saba, Vittorio AU - Saba V FAU - Scalise, Giorgio AU - Scalise G LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Glycopeptide) RN - 0 (Antibiotics, Peptide) RN - 11006-76-1 (Virginiamycin) RN - 126602-89-9 (quinupristin-dalfopristin) SB - IM MH - Animals MH - Antibiotics, Glycopeptide/*pharmacology MH - Antibiotics, Peptide/*therapeutic use MH - Blood Vessel Prosthesis/*adverse effects MH - Drug Resistance, Microbial MH - Male MH - Microbial Sensitivity Tests MH - Prosthesis-Related Infections/microbiology/*prevention & control MH - Rats MH - Rats, Wistar MH - Staphylococcal Infections/microbiology/*prevention & control MH - *Staphylococcus epidermidis MH - Vancomycin Resistance MH - Virginiamycin/*therapeutic use EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2885-8. PMID- 12183243 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - In vitro and in vivo synergy of fosmidomycin, a novel antimalarial drug, with clindamycin. PG - 2889-94 AB - Fosmidomycin acts through inhibition of 1-deoxy-D-xylulose 5-phosphate (DOXP) reductoisomerase, a key enzyme of the nonmevalonate pathway of isoprenoid biosynthesis. It possesses potent antimalarial activity in vitro and in murine malaria. In a recent clinical study, fosmidomycin was effective and well tolerated in the treatment of patients with acute uncomplicated Plasmodium falciparum malaria but resulted in an unacceptably high rate of recrudescence. In order to identify a potential combination partner, the interaction of fosmidomycin with a number of antimalarial drugs in current use was investigated in a series of in vitro experiments. Synergy was observed between fosmidomycin and the lincosamides, lincomycin and clindamycin. The efficacy of a combination of fosmidomycin and clindamycin was subsequently demonstrated in the Plasmodium vinckei mouse model. AD - Institute of Biochemistry, Academic Hospital Centre, Justus-Liebig-University, Giessen, Germany. Jochen.Wiesner@Jomaa.de FAU - Wiesner, Jochen AU - Wiesner J FAU - Henschker, Dajana AU - Henschker D FAU - Hutchinson, David B AU - Hutchinson DB FAU - Beck, Ewald AU - Beck E FAU - Jomaa, Hassan AU - Jomaa H LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Antimalarials) RN - 18323-44-9 (Clindamycin) RN - 23155-02-4 (Fosfomycin) RN - 66508-53-0 (fosmidomycin) SB - IM MH - Animals MH - Anti-Bacterial Agents/*pharmacology MH - Antimalarials/*pharmacology MH - Clindamycin/*pharmacology MH - Dose-Response Relationship, Drug MH - Drug Synergism MH - Fosfomycin/*analogs & derivatives/*pharmacology MH - Humans MH - Malaria/drug therapy/parasitology MH - Mice MH - Plasmodium falciparum/*drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2889-94. PMID- 12183244 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Ertapenem versus ceftriaxone followed by appropriate oral therapy for treatment of complicated urinary tract infections in adults: results of a prospective, randomized, double-blind multicenter study. PG - 2895-900 AB - The efficacy and safety of intravenous (i.v.) ertapenem (1 g once a day) with the option to switch to an oral agent for treatment of adults with complicated urinary tract infections (UTIs) were compared with that of i.v. ceftriaxone (1 g daily) with the same oral switch option in a multicenter, double-blind, prospective, randomized study. At entry, 592 patients were assigned to one of two strata: acute pyelonephritis or other complicated UTI without acute pyelonephritis. After a minimum of 3 days, patients could be switched to an oral antimicrobial agent. A total of 159 patients in the ertapenem group and 171 patients in the ceftriaxone group were microbiologically evaluable. Approximately 95% of patients in each treatment group were switched to oral therapy. The most common pathogens were Escherichia coli and Klebsiella pneumoniae. At the primary efficacy endpoint 5 to 9 days after treatment, 91.8% of patients who received ertapenem and 93.0% of those who received ceftriaxone had a favorable microbiological response (95% confidence interval for the difference, adjusting for strata, -7.6 to 5.1%), indicating that outcomes in the two treatment groups were equivalent. Microbiological success rates for the two treatment groups were similar when compared by stratum and also by severity of infection. The frequency and severity of drug-related adverse events were generally similar in both treatment groups. In this study, ertapenem was as effective as ceftriaxone for the initial treatment of complicated UTIs in adults, was generally well tolerated, and had a similar overall safety profile. AD - Alaska Clinical Research Center, Anchorage, Alaska, USA. FAU - Tomera, Kevin M AU - Tomera KM FAU - Burdmann, Emmanuel A AU - Burdmann EA FAU - Reyna, Oscar G Pamo AU - Reyna OG FAU - Jiang, Qi AU - Jiang Q FAU - Wimmer, Wendy M AU - Wimmer WM FAU - Woods, Gail L AU - Woods GL FAU - Gesser, Richard M AU - Gesser RM CN - Protocol 014 Study Group. LA - eng PT - Clinical Trial PT - Journal Article PT - Multicenter Study PT - Randomized Controlled Trial PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Anti-Infective Agents, Urinary) RN - 0 (Cephalosporins) RN - 0 (Lactams) RN - 0 (ertapenem) RN - 73384-59-5 (Ceftriaxone) SB - IM MH - Adult MH - Anti-Bacterial Agents/adverse effects/*therapeutic use MH - Anti-Infective Agents, Urinary/adverse effects/*therapeutic use MH - Ceftriaxone/adverse effects/*therapeutic use MH - Cephalosporins/adverse effects/*therapeutic use MH - Comparative Study MH - Double-Blind Method MH - Female MH - Humans MH - *Lactams MH - Male MH - Prospective Studies MH - Research Support, Non-U.S. Gov't MH - Urinary Tract Infections/*drug therapy/microbiology/urine EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2895-900. PMID- 12183245 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Method for estimation of low outer membrane permeability to beta-lactam antibiotics. PG - 2901-7 AB - The outer membrane of gram-negative bacteria plays a major role in beta-lactam resistance as it slows down antibiotic entry into the periplasm and therefore acts in synergy with beta-lactamases and efflux systems. Up to now, the quantitative estimation of low outer membrane permeability by the method of Zimmermann and Rosselet was difficult because of the secreted and cell surface-associated beta-lactamases. The method presented here uses the acylation of a highly sensitive periplasmic penicillin-binding protein (PBP) (BlaR-CTD) to assess the rate of beta-lactam penetration into the periplasm. The method is dedicated to measurement of low permeability and is only valid when the diffusion rate through the outer membrane is rate limiting. Cytoplasmic membrane associated PBPs do not interfere since they are acylated after the very sensitive BlaR-CTD. This method was used to measure the permeability of beta-lactamase-deficient strains of Enterobacter cloacae and Enterobacter aerogenes to benzylpenicillin, ampicillin, carbenicillin, cefotaxime, aztreonam, and cephacetrile. Except for that of cephacetrile, the permeability coefficients were equal to or below 10(-7) cm/s. For cephacetrile, carbenicillin, and benzylpenicillin, the outer membrane of E. cloacae was 20 to 60 times less permeable than that of Escherichia coli, whereas for cefotaxime, aztreonam, and ampicillin it was, respectively, 400, 1,000, and 700 times less permeable. The permeability coefficient for aztreonam is the lowest ever measured (P = 3.2 x 10(-9) cm/s). Using these values, the MICs for a beta-lactamase-overproducing strain of E. cloacae were successfully predicted, demonstrating the validity of the method. AD - Laboratoire d'Enzymologie and Centre d'Ingenierie des Proteines, Universite de Liege, Institut de Chimie, B-4000 Liege, Belgium. FAU - Lakaye, Bernard AU - Lakaye B FAU - Dubus, Alain AU - Dubus A FAU - Joris, Bernard AU - Joris B FAU - Frere, Jean-Marie AU - Frere JM LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Fluorescent Dyes) RN - 0 (Penicillins) RN - 0 (Plasmids) RN - 61-33-6 (Penicillin G) RN - EC 3.5.2.6 (beta-Lactamases) SB - IM MH - Acylation MH - Algorithms MH - Anti-Bacterial Agents/*metabolism MH - Bacteria/genetics/*metabolism MH - Electrophoresis, Polyacrylamide Gel MH - Enterobacter/drug effects MH - Fluorescent Dyes MH - Hydrolysis MH - Membranes/metabolism MH - Microbial Sensitivity Tests MH - Models, Biological MH - Penicillin G/metabolism MH - Penicillins/metabolism MH - Permeability MH - Plasmids MH - Research Support, Non-U.S. Gov't MH - beta-Lactamases/metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2901-7. PMID- 12183246 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20031114 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - High incidence of cefoxitin and clindamycin resistance among anaerobes in Taiwan. PG - 2908-13 AB - Susceptibilities to 16 antimicrobial agents were determined by measurement of MICs for 344 isolates of anaerobic bacteria recovered from patients with significant infections. Resistance rates varied among antimicrobial agents and the species tested. The beta-lactams were more active in gram-positive than in gram-negative anaerobes. Resistance to meropenem was low (<1%). For beta-lactam-beta-lactamase inhibitors, piperacillin-tazobactam was most active for all species (resistance, <6%). The rates of resistance to cefoxitin (31 to 65%) and clindamycin (50 to 70%) for non-Bacteroides fragilis species of the B. fragilis group were higher than those for B. fragilis (4% resistant to cefoxitin and 33% resistant to clindamycin). Among members of B. fragilis group, Bacteroides thetaiotaomicron was the most resistant to clindamycin (70%) and cefoxitin (65%). Rates of susceptibility to imipenem and metronidazole for B. fragilis continue to be high compared to those from a previous study 10 years ago. However, resistance to metronidazole was found recently in five strains of B. fragilis. We analyzed the genetic relationships among the metronidazole-resistant B. fragilis strains by pulsed-field gel electrophoresis. The metronidazole-resistant B. fragilis strains showed genotypic heterogeneity, excluding the dissemination of a single clone. AD - School of Medical Technology, National Taiwan University College of Medicine, Taipei, Taiwan. ljteng@ha.mc.ntu.edu.tw FAU - Teng, Lee-Jene AU - Teng LJ FAU - Hsueh, Po-Ren AU - Hsueh PR FAU - Tsai, Jui-Chang AU - Tsai JC FAU - Liaw, Shwu-Jen AU - Liaw SJ FAU - Ho, Shen-Wu AU - Ho SW FAU - Luh, Kwen-Tay AU - Luh KT LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Cephamycins) RN - 0 (DNA, Bacterial) RN - 18323-44-9 (Clindamycin) RN - 35607-66-0 (Cefoxitin) RN - 443-48-1 (Metronidazole) SB - IM MH - Anti-Bacterial Agents/*pharmacology MH - Bacteria, Anaerobic/*drug effects MH - Bacterial Infections/epidemiology/*microbiology MH - Bacteroides/drug effects MH - Bacteroides fragilis/drug effects MH - Cefoxitin/*pharmacology MH - Cephamycins/*pharmacology MH - Clindamycin/*pharmacology MH - DNA, Bacterial/drug effects/genetics MH - Drug Resistance, Microbial MH - Metronidazole/pharmacology MH - Microbial Sensitivity Tests MH - Taiwan/epidemiology MH - Time Factors EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2908-13. PMID- 12183247 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Characterization of sparsomycin resistance in Streptomyces sparsogenes. PG - 2914-9 AB - The antitumor antibiotic sparsomycin, produced by Streptomyces sparsogenes, is a universal translation inhibitor that blocks the peptide bond formation in ribosomes from all species. Sparsomycin-resistant strains were selected by transforming the sensitive Streptomyces lividans with an S. sparsogenes library. Resistance was linked to the presence of a plasmid containing an S. sparsogenes 5.9-kbp DNA insert. A restriction analysis of the insert traced down the resistance to a 3.6-kbp DNA fragment, which was sequenced. The analysis of the fragment nucleotide sequence together with the previous restriction data associate the resistance to srd, an open reading frame of 1,800 nucleotides. Ribosomes from S. sparsogenes and the S. lividans-resistant strains are equally sensitive to the inhibitor and bind the drug with similar affinity. Moreover, the drug was not modified by the resistant strains. However, resistant cells accumulated less antibiotic than the sensitive ones. In addition, membrane fractions from the resistant strains showed a higher capacity for binding the drug. The results indicate that resistance in the producer strain is not connected to either ribosome modification or drug inactivation, but it might be related to an alteration in the sparsomycin permeability barrier. AD - Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas y Universidad Autonoma de Madrid, Canto Blanco, 28049 Madrid, Spain. FAU - Lazaro, E AU - Lazaro E FAU - Sanz, E AU - Sanz E FAU - Remacha, M AU - Remacha M FAU - Ballesta, J P G AU - Ballesta JP LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Antineoplastic) RN - 0 (Culture Media) RN - 0 (DNA, Bacterial) RN - 1404-64-4 (Sparsomycin) RN - 63-91-2 (Phenylalanine) SB - IM MH - Antibiotics, Antineoplastic/metabolism/*pharmacology MH - Culture Media MH - DNA, Bacterial/genetics MH - Drug Resistance MH - Genomic Library MH - Kinetics MH - Microbial Sensitivity Tests MH - Phenylalanine/biosynthesis MH - Research Support, Non-U.S. Gov't MH - Ribosomes/genetics/metabolism MH - Sparsomycin/metabolism/*pharmacology MH - Streptomyces/*drug effects MH - Subcellular Fractions/drug effects/metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2914-9. PMID- 12183248 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Consumption of imipenem correlates with beta-lactam resistance in Pseudomonas aeruginosa. PG - 2920-5 AB - It is generally assumed that the antibiotic prescription policy of a hospital has a significant impact on bacterial resistance rates; however, few studies are available to support this concept with valid statistical data. During a 3-year period from 1997 to 2000, we monitored the consumption of beta-lactam and other antibiotics with known activity against Pseudomonas aeruginosa in a 600-bed community hospital. Monthly isolations of P. aeruginosa were assessed, and resistance rates were recorded. Partial correlation coefficients between consumption and resistance rates were determined, taking into account possible associations with other variables such as seasonal effects and transfers from other hospitals. A total of 30 +/- 7 novel P. aeruginosa strains per month were isolated without epidemic clustering. Prescriptions of imipenem varied significantly during the study period, while prescriptions of other antipseudomonal agents were stable, with the exception of an increase in piperacillin-tazobactam prescriptions. Rates of resistance of P. aeruginosa to the antimicrobial agents used showed a time course similar to figures for imipenem consumption. Monthly rates of resistance to imipenem (partial correlation coefficient [cc], 0.63), piperacillin-tazobactam (cc, 0.57), and ceftazidime (cc, 0.56) were significantly associated with imipenem prescription rates in the same or the preceding month, while consumption of ceftazidime or piperacillin-tazobactam had no apparent association with resistance. Among the variables investigated, imipenem consumption was identified as the major factor associated with both carbapenem and beta-lactam resistance in endemic P. aeruginosa. Periods of extensive imipenem use were associated with significant increases in resistance. Our data support the concept that a written antibiotic policy which balances the use of various antibiotic classes may help to avoid disturbances of a hospital's microbial sensitivity patterns. AD - Section of Hospital Hygiene, Department of Medical Microbiology and Hygiene, Ulm University Hospital, Germany. FAU - Lepper, Philipp M AU - Lepper PM FAU - Grusa, Eberhard AU - Grusa E FAU - Reichl, Helga AU - Reichl H FAU - Hogel, Josef AU - Hogel J FAU - Trautmann, Matthias AU - Trautmann M LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Thienamycins) RN - 64090-99-9 (thienamycin) RN - 74431-23-5 (Imipenem) SB - IM MH - Drug Utilization MH - Hospitals, Community/organization & administration MH - Humans MH - Hygiene MH - Imipenem/pharmacology/*therapeutic use MH - Prescriptions, Drug MH - Pseudomonas Infections/*epidemiology/*microbiology MH - Pseudomonas aeruginosa/*drug effects/*metabolism MH - Public Policy MH - Seasons MH - Thienamycins/pharmacology/*therapeutic use MH - *beta-Lactam Resistance EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2920-5. PMID- 12183249 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Human immunodeficiency virus type 1 genotypic and pharmacokinetic determinants of the virological response to lopinavir-ritonavir-containing therapy in protease inhibitor-experienced patients. PG - 2926-32 AB - The response to regimens including lopinavir-ritonavir (LPV/r) in patients who have received multiple protease (PR) inhibitors (PI) can be analyzed in terms of human immunodeficiency virus type 1 (HIV-1) genotypic and pharmacokinetic (pK) determinants. We studied these factors and the evolution of HIV-1 resistance in response to LPV/r in a prospective study of patients receiving LPV/r under a temporary authorization in Bordeaux, France. HIV-1 PR and reverse transcriptase sequences were determined at baseline LPV/r for all the patients and at month 3 (M3) and M6 in the absence of response to treatment. pK measurements were determined at M1 and M3. Virological failure (VF) was defined as a plasma viral load >or=400 copies/ml at M3. A multivariate analysis of the predictors of VF, including clinical and biological characteristics and the treatment history of the patients, was performed. The PR gene sequence at M0, including individual mutations or a previously defined LPV mutation score (D. J. Kempf, J. D. Isaacson, M. S. King, S. C. Brun, Y. Xu, K. Real, B. M. Bernstein, A. J. Japour, E. Sun, and R. A. Rode, J. Virol. 75:7262-7269, 2001), and the individual exposure to LPV were also included covariates. Sixty-eight patients were enrolled. Thirty-four percent had a virological response at M3. An LPV mutation score of >5 mutations, the presence of the PR I54V mutation at baseline, a high number of previous PIs, prior therapy with ritonavir or indinavir, absence of coprescription of efavirenz, and a lower exposure to LPV or lower LPV trough concentrations were independently associated with VF on LPV/r. Additional PI resistance mutations, including primary mutation I50V, could be selected in patients failing on LPV/r. Genotypic and pK parameters should be used to optimize the virological response to LPV/r in PI-experienced patients and to avoid further viral evolution. AD - Laboratoire de Virologie, Hopital Pellegrin, Bordeaux, France. bernard.masquelier@chu-bordeaux.fr FAU - Masquelier, Bernard AU - Masquelier B FAU - Breilh, Dominique AU - Breilh D FAU - Neau, Didier AU - Neau D FAU - Lawson-Ayayi, Sylvie AU - Lawson-Ayayi S FAU - Lavignolle, Valerie AU - Lavignolle V FAU - Ragnaud, Jean-Marie AU - Ragnaud JM FAU - Dupon, Michel AU - Dupon M FAU - Morlat, Philippe AU - Morlat P FAU - Dabis, F AU - Dabis F FAU - Fleury, H AU - Fleury H CN - Groupe d' Epidemiologie Clinique du SIDA en Aquitaine. LA - eng PT - Clinical Trial PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-HIV Agents) RN - 0 (Drug Combinations) RN - 0 (HIV Protease Inhibitors) RN - 0 (Pyrimidinones) RN - 0 (RNA, Viral) RN - 0 (Ritonavir) RN - 192725-17-0 (lopinavir) RN - EC 3.4.23.- (HIV Protease) SB - IM MH - Adult MH - Amino Acid Substitution MH - Anti-HIV Agents/*pharmacokinetics/*therapeutic use MH - Drug Combinations MH - Female MH - Genotype MH - HIV Infections/*drug therapy/*metabolism/virology MH - HIV Protease/genetics MH - HIV Protease Inhibitors/*pharmacokinetics/*therapeutic use MH - HIV-1/*drug effects/*genetics MH - Humans MH - Male MH - Models, Biological MH - Pyrimidinones/*pharmacokinetics/*therapeutic use MH - RNA, Viral/blood MH - Ritonavir/*pharmacokinetics/*therapeutic use MH - Salvage Therapy EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2926-32. PMID- 12183250 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20031114 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Comparative study of mechanisms of herpes simplex virus inactivation by sodium lauryl sulfate and n-lauroylsarcosine. PG - 2933-42 AB - The mechanisms of herpes simplex virus (HSV) inactivation by sodium lauryl sulfate (SLS) and n-lauroylsarcosine (LS), two anionic surfactants with protein denaturant potency, have been evaluated in cultured cells. Results showed that pretreatment of HSV type 1 (HSV-1) strain F and HSV-2 strain 333 with either surfactant inhibited, in a concentration- and time-dependent manner, their infectivities on Vero cells. SLS was a more potent inhibitor of HSV-2 strain 333 infectivity than LS with respect to the concentration (4.8-fold lower) and time (2.4-fold shorter) required to completely inactivate the virus. No inhibition of both herpesvirus strains infectivities was observed when Vero cells were pretreated with either surfactant. LS prevented the binding of HSV-2 strain 333 to cells without affecting the stable attachment and the rate of penetration into cells, whereas SLS exerted the opposite effect. Both SLS and LS inhibited, in a concentration-dependent manner, the HSV-2 strain 333-induced cytopathic effect, probably by affecting newly synthesized virions that come into contact with surfactant molecules present in culture medium. The pretreatment of HSV-2 strain 333 with specific combinations of SLS and LS concentrations inhibited the viral infectivity in a synergistic manner and resulted in only a small increase in their toxicities for exponentially growing Vero cells compared with that caused by each compound alone. Taken together, these results suggest that SLS and LS, alone or combined, could represent potent candidates as microbicides in topical vaginal formulations to prevent the transmission of herpes and possibly other pathogens that cause sexually transmitted diseases, including human immunodeficiency virus type 1. AD - Centre de Recherche en Infectiologie, Universite Laval, Quebec, Quebec, Canada. FAU - Piret, Jocelyne AU - Piret J FAU - Roy, Sylvie AU - Roy S FAU - Gagnon, Mylene AU - Gagnon M FAU - Landry, Sebastien AU - Landry S FAU - Desormeaux, Andre AU - Desormeaux A FAU - Omar, Rabeea F AU - Omar RF FAU - Bergeron, Michel G AU - Bergeron MG LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Receptors, Virus) RN - 0 (Surface-Active Agents) RN - 0 (Viral Proteins) RN - 107-97-1 (Sarcosine) RN - 151-21-3 (Sodium Dodecyl Sulfate) RN - 97-78-9 (sarkosyl) SB - IM MH - Animals MH - Cell Survival/drug effects MH - Cercopithecus aethiops MH - Comparative Study MH - Herpesvirus 1, Human/*drug effects MH - Herpesvirus 2, Human/*drug effects MH - Plaque Assay MH - Receptors, Virus/drug effects MH - Sarcosine/*analogs & derivatives/*pharmacology MH - Sodium Dodecyl Sulfate/*pharmacology MH - Surface-Active Agents/*pharmacology MH - Time Factors MH - Vero Cells MH - Viral Proteins/metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2933-42. PMID- 12183251 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Application of real-time PCR for determination of antiviral drug susceptibility of herpes simplex virus. PG - 2943-7 AB - A quantitative real-time PCR (TaqMan) assay was developed for determination of antiviral drug susceptibility of herpes simplex virus (HSV). After short-time culture of the virus, the antiviral drug susceptibility of HSV isolates for acyclovir (ACV) was determined by measuring the reduction of the HSV type 1 (HSV-1) DNA levels in culture supernatants using real-time PCR. The 50% inhibitory concentration was reported as the concentration of antiviral drug that reduced the number of HSV-1 DNA copies by 50%. A total of 15 well-characterized ACV-sensitive or -resistant strains and clinical isolates were used for assay evaluation. The new assay with real-time PCR readout permitted rapid (3 days), objective, and reproducible determination of HSV-1 drug susceptibilities with no need for stringent control of initial multiplicity of infection. Furthermore, the real-time PCR assay results showed good correlation (r = 0.86) with those for the plaque reduction assay. In conclusion, the real-time PCR assay described here is a suitable quantitative method for determination of antiviral susceptibility of HSV-1, amenable for use in the routine diagnostic virology laboratory. AD - Department of Virology, Eijkman-Winkler Center, University Medical Center Utrecht, The Netherlands. r.stranska@lab.azu.nl FAU - Stranska, Ruzena AU - Stranska R FAU - van Loon, Anton M AU - van Loon AM FAU - Polman, Merjo AU - Polman M FAU - Schuurman, Rob AU - Schuurman R LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antiviral Agents) RN - 0 (DNA, Viral) SB - IM MH - Animals MH - Antiviral Agents/*pharmacology MH - Cercopithecus aethiops MH - Computer Systems MH - DNA Replication/drug effects MH - DNA, Viral/analysis/biosynthesis MH - Herpesvirus 1, Human/drug effects MH - Kinetics MH - Plaque Assay MH - Reproducibility of Results MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction/*methods MH - Simplexvirus/*drug effects MH - Vero Cells EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2943-7. PMID- 12183253 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - In vitro selection of resistance in Haemophilus influenzae by amoxicillin-clavulanate, cefpodoxime, cefprozil, azithromycin, and clarithromycin. PG - 2956-62 AB - Abilities of amoxicillin-clavulanate, cefpodoxime, cefprozil, azithromycin, and clarithromycin to select resistant mutants of Haemophilus influenzae were tested by multistep and single-step methodologies. For multistep studies, 10 random strains were tested: 5 of these were beta-lactamase positive. After 50 daily subcultures in amoxicillin-clavulanate, MICs did not increase more than fourfold. However, cefprozil MICs increased eightfold for one strain. Clarithromycin and azithromycin gave a >4-fold increase in 8 and 10 strains after 14 to 46 and 20 to 50 days, respectively. Mutants selected by clarithromycin and azithromycin were associated with mutations in 23S rRNA and ribosomal proteins L4 and L22. Three mutants selected by clarithromycin or azithromycin had alterations in ribosomal protein L4, while five had alterations in ribosomal protein L22. Two mutants selected by azithromycin had mutations in the gene encoding 23S rRNA: one at position 2058 and the other at position 2059 (Escherichia coli numbering), with replacement of A by G. One clone selected by clarithromycin became hypersusceptible to macrolides. In single-step studies azithromycin and clarithromycin had the highest mutation rates, while amoxicillin-clavulanate had the lowest. All resistant clones were identical to parents as observed by pulsed-field gel electrophoresis. The MICs of azithromycin for azithromycin-resistant clones were 16 to >128 micro g/ml, and those of clarithromycin for clarithromycin-resistant clones were 32 to >128 micro g/ml in multistep studies. For strains selected by azithromycin, the MICs of clarithromycin were high and vice versa. After 50 daily subcultures in the presence of drugs, MICs of amoxicillin-clavulanate and cefpodoxime against H. influenzae did not rise more than fourfold, in contrast to cefprozil, azithromycin, and clarithromycin, whose MICs rose to variable degrees. AD - Department of Pathology, Hershey Medical Center, Hershey, Pennsylvania 17033, USA. FAU - Clark, Catherine AU - Clark C FAU - Bozdogan, Bulent AU - Bozdogan B FAU - Peric, Mihaela AU - Peric M FAU - Dewasse, Bonifacio AU - Dewasse B FAU - Jacobs, Michael R AU - Jacobs MR FAU - Appelbaum, Peter C AU - Appelbaum PC LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Antibiotics, Combined) RN - 0 (Cephalosporins) RN - 68401-81-0 (Ceftizoxime) RN - 74469-00-4 (Amoxicillin-Potassium Clavulanate Combination) RN - 81103-11-9 (Clarithromycin) RN - 82619-04-3 (cefpodoxime) RN - 83905-01-5 (Azithromycin) RN - 92665-29-7 (cefprozil) SB - IM MH - Amino Acid Sequence MH - Amoxicillin-Potassium Clavulanate Combination/*pharmacology MH - Anti-Bacterial Agents/*pharmacology MH - Antibiotics, Combined/*pharmacology MH - Azithromycin/*pharmacology MH - Ceftizoxime/*analogs & derivatives/*pharmacology MH - Cephalosporins/*pharmacology MH - Clarithromycin/*pharmacology MH - Drug Resistance MH - Genes, Bacterial/genetics MH - Haemophilus influenzae/*drug effects/genetics MH - Microbial Sensitivity Tests MH - Molecular Sequence Data MH - Mutation MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Transformation, Bacterial EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2956-62. PMID- 12183254 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Antistreptococcal activity of telithromycin compared with seven other drugs in relation to macrolide resistance mechanisms in Russia. PG - 2963-8 AB - The susceptibilities of 468 recent Russian clinical Streptococcus pneumoniae isolates and 600 Streptococcus pyogenes isolates, from 14 centers in Russia, to telithromycin, erythromycin, azithromycin, clarithromycin, clindamycin, levofloxacin, quinupristin-dalfopristin, and penicillin G were tested. Penicillin-nonsusceptible S. pneumoniae strains were rare except in Siberia, where their prevalence rate was 13.5%: most were penicillin intermediate, but for three strains (two from Smolensk and one from Novosibirsk) the MICs of penicillin G were 4 or 8 micro g/ml. Overall, 2.5% of S. pneumoniae isolates were resistant to erythromycin. Efflux was the prevalent resistance mechanism (five strains; 41.7%), followed by ribosomal methylation encoded by constitutive erm(B), which was found in four isolates. Ribosomal mutation was the mechanism of macrolide resistance in three isolates; one erythromycin-resistant S. pneumoniae isolate had an A2059G mutation in 23S rRNA, and two isolates had substitution of GTG by TPS at positions 69 to 71 in ribosomal protein L4. All S. pyogenes isolates were susceptible to penicillin, and 11% were erythromycin resistant. Ribosomal methylation was the most common resistance mechanism for S. pyogenes (89.4%). These methylases were encoded by erm(A) [subclass erm(TR)] genes, and their expression was inducible in 96.6% of isolates. The rest of the erythromycin-resistant Russian S. pyogenes isolates (7.6%) had an efflux resistance mechanism. Telithromycin was active against 100% of pneumococci and 99.2% of S. pyogenes, and levofloxacin and quinupristin-dalfopristin were active against all isolates of both species. AD - Institute of Antimicrobial Chemotherapy, Smolensk, Russia. FAU - Kozlov, Roman S AU - Kozlov RS FAU - Bogdanovitch, Tatiana M AU - Bogdanovitch TM FAU - Appelbaum, Peter C AU - Appelbaum PC FAU - Ednie, Lois AU - Ednie L FAU - Stratchounski, Leonid S AU - Stratchounski LS FAU - Jacobs, Michael R AU - Jacobs MR FAU - Bozdogan, Bulent AU - Bozdogan B LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Bacterial Proteins) RN - 0 (Ketolides) RN - 0 (Macrolides) RN - 0 (telithromycin) RN - EC 2.1.1. (Methyltransferases) RN - EC 2.1.1.48 (ErmA protein, Bacteria) SB - IM MH - Anti-Bacterial Agents/metabolism/*pharmacology MH - Bacterial Proteins/genetics MH - Comparative Study MH - Drug Resistance MH - Genes, Bacterial/genetics MH - *Ketolides MH - *Macrolides MH - Methylation MH - *Methyltransferases MH - Microbial Sensitivity Tests MH - Research Support, Non-U.S. Gov't MH - Ribosomes/metabolism MH - Russia/epidemiology MH - Streptococcal Infections/epidemiology/microbiology MH - Streptococcus/*drug effects/genetics/metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2963-8. PMID- 12183255 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Phase I dose escalation trial evaluating the pharmacokinetics, anti-human cytomegalovirus (HCMV) activity, and safety of 1263W94 in human immunodeficiency virus-infected men with asymptomatic HCMV shedding. PG - 2969-76 AB - 1263W94 [maribavir; 5,6-dichloro-2-(isopropylamino)-1,beta-L-ribofuranosyl-1-H-benzimidazole] is a novel benzimidazole compound for treatment of human cytomegalovirus (HCMV) infection and disease, with potent in vitro activity against HCMV and good oral bioavailability. A phase I study was conducted to determine the pharmacokinetics (PK), anti-HCMV activity, and safety of 1263W94 administered as multiple oral doses to human immunodeficiency virus type 1-infected adult male subjects with asymptomatic HCMV shedding. Subjects received one of six dosage regimens (100, 200, or 400 mg three times a day, or 600, 900, or 1,200 mg twice a day) or a placebo for 28 days. 1263W94 demonstrated linear PK, with steady-state plasma 1263W94 profiles predictable based on single-dose data. 1263W94 was rapidly absorbed following oral dosing, and values for the maximum concentration of the drug in plasma and the area under the concentration-time curve increased in proportion to the dose. 1263W94 demonstrated in vivo anti-HCMV activity in semen at all of the dosage regimens tested, with mean reductions in semen HCMV titers of 2.9 to 3.7 log(10) PFU/ml among the four regimens evaluated for anti-HCMV activity. 1263W94 was generally well tolerated; taste disturbance was the most frequently reported adverse event over the 28-day dosing period. AD - Quest Clinical Research, San Francisco, California, USA. FAU - Lalezari, Jacob P AU - Lalezari JP FAU - Aberg, Judith A AU - Aberg JA FAU - Wang, Laurene H AU - Wang LH FAU - Wire, Mary Beth AU - Wire MB FAU - Miner, Richard AU - Miner R FAU - Snowden, Wendy AU - Snowden W FAU - Talarico, Christine L AU - Talarico CL FAU - Shaw, Shuching AU - Shaw S FAU - Jacobson, Mark A AU - Jacobson MA FAU - Drew, W Lawrence AU - Drew WL LA - eng PT - Clinical Trial PT - Clinical Trial, Phase II PT - Journal Article PT - Randomized Controlled Trial PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Benzimidazoles) RN - 0 (Culture Media) RN - 0 (DNA, Viral) RN - 0 (Ribonucleosides) RN - 0 (maribavir) SB - IM MH - Adult MH - Area Under Curve MH - Benzimidazoles/adverse effects/*pharmacokinetics/*therapeutic use MH - Cells, Cultured MH - Culture Media MH - Cytomegalovirus Infections/*drug therapy/virology MH - DNA, Viral/biosynthesis/genetics MH - Dose-Response Relationship, Drug MH - HIV Infections/*drug therapy MH - Humans MH - Male MH - Plaque Assay MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Ribonucleosides/adverse effects/*pharmacokinetics/*therapeutic use MH - Semen/virology MH - Urine/virology MH - Virus Shedding EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2969-76. PMID- 12183256 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Characterization of a self-transferable plasmid from Salmonella enterica serotype typhimurium clinical isolates carrying two integron-borne gene cassettes together with virulence and drug resistance genes. PG - 2977-81 AB - An unusual self-transferable virulence-resistance plasmid (pUO-StVR2) was found in nine multidrug-resistant (ACSSuT phenotype) Salmonella enterica serotype Typhimurium clinical isolates that were assigned to four different phage types and a single and distinctive XbaI pulsed-field gel electrophoresis profile. pUO-StVR2 is an IncFII plasmid of about 140 kb in length carrying the spvA, spvB, and spvC (Salmonella plasmid virulence) and rck (resistance to complement killing) genes. It also carries the oxa1/aadA1a (ampicillin resistance and streptomycin-spectinomycin resistance) gene cassette configuration located within a class 1 integron with qacEDelta1/sul1 (ammonium antiseptics resistance and sulfadiazine resistance); the transposon genes merA, tnpA, and tnpR (mercury resistance, transposase, and resolvase of Tn21, respectively); and the catA1 (chloramphenicol resistance) and tet(B) (tetracycline resistance) genes. The insertion of resistance genes into a Salmonella virulence plasmid constitutes a new and interesting example of plasmid evolution and presents a serious public health problem. AD - Departamento de Biologia Funcional, Area de Microbiologia, Universidad de Oviedo, 33006 Oviedo, Principality of Asturias, Spain. FAU - Guerra, Beatriz AU - Guerra B FAU - Soto, Sara AU - Soto S FAU - Helmuth, Reiner AU - Helmuth R FAU - Mendoza, M Carmen AU - Mendoza MC LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (DNA, Bacterial) RN - 0 (Plasmids) RN - 0 (Virulence Factors) RN - EC 2.7.7.- (Tn21 resolvase) RN - EC 2.7.7.- (Transposon Resolvases) SB - IM MH - Bacteriophage Typing MH - DNA, Bacterial/biosynthesis/genetics/isolation & purification MH - Drug Resistance, Microbial MH - Genes, Bacterial/*genetics MH - Integrons/*genetics MH - Plasmids/*genetics MH - Research Support, Non-U.S. Gov't MH - Salmonella enterica/drug effects/*genetics/pathogenicity MH - Serotyping MH - *Transposon Resolvases MH - Virulence Factors EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2977-81. PMID- 12183257 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - In vitro interaction of flucytosine combined with amphotericin B or fluconazole against thirty-five yeast isolates determined by both the fractional inhibitory concentration index and the response surface approach. PG - 2982-9 AB - Combination therapy could be of benefit for the treatment of invasive yeast infections. However, in vitro interaction studies are relatively scarce and the interpretation of the fractional inhibitory concentration (FIC) index can be contradictory due to various definitions used; not all information on the interaction study is used in the index, and different MIC end points exist for different classes of drugs. Fitting an interaction model to the whole response surface and estimation of an interaction coefficient alpha (IC(alpha)) would overcome these objections and has the additional advantage that confidence intervals of the interaction are obtained. The efficacy of flucytosine (5FC) in combination with amphotericin B (AB) and fluconazole (FCZ) was studied against 35 yeast isolates in triplicate (Candida albicans [n = 9], Candida glabrata [n = 9], Candida krusei [n = 9], and Cryptococcus neoformans [n = 8]) using a broth microdilution checkerboard method and measuring growth after 48 h by a spectrophotometer. The FIC index and IC(alpha) were determined, the latter by estimation from the response surface approach described by Greco et al. (W. R. Greco, G. Bravo, and J. C. Parsons, Pharmacol. Rev. 47:331-385, 1995) by using a computer program developed for that purpose. For the 5FC-FCZ combination, the interactions determined by the IC(alpha) generally were in concordance with the interactions determined by the FIC index, but large discrepancies were found between both methods for the 5FC-AB combination. These could mainly be explained by shortcomings in the FIC approach. The in vitro interaction of 5FC-AB demonstrated variable results depending on the tested Candida isolate. In general, the 5FC-FCZ combination was antagonistic against Candida species, but for some Candida isolates synergism was found. For C. neoformans the interaction for both combinations was highly dependent on the tested isolate and the method used. Response surface approach is an alternative method for determining the interaction between antifungal agents. By using this approach, some of the problems encountered with the FIC were overcome. AD - Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands. FAU - Te Dorsthorst, D T A AU - Te Dorsthorst DT FAU - Verweij, P E AU - Verweij PE FAU - Meletiadis, J AU - Meletiadis J FAU - Bergervoet, M AU - Bergervoet M FAU - Punt, N C AU - Punt NC FAU - Meis, J F G M AU - Meis JF FAU - Mouton, J W AU - Mouton JW LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Antifungal) RN - 0 (Antifungal Agents) RN - 1397-89-3 (Amphotericin B) RN - 2022-85-7 (Flucytosine) RN - 86386-73-4 (Fluconazole) SB - IM MH - Algorithms MH - Amphotericin B/*pharmacology MH - Antibiotics, Antifungal/*pharmacology MH - Antifungal Agents/*pharmacology MH - Candida/*drug effects MH - Drug Interactions MH - Fluconazole/*pharmacology MH - Flucytosine/*pharmacology MH - Microbial Sensitivity Tests MH - Models, Biological EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2982-9. PMID- 12183258 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Pharmacodynamic assessment of ertapenem (MK-0826) against Streptococcus pneumoniae in a murine neutropenic thigh infection model. PG - 2990-5 AB - The objective of this study was to determine the susceptibility breakpoint of a new carbapenem, ertapenem (MK-0826), against Streptococcus pneumoniae strains based on bacterial density and survival studies in a murine thigh infection model. Sixteen S. pneumoniae isolates for which MICs ranged from 0.015 to 4.0 mg/liter were tested with neutropenic ICR mice. Animals were infected with bacteria at 10(5) to 10(6) CFU per thigh and were treated with ertapenem starting at 2 h postinfection for 4 days. Ertapenem was given subcutaneously at 50 mg/kg of body weight every 6 h, which simulates the human pharmacodynamic profile (in particular, the duration of time that the concentration of free drug remains above the MIC of 2 mg/liter). At 0 and 24 h postinfection, thighs were harvested for bacterial density determination. Survival was assessed during 4 days of therapy and 3 days after the therapy. A protein binding study was conducted with mice by use of the ultrafiltration method. Protein binding in mice was approximately 95%, which is comparable to that in humans. The average change in bacterial density ranged from -0.22 to -4.4 log CFU per thigh over 24 h compared to 0-h controls. The extent of microbial eradication was dependent on the MIC for the S. pneumoniae isolate. Substantial bactericidal activities (i.e., killing of approximately 2 log CFU per thigh) were consistently observed against isolates for which MICs were TTC, was present within this 361-bp fragment. This triple-base-pair substitution, present in both copies of the 16S rRNA gene of all our Tet(r) H. pylori transformants, resulted in an increased MIC of tetracycline that was identical to that for the Tet(r) strain 181. AD - Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Center Rotterdam, The Netherlands. FAU - Gerrits, Monique M AU - Gerrits MM FAU - de Zoete, Marcel R AU - de Zoete MR FAU - Arents, Niek L A AU - Arents NL FAU - Kuipers, Ernst J AU - Kuipers EJ FAU - Kusters, Johannes G AU - Kusters JG LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (RNA, Ribosomal, 16S) RN - 0 (Repressor Proteins) RN - 0 (tetracycline resistance-encoding transposon repressor protein) SB - IM MH - Aged MH - Amino Acid Substitution/genetics MH - Binding Sites MH - Helicobacter Infections/microbiology MH - Helicobacter pylori/drug effects/*genetics MH - Humans MH - Male MH - Mutation/*genetics MH - RNA, Ribosomal, 16S/*genetics MH - Repressor Proteins/genetics MH - Reverse Transcriptase Polymerase Chain Reaction MH - Tetracycline Resistance/*genetics MH - Transformation, Bacterial EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):2996-3000. PMID- 12183260 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041209 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - The antifungal echinocandin caspofungin acetate kills growing cells of Aspergillus fumigatus in vitro. PG - 3001-12 AB - Caspofungin acetate is an antifungal antibiotic that inhibits synthesis of 1,3-beta-D-glucan, an essential component of the fungal cell wall. While caspofungin causes cell death in yeasts and dimorphic fungi such as Candida albicans, its effect on Aspergillus fumigatus is less well understood. We used the fluorescent dyes 5,(6)-carboxyfluorescein diacetate (CFDA) and bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC), which stain live and dead cells, respectively, to further characterize the antifungal activity of caspofungin. For comparison, compounds whose mode of action was either fungistatic (fluconazole, itraconazole) or fungicidal (amphotericin B) were also evaluated. A correlation between caspofungin-induced loss of viability, decreased CFDA staining, and increased DiBAC staining was established first with C. albicans. For A. fumigatus, caspofungin caused similar dye-staining changes, which were quantified by fluorimetric analysis of stained hyphae grown in a medium that promoted dispersed growth. The minimum concentration of caspofungin required to produce these changes also decreased the level of growth-dependent reduction of the indicator dye Alamar Blue. We observed a differential effect of caspofungin as a function of cell position: 88% of apical cells and 61% of subapical branching cells failed to stain with the viable dye CFDA, but only 24% of subapical cells were unstained. Complementary results were seen with germlings from DiBAC-stained, caspofungin-treated cultures. Extended incubation of A. fumigatus with a single dose of caspofungin affected the same proportion of apical and subapical branching cells for up to 72 h. The dye-staining patterns illustrate that the cells at the active centers for new cell wall synthesis within A. fumigatus hyphae are killed when they are exposed to caspofungin. AD - Department of Human and Animal Infectious Disease Research, Merck Research Laboratories, Rahway, New Jersey 07065-0900, USA. FAU - Bowman, J C AU - Bowman JC FAU - Hicks, P Scott AU - Hicks PS FAU - Kurtz, M B AU - Kurtz MB FAU - Rosen, H AU - Rosen H FAU - Schmatz, D M AU - Schmatz DM FAU - Liberator, P A AU - Liberator PA FAU - Douglas, C M AU - Douglas CM LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Antifungal) RN - 0 (Antibiotics, Peptide) RN - 0 (Fluorescent Dyes) RN - 0 (Peptides) RN - 0 (Peptides, Cyclic) RN - 0 (caspofungin) SB - IM MH - Antibiotics, Antifungal/*pharmacology MH - Antibiotics, Peptide/*pharmacology MH - Aspergillus fumigatus/*drug effects/growth & development MH - Cell Count MH - Dose-Response Relationship, Drug MH - Fluorescent Dyes MH - Fluorometry MH - Microbial Sensitivity Tests MH - *Peptides MH - *Peptides, Cyclic EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3001-12. PMID- 12183261 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Pharmacokinetic and pharmacodynamic profiles of danofloxacin administered by two dosing regimens in calves infected with Mannheimia (Pasteurella) haemolytica. PG - 3013-9 AB - The pharmacokinetics and pharmacodynamics of danofloxacin in calves with induced Mannheimia (Pasteurella) haemolytica pneumonia were evaluated. Calves received either saline as an intravenous (IV) bolus or danofloxacin (0.738 mg/kg of body weight) administered as either a single IV bolus or a 36-h continuous IV infusion. Blood samples and bronchial secretions were collected before and at predetermined times over 48 h following the start of treatment. Calves were assessed clinically throughout, and lung consolidation was assessed at necropsy. Bronchial secretions and lung tissue were cultured for M. haemolytica. Bolus administration of danofloxacin produced a high maximum drug concentration-to-MIC ratio (C(max):MIC) of 14.5 and a time period of 9.1 h when plasma danofloxacin concentrations exceeded the MIC (T>MIC). Following danofloxacin infusion, the C(max):MIC was low (2.3), with a long T>MIC (33.3 h). The area under the curve-to-MIC ratios were 43.3 and 49.1 for the bolus and infusion administrations, respectively. The single bolus of danofloxacin was more effective than the same dose administered by continuous infusion, as indicated by a significantly lower (P < 0.05) number of animals with M. haemolytica in bronchial secretions after treatment and lower rectal temperatures in the 24 h after the start of treatment. Thus, danofloxacin exhibited concentration-dependent antimicrobial activity in cattle with respiratory disease caused by M. haemolytica. AD - Ondax Scientific, 20280 Hondarribia, Gipuzkoa, Spain. FAU - Sarasola, Patxi AU - Sarasola P FAU - Lees, Peter AU - Lees P FAU - AliAbadi, Fariborz Shojaee AU - AliAbadi FS FAU - McKellar, Quintin A AU - McKellar QA FAU - Donachie, William AU - Donachie W FAU - Marr, Kate A AU - Marr KA FAU - Sunderland, Simon J AU - Sunderland SJ FAU - Rowan, Tim G AU - Rowan TG LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Infective Agents) RN - 0 (Fluoroquinolones) RN - 112398-08-0 (danofloxacin) SB - IM MH - Animals MH - Anti-Infective Agents/administration & dosage/*pharmacokinetics/*therapeutic use MH - Area Under Curve MH - Dogs MH - *Fluoroquinolones MH - Infusions, Intravenous MH - Injections, Intravenous MH - Lung/microbiology MH - Male MH - Mannheimia haemolytica/*drug effects MH - Pasteurella Infections/*drug therapy/microbiology/physiopathology MH - Pneumonia, Bacterial/drug therapy/microbiology/physiopathology MH - Respiratory Mechanics/drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3013-9. PMID- 12183262 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Mutation in 23S rRNA associated with macrolide resistance in Neisseria gonorrhoeae. PG - 3020-5 AB - Fifty-six azithromycin-resistant (MICs, 2.0 to 4.0 micro g/ml) Neisseria gonorrhoeae strains with cross-resistance to erythromycin (MICs, 2.0 to 64.0 micro g/ml), isolated in Canada between 1997 and 1999, were characterized, and their mechanisms of azithromycin resistance were determined. Most (58.9%) of them belonged to auxotype-serotype class NR/IB-03, with a 2.6-mDa plasmid. Based on resistance to crystal violet (MICs >or= 1 micro g/ml), 96.4% of these macrolide-resistant strains appeared to have increased efflux. Nine of the eleven strains selected for further characterization were found to have a promoter region mtrR mutation, a single-base-pair (A) deletion in the 13-bp inverted repeat, which is believed to cause overexpression of the mtrCDE-encoded efflux pump. The two remaining macrolide-resistant strains (erythromycin MIC, 64.0 micro g/ml; azithromycin MIC, 4.0 micro g/ml), which did not have the mutation in the mtrR promoter region, were found to have a C2611T mutation (Escherichia coli numbering) in the peptidyltransferase loop in domain V of the 23S rRNA alleles. Although mutations in domain V of 23S rRNA alleles had been reported in other bacteria, including E. coli, Streptococcus pneumoniae, and Helicobacter pylori, this is the first observation of these mutations associated with macrolide resistance in N. gonorrhoeae. AD - National Laboratory for Sexually Transmitted Diseases, National Microbiology Laboratory, Population and Public Health Branch, Health Canada, Winnipeg, Manitoba, Canada. Lai_King_Ng@hc-sc.gc.ca FAU - Ng, Lai-King AU - Ng LK FAU - Martin, Irene AU - Martin I FAU - Liu, Gary AU - Liu G FAU - Bryden, Louis AU - Bryden L LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (DNA, Bacterial) RN - 0 (Oligonucleotide Probes) RN - 0 (RNA, Ribosomal, 23S) RN - 114-07-8 (Erythromycin) RN - 83905-01-5 (Azithromycin) RN - EC 2.3.2.12 (Peptidyl Transferases) SB - IM MH - Anti-Bacterial Agents/*pharmacology MH - Azithromycin/pharmacology MH - Canada/epidemiology MH - DNA, Bacterial/drug effects/genetics MH - Drug Resistance MH - Erythromycin/pharmacology MH - Gonorrhea/epidemiology/microbiology MH - Humans MH - Microbial Sensitivity Tests MH - Mutation/genetics MH - Neisseria gonorrhoeae/*drug effects MH - Nucleic Acid Conformation MH - Oligonucleotide Probes MH - Peptidyl Transferases/genetics MH - RNA, Ribosomal, 23S/*genetics MH - Reverse Transcriptase Polymerase Chain Reaction MH - Transformation, Bacterial EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3020-5. PMID- 12183263 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Antibiotic pharmacodynamics in surgical prophylaxis: an association between intraoperative antibiotic concentrations and efficacy. PG - 3026-30 AB - The objective of this study was to characterize the relationship between gentamicin concentrations during surgery and the development of wound infection following colorectal operations. Despite decades of research in surgical prophylaxis, the relationship between intraoperative antibiotic concentrations and postoperative infection and the concentrations required for effective prophylaxis have not been established. A pharmacodynamic analysis was conducted using data from a previous prospective, randomized, double-blind clinical study which compared two dosage regimens of gentamicin plus metronidazole for prophylaxis in connection with elective colorectal surgery. Univariate and multivariate analyses of risk factors for postoperative wound infection were conducted, and the relationship between intraoperative gentamicin concentrations and surgical outcome was characterized. The gentamicin concentration at the time of surgical closure was one of the strongest independent risk factors for infection (P = 0.02), along with the presence of diabetes mellitus (P = 0.02), stoma (P = 0.04), and advanced age (P = 0.05). Gentamicin concentrations at closure of less than 0.5 mg/liter were associated with an infection rate of 80% (representing 8 of 10 patients with concentrations below that level) (P = 0.003). Receiver operating characteristic curve analysis identified a critical closure concentration of 1.6 mg/liter for effective surgical prophylaxis (P = 0.002; sensitivity, 70.8%; specificity, 65.9%). This study provides new and important information on antibiotic pharmacodynamics in surgical prophylaxis. It demonstrates the critical effect of the antibiotic concentration at closure on wound infection and suggests a significant association between the concentration and other well-established risk factors, like the timing of preoperative antibiotic administration and surgery duration. AD - Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada. zelenits@ms.umanitoba.ca FAU - Zelenitsky, Sheryl A AU - Zelenitsky SA FAU - Ariano, Robert E AU - Ariano RE FAU - Harding, Godfrey K M AU - Harding GK FAU - Silverman, Richard E AU - Silverman RE LA - eng PT - Clinical Trial PT - Journal Article PT - Randomized Controlled Trial PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Gentamicins) SB - IM MH - Algorithms MH - Anti-Bacterial Agents/blood/*pharmacokinetics/therapeutic use MH - *Antibiotic Prophylaxis MH - Area Under Curve MH - Colon/surgery MH - Double-Blind Method MH - Female MH - Gentamicins/blood/pharmacokinetics/therapeutic use MH - Humans MH - Intraoperative Period MH - Male MH - Middle Aged MH - Rectum/surgery MH - Risk Factors MH - Surgical Wound Infection/epidemiology/microbiology/prevention & control EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3026-30. PMID- 12183264 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Prospective survey of beta-lactamases produced by ceftazidime- resistant Pseudomonas aeruginosa isolated in a French hospital in 2000. PG - 3031-4 AB - In 2000, at the Universite d'Auvergne teaching hospital in Clermont-Ferrand, France, 44 (6.2%) strains of Pseudomonas aeruginosa were found to be resistant to ceftazidime. After genotyping, 34 strains were selected. Nine had an additional beta-lactamase: OXA-21 (n = 6), PSE-1 (CARB-2) (n = 2), or PER-1 (n = 1). Ceftazidime resistance was related solely to the overproduction of the cephalosporinase in 30 strains. Sequencing of five bla(AmpC) genes encoding cephalosporinases with different pIs showed 99% identity with the ampC gene of P. aeruginosa PAO1. AD - Service de Bacteriologie, Faculte de Medecine, Universite d'Auvergne, 63001 Clermont-Ferrand, France. christophe.dechamps@u-clermont1.fr FAU - De Champs, Christophe AU - De Champs C FAU - Poirel, Laurent AU - Poirel L FAU - Bonnet, Richard AU - Bonnet R FAU - Sirot, Danielle AU - Sirot D FAU - Chanal, Catherine AU - Chanal C FAU - Sirot, Jacques AU - Sirot J FAU - Nordmann, Patrice AU - Nordmann P LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Cephalosporins) RN - 78439-06-2 (Ceftazidime) RN - EC 3.5.2.- (Cephalosporinase) RN - EC 3.5.2.6 (beta-Lactamases) SB - IM MH - Ceftazidime/*pharmacology/therapeutic use MH - Cephalosporinase/antagonists & inhibitors/genetics/metabolism MH - Cephalosporins/*pharmacology/therapeutic use MH - Data Collection MH - Drug Resistance, Microbial MH - France/epidemiology MH - Genes, Bacterial MH - Genotype MH - Humans MH - Microbial Sensitivity Tests MH - Prospective Studies MH - Pseudomonas Infections/*epidemiology/*microbiology MH - Pseudomonas aeruginosa/*drug effects/*enzymology MH - Research Support, Non-U.S. Gov't MH - beta-Lactamases/antagonists & inhibitors/genetics/*metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3031-4. PMID- 12183265 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Predicting evolution by in vitro evolution requires determining evolutionary pathways. PG - 3035-8 AB - In an early example of DNA shuffling, Stemmer (W. P. C. Stemmer, Nature 370:389-390, 1994) demonstrated a dramatic improvement in the activity of the TEM-1 beta-lactamase toward cefotaxime as the consequence of six amino acid substitutions. It has been pointed out (B. G. Hall, FEMS Microbiol. Lett. 178:1-6, 1999; M. C. Orencia, J. S. Yoon, J. E. Ness, W. P. Stemmer, and R. C. Stevens, Nat. Struct. Biol. 8:238-242, 2001) that the power of DNA shuffling might be applied to the problem of predicting evolution in nature from in vitro evolution in the laboratory. As a predictor of natural evolutionary processes, that power may be misleading because in nature mutations almost always arise one at a time, and each advantageous mutation must be fixed into the population by an evolutionary pathway that leads from the wild type to the fully evolved sequence. Site-directed mutagenesis was used to introduce each of Stemmer's six substitutions into TEM-1, the best single mutant was chosen, and each of the remaining five substitutions was introduced. Repeated rounds of site-directed mutagenesis and selection of the best mutant were used in an attempt to construct a pathway between the wild-type TEM-1 and Stemmer's mutant with six mutations. In the present study it is shown (i) that no such pathway exists between the wild-type TEM-1 and the supereffective cefotaxime-hydrolyzing mutant that was generated by six amino acid substitutions via DNA shuffling (Stemmer, Nature 370:389-390, 1994) but that a pathway to a fourfold more efficient enzyme resulting from four of the same substitutions does exist, and (ii) that the more efficient enzyme is likely to arise in nature as the result of a single mutation in the naturally occurring TEM-52 allele. AD - Biology Department, University of Rochester, Rochester, New York 14627-0211, USA. drbh@mail.rochester.edu FAU - Hall, Barry G AU - Hall BG LA - eng GR - GM60761/GM/NIGMS PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (DNA, Bacterial) RN - EC 3.5.2.6 (beta-Lactamases) SB - IM MH - Alleles MH - Amino Acid Substitution/genetics MH - *DNA Shuffling MH - DNA, Bacterial/genetics MH - Drug Resistance, Microbial/*genetics MH - *Evolution MH - Mutation/genetics MH - Research Support, U.S. Gov't, P.H.S. MH - beta-Lactamases/*genetics/metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3035-8. PMID- 12183266 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041209 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - In vitro interaction of caspofungin acetate with voriconazole against clinical isolates of Aspergillus spp. PG - 3039-41 AB - The interaction between caspofungin acetate and voriconazole was studied in vitro by using 48 clinical Aspergillus spp. isolates obtained from patients with invasive aspergillosis. MICs were determined by the NCCLS broth microdilution method. Synergy, defined as a fractional inhibitory concentration (FIC) index of <1, was detected in 87.5% of the interactions; an additive effect, defined as an FIC index of 1.0, was observed in 4.2% of the interactions; and a subadditive effect, defined as an FIC index of 1.0 to 2.0, was found in 8.3% of the interactions. No antagonism was observed. Animal models are required to validate the in vivo significance of these in vitro data presented for the combination of caspofungin and voriconazole. AD - Department of Medicine, Division of Infectious Diseases, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900, USA. perea@uthscsa.edu FAU - Perea, Sofia AU - Perea S FAU - Gonzalez, Gloria AU - Gonzalez G FAU - Fothergill, Annette W AU - Fothergill AW FAU - Kirkpatrick, William R AU - Kirkpatrick WR FAU - Rinaldi, Michael G AU - Rinaldi MG FAU - Patterson, Thomas F AU - Patterson TF LA - eng GR - 5 R01 DE11381/DE/NIDCR GR - M01-RR-01346/RR/NCRR PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Peptide) RN - 0 (Antifungal Agents) RN - 0 (Peptides) RN - 0 (Peptides, Cyclic) RN - 0 (Pyrimidines) RN - 0 (Triazoles) RN - 0 (caspofungin) RN - 0 (voriconazole) SB - IM MH - Antibiotics, Peptide/*pharmacology MH - Antifungal Agents/*pharmacology MH - Aspergillosis/*microbiology MH - Aspergillus/*drug effects MH - Drug Interactions MH - Humans MH - Microbial Sensitivity Tests MH - *Peptides MH - *Peptides, Cyclic MH - Pyrimidines/*pharmacology MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Triazoles/*pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3039-41. PMID- 12183267 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Surveillance for antiviral-agent-resistant herpes simplex virus in the general population with recurrent herpes labialis. PG - 3042-4 AB - In a general population survey in the United States, the prevalence of antiviral-agent-resistant herpes simplex virus was very low among more than 1,000 isolates from individuals with an episode of recurrent herpes labialis not treated with topical antiviral agents. Two isolates had borderline resistance to acyclovir (0.2%), and all were susceptible to penciclovir. AD - GlaxoSmithKline Consumer Healthcare, Weybridge, Surrey KT15 0DE, United Kingdom. teresa.h.bacon@gsk.com FAU - Bacon, Teresa H AU - Bacon TH FAU - Boon, Ron J AU - Boon RJ FAU - Schultz, Margaret AU - Schultz M FAU - Hodges-Savola, Cheryl AU - Hodges-Savola C LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antiviral Agents) RN - 39809-25-1 (penciclovir) RN - 59277-89-3 (Acyclovir) SB - IM MH - Acyclovir/administration & dosage/*analogs & derivatives/pharmacology/therapeutic use MH - Administration, Topical MH - Adult MH - Animals MH - Antiviral Agents/*pharmacology MH - Cells, Cultured MH - Drug Resistance, Viral MH - Female MH - Herpes Labialis/*drug therapy/epidemiology/virology MH - Herpesvirus 1, Human/drug effects MH - Humans MH - Male MH - Population MH - Rabbits MH - Recurrence MH - United States/epidemiology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3042-4. PMID- 12183268 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Beta-lactamases of Kluyvera ascorbata, probable progenitors of some plasmid-encoded CTX-M types. PG - 3045-9 AB - Kluyvera ascorbata produces a beta-lactamase that results in an atypical susceptibility pattern, including low-level resistance to penicillins, cephalothin, and cefuroxime, but this resistance is reversed by clavulanate. Ten nucleotide sequences of the corresponding gene, bla(KLUA), were obtained and were found to have minor variations (96 to 100%). Otherwise, bla(KLUA) was found to be similar (95 to 100%) to some plasmid-encoded CTX-M-type beta-lactamases. Finally, mobilization of bla(KLUA) on a plasmid was found to be mediated probably by a genetic mobile element like ISEcp1. AD - Service de Bacteriologie, CHU Cochin, Paris, France. FAU - Humeniuk, Christel AU - Humeniuk C FAU - Arlet, Guillaume AU - Arlet G FAU - Gautier, Valerie AU - Gautier V FAU - Grimont, Patrick AU - Grimont P FAU - Labia, Roger AU - Labia R FAU - Philippon, Alain AU - Philippon A LA - eng SI - GENBANK/AF252622 SI - GENBANK/AF252623 SI - GENBANK/AF311345 SI - GENBANK/AJ272538 PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Plasmids) RN - EC 3.5.2.6 (beta-Lactamases) RN - EC 3.5.2.6 (beta-lactamase bla(KLUA), Klyvera ascorbata) SB - IM MH - Enterobacteriaceae/drug effects/*enzymology/genetics MH - Genes, Bacterial/genetics MH - Genotype MH - Kinetics MH - Microbial Sensitivity Tests MH - Molecular Sequence Data MH - Plasmids/*genetics MH - Research Support, Non-U.S. Gov't MH - beta-Lactamases/*genetics/*metabolism EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3045-9. PMID- 12183269 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20031114 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Functional characterization of Brucella melitensis NorMI, an efflux pump belonging to the multidrug and toxic compound extrusion family. PG - 3050-3 AB - Two putative proteins (NorMI and NorMII) similar to the multidrug efflux protein NorM of Vibrio parahaemolyticus are encoded by the Brucella melitensis 16 M genome. We show that a drug-hypersusceptible Escherichia coli strain overexpressing NorMI displays increased resistance to norfloxacin, ciprofloxacin, gentamicin, tetraphenylphosphonium ion, acriflavine, and berberine. This elevated resistance was proven to be mediated by an energy-dependent efflux mechanism. NorMI belongs to the multidrug and toxic compound extrusion family and is the first multidrug efflux protein identified in Brucella spp. AD - Unite de Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, 37380 Nouzilly, France. FAU - Braibant, Martine AU - Braibant M FAU - Guilloteau, Laurence AU - Guilloteau L FAU - Zygmunt, Michel S AU - Zygmunt MS LA - eng SI - GENBANK/AE008917 SI - GENBANK/AE008918 PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Membrane Transport Proteins) SB - IM MH - Anti-Bacterial Agents/*metabolism/*pharmacology MH - Biological Transport, Active MH - Brucella melitensis/*genetics/metabolism MH - Drug Resistance, Multiple, Bacterial/genetics MH - Energy Metabolism/physiology MH - Escherichia coli/drug effects MH - Genome, Bacterial MH - Kinetics MH - Membrane Transport Proteins/*metabolism MH - Molecular Sequence Data EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3050-3. PMID- 12183270 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Clinical isolates of Staphylococcus aureus with ribosomal mutations conferring resistance to macrolides. PG - 3054-6 AB - Six strains of Staphylococcus aureus isolated from cystic fibrosis patients after treatment with azithromycin were cross-resistant to azithromycin and erythromycin. None of the isolates contained erm or msr(A) genes, but they all carried either A2058G/U or A2059G mutations within the rrl genes, with a majority of the rRNA copies bearing the mutation. One strain displayed an additional mutation in the rplV gene, encoding the L22 ribosomal protein. AD - Service de Microbiologie, CHU Cote de Nacre, Caen, France. FAU - Prunier, Anne-Laure AU - Prunier AL FAU - Malbruny, Brigitte AU - Malbruny B FAU - Tande, Didier AU - Tande D FAU - Picard, Bertrand AU - Picard B FAU - Leclercq, Roland AU - Leclercq R LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Bacterial Proteins) RN - 0 (DNA, Bacterial) RN - 0 (Membrane Transport Proteins) RN - 0 (Oligonucleotide Probes) RN - 0 (msrA protein, Staphylococcus epidermidis) RN - 114-07-8 (Erythromycin) RN - 83905-01-5 (Azithromycin) RN - EC 2.1.1. (Methyltransferases) RN - EC 2.1.1.48 (ErmA protein, Bacteria) SB - IM MH - Anti-Bacterial Agents/*pharmacology MH - Azithromycin/pharmacology MH - Bacterial Proteins/genetics MH - Cystic Fibrosis/microbiology MH - DNA, Bacterial/genetics MH - Drug Resistance MH - Erythromycin/pharmacology MH - Genes, Bacterial/genetics MH - Humans MH - *Membrane Transport Proteins MH - *Methyltransferases MH - Microbial Sensitivity Tests MH - Mutation/*genetics MH - Oligonucleotide Probes MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Ribosomes/*genetics MH - Staphylococcal Infections/*microbiology MH - Staphylococcus aureus/*genetics EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3054-6. PMID- 12183271 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Phenylpropenamide derivatives AT-61 and AT-130 inhibit replication of wild-type and lamivudine-resistant strains of hepatitis B virus in vitro. PG - 3057-60 AB - The phenylpropenamide derivatives AT-61 and AT-130 are nonnucleoside analogue inhibitors of hepatitis B virus (HBV) replication. They inhibited the replication of wild-type HBV with 50% inhibitory concentrations of 21.2 +/- 9.5 and 2.40 +/- 0.92 micro M, respectively, compared to 0.064 +/- 0.020 micro M lamivudine. There were no significant differences in sensitivity between wild-type and nucleoside analogue-resistant (rtL180M, rtM204I, and rtL180M + rtM204V) HBV. AD - Victorian Infectious Diseases Reference Laboratory, North Melbourne, Victoria 3051, Australia. FAU - Delaney, William E 4th AU - Delaney WE 4th FAU - Edwards, Ros AU - Edwards R FAU - Colledge, Danni AU - Colledge D FAU - Shaw, Tim AU - Shaw T FAU - Furman, Phil AU - Furman P FAU - Painter, George AU - Painter G FAU - Locarnini, Stephen AU - Locarnini S LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (AT 130) RN - 0 (AT 61) RN - 0 (Amides) RN - 0 (Anti-HIV Agents) RN - 0 (Benzamides) RN - 0 (Benzene Derivatives) RN - 134678-17-4 (Lamivudine) SB - IM MH - Amides/*pharmacology MH - Anti-HIV Agents/*pharmacology MH - Benzamides/*pharmacology MH - Benzene Derivatives/*pharmacology MH - Drug Resistance, Viral MH - Hepatitis B virus/*drug effects MH - Lamivudine/*pharmacology MH - Plaque Assay MH - Research Support, Non-U.S. Gov't MH - Virus Replication/*drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3057-60. PMID- 12183272 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Efficacies of quinupristin-dalfopristin combined with vancomycin in vitro and in experimental endocarditis due to methicillin-resistant Staphylococcus aureus in relation to cross-resistance to macrolides, lincosamides, and streptogramin B- type antibiotics. PG - 3061-4 AB - A beneficial effect of the combination of quinupristin-dalfopristin and vancomycin was observed against two methicillin-resistant strains of Staphylococcus aureus harboring or not harboring the ermC gene, which codes for constitutive macrolide, lincosamide, and streptogramin B resistance. The beneficial effect was observed in time-kill studies, in which the drugs were used at inhibitory concentrations, and in a rabbit model of endocarditis, in which the combination was highly bactericidal and more active than monotherapies. AD - Institut National de la Sante et de la Recherche Medicale (EMI 9933), Faculte de Medecine Xavier Bichat, Paris. FAU - Pavie, Juliette AU - Pavie J FAU - Lefort, Agnes AU - Lefort A FAU - Zarrouk, Virginie AU - Zarrouk V FAU - Chau, Francoise AU - Chau F FAU - Garry, Louis AU - Garry L FAU - Leclercq, Roland AU - Leclercq R FAU - Fantin, Bruno AU - Fantin B LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Antibiotics, Combined) RN - 0 (Antibiotics, Glycopeptide) RN - 0 (Culture Media) RN - 0 (Macrolides) RN - 11006-76-1 (Virginiamycin) RN - 126602-89-9 (quinupristin-dalfopristin) RN - 1404-90-6 (Vancomycin) RN - 3131-03-1 (Streptogramin B) RN - 80738-43-8 (lincosamide) SB - IM MH - Animals MH - Anti-Bacterial Agents/pharmacology MH - Antibiotics, Combined/*pharmacology/*therapeutic use MH - Antibiotics, Glycopeptide/*pharmacology/*therapeutic use MH - Culture Media MH - Drug Resistance MH - Endocarditis, Bacterial/*drug therapy/microbiology MH - *Macrolides MH - Methicillin Resistance MH - Microbial Sensitivity Tests MH - Rabbits MH - Research Support, Non-U.S. Gov't MH - Staphylococcus aureus/*drug effects MH - Streptogramin B/pharmacology MH - Time Factors MH - Vancomycin/*pharmacology/*therapeutic use MH - Virginiamycin/*pharmacology/*therapeutic use EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3061-4. PMID- 12183273 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Antibiotic susceptibilities of Parachlamydia acanthamoeba in amoebae. PG - 3065-7 AB - Parachlamydia acanthamoeba are intracellular bacteria of amoebae and are considered potential etiological agents of human pneumonia. We have determined the in vitro antibiotic susceptibilities of two strains (strain Bn(9) and Hall's coccus) in Acanthamoeba polyphaga. The two strains were susceptible to tetracyclines, macrolides, and rifampin, but resistant to fluoroquinolones. AD - Unite des Rickettsies, CNRS UPRES A 6020, Faculte de Medecine, Universite de la Mediterranee, 13385 Marseille Cedex 05, France. FAU - Maurin, M AU - Maurin M FAU - Bryskier, A AU - Bryskier A FAU - Raoult, D AU - Raoult D LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Bacterial Agents) RN - 0 (Anti-Infective Agents) RN - 0 (Antibiotics, Antitubercular) RN - 0 (DNA, Bacterial) RN - 0 (Fluoroquinolones) RN - 0 (Macrolides) RN - 0 (Tetracyclines) RN - 13292-46-1 (Rifampin) SB - IM MH - Acanthamoeba/drug effects/*microbiology MH - Animals MH - Anti-Bacterial Agents/*pharmacology MH - Anti-Infective Agents/pharmacology MH - Antibiotics, Antitubercular/pharmacology MH - Bacteria/*drug effects MH - DNA, Bacterial/genetics MH - Drug Resistance, Bacterial MH - Fluoroquinolones MH - Humans MH - Macrolides MH - Microbial Sensitivity Tests MH - Pneumonia, Bacterial/microbiology MH - Rifampin/pharmacology MH - Tetracyclines/pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3065-7. PMID- 12183274 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - In vitro activities of garenoxacin (BMS-284756) against 170 clinical isolates of nine Pasteurella species. PG - 3068-70 AB - The in vitro susceptibilities of 170 clinical isolates plus 12 American Type Culture Collection strains of Pasteurella species comprising nine species and three Pasteurella multocida subspecies were studied by an agar dilution method. Garenoxacin (BMS-284756), a new des-fluoro(6) quinolone, was active at 90% of the strains susceptible to or=8-fold more active against QRSA than the other fluoroquinolones. And the 50% protective doses for DW286 were correspondent with the in vitro activities. AD - College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 151-742, Korea. FAU - Yun, Hee-Jeong AU - Yun HJ FAU - Min, Yu-Hong AU - Min YH FAU - Lim, Jung-A AU - Lim JA FAU - Kang, Jin-Wook AU - Kang JW FAU - Kim, So-Young AU - Kim SY FAU - Kim, Mi-Jeong AU - Kim MJ FAU - Jeong, Jae-Hee AU - Jeong JH FAU - Choi, Yun-Jeong AU - Choi YJ FAU - Kwon, Hyun-Jin AU - Kwon HJ FAU - Jung, Yong-Ho AU - Jung YH FAU - Shim, Mi-Ja AU - Shim MJ FAU - Choi, Eung-Chil AU - Choi EC LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (7-(3-(aminomethyl)-4-(methoxyimino)-3-methyltetrahydro-1H-1-pyrrolyl)-1-c yclopropyl-6-fluoro-4-oxo-1,4-dihydro(1,8)naphthyridine-3-carboxylic acid) RN - 0 (Anti-Bacterial Agents) RN - 0 (Anti-Infective Agents) RN - 0 (Fluoroquinolones) RN - 0 (Naphthyridines) SB - IM MH - Animals MH - Anti-Bacterial Agents/*pharmacology/*therapeutic use MH - Anti-Infective Agents/pharmacology/therapeutic use MH - Bacteria/*drug effects MH - Bacterial Infections/*drug therapy/microbiology MH - Comparative Study MH - Dogs MH - Fluoroquinolones MH - Mice MH - Microbial Sensitivity Tests MH - Naphthyridines/*pharmacology MH - Rats MH - Research Support, Non-U.S. Gov't EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3071-4. PMID- 12183276 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Prevalence of protease and reverse transcriptase drug resistance mutations over time in drug-naive human immunodeficiency virus type 1-positive individuals in Rio de Janeiro, Brazil. PG - 3075-9 AB - The prevalence of mutations that confer resistance to protease inhibitors and to nucleoside and nonnucleoside reverse transcriptase inhibitors in 49 blood samples from drug-naive human immunodeficiency virus type 1-infected blood donors living in Rio de Janeiro state, Brazil, in 1998 was evaluated genotypically and phenotypically. AD - Unidade de Genetica, Departamento de Ciencias Morfologicas, Instituto Biomedico, Universidade do Rio de Janeiro, Brazil. FAU - Dumans, Ana T AU - Dumans AT FAU - Soares, Marcelo A AU - Soares MA FAU - Pieniazek, Danuta AU - Pieniazek D FAU - Kalish, Marcia L AU - Kalish ML FAU - De Vroey, Veronique AU - De Vroey V FAU - Hertogs, Kurt AU - Hertogs K FAU - Tanuri, Amilcar AU - Tanuri A LA - eng SI - GENBANK/AF413811 SI - GENBANK/AF413812 SI - GENBANK/AF413813 SI - GENBANK/AF413814 SI - GENBANK/AF413815 SI - GENBANK/AF413816 SI - GENBANK/AF413817 SI - GENBANK/AF413818 SI - GENBANK/AF413819 SI - GENBANK/AF413820 SI - GENBANK/AF413821 SI - GENBANK/AF413822 SI - GENBANK/AF413823 SI - GENBANK/AF413824 SI - GENBANK/AF413825 SI - GENBANK/AF413826 SI - GENBANK/AF413827 SI - GENBANK/AF413828 SI - GENBANK/AF413829 SI - GENBANK/AF413830 SI - GENBANK/AF413831 SI - GENBANK/AF413832 SI - GENBANK/AF413833 SI - GENBANK/AF413834 SI - GENBANK/AF413835 SI - GENBANK/AF413836 SI - GENBANK/AF413837 SI - GENBANK/AF413838 SI - GENBANK/AF413839 SI - GENBANK/AF413840 SI - GENBANK/AF413841 SI - GENBANK/AF413842 SI - GENBANK/AF413843 SI - GENBANK/AF413844 SI - GENBANK/AF413845 SI - GENBANK/AF413846 SI - GENBANK/AF413847 SI - GENBANK/AF413848 SI - GENBANK/AF413849 SI - GENBANK/AF413850 SI - GENBANK/AF413851 SI - GENBANK/AF413852 SI - GENBANK/AF413853 SI - GENBANK/AF413854 SI - GENBANK/AF413855 SI - GENBANK/AF413856 SI - GENBANK/AF413857 SI - GENBANK/AF413858 SI - GENBANK/AF413859 SI - GENBANK/AF413860 SI - GENBANK/AF413861 SI - GENBANK/AF413862 SI - GENBANK/AF413863 SI - GENBANK/AF413864 SI - GENBANK/AF413865 SI - GENBANK/AF413866 SI - GENBANK/AF413867 SI - GENBANK/AF413868 SI - GENBANK/AF413869 SI - GENBANK/AF413870 SI - GENBANK/AF413871 SI - GENBANK/AF413872 SI - GENBANK/AF413873 SI - GENBANK/AF413874 SI - GENBANK/AF413875 SI - GENBANK/AF413876 SI - GENBANK/AF413877 SI - GENBANK/AF413878 SI - GENBANK/AF413879 SI - GENBANK/AF413880 SI - GENBANK/AF413881 SI - GENBANK/AF413882 SI - GENBANK/AF413883 SI - GENBANK/AF413884 SI - GENBANK/AF413885 SI - GENBANK/AF413886 SI - GENBANK/AF413887 SI - GENBANK/AF413888 SI - GENBANK/AF413889 SI - GENBANK/AF413890 SI - GENBANK/AF413891 SI - GENBANK/AF413892 SI - GENBANK/AF413893 SI - GENBANK/AF413894 SI - GENBANK/AF413895 SI - GENBANK/AF413896 SI - GENBANK/AF413897 SI - GENBANK/AF413898 SI - GENBANK/AF413899 SI - GENBANK/AF413900 SI - GENBANK/AF413901 SI - GENBANK/AF413902 SI - GENBANK/AF413903 SI - GENBANK/AF413904 SI - GENBANK/AF413905 SI - GENBANK/AF413906 SI - GENBANK/AF413907 SI - GENBANK/AF413908 SI - GENBANK/AF413909 SI - GENBANK/AF413910 SI - GENBANK/AF413911 SI - GENBANK/AF413912 PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (HIV Protease Inhibitors) RN - 0 (Reverse Transcriptase Inhibitors) RN - EC 2.7.7.- (HIV-1 Reverse Transcriptase) RN - EC 3.4.23.- (HIV Protease) SB - IM MH - Amino Acid Substitution MH - Brazil/epidemiology MH - Drug Resistance, Viral MH - Genotype MH - HIV Infections/drug therapy/epidemiology MH - HIV Protease/metabolism MH - HIV Protease Inhibitors/*pharmacology MH - HIV Seropositivity/enzymology/metabolism MH - HIV-1/*drug effects/enzymology MH - HIV-1 Reverse Transcriptase/metabolism MH - Humans MH - Molecular Sequence Data MH - Mutation/genetics MH - Phenotype MH - Phylogeny MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Inhibitors/*pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3075-9. PMID- 12183277 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Effect of lamivudine on transmission of human T-cell lymphotropic virus type 1 to adult peripheral blood mononuclear cells in vitro. PG - 3080-3 AB - The effects of lamivudine (3TC) on in vitro infection of peripheral blood mononuclear cells (PBMC) from healthy donors with human T-cell lymphotropic virus type 1 (HTLV-1) were investigated. Direct measures of viral replication (viral DNA, RNA, and protein) all gave similar, very high 50% inhibitory concentrations in comparison with those previously reported for zidovudine. Nevertheless, 3TC inhibited HTLV-1-driven long-term growth of infected PBMC in vitro at concentrations (6.25 micro M) which had poor or no direct antiviral effects, suggesting that another mechanism may be playing a role. AD - Department of Microbiological, Genetic and Molecular Science, University of Messina, Messina, Italy. FAU - Balestrieri, Emanuela AU - Balestrieri E FAU - Forte, Giancarlo AU - Forte G FAU - Matteucci, Claudia AU - Matteucci C FAU - Mastino, Antonio AU - Mastino A FAU - Macchi, Beatrice AU - Macchi B LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-HIV Agents) RN - 0 (DNA, Viral) RN - 134678-17-4 (Lamivudine) SB - IM CIN - Antimicrob Agents Chemother. 2003 May;47(5):1774;author reply 1774-5. PMID: 12709359 MH - Anti-HIV Agents/*pharmacology MH - Coculture Techniques MH - DNA, Viral/biosynthesis/genetics MH - Deltaretrovirus Infections/prevention & control/*transmission/*virology MH - Human T-lymphotropic virus 1/*drug effects MH - Humans MH - Lamivudine/*pharmacology MH - Monocytes/*virology MH - Research Support, Non-U.S. Gov't MH - Virus Replication/drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3080-3. PMID- 12183278 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041209 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Comparative evaluation of disk diffusion with microdilution assay in susceptibility testing of caspofungin against Aspergillus and Fusarium isolates. PG - 3084-7 AB - We compared the disk diffusion and broth microdilution methods for susceptibility testing of caspofungin against Aspergillus (n = 78) and Fusarium (n = 22) isolates. Microdilution testing followed the NCCLS M-38P guidelines but was performed in antibiotic medium 3 supplemented to 2% glucose (AM3). Disk diffusion assays were performed on AM3 agar plates with a 2- micro g caspofungin disk. By both methods, caspofungin showed favorable activity against Aspergillus isolates and no activity against Fusarium isolates. In the disk-based format, intrazonal growth that was not influenced by the drug concentration gradient was consistently observed for all of the Aspergillus isolates tested. AD - Division of Infectious Diseases, Department of Internal Medicine, Center for the Study of Emerging and Reemerging Pathogens, University of Texas Medical School, Houston, Texas 77030, USA. sarikan@metu.edu.tr FAU - Arikan, Sevtap AU - Arikan S FAU - Paetznick, Victor AU - Paetznick V FAU - Rex, John H AU - Rex JH LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Peptide) RN - 0 (Peptides) RN - 0 (Peptides, Cyclic) RN - 0 (caspofungin) SB - IM MH - Antibiotics, Peptide/*pharmacology MH - Aspergillosis/microbiology MH - Aspergillus/*drug effects MH - Comparative Study MH - Fusarium/*drug effects MH - Microbial Sensitivity Tests MH - Mycoses/microbiology MH - *Peptides MH - *Peptides, Cyclic EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3084-7. PMID- 12183279 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Incidence of high-level evernimicin resistance in Enterococcus faecium among food animals and humans. PG - 3088-90 AB - Six high-level evernimicin-resistant Enterococcus faecium isolates were identified among 304 avilamycin-resistant E. faecium isolates from animals and 404 stool samples from humans with diarrhea. All four animal isolates, and one of the human isolates, were able to transfer resistance to a susceptible E. faecium strain. The resulting transconjugants all tested positive for the presence of emtA, a gene encoding a methyltransferase previously linked with high-level evernimicin resistance. The four transconjugants derived from animal isolates all carried the same plasmid, while a differently sized plasmid was found in the isolate from humans. This study demonstrated a low incidence of high-level evernimicin resistance mediated by the emtA gene in different E. faecium isolates of animal and human origin. AD - Danish Veterinary Institute, DK-1790 Copenhagen V, Denmark. faa@vetinst.dk FAU - Aarestrup, Frank Moller AU - Aarestrup FM FAU - McNicholas, Paul M AU - McNicholas PM LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Aminoglycosides) RN - 0 (Anti-Bacterial Agents) RN - 0 (Oligosaccharides) RN - 11051-71-1 (avilamycin) RN - 53024-98-9 (everninomicin) RN - EC 2.1.1. (Methyltransferases) SB - IM MH - *Aminoglycosides MH - Animal Feed/analysis MH - Animals MH - Anti-Bacterial Agents/*pharmacology MH - Cattle MH - Chickens/*microbiology MH - Conjugation, Genetic MH - Denmark/epidemiology MH - Diarrhea/microbiology MH - Drug Resistance MH - Enterococcus faecium/*drug effects MH - Humans MH - Methyltransferases/genetics MH - Oligosaccharides/pharmacology MH - Reverse Transcriptase Polymerase Chain Reaction MH - Swine/*microbiology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3088-90. PMID- 12183280 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Voriconazole inhibition of the metabolism of tacrolimus in a liver transplant recipient and in human liver microsomes. PG - 3091-3 AB - The purpose of this study was to assess the effect of voriconazole on the blood tacrolimus concentration in a liver transplant recipient and to examine the interaction between voriconazole and tacrolimus by using human liver microsomes. Two subjects were enrolled in the clinical study: one received voriconazole, and the other received a placebo. Tacrolimus metabolism was evaluated in human liver microsomes at various concentrations in the absence and presence of various concentrations of voriconazole. Coadministration of voriconazole and tacrolimus resulted in elevated (nearly 10-fold-higher) trough tacrolimus blood concentrations in the liver transplant patient. In the in vitro study, voriconazole at a concentration of 10.4 +/- 4.3 micro g/ml inhibited the metabolism of tacrolimus by 50%. Clinically relevant concentrations of voriconazole inhibited the metabolism of tacrolimus in human liver microsomes. Close monitoring of the blood concentration and adjustment in the dose of tacrolimus are warranted in transplant recipients treated with voriconazole. AD - School of Pharmacy, University of Pittsburgh, and Veterans Affairs Medical Center, Pittsburgh, Pennsylvania 15240, USA. FAU - Venkataramanan, Raman AU - Venkataramanan R FAU - Zang, Shimin AU - Zang S FAU - Gayowski, Timothy AU - Gayowski T FAU - Singh, Nina AU - Singh N LA - eng PT - Clinical Trial PT - Journal Article PT - Randomized Controlled Trial PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antifungal Agents) RN - 0 (Enzyme Inhibitors) RN - 0 (Immunosuppressive Agents) RN - 0 (Pyrimidines) RN - 0 (Triazoles) RN - 0 (voriconazole) RN - 109581-93-3 (Tacrolimus) RN - 9035-51-2 (Cytochrome P-450 Enzyme System) RN - EC 1.14.14.1 (CYP3A protein, human) SB - IM MH - Antifungal Agents/*pharmacology MH - Cytochrome P-450 Enzyme System/antagonists & inhibitors MH - Drug Interactions MH - Enzyme Inhibitors/pharmacology MH - Humans MH - Immunosuppressive Agents/*metabolism MH - In Vitro MH - Kinetics MH - Liver Transplantation/*physiology MH - Microsomes, Liver/drug effects/*metabolism MH - Pyrimidines/*pharmacology MH - Tacrolimus/*metabolism MH - Triazoles/*pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3091-3. PMID- 12183281 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Comparative activities of the oxazolidinone AZD2563 and linezolid against selected recent North American isolates of Streptococcus pneumoniae. PG - 3094-5 AB - The activity of AZD2563 against 250 highly resistant pneumococci and 267 drug-susceptible isolates was determined. The AZD2563 MICs for 50 and 90% of the strains tested were 1 and 2 micro g/ml and 0.5 and 1 micro g/ml, respectively, for the two isolate groups. These MICs were within 1 log(2) dilution of those of linezolid. AD - Infectious Disease Service of Brooke Army Medical Center, San Antonio, Texas, USA. FAU - Baum, Sue E AU - Baum SE FAU - Crawford, Sharon A AU - Crawford SA FAU - McElmeel, M L AU - McElmeel ML FAU - Whitney, Cynthia G AU - Whitney CG FAU - Jorgensen, James H AU - Jorgensen JH LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (AZD2563) RN - 0 (Acetamides) RN - 0 (Anti-Bacterial Agents) RN - 0 (Oxazolidinones) RN - 165800-03-3 (linezolid) SB - IM MH - Acetamides/*pharmacology MH - Anti-Bacterial Agents/*pharmacology MH - Comparative Study MH - Drug Resistance, Microbial MH - Humans MH - Microbial Sensitivity Tests MH - North America MH - Oxazolidinones/*pharmacology MH - Pneumococcal Infections/microbiology MH - Research Support, Non-U.S. Gov't MH - Streptococcus pneumoniae/*drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3094-5. PMID- 12183282 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041209 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Effect of the echinocandin caspofungin on expression of Candida albicans secretory aspartyl proteinases and phospholipase in vitro. PG - 3096-100 AB - Although the echinocandin caspofungin primarily inhibits the synthesis of cell wall 1,3-beta-D-glucan, its fungicidal activity could also potentially perturb the expression of virulence factors involved in the ability of Candida albicans to cause infection. Expression of the C. albicans secretory aspartyl proteinase (SAP) and phospholipase B (PLB) virulence genes was determined by reverse transcription-PCR after the addition of caspofungin to cells grown for 15 h in Sabouraud dextrose broth. In cells that remained viable, expression of SAP1 to SAP3, SAP7 to SAP9, and PLB1 was unaltered after exposure to fungicidal concentrations (4 to 16 micro g/ml) of caspofungin over a period of 7 h. However, expression of SAP5 increased steadily beginning 1 h after exposure to caspofungin. These results indicate that caspofungin is rapidly fungicidal against C. albicans, before any suppression of SAP or PLB1 gene expression can occur. AD - Department of Microbiology and Immunology, Faculty of Medicine, University of Montreal, Quebec, Canada. FAU - Ripeau, Jean-Sebastien AU - Ripeau JS FAU - Aumont, Francine AU - Aumont F FAU - Belhumeur, Pierre AU - Belhumeur P FAU - Ostrosky-Zeichner, Luis AU - Ostrosky-Zeichner L FAU - Rex, John H AU - Rex JH FAU - de Repentigny, Louis AU - de Repentigny L LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Antibiotics, Antifungal) RN - 0 (Antibiotics, Peptide) RN - 0 (Peptides) RN - 0 (Peptides, Cyclic) RN - 0 (RNA, Fungal) RN - 0 (RNA, Messenger) RN - 0 (caspofungin) RN - EC 3.1.- (Phospholipases) RN - EC 3.4.23 (Aspartic Endopeptidases) SB - IM MH - Antibiotics, Antifungal/*pharmacology MH - Antibiotics, Peptide/*pharmacology MH - Aspartic Endopeptidases/*biosynthesis/genetics MH - Candida albicans/drug effects/*enzymology/genetics MH - Gene Expression Regulation, Enzymologic/*drug effects MH - Genes, Fungal/genetics MH - Microbial Sensitivity Tests MH - *Peptides MH - *Peptides, Cyclic MH - Phospholipases/*biosynthesis/genetics MH - RNA, Fungal/biosynthesis/genetics MH - RNA, Messenger/biosynthesis/genetics MH - Research Support, Non-U.S. Gov't MH - Reverse Transcriptase Polymerase Chain Reaction MH - Time Factors EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3096-100. PMID- 12183283 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Development of a yeast assay for rapid screening of inhibitors of human-derived Pneumocystis carinii dihydrofolate reductase. PG - 3101-3 AB - Human-derived Pneumocystis carinii dihydrofolate reductase (DHFR) was expressed in a Saccharomyces cerevisiae strain whose growth depends on complementation by this enzyme. We utilized a quantitative assay to measure the sensitivity of this yeast strain to DHFR inhibitors. This assay should be useful for identifying new inhibitors of human-derived P. carinii DHFR. AD - Critical Care Medicine Department, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland 20892-1662, USA. FAU - Ma, Liang AU - Ma L FAU - Jia, Qiuyao AU - Jia Q FAU - Kovacs, Joseph A AU - Kovacs JA LA - eng PT - Journal Article PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Folic Acid Antagonists) RN - 0 (Triazines) RN - 47326-86-3 (BRL 6231) RN - 52128-35-5 (Trimetrexate) RN - 58-14-0 (Pyrimethamine) RN - 738-70-5 (Trimethoprim) RN - EC 1.5.1.3 (Tetrahydrofolate Dehydrogenase) SB - IM MH - Animals MH - Drug Evaluation, Preclinical MH - Folic Acid Antagonists/*pharmacology MH - Humans MH - Pneumocystis/*enzymology MH - Pneumocystis Infections/enzymology/microbiology MH - Pyrimethamine/pharmacology MH - Rats MH - Saccharomyces cerevisiae/drug effects/*enzymology MH - Tetrahydrofolate Dehydrogenase/*metabolism MH - Triazines/pharmacology MH - Trimethoprim/pharmacology MH - Trimetrexate/pharmacology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3101-3. PMID- 12183284 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Prevalence of oxacillin resistance in Staphylococcus aureus among inpatients and outpatients in the United States during 2000. PG - 3104-5 FAU - Jones, Mark E AU - Jones ME FAU - Mayfield, David C AU - Mayfield DC FAU - Thornsberry, Clyde AU - Thornsberry C FAU - Karlowsky, James A AU - Karlowsky JA FAU - Sahm, Daniel F AU - Sahm DF FAU - Peterson, Dan AU - Peterson D LA - eng PT - Letter PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Penicillins) RN - 66-79-5 (Oxacillin) SB - IM MH - Databases, Factual MH - Humans MH - Methicillin Resistance MH - Oxacillin/*pharmacology MH - Penicillin Resistance MH - Penicillins/*pharmacology MH - Staphylococcal Infections/*epidemiology/*microbiology MH - Staphylococcus aureus/*drug effects MH - United States/epidemiology EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3104-5. PMID- 12183285 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Relationship between antibiotic resistance in Streptococcus pneumoniae and that in Haemophilus influenzae: evidence for common selective pressure. PG - 3106-7 FAU - Jones, Mark E AU - Jones ME FAU - Karlowsky, James A AU - Karlowsky JA FAU - Blosser-Middleton, Renee AU - Blosser-Middleton R FAU - Critchley, Ian AU - Critchley I FAU - Thornsberry, Clyde AU - Thornsberry C FAU - Sahm, Daniel F AU - Sahm DF LA - eng PT - Letter PL - United States TA - Antimicrob Agents Chemother JID - 0315061 SB - IM MH - *Drug Resistance MH - Drug Utilization MH - Haemophilus influenzae/*drug effects MH - Microbial Sensitivity Tests MH - Research Support, Non-U.S. Gov't MH - Selection (Genetics) MH - Species Specificity MH - Streptococcus pneumoniae/*drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3106-7. PMID- 12183286 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - gyrA Mutations associated with nalidixic acid-resistant salmonellae from wild birds. PG - 3108-9 FAU - Reche, M Paloma AU - Reche MP FAU - Garcia de los Rios, Jose E AU - Garcia de los Rios JE FAU - Jimenez, Pedro A AU - Jimenez PA FAU - Rojas, Ana M AU - Rojas AM FAU - Rotger, Rafael AU - Rotger R LA - eng SI - GENBANK/AF447053 SI - GENBANK/AF447054 SI - GENBANK/AF447055 SI - GENBANK/AF447056 SI - GENBANK/AF447057 SI - GENBANK/AF447058 SI - GENBANK/AF447059 SI - GENBANK/X78977 PT - Letter PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-Infective Agents) RN - 389-08-2 (Nalidixic Acid) RN - EC 5.99.1.- (DNA Gyrase) SB - IM MH - Animals MH - Anti-Infective Agents/*pharmacology MH - Birds/*microbiology MH - DNA Gyrase/*genetics MH - Drug Resistance, Bacterial MH - Feces/microbiology MH - Microbial Sensitivity Tests MH - Molecular Sequence Data MH - Mutation/*genetics MH - Nalidixic Acid/*pharmacology MH - Research Support, Non-U.S. Gov't MH - Salmonella/*drug effects EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3108-9. PMID- 12183287 OWN - NLM STAT- MEDLINE DA - 20020816 DCOM- 20030213 LR - 20041117 PUBM- Print IS - 0066-4804 VI - 46 IP - 9 DP - 2002 Sep TI - Amino acid substitutions at position 69 of the reverse transcriptase of human immunodeficiency virus type 1 are frequent in zalcitabine-naive antiretroviral-drug-experienced patients. PG - 3110-1 FAU - Montes, Brigitte AU - Montes B FAU - Segondy, Michel AU - Segondy M LA - eng PT - Comment PT - Letter PL - United States TA - Antimicrob Agents Chemother JID - 0315061 RN - 0 (Anti-HIV Agents) RN - 0 (Reverse Transcriptase Inhibitors) RN - 7481-89-2 (Zalcitabine) RN - EC 2.7.7.- (HIV-1 Reverse Transcriptase) SB - IM CON - Antimicrob Agents Chemother. 2001 Aug;45(8):2276-9. PMID: 11451685 MH - Amino Acid Substitution/*genetics MH - Anti-HIV Agents/*therapeutic use MH - *Antiretroviral Therapy, Highly Active MH - Drug Resistance, Viral MH - HIV-1/*enzymology/*genetics MH - HIV-1 Reverse Transcriptase/*genetics MH - Humans MH - Logistic Models MH - Reverse Transcriptase Inhibitors/*therapeutic use MH - Zalcitabine/*therapeutic use EDAT- 2002/08/17 10:00 MHDA- 2003/02/14 04:00 PST - ppublish SO - Antimicrob Agents Chemother 2002 Sep;46(9):3110-1. PMID- 12184817 OWN - NLM STAT- MEDLINE DA - 20030415 DCOM- 20030512 LR - 20041117 PUBM- Print-Electronic IS - 1472-6963 VI - 2 IP - 1 DP - 2002 Aug 16 TI - Low agreement for assessing the risk of postoperative deep venous thrombosis when deciding prophylaxis strategies: a study using clinical vignettes. PG - 16 AB - BACKGROUND: Several clinical practice guidelines (CPG) on antithrombotic prophylaxis in surgical patients help to decide about the prophylaxis strategy based on the patient risk of deep venous thrombosis (DVT). However, the physician risk estimates of DVT could have little inter-observer reproducibility, which could lead to different individual prophylaxis practices. METHODS: Physicians were asked to evaluate DVT risk in eight clinical vignettes, describing actual patients cared for in our hospital. The vignettes included all possible levels of DVT risk. RESULTS: The degree of prophylaxis strategies accuracy was 63% (95% CI 523-75%). Overall agreement was 0.32 (z = 7.61, p < 0.001) and for each level of risk kappa was 0.38 (z = 6.50, p < 0.001); 0.1 (z = 1.65, p < 0.049) and 0.5 (z = 8.45, p < 0.001) for small, moderate and high risk group respectively CONCLUSIONS: Our results showed that there is poor agreement when physicians have to evaluate the risk for postoperative DVT, and in the cases of low and moderate risks of DVT there is the smallest agreement. In addition, the data also showed that the overall accuracy of DVT prophylaxis strategy was only moderate and the risk evaluation did not correlate to the selection of the strategy. The issue of inter-observers variability should be taken into account when CPG performance are analysed, especially when considering the risk-evaluation to choose the appropriate actions. AD - Departamento de Medicina, Hospital Universitario Austral. moflaherty@cas.austral.edu.ar FAU - O'Flaherty, Martin AU - O'Flaherty M FAU - Lerum, Kaja AU - Lerum K FAU - Martin, Paula AU - Martin P FAU - Grassi, Daniel AU - Grassi D LA - eng PT - Journal Article DEP - 20020816 PL - England TA - BMC Health Serv Res JID - 101088677 RN - 0 (Nadroparin) RN - 9005-49-6 (Heparin) SB - IM MH - Adult MH - Aged MH - Aged, 80 and over MH - Argentina/epidemiology MH - Bandages/utilization MH - Decision Making MH - Early Ambulation/utilization MH - Female MH - Heparin/therapeutic use MH - Hospitals, University/standards MH - Humans MH - Male MH - Medical Audit MH - Middle Aged MH - Nadroparin/therapeutic use MH - Observer Variation MH - Postoperative Complications/classification/*epidemiology/*prevention & control MH - *Practice Guidelines MH - Premedication/utilization MH - Risk Assessment/*statistics & numerical data MH - Venous Thrombosis/*epidemiology/etiology/*prevention & control EDAT- 2002/08/20 10:00 MHDA- 2003/05/13 05:00 PHST- 2002/04/19 [received] PHST- 2002/08/16 [accepted] PHST- 2002/08/16 [aheadofprint] PST - epublish SO - BMC Health Serv Res 2002 Aug 16;2(1):16. PMID- 12185243 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041203 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Stimulation of the cell cycle and maize transformation by disruption of the plant retinoblastoma pathway. PG - 11975-80 AB - The genome of the Mastreviruses encodes a replication-associated protein (RepA) that interacts with members of the plant retinoblastoma-related protein family, which are putative cell cycle regulators. Expression of ZmRb1, a maize retinoblastoma-related gene, and RepA inhibited and stimulated, respectively, cell division in tobacco cell cultures. The effect of RepA was mitigated by over-expression of ZmRb1. RepA increased transformation frequency and callus growth rate of high type II maize germplasm. RepA-containing transgenic maize calli remained embryogenic, were readily regenerable, and produced fertile plants that transmitted transgene expression in a Mendelian fashion. In high type II, transformation frequency increased with the strength of the promoter driving RepA expression. When a construct in which RepA was expressed behind its native LIR promoter was used, primary transformation frequencies did not improve for two elite Pioneer maize inbreds. However, when LIR:RepA-containing transgenic embryos were used in subsequent rounds of transformation, frequencies were higher in the RepA+ embryos. These data demonstrate that RepA can stimulate cell division and callus growth in culture, and improve maize transformation. AD - Pioneer Hi-Bred, International, Inc., Johnston, IA 50131, USA. william.gordon-kamm@pioneer.com FAU - Gordon-Kamm, William AU - Gordon-Kamm W FAU - Dilkes, Brian P AU - Dilkes BP FAU - Lowe, Keith AU - Lowe K FAU - Hoerster, George AU - Hoerster G FAU - Sun, Xifan AU - Sun X FAU - Ross, Margit AU - Ross M FAU - Church, Laura AU - Church L FAU - Bunde, Chris AU - Bunde C FAU - Farrell, Jeff AU - Farrell J FAU - Hill, Patrea AU - Hill P FAU - Maddock, Sheila AU - Maddock S FAU - Snyder, Jane AU - Snyder J FAU - Sykes, Louisa AU - Sykes L FAU - Li, Zhongsen AU - Li Z FAU - Woo, Young-min AU - Woo YM FAU - Bidney, Dennis AU - Bidney D FAU - Larkins, Brian A AU - Larkins BA LA - eng SI - GENBANK/AY138520 SI - GENBANK/AY138521 PT - Journal Article DEP - 20020816 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (DNA-Binding Proteins) RN - 0 (Proteins) RN - 0 (Retinoblastoma Protein) RN - 0 (Trans-Activators) RN - 0 (replication initiator protein) RN - EC 5.99.- (DNA Helicases) SB - IM MH - *Cell Cycle MH - Cell Division MH - *DNA Helicases MH - *DNA-Binding Proteins MH - Molecular Sequence Data MH - Plants, Genetically Modified MH - Proteins/physiology MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Retinoblastoma Protein/*metabolism MH - *Trans-Activators MH - Zea mays/*cytology/metabolism EDAT- 2002/08/20 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/16 [aheadofprint] AID - 10.1073/pnas.142409899 [doi] AID - 142409899 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11975-80. Epub 2002 Aug 16. PMID- 12185244 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Formation of geometrically complex lipid nanotube-vesicle networks of higher-order topologies. PG - 11573-8 AB - We present a microelectrofusion method for construction of fluid-state lipid bilayer networks of high geometrical complexity up to fully connected networks with genus = 3 topology. Within networks, self-organizing branching nanotube architectures could be produced where intersections spontaneously arrange themselves into three-way junctions with an angle of 120 degrees between each nanotube. Formation of branching nanotube networks appears to follow a minimum-bending energy algorithm that solves for pathway minimization. It is also demonstrated that materials can be injected into specific containers within a network by nanotube-mediated transport of satellite vesicles having defined contents. Using a combination of microelectrofusion, spontaneous nanotube pattern formation, and satellite-vesicle injection, complex networks of containers and nanotubes can be produced for a range of applications in, for example, nanofluidics and artificial cell design. In addition, this electrofusion method allows integration of biological cells into lipid nanotube-vesicle networks. AD - Department of Physical Chemistry and Microtechnology Center, Chalmers University of Technology, SE-412 96 Goteborg, Sweden. FAU - Karlsson, Mattias AU - Karlsson M FAU - Sott, Kristin AU - Sott K FAU - Davidson, Maximillian AU - Davidson M FAU - Cans, Ann-Sofie AU - Cans AS FAU - Linderholm, Pontus AU - Linderholm P FAU - Chiu, Daniel AU - Chiu D FAU - Orwar, Owe AU - Orwar O LA - eng PT - Journal Article DEP - 20020816 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (Lipids) SB - IM MH - Lipids/*chemistry MH - Microscopy, Fluorescence MH - Molecular Structure MH - Research Support, Non-U.S. Gov't EDAT- 2002/08/20 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/16 [aheadofprint] AID - 10.1073/pnas.172183699 [doi] AID - 172183699 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11573-8. Epub 2002 Aug 16. PMID- 12185245 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Loss of microsatellite diversity and low effective population size in an overexploited population of New Zealand snapper (Pagrus auratus). PG - 11742-7 AB - Although the effects of overfishing on species diversity and abundance are well documented, threats to the genetic diversity of marine fish populations have so far been largely neglected. Indeed, there seems to be little cause for concern, as even "collapsed" stocks usually consist of several million individuals, whereas population genetics theory suggests that only very small populations suffer significant loss of genetic diversity. On the other hand, in many marine species the genetically effective population size (N(e)), which determines the genetic properties of a population, may be orders of magnitude smaller than the census population size (N). Here, microsatellite analyses of a time series of archived scales demonstrated a significant decline in genetic diversity in a New Zealand snapper population during its exploitation history. Effective population sizes estimated both from the decline in heterozygosity and from temporal fluctuations in allele frequency were five orders of magnitude smaller than census population sizes from fishery data. If such low N(e)/N ratios are commonplace in marine species, many exploited marine fish stocks may be in danger of losing genetic variability, potentially resulting in reduced adaptability, population persistence, and productivity. AD - Molecular Ecology and Fisheries Genetics Laboratory, Department of Biological Sciences, University of Hull, Hull HU6 7RX, United Kingdom. lhauser@u.washington.edu FAU - Hauser, Lorenz AU - Hauser L FAU - Adcock, Greg J AU - Adcock GJ FAU - Smith, Peter J AU - Smith PJ FAU - Ramirez, Julio H Bernal AU - Ramirez JH FAU - Carvalho, Gary R AU - Carvalho GR LA - eng PT - Journal Article DEP - 20020816 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 SB - IM MH - Animals MH - *Evolution MH - Microsatellite Repeats/*genetics MH - Perciformes/*genetics MH - Population Density MH - Research Support, Non-U.S. Gov't MH - Species Specificity EDAT- 2002/08/20 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/16 [aheadofprint] AID - 10.1073/pnas.172242899 [doi] AID - 172242899 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11742-7. Epub 2002 Aug 16. PMID- 12186657 OWN - NLM STAT- MEDLINE DA - 20021119 DCOM- 20021216 LR - 20041117 PUBM- Print-Electronic IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Aug 19 TI - A approximately 35 kDa polypeptide from insect cells binds to yeast ACS like elements in the presence of ATP. PG - 23 AB - BACKGROUND: The S. cerevisiae origin recognition complex binds to the ARS consensus sequence in an ATP dependent fashion. Recently, the yeast Cdc6 has been reported to have DNA binding activity. Conservation of replication proteins among different species strongly supports their functional similarity. Here we report the results of an investigation into the DNA binding activity of human Cdc6 protein. Cdc6 was expressed and purified from baculovirus infected Sf9 (Spodoptera frugiperda) insect cells as GST fusion protein (GST-Cdc6) and its DNA binding activity was tested. RESULTS: Partially purified fractions containing GSTCdc6 or GST showed an ACS binding activity in an ATP dependent manner. However, further purification revealed the presence of a putative 35 kDa insect cell protein (p35) which was found responsible for the DNA binding activity. A close match to the 9/11 bases of the ARS consensus sequence was sufficient for p35 binding activity. A DNA fragment from the human c-myc origin region containing yeast ACS like elements also showed p35 binding activity. CONCLUSIONS: We have identified a Spodoptera frugiperda protein with ATP dependent DNA binding activity to ACS like elements. ACS like elements have been reported to be essential for ORC binding and replication initiation in yeast but their role in higher eukaryotes still remains elusive. Like the ARS consensus sequence elements of yeast, ACS like elements found in c-myc and lamin beta 2 origin regions may play similar roles in replication and indicate a conserved role for this DNA motif among eukaryotes. AD - Special Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi-110067, India. skdhar2002@yahoo.co.in FAU - Dhar, Suman K AU - Dhar SK FAU - Mondal, Neelima AU - Mondal N FAU - Soni, Rajesh K AU - Soni RK FAU - Mukhopadhyay, Gauranga AU - Mukhopadhyay G LA - eng PT - Journal Article DEP - 20020819 PL - England TA - BMC Biochem JID - 101084098 RN - 0 (Cell Cycle Proteins) RN - 0 (DNA, Fungal) RN - 0 (DNA-Binding Proteins) RN - 0 (Peptides) RN - 0 (Proto-Oncogene Proteins c-myc) RN - 0 (Recombinant Fusion Proteins) RN - 0 (Saccharomyces cerevisiae Proteins) RN - 122544-18-7 (CDC6 protein, S cerevisiae) RN - 56-65-5 (Adenosine Triphosphate) RN - 60-00-4 (Edetic Acid) RN - 7647-14-5 (Sodium Chloride) RN - EC 2.5.1.18 (Glutathione Transferase) SB - IM MH - Adenosine Triphosphate/*pharmacology MH - Animals MH - Binding Sites/genetics MH - Cell Cycle Proteins/chemistry/genetics/metabolism MH - Cell Line MH - DNA, Fungal/genetics/metabolism MH - DNA-Binding Proteins/chemistry/genetics/*metabolism MH - Edetic Acid/pharmacology MH - Glutathione Transferase/genetics/metabolism MH - Humans MH - Molecular Weight MH - Mutation MH - Peptides/chemistry/genetics/*metabolism MH - Protein Binding/drug effects MH - Proto-Oncogene Proteins c-myc/genetics MH - Recombinant Fusion Proteins/genetics/metabolism MH - Replication Origin/*genetics MH - Research Support, Non-U.S. Gov't MH - Saccharomyces cerevisiae/genetics MH - *Saccharomyces cerevisiae Proteins MH - Sodium Chloride/pharmacology MH - Spodoptera MH - Temperature EDAT- 2002/08/21 10:00 MHDA- 2002/12/18 04:00 PHST- 2002/03/28 [received] PHST- 2002/08/19 [accepted] PHST- 2002/08/19 [aheadofprint] PST - epublish SO - BMC Biochem 2002 Aug 19;3(1):23. PMID- 12186975 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - SALSA, a variant of yeast SAGA, contains truncated Spt7, which correlates with activated transcription. PG - 11622-7 AB - Spt-Ada-Gcn5 acetyltransferase (SAGA) is a previously described histone acetyltransferase/transcriptional coactivator complex in yeast. At promoters of certain genes (HIS3 and TRP3), SAGA has an inhibitory function involving a nonproductive TATA-binding protein interaction mediated by the Spt3 and Spt8 subunits. Related to this, Spt8-less SAGA is a major form of the complex under activating conditions for these genes. In the present study, we purify this activation-specific complex, called SALSA (SAGA altered, Spt8 absent). Besides lacking Spt8, SALSA contains Spt7 subunit that is truncated. Examining the role of this subunit, we find that C-terminally truncated SPT7 resulted in derepressed HIS3 transcription. Furthermore, when grown in rich media (repressing conditions), wild-type cells yielded predominantly SAGA, but Spt7 C-terminal truncations resulted primarily in a form of complex similar to SALSA. Thus, SALSA-like structure and activating function can be partially recapitulated in yeast by truncating the C terminus of Spt7. Overall, these results lead to a model that for a subset of promoters SAGA is inhibitory through Spt3, Spt8, and an Spt8-interacting subdomain of Spt7, whereas SALSA is a form of complex for positive transcriptional regulation. These data clarify a mechanism by which a transcriptional regulatory complex can switch between positive and negative modulation. AD - The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA. FAU - Sterner, David E AU - Sterner DE FAU - Belotserkovskaya, Rimma AU - Belotserkovskaya R FAU - Berger, Shelley L AU - Berger SL LA - eng PT - Journal Article DEP - 20020819 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (Fungal Proteins) RN - 0 (Plasmids) RN - 0 (SPT7 protein, S cerevisiae) RN - 0 (Saccharomyces cerevisiae Proteins) RN - 0 (Transcription Factors) SB - IM MH - Amino Acid Sequence MH - Base Sequence MH - Blotting, Western MH - Fungal Proteins/*metabolism MH - Molecular Sequence Data MH - Plasmids MH - Precipitin Tests MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Research Support, U.S. Gov't, P.H.S. MH - Saccharomyces cerevisiae/*metabolism MH - *Saccharomyces cerevisiae Proteins MH - Transcription Factors/chemistry/genetics/metabolism/*physiology MH - *Transcription, Genetic EDAT- 2002/08/21 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/19 [aheadofprint] AID - 10.1073/pnas.182021199 [doi] AID - 182021199 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11622-7. Epub 2002 Aug 19. PMID- 12186978 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Essential myosin light chain as a target for caspase-3 in failing myocardium. PG - 11860-5 AB - Programmed cell death involves the activation of caspase proteases that can mediate the cleavage of vital cytoskeletal proteins. We have recently reported that, in failing cardiac myocytes, caspase-3 activation is associated with a reduction in contractile performance. In this study we used a modified yeast two-hybrid system to screen for caspase-3 interacting proteins of the cardiac cytoskeleton. We identified ventricular essential myosin light chain (vMLC1) as a target for caspase-3. By sequencing and site-directed mutagenesis, a noncanonical cleavage site for caspase-3 was mapped to the C-terminal DFVE(135)G motif. We demonstrated that vMLC1 cleavage in failing myocardium in vivo is associated with a morphological disruption of the organized vMLC1 staining of sarcomeres, and with a reduction in myocyte contractile performance. Adenoviral gene transfer of the caspase inhibitor p35 in vivo prevented caspase-3 activation and vMLC1 cleavage, with positive impact on contractility. These data suggest that direct cleavage of vMLC1 by activated caspase-3 may contribute to depression of myocyte function by altering cross-bridge interaction between myosin and actin molecules. Therefore, activation of apoptotic pathways in the heart may lead to contractile dysfunction before cell death. AD - I. Medizinische Klinik and Deutsches Herzzentrum, D-81675 Munich, Germany. FAU - Moretti, Alessandra AU - Moretti A FAU - Weig, Hans-Jorg AU - Weig HJ FAU - Ott, Thomas AU - Ott T FAU - Seyfarth, Melchior AU - Seyfarth M FAU - Holthoff, Hans-Peter AU - Holthoff HP FAU - Grewe, Diana AU - Grewe D FAU - Gillitzer, Angelika AU - Gillitzer A FAU - Bott-Flugel, Lorenz AU - Bott-Flugel L FAU - Schomig, Albert AU - Schomig A FAU - Ungerer, Martin AU - Ungerer M FAU - Laugwitz, Karl-Ludwig AU - Laugwitz KL LA - eng PT - Journal Article DEP - 20020819 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (Myosin Light Chains) RN - EC 3.4.22.- (Caspases) RN - EC 3.4.22.- (caspase-3) SB - IM MH - Animals MH - COS Cells MH - Caspases/*metabolism MH - Heart/*physiopathology MH - Hydrolysis MH - Kinetics MH - Mutagenesis, Site-Directed MH - Myocardium/enzymology/*metabolism MH - Myosin Light Chains/genetics/*metabolism MH - Precipitin Tests MH - Rabbits MH - Research Support, Non-U.S. Gov't MH - Sarcomeres/metabolism MH - Substrate Specificity EDAT- 2002/08/21 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/19 [aheadofprint] AID - 10.1073/pnas.182373099 [doi] AID - 182373099 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11860-5. Epub 2002 Aug 19. PMID- 12186979 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Evidence for an ancient selective sweep in the MHC class I gene repertoire of chimpanzees. PG - 11748-53 AB - MHC class I molecules play an essential role in the immune defense against intracellular infections. The hallmark of the MHC is its extensive degree of polymorphism at the population level. However, the present comparison of MHC class I gene intron variation revealed that chimpanzees have experienced a severe repertoire reduction at the orthologues of the HLA-A, -B, and -C loci. The loss of variability predates the (sub)speciation of chimpanzees and did not effect other known gene systems. Therefore the selective sweep in the MHC class I gene may have resulted from a widespread viral infection. Based on the present results and the fact that chimpanzees have a natural resistance to the development of AIDS, we hypothesize that the selective sweep was caused by the chimpanzee-derived simian immunodeficiency virus (SIVcpz), the closest relative of HIV-1, or a closely related retrovirus. Hence, the contemporary chimpanzee populations represent the offspring of AIDS-resistant animals, the survivors of a HIV-like pandemic that took place in the distant past. AD - Department of Immunobiology, Biomedical Primate Research Centre, P.O. Box 3306, 2280 GH, Rijswijk, The Netherlands. FAU - de Groot, Natasja G AU - de Groot NG FAU - Otting, Nel AU - Otting N FAU - Doxiadis, Gaby G M AU - Doxiadis GG FAU - Balla-Jhagjhoorsingh, Sunita S AU - Balla-Jhagjhoorsingh SS FAU - Heeney, Jonathan L AU - Heeney JL FAU - van Rood, Jon J AU - van Rood JJ FAU - Gagneux, Pascal AU - Gagneux P FAU - Bontrop, Ronald E AU - Bontrop RE LA - eng SI - GENBANK/AJ313335 SI - GENBANK/AJ313336 SI - GENBANK/AJ313337 SI - GENBANK/AJ313338 SI - GENBANK/AJ313339 SI - GENBANK/AJ313340 SI - GENBANK/AJ313341 SI - GENBANK/AJ313342 SI - GENBANK/AJ313343 SI - GENBANK/AJ313344 SI - GENBANK/AJ313345 SI - GENBANK/AJ313346 SI - GENBANK/AJ313347 SI - GENBANK/AJ313348 SI - GENBANK/AJ313349 SI - GENBANK/AJ313350 SI - GENBANK/AJ313351 SI - GENBANK/AJ313352 SI - GENBANK/AJ313353 SI - GENBANK/AJ313354 SI - GENBANK/AJ313355 SI - GENBANK/AJ313356 SI - GENBANK/AJ313357 SI - GENBANK/AJ313358 SI - GENBANK/AJ313359 PT - Journal Article DEP - 20020819 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 9007-49-2 (DNA) SB - IM MH - Animals MH - Base Sequence MH - DNA MH - *Evolution, Molecular MH - *Genes, MHC Class I MH - Introns MH - Molecular Sequence Data MH - Pan troglodytes/*genetics MH - Phylogeny MH - Research Support, Non-U.S. Gov't MH - Sequence Homology, Nucleic Acid EDAT- 2002/08/21 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/19 [aheadofprint] AID - 10.1073/pnas.182420799 [doi] AID - 182420799 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11748-53. Epub 2002 Aug 19. PMID- 12192093 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Functional plasticity of an antigen-specific memory CD4 T cell population. PG - 11802-7 AB - The protective nature of memory immune responses is attributed largely to terminally differentiated memory T cells that retain memory of the antigen via the antigen receptor and memory of the effector functions that initially cleared the pathogen. It is not known whether a given population of antigen-specific memory T cells is endowed with functional flexibility to provide protective responses against antigens reencountered in different immunological contexts. Here, we examine functional properties of influenza hemagglutinin (HA)-specific memory CD4 T cells recovered from adoptive hosts that received in vitro-activated HA-specific T cell receptor-transgenic CD4 T cells 2 months to 1 year previously. We demonstrate that this HA-specific memory CD4 T cell population bearing a clonal T cell receptor can produce predominantly T helper 1 or T helper 2 effector cytokines depending on the nature of the recall stimulus. Our findings reveal remarkable functional plasticity within an antigen-specific memory T cell population and have direct implications for modulating memory T cell function in vaccine design and treatments for autoimmune diseases. AD - Department of Surgery, University of Maryland School of Medicine, MSTF Building, Room 400, 685 West Baltimore Street, Baltimore, MD 21201, USA. FAU - Ahmadzadeh, Mojgan AU - Ahmadzadeh M FAU - Farber, Donna L AU - Farber DL LA - eng GR - AI42092/AI/NIAID PT - Journal Article DEP - 20020821 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (Antigens) RN - 0 (Cytokines) SB - IM MH - Adoptive Transfer MH - Animals MH - Antigens/*immunology MH - CD4-Positive T-Lymphocytes/cytology/*immunology MH - Cell Differentiation MH - Cytokines/biosynthesis MH - *Immunologic Memory MH - Mice MH - Mice, Inbred BALB C MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2002/08/23 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/21 [aheadofprint] AID - 10.1073/pnas.192263099 [doi] AID - 192263099 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11802-7. Epub 2002 Aug 21. PMID- 12193273 OWN - NLM STAT- Publisher DA - 20020823 PUBM- Print IS - 1471-2091 VI - 3 IP - 1 DP - 2002 Aug 23 TI - Delineation of RAID1, the RACK1 interaction domain located within the unique N-terminal region of the cAMP-specific phosphodiesterase, PDE4D5. PG - 24 AB - Background The cyclic AMP specific phosphodiesterase, PDE4D5 interacts with the beta-propeller protein RACK1 to form a signaling scaffold complex in cells. Two-hybrid analysis of truncation and mutant constructs of the unique N-terminal region of the cAMP-specific phosphodiesterase, PDE4D5 was used to define a domain conferring interaction with the signaling scaffold protein, RACK1. Results Truncation and mutagenesis approaches showed that the RACK1-interacting domain on PDE4D5 comprised a cluster of residues provided by Asn-22/Pro-23/Trp-24/Asn-26 together with a series of hydrophobic amino acids, namely Leu-29, Val-30, Leu-33, Leu-37 and Leu-38 in a 'Leu-Xaa-Xaa-Xaa-Leu' repeat. This was done by 2-hybrid analyses and then confirmed in biochemical pull down analyses using GST-RACK1 and mutant PDE4D5 forms expressed in COS cells. Mutation of Arg-34, to alanine, in PDE4D5 attenuated its interaction with RACK1 both in 2-hybrid screens and in pull down analyses. A 38-mer peptide, whose sequence reflected residues 12 through 49 of PDE4D5, bound to RACK1 with similar affinity to native PDE4D5 itself (Ka circa 6 nM). Conclusion The RACK1 Interaction Domain on PDE4D5, that we here call RAID1, is proposed to form an amphipathic helical structure that we suggest may interact with the C-terminal beta-propeller blades of RACK1 in a manner akin to the interaction of the helical G-gamma signal transducing protein with the beta-propeller protein, G-beta. AU - Bolger GB AU - McCahill A AU - Yarwood SJ AU - Steele MS AU - Warwicker J AU - Houslay MD LA - ENG PT - JOURNAL ARTICLE TA - BMC Biochem JID - 101084098 EDAT- 2002/08/24 10:00 MHDA- 2002/08/24 10:00 PHST- 2002/06/06 [received] PHST- 2002/08/23 [accepted] PHST- 2002/08/23 [aheadofprint] PST - aheadofprint SO - BMC Biochem 2002 Aug 23;3(1):24. PMID- 12193647 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Using mechanical force to probe the mechanism of pausing and arrest during continuous elongation by Escherichia coli RNA polymerase. PG - 11682-7 AB - Escherichia coli RNA polymerase translocates along the DNA discontinuously during the elongation phase of transcription, spending proportionally more time at some template positions, known as pause and arrest sites, than at others. Current models of elongation suggest that the enzyme backtracks at these locations, but the dynamics are unresolved. Here, we study the role of lateral displacement in pausing and arrest by applying force to individually transcribing molecules. We find that an assisting mechanical force does not alter the translocation rate of the enzyme, but does reduce the efficiency of both pausing and arrest. Moreover, arrested molecules cannot be rescued by force, suggesting that arrest occurs by a bipartite mechanism: the enzyme backtracks along the DNA followed by a conformational change of the ternary complex (RNA polymerase, DNA and transcript), which cannot be reversed mechanically. AD - The Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA. FAU - Forde, Nancy R AU - Forde NR FAU - Izhaky, David AU - Izhaky D FAU - Woodcock, Glenna R AU - Woodcock GR FAU - Wuite, Gijs J L AU - Wuite GJ FAU - Bustamante, Carlos AU - Bustamante C LA - eng GR - 5R37GM032543-21/GM/NIGMS PT - Journal Article DEP - 20020822 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - EC 2.7.7.6 (DNA-Directed RNA Polymerases) SB - IM MH - DNA-Directed RNA Polymerases/chemistry/*metabolism MH - Escherichia coli/*enzymology MH - Kinetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Research Support, U.S. Gov't, P.H.S. MH - Transcription, Genetic EDAT- 2002/08/24 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/22 [aheadofprint] AID - 10.1073/pnas.142417799 [doi] AID - 142417799 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11682-7. Epub 2002 Aug 22. PMID- 12193650 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - Sleep forms memory for finger skills. PG - 11987-91 AB - Practicing a motor skill triggers a process of memory consolidation that continues for hours after practice has ended, and becomes manifest in an improved skill at later testing. We used a sequential motor task (finger-to-thumb opposition task) to show that, in humans, the formation of motor skill memories essentially benefits from sleep. Independent of whether placed during daytime or nighttime, sleep after practice enhanced speed of sequence performance on average by 33.5% and reduced error rate by 30.1% as compared with corresponding intervals of wakefulness. The effect of sleep after learning proved to be stable when retesting was postponed for another night, to exclude effects of sleep loss and to assure that all subjects had sufficient sleep before retrieval testing. Also, the consolidating effect of sleep was specific for the motor sequence learned. It did not generalize to a similar sequence containing identical movement segments in a different order. Retention periods of wakefulness improved performance only moderately and only if placed during daytime. The observations demonstrate a critical role of sleep for storing and optimizing motor skills. AD - Department of Neuroendocrinology, University of Lubeck, D-23538 Lubeck, Germany. FAU - Fischer, Stefan AU - Fischer S FAU - Hallschmid, Manfred AU - Hallschmid M FAU - Elsner, Anna Lisa AU - Elsner AL FAU - Born, Jan AU - Born J LA - eng PT - Journal Article DEP - 20020822 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 SB - IM MH - Adolescent MH - Adult MH - Fingers/*physiology MH - Humans MH - *Memory MH - *Motor Activity MH - Research Support, Non-U.S. Gov't MH - Sleep/*physiology EDAT- 2002/08/24 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/22 [aheadofprint] AID - 10.1073/pnas.182178199 [doi] AID - 182178199 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11987-91. Epub 2002 Aug 22. PMID- 12195021 OWN - NLM STAT- MEDLINE DA - 20020904 DCOM- 20020927 LR - 20041117 PUBM- Print-Electronic IS - 0027-8424 VI - 99 IP - 18 DP - 2002 Sep 3 TI - SAGE Genie: a suite with panoramic view of gene expression. PG - 11547-8 AD - Department of Cancer Biology, 658 MRB II, Vanderbilt-Ingram Cancer Center, Nashville, TN 37232, USA. peng.liang@vanderbilt.edu FAU - Liang, Peng AU - Liang P LA - eng PT - Comment PT - Journal Article DEP - 20020823 PL - United States TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (DNA, Complementary) RN - 0 (RNA, Messenger) SB - IM CON - Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11287-92. PMID: 12119410 MH - Brain/metabolism MH - Brain Neoplasms/genetics MH - DNA, Complementary MH - Expressed Sequence Tags MH - *Gene Expression Profiling MH - Humans MH - RNA, Messenger/genetics EDAT- 2002/08/27 10:00 MHDA- 2002/09/28 04:00 PHST- 2002/08/23 [aheadofprint] AID - 10.1073/pnas.192436299 [doi] AID - 192436299 [pii] PST - ppublish SO - Proc Natl Acad Sci U S A 2002 Sep 3;99(18):11547-8. Epub 2002 Aug 23. PMID- 12202540 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Laboratory diagnosis of lower respiratory tract infections: controversy and conundrums. PG - 3115-20 AD - University of Utah School of Medicine and Diagnostic Infectious Diseases Laboratories, ARUP Laboratories, Inc., Salt Lake City, Utah, USA. kcarrol7@jhmi.edu FAU - Carroll, Karen C AU - Carroll KC LA - eng PT - Journal Article PT - Review PT - Review, Tutorial PL - United States TA - J Clin Microbiol JID - 7505564 SB - IM MH - Humans MH - Laboratories MH - *Laboratory Techniques and Procedures MH - Microbiology MH - Respiratory Tract Infections/*diagnosis/*etiology RF - 26 EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3115-20. PMID- 12202541 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041118 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Emergence of Klebsiella pneumoniae isolates producing inducible DHA-1 beta-lactamase in a university hospital in Taiwan. PG - 3121-6 AB - Ten nonrepetitive clinical isolates of Klebsiella pneumoniae exhibiting an unusual inducible beta-lactam resistance phenotype were identified between January 1999 and September 2001 in a university hospital in Taiwan. In the presence of 2 micro g of clavulanic acid, the isolates showed a one to four twofold concentration increase in the MICs of ceftazidime, cefotaxime, and aztreonam but remained susceptible to cefepime (MICs, /=64 micro g/ml). C. dubliniensis was not detected at end of study in any of these patients. Of the remaining 30 patients, eight (27%) demonstrated a replacement of C. albicans by C. dubliniensis when a comparison of isolates obtained at baseline and those from the last culture was done. For the 22 of these 30 patients in whom no switch in species was detected, the fluconazole MICs for initial and end-of-study C. albicans isolates ranged from 0.125 to 2.0 micro g/ml. For the eight patients in whom a switch to C. dubliniensis was detected, the fluconazole MICs for C. dubliniensis isolates at end of study ranged from 0.25 to 64 micro g/ml: the fluconazole MICs for isolates from six patients were 0.25 to 2.0 micro g/ml and those for the other two were 32 and 64 micro g/ml, respectively. In conclusion, a considerable number of patients initially infected with C. albicans strains that failed to develop fluconazole resistance demonstrated a switch to C. dubliniensis. C. dubliniensis in this setting may be underestimated due to lack of identification and may occur due to the impact of fluconazole on the ecology of oral yeast species. AD - Department of Medicine, Division of Infectious Diseases, The University of Texas Health Science Center at San Antonio, 78245, USA. FAU - Martinez, Marcos AU - Martinez M FAU - Lopez-Ribot, Jose L AU - Lopez-Ribot JL FAU - Kirkpatrick, William R AU - Kirkpatrick WR FAU - Coco, Brent J AU - Coco BJ FAU - Bachmann, Stefano P AU - Bachmann SP FAU - Patterson, Thomas F AU - Patterson TF LA - eng GR - 5 R01 DE11381-04A2/DE/NIDCR GR - M01-RR-01346/RR/NCRR GR - R29 AI42401/AI/NIAID PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Antifungal Agents) RN - 86386-73-4 (Fluconazole) SB - IM MH - AIDS-Related Opportunistic Infections/*drug therapy/microbiology MH - Antifungal Agents/pharmacology/*therapeutic use MH - Candida/*classification/drug effects/genetics/isolation & purification MH - Candida albicans/*classification/drug effects/genetics/isolation & purification MH - Candidiasis, Oral/*drug therapy/microbiology MH - DNA Fingerprinting MH - Drug Resistance, Fungal MH - Fluconazole/pharmacology/*therapeutic use MH - HIV Infections/complications MH - Humans MH - Karyotyping/methods MH - Microbial Sensitivity Tests MH - Mycological Typing Techniques MH - Oropharynx/microbiology MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3135-9. PMID- 12202544 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Weak association between SEN virus viremia and liver disease. PG - 3140-5 AB - Recently, a novel DNA virus designated SEN virus (SEN-V), which is thought to be related to posttransfusion hepatitis, was discovered. The aim of the present study was to clarify the relationship between SEN-V infection and the development of liver disease. We examined SEN-V from the sera of 21 patients with non-B, non-C hepatocellular carcinoma (HCC) and 13 patients with non-B, non-C chronic liver disease (CLD) without HCC who were admitted to our hospital between 1995 and 1997. Thirty-two patients without liver disease served as controls and were also examined for SEN-V. SEN-V DNA was detected by the nested PCR method after extraction of DNA from serum. SEN-V DNA was detected in 74% (25 of 34) of patients with CLD with or without HCC who were negative for both hepatitis B virus surface antigen and anti-hepatitis C virus antibody. SEN-V DNA was detected in 69% (9 of 13) of CLD patients without HCC and in 76% (16 of 21) of HCC patients. The prevalence of SEN-V was no higher in patients with liver disease than in patients without liver disease (24 of 32; 75%). There were no significant differences in age, sex, liver function, history of blood transfusion, or amount of alcohol intake between SEN-V-positive and SEN-V-negative CLD and HCC patients. Genetic analysis suggested that SEN-V is closely related to the TT virus family. SEN-V was detected at almost the same frequency in patients with and without liver disease. SEN-V does not seem to contribute either to the pathogenesis of liver disease or to the development of HCC from chronic liver disease. AD - Department of Gastroenterology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. ch2h-ysd@asahi-net.or.jp FAU - Yoshida, Hideo AU - Yoshida H FAU - Kato, Naoya AU - Kato N FAU - Shiratori, Yasushi AU - Shiratori Y FAU - Shao, Runxuan AU - Shao R FAU - Wang, Yue AU - Wang Y FAU - Shiina, Shuichiro AU - Shiina S FAU - Omata, Masao AU - Omata M LA - eng SI - GENBANK/AB059353 SI - GENBANK/AB059532 PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (DNA, Viral) SB - IM MH - Aged MH - Base Sequence MH - Carcinoma, Hepatocellular/*epidemiology/virology MH - DNA Virus Infections/*epidemiology/virology MH - DNA Viruses/genetics/*isolation & purification MH - DNA, Viral/blood MH - Female MH - Hepatitis, Viral, Human/virology MH - Humans MH - Incidence MH - Liver Diseases/*epidemiology/virology MH - Male MH - Middle Aged MH - Molecular Sequence Data MH - Polymerase Chain Reaction/methods MH - Research Support, Non-U.S. Gov't MH - Sequence Analysis, DNA MH - Viremia/*epidemiology/virology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3140-5. PMID- 12202545 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Cavitary pneumonia in an AIDS patient caused by an unusual Bordetella bronchiseptica variant producing reduced amounts of pertactin and other major antigens. PG - 3146-54 AB - Although Bordetella bronchiseptica can infect and colonize immunocompromised humans, its role as a primary pathogen in pneumonia and other respiratory processes affecting those patients remains controversial. A case of cavitary pneumonia caused by B. bronchiseptica in an AIDS patient is presented, and the basis of the seemingly enhanced pathogenic potential of this isolate (designated 814) is investigated. B. bronchiseptica was the only microorganism recovered from sputum, bronchoalveolar lavage fluid, and samples taken through the protected brush catheter. Unlike previous work reporting the involvement of B. bronchiseptica in cases of pneumonia, antibiotic treatment selected on the basis of in vitro antibacterial activity resulted in clearance of the infection and resolution of the pulmonary infiltrate. Although isolate 814 produced reduced amounts of several major antigens including at least one Bvg-activated factor (pertactin), the molecular basis of this deficiency was found to be BvgAS independent since the defect persisted after the bvgAS locus of isolate 814 was replaced with a wild-type bvgAS allele. Despite its prominent phenotype, isolate 814 displayed only a modest yet a significant deficiency in its ability to colonize the respiratory tracts of immunocompetent rats at an early time point. Interestingly, the antibody response elicited by isolate 814 in these animals was almost undetectable. We propose that isolate 814 may be more virulent in immunocompromised patients due, at least in part, to its innate ability to produce low amounts of immunogenic factors which may be required at only normal levels for the interaction of this pathogen with its immunocompetent natural hosts. AD - Departamento de Microbiologia y Parasitologia, Universidad de Navarra, 31080 Pamplona, Spain. FAU - Lorenzo-Pajuelo, Benito AU - Lorenzo-Pajuelo B FAU - Villanueva, Jose Luis AU - Villanueva JL FAU - Rodriguez-Cuesta, Juan AU - Rodriguez-Cuesta J FAU - Vergara-Irigaray, Nuria AU - Vergara-Irigaray N FAU - Bernabeu-Wittel, Maximo AU - Bernabeu-Wittel M FAU - Garcia-Curiel, Andres AU - Garcia-Curiel A FAU - Martinez de Tejada, Guillermo AU - Martinez de Tejada G LA - eng PT - Case Reports PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Antigens, Bacterial) RN - 0 (Bacterial Outer Membrane Proteins) RN - 0 (Virulence Factors, Bordetella) RN - 0 (pertactin) SB - IM MH - AIDS-Related Opportunistic Infections/*microbiology/physiopathology/radiography MH - Adult MH - Animals MH - Antigens, Bacterial/genetics/metabolism MH - Bacterial Outer Membrane Proteins/genetics/metabolism MH - Bordetella Infections/*microbiology/physiopathology/radiography MH - Bordetella bronchiseptica/classification/genetics/*isolation & purification/pathogenicity MH - Female MH - Humans MH - Male MH - Phenotype MH - Pneumonia, Bacterial/*microbiology/physiopathology/radiography MH - Rats MH - Rats, Wistar MH - Research Support, Non-U.S. Gov't MH - *Variation (Genetics) MH - Virulence MH - Virulence Factors, Bordetella/genetics/metabolism EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3146-54. PMID- 12202546 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Detection of human papillomavirus DNA in urine specimens from human immunodeficiency virus-positive women. PG - 3155-61 AB - Human immunodeficiency virus (HIV)-positive women may represent one of the fastest-growing populations at risk for acquiring cervical cancer and thus require frequent screening. The purpose of the present studies was to validate a PCR-based urine assay by comparing detection and genotyping of human papillomavirus (HPV) DNA in urine samples and matching cervical swab specimens of HIV-positive women. Despite a difference in amplifiability, the prevalence of any HPV genotype (58% for the cervical swab specimens and 48% for the urine specimens) was not significantly different in this population. The levels of concordance were 70, 71, and 78% for detection of any HPV type, any high-risk HPV type, or any low-risk HPV type in the two specimen types, respectively. While instances of discordant detection were greater for the cervical swab specimens than for the urine specimens, this was not statistically significant. The distributions of HPV genotypes were similar in the cervix and the urine for the majority of types examined. Importantly, detection of HPV DNA in urine was associated with an abnormal Papanicolaou smear to the same extent that detection of HPV DNA in a cervical swab specimen was. These data provide preliminary support for the proposal to use urine testing as a primary or secondary screening tool for cervical cancer in HIV-positive women or as an epidemiological tool. Additional studies with larger sample sizes must be conducted in order to further verify these findings. AD - Department of Microbiology. Department of Medicine, Louisiana State University Health Sciences Center, New Orleans, Louisiana 70112-2822, USA. jbrink@lsuhsc.edu FAU - Brinkman, Joeli A AU - Brinkman JA FAU - Jones, W Elizabeth AU - Jones WE FAU - Gaffga, Ann M AU - Gaffga AM FAU - Sanders, Jonathan A AU - Sanders JA FAU - Chaturvedi, Anil K AU - Chaturvedi AK FAU - Slavinsky III, Joseph AU - Slavinsky III J FAU - Clayton, John L AU - Clayton JL FAU - Dumestre, Jeanne AU - Dumestre J FAU - Hagensee, Michael E AU - Hagensee ME LA - eng GR - CA86378-01/CA/NCI PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (DNA, Viral) SB - IM MH - Adult MH - Cervical Intraepithelial Neoplasia MH - Cervix Neoplasms/virology MH - Cervix Uteri/virology MH - Comparative Study MH - DNA, Viral/*urine MH - Female MH - *HIV Seropositivity MH - Humans MH - Papillomavirus, Human/*classification/genetics/*isolation & purification MH - Papovaviridae Infections/*virology MH - Polymerase Chain Reaction/methods MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Specimen Handling MH - Tumor Virus Infections/*virology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3155-61. PMID- 12202547 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Reactive nitrogen intermediates have a bacteriostatic effect on Mycobacterium tuberculosis in vitro. PG - 3162-6 AB - Susceptibility of six isolates of Mycobacterium tuberculosis (CB3.3, CDC1551, RJ2E, C.C.13, H37Rv, and H37Ra) and two isolates of Mycobacterium bovis (Ravenel and BCG) to reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) was determined by standard in vitro survival assays. After 21 days of incubation, the survival of most strains exposed to either acidified sodium nitrite (ASN) or hydrogen peroxide (H(2)O(2)) was significantly lower than the same strains unexposed to these RNI or ROI products. However, after 50 days of incubation, these differences in susceptibility became less apparent for strains exposed to ASN but not for strains exposed to H(2)O(2). The recovery of these strains after exposure to RNI suggests that the effect of RNI on M. tuberculosis is bacteriostatic. The in vitro concentrations of ROI and RNI used in these assays were higher than those expected in vivo. These observations suggest that, in vivo, RNI expression at physiologically achievable concentrations may keep M. tuberculosis from proliferating but that removal of RNI may allow the organisms to proliferate. Furthermore, the ability of some M. tuberculosis strains to cause rapidly progressive disease may relate to their intrinsic levels of RNI and ROI resistance. AD - School of Public Health, Division of Infectious Diseases, University of California at Berkeley, Berkeley, California 94720, USA. FAU - Firmani, Marcia A AU - Firmani MA FAU - Riley, Lee W AU - Riley LW LA - eng GR - F31 DA05874/DA/NIDA GR - HL51967/HL/NHLBI PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Reactive Nitrogen Species) RN - 0 (Reactive Oxygen Species) RN - 7632-00-0 (Sodium Nitrite) RN - 7722-84-1 (Hydrogen Peroxide) SB - IM MH - Humans MH - Hydrogen Peroxide/pharmacology MH - Microbial Sensitivity Tests/methods MH - Mycobacterium tuberculosis/*drug effects/growth & development MH - Reactive Nitrogen Species/*pharmacology MH - Reactive Oxygen Species/pharmacology MH - Research Support, U.S. Gov't, P.H.S. MH - Sodium Nitrite/*pharmacology MH - Tuberculosis, Pulmonary/microbiology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3162-6. PMID- 12202548 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Detection of simian immunodeficiency virus in diverse species and of human immunodeficiency virus Type 2 by using consensus primers within the pol region. PG - 3167-71 AB - Human immunodeficiency virus type 2 (HIV-2) is the result of cross-species transmission of simian immunodeficiency virus (SIV) from sooty mangabey monkeys to humans. Primer pairs (intHIV-2/SIV) based on a region of integrase that has considerable homology across HIV-2 and SIV lineages were designed to develop a broadly cross-reactive molecular assay to detect lentivirus infection in primates. The intHIV-2/SIV primers detect HIV-2 and simian viruses SIVcpz, SIVsmm, SIVsyk, SIVagm, and SIVmnd. The primers are also capable of amplifying some HIV-1 strains. Additionally, sequences from the integrase amplicons were of sufficient genetic diversity to permit not only phylogenetic clustering of all simian viruses to their respective lineages but also HIV type and group classification. Thus, the primers described here provide a method to detect primate lentiviruses from diverse species of nonhuman primates, as well as from persons infected with HIV-1 and HIV-2. AD - HIV Immunology and Diagnostics Branch, Division of AIDS, Sexually Transmitted Diseases, and Tuberculosis Laboratory Research, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. FAU - Masciotra, Silvina AU - Masciotra S FAU - Yang, Chunfu AU - Yang C FAU - Pieniazek, Danuta AU - Pieniazek D FAU - Thomas, Chanda AU - Thomas C FAU - Owen, Sherry M AU - Owen SM FAU - McClure, Harold M AU - McClure HM FAU - Lal, Renu B AU - Lal RB LA - eng SI - GENBANK/AF395546 SI - GENBANK/AF395547 SI - GENBANK/AF395548 SI - GENBANK/AF395549 SI - GENBANK/AF395550 SI - GENBANK/AF395551 SI - GENBANK/AF395552 SI - GENBANK/AF395553 SI - GENBANK/AF395554 SI - GENBANK/AF395555 SI - GENBANK/AF395556 SI - GENBANK/AF395557 SI - GENBANK/AF395558 SI - GENBANK/AF395559 SI - GENBANK/AF395560 SI - GENBANK/AF395561 SI - GENBANK/AF395562 SI - GENBANK/AF395563 SI - GENBANK/AF395564 SI - GENBANK/AF395565 SI - GENBANK/AF395566 SI - GENBANK/AF395567 SI - GENBANK/AF395568 SI - GENBANK/AF395569 SI - GENBANK/AF395570 SI - GENBANK/AF395571 GR - RR0016/RR/NCRR PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (DNA Primers) RN - 0 (DNA, Viral) RN - 0 (Gene Products, pol) RN - EC 2.7.7.- (HIV Integrase) SB - IM MH - Animals MH - Cercopithecinae MH - DNA Primers MH - DNA, Viral/analysis MH - Gene Products, pol/*genetics MH - HIV Infections/virology MH - HIV Integrase/genetics MH - HIV-1/classification/genetics/isolation & purification MH - HIV-2/classification/genetics/*isolation & purification MH - Humans MH - Molecular Sequence Data MH - Monkey Diseases/virology MH - Phylogeny MH - Polymerase Chain Reaction MH - Research Support, U.S. Gov't, P.H.S. MH - SIV/classification/genetics/*isolation & purification MH - Sequence Analysis, DNA MH - Simian Acquired Immunodeficiency Syndrome/virology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3167-71. PMID- 12202549 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - groESL sequence determination, phylogenetic analysis, and species differentiation for viridans group streptococci. PG - 3172-8 AB - The full-length sequences of the groESL genes (also known as cpn10/60) of Streptococcus anginosus, Streptococcus constellatus, Streptococcus gordonii, and Streptococcus sanguis and the near full-length sequence of the groESL genes of Streptococcus intermedius, Streptococcus bovis, Streptococcus mitis, Streptococcus mutans, Streptococcus oralis, and Streptococcus salivarius were determined. The lengths of the groES genes from the 10 species listed above ranged from 282 to 288 bp, and the full-length sequences of groEL determined for 4 species (S. anginosus, S. constellatus, S. gordonii, and S. sanguis) revealed that each was 1,623 bp. The intergenic region (spacer) between the groES and groEL genes varies in size (15 to 111 bp) and sequence between species. The variation of the groES sequences among the species tested was greater (62.1 to 95.1% nucleotide sequence identities) than that of the groEL sequences (77.2 to 95.2% nucleotide sequence identities). Phylogenetic analysis of the groES and groEL genes yielded evolutionary trees similar to the tree constructed by use of the 16S rRNA gene. The intraspecies variation of the spacer was minimal for clinical isolates of some species. The groESL sequence data provide an additional parameter for identification of viridans group streptococcal species. AD - School of Medical Technology Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan. ljteng@ha.mc.ntu.edu.tw FAU - Teng, Lee-Jene AU - Teng LJ FAU - Hsueh, Po-Ren AU - Hsueh PR FAU - Tsai, Jui-Chang AU - Tsai JC FAU - Chen, Pin-Wun AU - Chen PW FAU - Hsu, Jia-Chuan AU - Hsu JC FAU - Lai, Hsin-Chih AU - Lai HC FAU - Lee, Chun-Nan AU - Lee CN FAU - Ho, Shen-Wu AU - Ho SW LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Bacterial Proteins) RN - 0 (Chaperonins) RN - 0 (GroESL protein, Bacteria) SB - IM MH - Bacterial Proteins/*genetics MH - Bacterial Typing Techniques MH - Base Sequence MH - Chaperonins/*genetics MH - Humans MH - Molecular Sequence Data MH - *Phylogeny MH - Polymerase Chain Reaction MH - Research Support, Non-U.S. Gov't MH - *Sequence Analysis, DNA MH - Streptococcal Infections/*microbiology MH - Streptococcus/*classification/genetics MH - Variation (Genetics) EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3172-8. PMID- 12202550 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Development and evaluation of rapid urinary antigen detection tests for diagnosis of penicilliosis marneffei. PG - 3179-83 AB - Penicilliosis, caused by the dimorphic fungus Penicillium marneffei, is an important opportunistic systemic fungal infection affecting immunocompromised individuals living in areas where penicilliosis is endemic. We have demonstrated previously that a urinary enzyme-linked immunosorbent assay (ELISA) with purified rabbit polyclonal antibody against killed whole-fission-form arthroconidia of P. marneffei was specific and highly sensitive for the diagnosis of penicilliosis. In this study, a dot blot ELISA and a latex agglutination (LA) test were developed with the same polyclonal antibody and compared with the ELISA for the detection of P. marneffei urinary antigen. Urine specimens from 37 patients with culture-proven penicilliosis and 300 controls (52 healthy subjects and 248 hospitalized patients without penicilliosis) were tested. Antigen was detected in urine from all 37 (100%) penicilliosis patients by the LA test, 35 (94.6%) penicilliosis patients by the dot blot ELISA, and 36 (97.3%) penicilliosis patients by the ELISA. False-positive results were found by the three assays for 2 (0.7%), 8 (2.7%), and 6 (2%) of 300 controls, respectively. The overall sensitivities of the diagnostic tests were as follows: dot blot ELISA, 94.6%; ELISA, 97.3%; and LA test, 100% (specificities, 97.3, 98, and 99.3%, respectively). The LA test is simple, robust, rapid, and convenient and should prove to be an important addition to the existing diagnostic tests for penicilliosis. AD - Clinical Infectious Diseases Research Unit, Department of Clinical Tropical Medicine, Faculty of Tropical Medicine Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. FAU - Desakorn, Varunee AU - Desakorn V FAU - Simpson, Andrew J H AU - Simpson AJ FAU - Wuthiekanun, Vanaporn AU - Wuthiekanun V FAU - Sahassananda, Duangjai AU - Sahassananda D FAU - Rajanuwong, Adul AU - Rajanuwong A FAU - Pitisuttithum, Punnee AU - Pitisuttithum P FAU - Howe, Paul A AU - Howe PA FAU - Smith, Michael D AU - Smith MD FAU - White, Nicholas J AU - White NJ LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Antigens, Fungal) SB - IM MH - AIDS-Related Opportunistic Infections/*diagnosis/microbiology MH - Antigens, Fungal/*urine MH - Enzyme-Linked Immunosorbent Assay/methods MH - Humans MH - Latex Fixation Tests MH - Mycoses/*diagnosis/microbiology MH - Penicillium/*isolation & purification MH - Research Support, Non-U.S. Gov't MH - Sensitivity and Specificity MH - Time Factors EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3179-83. PMID- 12202551 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20031114 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Molecular characterization of multiresistant d-tartrate-positive Salmonella enterica serovar paratyphi B isolates. PG - 3184-91 AB - Since 1996, the National Salmonella Reference Laboratory of Germany has received an increasing number of Salmonella enterica subsp. enterica serovar Paratyphi B isolates. Nearly all of these belonged to the dextrorotatory tartrate-positive variant (S. enterica subsp. enterica serovar Paratyphi B dT(+)), formerly called S. enterica subsp. enterica serovar Java. A total of 55 selected contemporary and older S. enterica subsp. enterica serovar Paratyphi B dT(+) isolates were analyzed by plasmid profiling, antimicrobial resistance testing, pulsed-field gel electrophoresis, IS200 profiling, and PCR-based detection of integrons. The results showed a high genetic heterogeneity among 10 old strains obtained from 1960 to 1993. In the following years, however, new distinct multiresistant S. enterica subsp. enterica serovar Paratyphi B dT(+) clones emerged, and one clonal lineage successfully displaced the older ones. Since 1994, 88% of the isolates investigated were multiple drug resistant. Today, a particular clone predominates in some German poultry production lines, poultry products, and various other sources. It was also detected in contemporary isolates from two neighboring countries as well. AD - National Salmonella Reference Laboratory, Federal Institute for Health Protection of Consumers and Veterinary Medicine, 12277 Berlin, Germany. FAU - Miko, Angelika AU - Miko A FAU - Guerra, Beatriz AU - Guerra B FAU - Schroeter, Andreas AU - Schroeter A FAU - Dorn, Christina AU - Dorn C FAU - Helmuth, Reiner AU - Helmuth R LA - eng PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Anti-Bacterial Agents) RN - 0 (DNA Transposable Elements) RN - 0 (Plasmids) RN - 0 (Tartrates) RN - EC 2.7.7.- (Integrases) SB - IM MH - Animals MH - Anti-Bacterial Agents/pharmacology MH - *Bacterial Typing Techniques MH - DNA Transposable Elements MH - *Drug Resistance, Multiple, Bacterial MH - Electrophoresis, Gel, Pulsed-Field MH - Germany MH - Integrases/genetics MH - Microbial Sensitivity Tests MH - Plasmids/genetics MH - Polymerase Chain Reaction MH - Poultry MH - Poultry Diseases/microbiology MH - Poultry Products/microbiology MH - Salmonella Infections, Animal/microbiology MH - Salmonella enterica/*classification/drug effects/*genetics/isolation & purification MH - Serotyping MH - Tartrates/*metabolism EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3184-91. PMID- 12202552 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20031114 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Identification of Anaplasma phagocytophila (formerly Ehrlichia phagocytophila) variants in blood from sheep in Norway. PG - 3192-7 AB - A total of 41 blood samples were collected from 40 Anaplasma phagocytophila-infected sheep in 11 sheep flocks from four different counties of southern Norway. The presence and nature of the Anaplasma species were identified by microscopic detection of morulae, PCR, reverse line blot hybridization, and 16S rRNA gene sequencing. A. phagocytophila was identified in all of the samples, and sequencing of the 16S rRNA gene revealed the presence of four variants of A. phagocytophila. Two of these variants have been described before, but two were newly identified 16S rRNA variants of this species. A. phagocytophila variant 1 was found in nine flocks, A. phagocytophila variant 2 was found in four flocks, the A. phagocytophila prototype was found in two flocks, and A. phagocytophila variant 5 was found in one flock. In two flocks, some sheep were infected with A. phagocytophila variant 1, whereas others were infected with A. phagocytophila variant 2, and in three animals a double infection with two variants was registered. Analyses of the blood samples revealed that blood from sheep infected with A. phagocytophila variant 2 contained nearly twice as many neutrophils and eight times as many Anaplasma-infected neutrophils as blood from sheep infected with the A. phagocytophila variant 1. Furthermore, only 43% of the A. phagocytophila variant 2-infected sheep displayed antibody responses in an immune fluorescence assay, whereas 93% of the sheep with the A. phagocytophila variant 1-infected sheep were seropositive. AD - Department of Sheep and Goat Research, Norwegian School of Veterinary Science, Sandnes, Norway. snoore.Stuen@veths.no FAU - Stuen, Snorre AU - Stuen S FAU - Van De Pol, Ingrid AU - Van De Pol I FAU - Bergstrom, Karin AU - Bergstrom K FAU - Schouls, Leo M AU - Schouls LM LA - eng PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Culture Media) RN - 0 (DNA, Ribosomal) RN - 0 (RNA, Ribosomal, 16S) SB - IM MH - Anaplasma/*classification/genetics/isolation & purification MH - Anaplasmosis/*microbiology MH - Animals MH - Bacterial Typing Techniques MH - Base Sequence MH - Blood/*microbiology MH - Culture Media MH - DNA, Ribosomal/analysis MH - Molecular Sequence Data MH - Norway MH - Nucleic Acid Hybridization/methods MH - Polymerase Chain Reaction MH - RNA, Ribosomal, 16S/genetics MH - Sequence Analysis, DNA MH - Sheep MH - Sheep Diseases/*microbiology MH - *Variation (Genetics) EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3192-7. PMID- 12202553 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Comparison of rapid, automated ribotyping and DNA macrorestriction analysis of Burkholderia pseudomallei. PG - 3198-203 AB - An automated ribotyping device (RiboPrinter) was used to determine the ribotypes of a collection of Burkholderia pseudomallei isolates. In a preliminary evaluation with the restriction enzymes BamHI and EcoRI, the protocol with EcoRI was more discriminating. The reproducibilities of the ribotypes obtained with EcoRI (EcoRI ribotypes) were determined by testing three levels of bacterial loads. The performance of the manufacturer's software was assessed by comparing the machine-optimized ribotypes with the type determined from the original gel image analyzed with Bionumerics software. The library of B. pseudomallei EcoRI ribotypes was then compared with the ribotypes obtained by DNA macrorestriction analysis of XbaI digests by pulsed-field gel electrophoresis. The typeability of B. pseudomallei by EcoRI ribotyping was 100%, and the discrimination index was 0.94. The slightly greater discrimination provided by DNA macrorestriction analysis (0.96) was achieved at the expense of a significantly longer processing time of 6 days, although the method was only half the cost of automated ribotyping. Typeability by macrorestriction analysis was lower (97%) unless a thiourea step was added to neutralize the action of Tris-dependent endonucleases. The digital record of B. pseudomallei isolates analyzed thus far provides a useful resource for future epidemiological studies and will help shorten the response time in the event of a further melioidosis outbreak or the deliberate release of B. pseudomallei as a biohazard. AD - Division of Microbiology and Infectious Diseases, Western Australian Centre for Pathology, Nedlands, Australia. tim.inglis@health.wa.gov.au FAU - Inglis, Timothy J J AU - Inglis TJ FAU - O'Reilly, Lyn AU - O'Reilly L FAU - Foster, Niki AU - Foster N FAU - Clair, Adele AU - Clair A FAU - Sampson, Judy AU - Sampson J LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - EC 3.1.21.- (Deoxyribonuclease BamHI) RN - EC 3.1.21.- (Deoxyribonuclease EcoRI) SB - IM MH - Automation MH - Bacterial Typing Techniques MH - Burkholderia pseudomallei/*classification/genetics MH - Comparative Study MH - Deoxyribonuclease BamHI MH - Deoxyribonuclease EcoRI MH - Electrophoresis, Gel, Pulsed-Field MH - Humans MH - Melioidosis/*microbiology MH - Research Support, Non-U.S. Gov't MH - Restriction Mapping/*methods MH - *Ribotyping EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3198-203. PMID- 12202554 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Testing conditions for determination of minimum fungicidal concentrations of new and established antifungal agents for Aspergillus spp.: NCCLS collaborative study. PG - 3204-8 AB - Standard conditions are not available for evaluating the minimum fungicidal concentrations (MFCs) of antifungal agents. This multicenter collaborative study investigated the reproducibility in three laboratories of itraconazole, posaconazole, ravuconazole, voriconazole, and amphotericin B MFCs for 15 selected isolates of Aspergillus spp. After MIC determinations for the 15 isolates in each center by the NCCLS M38-A broth microdilution method with four media, standard RPMI 1640 (RPMI), RPMI with 2% dextrose, antibiotic medium 3 (M3), and M3 with 2% dextrose, MFCs were determined for each isolate-medium-drug combination. MFCs were defined as the lowest drug dilutions that yielded <3 colonies (approximately 99 to 99.5% killing activity). The highest reproducibility (96 to 100%) was for amphotericin B MFCs with the four media. Although reproducibility was more variable and medium dependent for the azoles (91 to 98%), agreement was good to excellent for itraconazole, ravuconazole, and voriconazole MFCs with RPMI and M3 (93 to 98%). For posaconazole, the agreement was higher with M3 media (91 to 96%) than with RPMI media (91%). These data extend the refinement of testing guidelines for susceptibility testing of Aspergillus spp. and warrant consideration for introduction into future versions of the M38 document. The role of the MFC under these standardized testing conditions as a predictor of clinical outcome needs to be established in clinical trials. AD - Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia 23298-0049, USA. avingrof@hsc.vcu.edu FAU - Espinel-Ingroff, A AU - Espinel-Ingroff A FAU - Fothergill, A AU - Fothergill A FAU - Peter, J AU - Peter J FAU - Rinaldi, M G AU - Rinaldi MG FAU - Walsh, T J AU - Walsh TJ LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Antifungal Agents) RN - 0 (Culture Media) SB - IM MH - Antifungal Agents/*pharmacology MH - Aspergillus/*classification/*drug effects MH - Culture Media MH - Drug Resistance, Fungal MH - Humans MH - Microbial Sensitivity Tests/methods/*standards MH - Reference Standards MH - Reproducibility of Results MH - Research Support, Non-U.S. Gov't EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3204-8. PMID- 12202555 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Polyphyletic strains of hepatitis E virus are responsible for sporadic cases of acute hepatitis in Japan. PG - 3209-18 AB - Among 87 patients who were previously treated for acute hepatitis of unknown etiology between 1992 and 2001 at five hospitals in Japan, 11 (13%) patients were positive for immunoglobulin M-class antibodies to hepatitis E virus (HEV) by enzyme immunoassay and had detectable HEV RNA by reverse transcription-PCR with two independent sets of primers derived from well-conserved genomic areas in open reading frames 1 and 2. Clinical HEV infection was significantly associated with male sex (9 of 11 versus 29 of 76 patients [P < 0.01]) and older age (52 +/- 11 [mean +/- standard deviation] versus 41 +/- 17 years [P < 0.05]), and its prevalence differed by geographic region (6 to 25%), with a higher rate in the northern part of Japan. At admission, the 11 patients with HEV-associated hepatitis had elevated alanine aminotransferase levels of 914 to 4,850 IU/liter, and all but 1 had elevated bilirubin levels of 1.5 to 24.0 mg/dl. The 11 HEV isolates were of genotype III or IV and were segregated into three groups with intergroup nucleotide differences of 9.5 to 22.0%. Phylogenetic analysis revealed that four isolates of genotype III were closely related to a Japanese isolate, while the other four isolates of the same genotype were nearest those from the United States. The remaining three isolates were close to known isolates of genotype IV in China and Taiwan but shared less than 88% identity with them. These results indicate that multiple genotypes of HEV cocirculate in Japan and contribute to the development of sporadic acute hepatitis, with the prevalence differing by age, sex, and geographic region. AD - Department of Internal Medicine, Kin-ikyo Chuo Hospital, Hokkaido 007-0870, Japan. FAU - Mizuo, Hitoshi AU - Mizuo H FAU - Suzuki, Kazuyuki AU - Suzuki K FAU - Takikawa, Yasuhiro AU - Takikawa Y FAU - Sugai, Yoshiki AU - Sugai Y FAU - Tokita, Hajime AU - Tokita H FAU - Akahane, Yoshihiro AU - Akahane Y FAU - Itoh, Keiichi AU - Itoh K FAU - Gotanda, Yuhko AU - Gotanda Y FAU - Takahashi, Masaharu AU - Takahashi M FAU - Nishizawa, Tsutomu AU - Nishizawa T FAU - Okamoto, Hiroaki AU - Okamoto H LA - eng SI - GENBANK/AB082545 SI - GENBANK/AB082546 SI - GENBANK/AB082547 SI - GENBANK/AB082548 SI - GENBANK/AB082549 SI - GENBANK/AB082550 SI - GENBANK/AB082551 SI - GENBANK/AB082552 SI - GENBANK/AB082553 SI - GENBANK/AB082554 SI - GENBANK/AB082555 SI - GENBANK/AB082556 SI - GENBANK/AB082557 SI - GENBANK/AB082558 SI - GENBANK/AB082559 SI - GENBANK/AB082560 SI - GENBANK/AB082561 SI - GENBANK/AB082562 SI - GENBANK/AB082563 SI - GENBANK/AB082564 SI - GENBANK/AB082565 SI - GENBANK/AB082566 SI - GENBANK/AB082567 PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (DNA, Viral) RN - 0 (Hepatitis Antibodies) RN - 0 (Immunoglobulin G) RN - 0 (Immunoglobulin M) RN - 0 (RNA, Viral) SB - IM MH - Acute Disease MH - Adult MH - Aged MH - DNA, Viral/analysis MH - Female MH - Hepatitis Antibodies/blood MH - Hepatitis E/*epidemiology/*virology MH - Hepatitis E virus/*classification/genetics/*immunology/isolation & purification MH - Hepatitis, Viral, Human/epidemiology/virology MH - Humans MH - Immunoglobulin G/blood MH - Immunoglobulin M/blood MH - Japan/epidemiology MH - Male MH - Middle Aged MH - Molecular Sequence Data MH - Phylogeny MH - Prevalence MH - RNA, Viral/blood MH - Research Support, Non-U.S. Gov't MH - Sequence Analysis, DNA MH - *Variation (Genetics) EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3209-18. PMID- 12202556 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Comparison of C(18)-carboxypropylbetaine and standard N-acetyl-L-cysteine-NaOH processing of respiratory specimens for increasing tuberculosis smear sensitivity in Brazil. PG - 3219-22 AB - Techniques to improve the sensitivity of smear microscopy would facilitate early tuberculosis (TB) diagnosis and disease control, especially in low-income countries where the positive predictive value is high. C(18)-carboxypropylbetaine (CB-18) is a zwitterionic detergent that helps to compensate for the innate buoyancy of mycobacteria, potentially enhancing recovery by centrifugation. Previous data suggest that CB-18 may increase the sensitivity of smear, culture, and molecular amplification diagnostic testing. The goal of the present study was to evaluate if the sensitivity of the smear technique using light microscopy could be improved by treating respiratory samples with CB-18. In the first phase, respiratory specimens were collected consecutively from patients with suspected pulmonary tuberculosis in a tertiary-care hospital in Rio de Janeiro, Brazil (236 specimens were analyzed). After protocol modifications, another 120 respiratory specimens were evaluated. The standard technique was N-acetyl-L-cysteine with sodium hydroxide (NALC-NaOH) treatment, smear concentration with centrifugation, and Ziehl-Neelsen staining. Culture on Lowenstein-Jensen slants was performed on all specimens for use as the "gold standard." No specimens from patients undergoing active TB treatment were included. The initial protocol for CB-18 processing resulted in a sensitivity of 59.6% and specificity of 96.8% compared to standard processing with a sensitivity of 66.0% and specificity of 96.8%. Using the modified protocol, the sensitivity of CB-18 increased to 71.4% with a specificity of 97.0% versus standard processing with a sensitivity of 61.9% and a specificity of 99.0%. The diagnostic yield of acid-fast bacillus smear with CB-18 in the absence of fluorescence microscopy and PCR compared to standard processing with NALC-NaOH was not significantly different, although the power to detect a difference by the modified assay was low. AD - Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland, USA. FAU - Scott, Cherise P AU - Scott CP FAU - Dos Anjos Filho, Luciano AU - Dos Anjos Filho L FAU - De Queiroz Mello, Fernanda Carvalho AU - De Queiroz Mello FC FAU - Thornton, Charles G AU - Thornton CG FAU - Bishai, William R AU - Bishai WR FAU - Fonseca, Leila S AU - Fonseca LS FAU - Kritski, AfrAnio L AU - Kritski AL FAU - Chaisson, Richard E AU - Chaisson RE FAU - Manabe, Yukari C AU - Manabe YC LA - eng GR - 2D43TW000010/TW/FIC GR - AI 45432/AI/NIAID GR - K24 AI 01637/AI/NIAID PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (C(18)-carboxypropylbetaine) RN - 0 (Culture Media) RN - 107-43-7 (Betaine) RN - 1310-73-2 (Sodium Hydroxide) RN - 616-91-1 (Acetylcysteine) SB - IM MH - *Acetylcysteine MH - Bacteriological Techniques MH - *Betaine/*analogs & derivatives MH - Brazil MH - Comparative Study MH - Culture Media MH - Humans MH - Mycobacterium tuberculosis/isolation & purification MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. MH - Sensitivity and Specificity MH - Sodium Hydroxide MH - Specimen Handling/*methods MH - Sputum/*microbiology MH - Staining and Labeling/methods MH - Tuberculosis, Pulmonary/*diagnosis/microbiology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3219-22. PMID- 12202557 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - PCR-based identification of bacteria associated with endodontic infections. PG - 3223-31 AB - PCR primers that target the bacterial 16S rRNA genes (or the tuf gene for the genus Enterococcus) were used to identify 10 putative bacterial pathogens in root canals with necrotic pulp. In addition, the associations of these microorganisms with symptoms and a history of diabetes mellitus were investigated. Microbial samples from the root canals of 24 teeth with necrotic pulp were included in the study. PCR with universal bacterial primers identified bacterial DNA in 22 specimens; the remaining 2 specimens were from intact teeth that had been traumatized 6 months prior to treatment. PCR with specific primers showed that preoperative symptoms were significantly associated with the presence of Streptococcus spp. (P < 0.001 by chi-square analysis). There was also a nonsignificant trend for symptoms to be associated with Fusobacterium nucleatum and Porphyromonas gingivalis (odds ratio, >2) and for diabetes mellitus to be associated with P. gingivalis and Porphyromonas endodontalis (odds ratio, >2). Cloning and sequencing of the universal PCR product in one specimen revealed the presence of an organism related to the genus Olsenella, which has not previously been described in endodontic infections. AD - Department of Endodontology, School of Dental Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030, U.S.A. fouad@nso.uchc.edu FAU - Fouad, Ashraf F AU - Fouad AF FAU - Barry, Jody AU - Barry J FAU - Caimano, Melissa AU - Caimano M FAU - Clawson, Michael AU - Clawson M FAU - Zhu, Qiang AU - Zhu Q FAU - Carver, Rachaele AU - Carver R FAU - Hazlett, Karsten AU - Hazlett K FAU - Radolf, Justin D AU - Radolf JD LA - eng SI - GENBANK/AF426827 GR - AI-26756/AI/NIAID GR - AI-29735/AI/NIAID GR - M01RR06192/RR/NCRR PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (DNA, Ribosomal) RN - 0 (RNA, Ribosomal, 16S) SB - IM MH - Bacteria/*classification/genetics/isolation & purification MH - Bacterial Infections/*microbiology MH - Bacterial Typing Techniques MH - DNA, Ribosomal/analysis MH - Dental Pulp Cavity/*microbiology MH - Dental Pulp Necrosis/*microbiology MH - Diabetes Mellitus/microbiology MH - Molecular Sequence Data MH - Periapical Periodontitis/*microbiology MH - Polymerase Chain Reaction/*methods MH - RNA, Ribosomal, 16S/genetics MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, P.H.S. EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3223-31. PMID- 12202558 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Sensitivity of three urinary antigen tests associated with clinical severity in a large outbreak of Legionnaires' disease in The Netherlands. PG - 3232-6 AB - In 1999 an outbreak involving 188 patients with Legionnaires' disease (LD) occurred among visitors to a flower show in the Netherlands. Two enzyme immunoassays (Binax and Biotest) and one immunochromatographic assay (Binax NOW) were tested, using urine samples from LD patients from the 1999 outbreak. Sensitivity was calculated using positive culture and/or seroconversion as the "gold standard" in outbreak-related patients with radiographically confirmed pneumonia who fulfilled the epidemiological critera. The Binax EIA, Biotest EIA, and Binax NOW assay showed overall sensitivities of 69, 71, and 72%, respectively. When the tests were performed with concentrated urine samples, the overall sensitivities increased to 79, 74, and 81%, respectively. Using multiple logistic regression analysis with backward elimination, a statistically significant association was found between clinical severity and test sensitivity for all tests. For patients with mild LD, the test sensitivities ranged from 40 to 53%, whereas for patients with severe LD who needed immediate special medical care, the sensitivities reached 88 to 100%. These findings have major implications for the diagnostic process in patients with mild pneumonia and suggest that patients with mild pneumonia may go underdiagnosed if urine antigen tests alone are used. AD - Regional Laboratory of Public Health Haarlem, The Netherlands. e.yzerman@streeklabhaarlem.nl FAU - Yzerman, Ed P F AU - Yzerman EP FAU - den Boer, Jeroen W AU - den Boer JW FAU - Lettinga, Kamilla D AU - Lettinga KD FAU - Schellekens, Joop AU - Schellekens J FAU - Dankert, Jacob AU - Dankert J FAU - Peeters, Marcel AU - Peeters M LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Antigens, Bacterial) RN - 0 (Reagent Kits, Diagnostic) SB - IM MH - Aged MH - Antigens, Bacterial/*urine MH - Chromatography/methods MH - *Disease Outbreaks MH - Female MH - Humans MH - Immunoenzyme Techniques MH - Legionella pneumophila/*isolation & purification MH - Legionnaires' Disease/diagnosis/*epidemiology/microbiology/physiopathology MH - Male MH - Middle Aged MH - Netherlands/epidemiology MH - Reagent Kits, Diagnostic MH - Sensitivity and Specificity MH - *Severity of Illness Index EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3232-6. PMID- 12202559 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Detection and genotyping of oocysts of Cryptosporidium parvum by real-time PCR and melting curve analysis. PG - 3237-44 AB - Several real-time PCR procedures for the detection and genotyping of oocysts of Cryptosporidium parvum were evaluated. A 40-cycle amplification of a 157-bp fragment from the C. parvum beta-tubulin gene detected individual oocysts which were introduced into the reaction mixture by micromanipulation. SYBR Green I melting curve analysis was used to confirm the specificity of the method when DNA extracted from fecal samples spiked with oocysts was analyzed. Because C. parvum isolates infecting humans comprise two distinct genotypes, designated type 1 and type 2, real-time PCR methods for discriminating C. parvum genotypes were developed. The first method used the same beta-tubulin amplification primers and two fluorescently labeled antisense oligonucleotide probes spanning a 49-bp polymorphic sequence diagnostic for C. parvum type 1 and type 2. The second genotyping method used SYBR Green I fluorescence and targeted a polymorphic coding region within the GP900/poly(T) gene. Both methods discriminated between type 1 and type 2 C. parvum on the basis of melting curve analysis. To our knowledge, this is the first report describing the application of melting curve analysis for genotyping of C. parvum oocysts. AD - Department of Pediatric Hematology-Oncology, School of Medicine, Cukurova University, 01330 Adana, Turkey. FAU - Tanriverdi, Sultan AU - Tanriverdi S FAU - Tanyeli, Atila AU - Tanyeli A FAU - Baslamisli, Fikri AU - Baslamisli F FAU - Koksal, Fatih AU - Koksal F FAU - Kilinc, Yurdanur AU - Kilinc Y FAU - Feng, Xiaochuan AU - Feng X FAU - Batzer, Glenda AU - Batzer G FAU - Tzipori, Saul AU - Tzipori S FAU - Widmer, Giovanni AU - Widmer G LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Fluorescent Dyes) RN - 0 (Organic Chemicals) RN - 0 (Tubulin) RN - 163795-75-3 (SYBR Green I) SB - IM MH - Animals MH - Cattle MH - Cryptosporidiosis/*parasitology MH - Cryptosporidium parvum/*classification/genetics/growth & development/*isolation & purification MH - Fluorescent Dyes MH - Genotype MH - Humans MH - *Organic Chemicals MH - Polymerase Chain Reaction/*methods MH - Research Support, Non-U.S. Gov't MH - Research Support, U.S. Gov't, Non-P.H.S. MH - Sensitivity and Specificity MH - *Temperature MH - Tubulin/genetics EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3237-44. PMID- 12202560 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Dichotomy of glycoprotein g gene in herpes simplex virus type 1 isolates. PG - 3245-51 AB - Herpes simplex virus type 1 (HSV-1) encodes 11 envelope glycoproteins, of which glycoprotein G-1 (gG-1) induces a type-specific antibody response. Variability of the gG-1 gene among wild-type strains may be a factor of importance for a reliable serodiagnosis and typing of HSV-1 isolates. Here, we used a gG-1 type-specific monoclonal antibody (MAb) to screen for mutations in the immunodominant region of this protein in 108 clinical HSV-1 isolates. Of these, 42 isolates showed no reactivity to the anti-gG-1 MAb. One hundred five strains were further examined by DNA sequencing of the middle part of the gG-1 gene, encompassing 106 amino acids including the immunodominant region and epitope of the anti-gG-1 MAb. By phylogenetic comparisons based on the sequence data, we observed two (main) genetic variants of the gG-1 gene among the clinical isolates corresponding to reactivity or nonreactivity to the anti-gG-1 MAb. Furthermore, four strains appeared to be recombinants of the two gG-1 variants. In addition, one strain displayed a gG-1-negative phenotype due to a frameshift mutation, in the form of insertion of a cytosine nucleotide. When immunoglobulin G reactivity to HSV-1 in sera from patients infected with either of the two variants was investigated, no significant differences were found between the two groups, either in a type-common enzyme-linked immunosorbent assay (ELISA) or in a type-specific gG-1 antigen-based ELISA. Despite the here-documented existence of two variants of the gG-1 gene affecting the immunodominant region of the protein, other circumstances, such as early phase of infection, might be sought for explaining the seronegativity to gG-1 commonly found in a proportion of the HSV-1-infected patients. AD - Department of Clinical Virology, Goteborg University, Goteborg, Sweden. elham.rekabdar@microbio.gu.se FAU - Rekabdar, Elham AU - Rekabdar E FAU - Tunback, Petra AU - Tunback P FAU - Liljeqvist, Jan-Ake AU - Liljeqvist JA FAU - Lindh, Magnus AU - Lindh M FAU - Bergstrom, Tomas AU - Bergstrom T LA - eng SI - GENBANK/AF513114 SI - GENBANK/AF513115 SI - GENBANK/AF513116 SI - GENBANK/AF513117 SI - GENBANK/AF513118 SI - GENBANK/AF513119 SI - GENBANK/AF513120 SI - GENBANK/AF513121 SI - GENBANK/AF513122 SI - GENBANK/AF513123 SI - GENBANK/AF513124 SI - GENBANK/AF513125 SI - GENBANK/AF513126 SI - GENBANK/AF513127 SI - GENBANK/AF513128 SI - GENBANK/AF513129 SI - GENBANK/AF513130 SI - GENBANK/AF513131 SI - GENBANK/AF513132 SI - GENBANK/AF513133 SI - GENBANK/AF513134 SI - GENBANK/AF513135 SI - GENBANK/AF513136 SI - GENBANK/AF513137 SI - GENBANK/AF513138 SI - GENBANK/AF513139 SI - GENBANK/AF513140 SI - GENBANK/AF513141 SI - GENBANK/AF513142 SI - GENBANK/AF513143 SI - GENBANK/AF513144 SI - GENBANK/AF513145 SI - GENBANK/AF513146 SI - GENBANK/AF513147 SI - GENBANK/AF513148 SI - GENBANK/AF513149 SI - GENBANK/AF513150 SI - GENBANK/AF513151 SI - GENBANK/AF513152 SI - GENBANK/AF513153 SI - GENBANK/AF513154 SI - GENBANK/AF513155 SI - GENBANK/AF513156 SI - GENBANK/AF513157 SI - GENBANK/AF513158 SI - GENBANK/AF513159 SI - GENBANK/AF513160 SI - GENBANK/AF513161 SI - GENBANK/AF513162 SI - GENBANK/AF513163 SI - GENBANK/AF513164 SI - GENBANK/AF513165 SI - GENBANK/AF513166 SI - GENBANK/AF513167 SI - GENBANK/AF513168 SI - GENBANK/AF513169 SI - GENBANK/AF513170 SI - GENBANK/AF513171 SI - GENBANK/AF513172 SI - GENBANK/AF513173 SI - GENBANK/AF513174 SI - GENBANK/AF513175 SI - GENBANK/AF513176 SI - GENBANK/AF513177 SI - GENBANK/AF513178 SI - GENBANK/AF513179 SI - GENBANK/AF513180 SI - GENBANK/AF513181 SI - GENBANK/AF513182 SI - GENBANK/AF513183 SI - GENBANK/AF513184 SI - GENBANK/AF513185 SI - GENBANK/AF513186 SI - GENBANK/AF513187 SI - GENBANK/AF513188 SI - GENBANK/AF513189 SI - GENBANK/AF513190 SI - GENBANK/AF513191 SI - GENBANK/AF513192 SI - GENBANK/AF513193 SI - GENBANK/AF513194 SI - GENBANK/AF513195 SI - GENBANK/AF513196 SI - GENBANK/AF513197 SI - GENBANK/AF513198 SI - GENBANK/AF513199 SI - GENBANK/AF513200 SI - GENBANK/AF513201 SI - GENBANK/AF513202 SI - GENBANK/AF513203 SI - GENBANK/AF513204 SI - GENBANK/AF513205 SI - GENBANK/AF513206 SI - GENBANK/AF513207 SI - GENBANK/AF513208 SI - GENBANK/AF513209 SI - GENBANK/AF513210 SI - GENBANK/AF513211 SI - GENBANK/AF513212 SI - GENBANK/AF513213 SI - GENBANK/AF513214 SI - GENBANK/AF513215 SI - GENBANK/AF513216 SI - GENBANK/AF513217 SI - GENBANK/AF513218 PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Antibodies, Monoclonal) RN - 0 (Antibodies, Viral) RN - 0 (Immunodominant Epitopes) RN - 0 (Viral Envelope Proteins) RN - 0 (glycoprotein gG-1, herpes simplex virus type 1) SB - IM MH - Amino Acid Sequence MH - Antibodies, Monoclonal/*immunology MH - Antibodies, Viral/immunology MH - Cell Line MH - Enzyme-Linked Immunosorbent Assay MH - Frameshift Mutation MH - Herpes Simplex/*virology MH - Herpesvirus 1, Human/*classification/*genetics/immunology MH - Humans MH - Immunodominant Epitopes MH - Molecular Sequence Data MH - Research Support, Non-U.S. Gov't MH - Sequence Analysis, DNA MH - *Variation (Genetics) MH - Viral Envelope Proteins/chemistry/*genetics/immunology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3245-51. PMID- 12202561 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Detection and characterization of hepatitis C virus RNA in seminal plasma and spermatozoon fractions of semen from patients attempting medically assisted conception. PG - 3252-5 AB - To investigate the risk of transmission of hepatitis C virus (HCV) via semen in assisted reproduction techniques, semen samples from 32 men chronically infected with HCV attending a center for assisted procreation were tested for HCV RNA by a reverse transcription-PCR protocol by using a modified version of the Cobas AMPLICOR HCV assay (version 2.0; Roche Diagnostics). The sensitivity of the test was 40 copies/ml. Four of 32 seminal plasma samples (12.5%) were found to be positive for the presence of HCV RNA. The median HCV load in blood was significantly higher in patients who were found to be positive for the presence of HCV RNA in semen than in those who tested negative (P = 0.02). In one man, seven consecutive seminal plasma samples tested positive for HCV RNA, as did two consecutive motile spermatozoon fractions; the corresponding fractions obtained after migration of the spermatozoa remained negative. Despite the absence of the proven infectivity of virus in semen samples that test positive for HCV RNA, these findings highlight the fact that seminal fluid may exhibit prolonged HCV RNA excretion. The usefulness of HCV RNA detection in both seminal plasma and spermatozoon fractions before the start of a program of medically assisted reproduction in couples in whom the male partner is chronically infected with HCV would need to be evaluated prospectively with a larger population of subjects exhibiting HCV RNA in their semen. AD - Laboratoire de Bacteriologie-Virologie, GIMAP, Faculty of Medicine of Saint-Etienne, France. FAU - Bourlet, Thomas AU - Bourlet T FAU - Levy, Rachel AU - Levy R FAU - Maertens, Anne AU - Maertens A FAU - Tardy, Jean-Claude AU - Tardy JC FAU - Grattard, Florence AU - Grattard F FAU - Cordonier, Helene AU - Cordonier H FAU - Laurent, Jean-Louis AU - Laurent JL FAU - Guerin, Jean-Francois AU - Guerin JF FAU - Pozzetto, Bruno AU - Pozzetto B LA - eng PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (RNA, Viral) SB - IM MH - Adult MH - Female MH - Genotype MH - Hepacivirus/*classification/genetics/*isolation & purification MH - Hepatitis C, Chronic/*virology MH - Humans MH - Male MH - Middle Aged MH - RNA, Viral/analysis/blood MH - *Reproductive Techniques, Assisted MH - Reverse Transcriptase Polymerase Chain Reaction MH - Semen/*virology MH - Sensitivity and Specificity MH - Spermatozoa/*virology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3252-5. PMID- 12202562 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. PG - 3256-60 AB - A real-time reverse transcriptase PCR (RRT-PCR) assay based on the avian influenza virus matrix gene was developed for the rapid detection of type A influenza virus. Additionally, H5 and H7 hemagglutinin subtype-specific probe sets were developed based on North American avian influenza virus sequences. The RRT-PCR assay utilizes a one-step RT-PCR protocol and fluorogenic hydrolysis type probes. The matrix gene RRT-PCR assay has a detection limit of 10 fg or approximately 1,000 copies of target RNA and can detect 0.1 50% egg infective dose of virus. The H5- and H7-specific probe sets each have a detection limit of 100 fg of target RNA or approximately 10(3) to 10(4) gene copies. The sensitivity and specificity of the real-time PCR assay were directly compared with those of the current standard for detection of influenza virus: virus isolation (VI) in embryonated chicken eggs and hemagglutinin subtyping by hemagglutination inhibition (HI) assay. The comparison was performed with 1,550 tracheal and cloacal swabs from various avian species and environmental swabs obtained from live-bird markets in New York and New Jersey. Influenza virus-specific RRT-PCR results correlated with VI results for 89% of the samples. The remaining samples were positive with only one detection method. Overall the sensitivity and specificity of the H7- and H5-specific RRT-PCR were similar to those of VI and HI. AD - Southeast Poultry Research Laboratory, USDA Agricultural Research Service, Athens, Georgia 30605, USA. FAU - Spackman, Erica AU - Spackman E FAU - Senne, Dennis A AU - Senne DA FAU - Myers, T J AU - Myers TJ FAU - Bulaga, Leslie L AU - Bulaga LL FAU - Garber, Lindsey P AU - Garber LP FAU - Perdue, Michael L AU - Perdue ML FAU - Lohman, Kenton AU - Lohman K FAU - Daum, Luke T AU - Daum LT FAU - Suarez, David L AU - Suarez DL LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Fluorescent Dyes) RN - 0 (Hemagglutinin Glycoproteins, Influenza Virus) SB - IM MH - Animals MH - Chick Embryo MH - Fluorescent Dyes MH - Hemagglutination Inhibition Tests MH - Hemagglutinin Glycoproteins, Influenza Virus/*genetics MH - Influenza A Virus, Avian/classification/genetics/*isolation & purification MH - Influenza, Avian/*virology MH - Poultry MH - Poultry Diseases/*virology MH - Research Support, U.S. Gov't, Non-P.H.S. MH - *Reverse Transcriptase Polymerase Chain Reaction MH - Sensitivity and Specificity EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3256-60. PMID- 12202563 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Distribution of environmentally regulated genes of Streptococcus suis serotype 2 among S. suis serotypes and other organisms. PG - 3261-8 AB - The occurrence of 36 environmentally regulated genes of Streptococcus suis strain 10 among all 35 S. suis serotypes was determined by using hybridization with the amplified genes as probes. In addition, the distribution of these genes among the virulence phenotypes of serotypes 1 and 2 was assessed. Hybridization was also performed with various other streptococcal species and nonstreptococcal bacterial species which may be present in pigs. Interestingly, probe ivs-25/iri-1, similar to agrA and sapR, hybridized only with S. suis serotype 1 and 2 strains with virulent phenotypes and is therefore suitable as a diagnostic parameter. Only one probe was specific for S. suis. This probe's sequence was identical to the epf gene, a putative virulence factor of S. suis. Probe ivs-31 was similar to a virulence factor of S. suis, namely, a gene encoding a fibronectin- and fibrinogen-binding protein. This probe hybridized only with oral streptococci. Nearly half of the probes (45%) hybridized with the oral streptococci (S. oralis, S. milleri, S. sanguis, S. gordonii, and S. mitis) and with Streptococcus pneumoniae. This indicates a close relationship between S. suis, the oral streptococci, and S. pneumoniae with respect to the selected environmentally regulated genes. One probe only hybridized with gram-negative species and therefore seems to be obtained by S. suis from a gram-negative organism by horizontal transfer. AD - Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, 1100 DD, The Netherlands. a.degreeff@id.wag-ur.nl FAU - De Greeff, Astrid AU - De Greeff A FAU - Buys, Herma AU - Buys H FAU - Verhaar, Robin AU - Verhaar R FAU - Van Alphen, Loek AU - Van Alphen L FAU - Smith, Hilde E AU - Smith HE LA - eng PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Bacterial Proteins) RN - 0 (DNA Probes) SB - IM MH - Animals MH - Bacteria/genetics MH - Bacterial Proteins/*genetics MH - DNA Probes/genetics MH - *Environment MH - *Gene Expression Regulation, Bacterial MH - Humans MH - Serotyping MH - Streptococcal Infections/*microbiology/veterinary MH - Streptococcus suis/classification/genetics/*pathogenicity MH - Swine MH - Swine Diseases/*microbiology MH - Virulence/genetics EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3261-8. PMID- 12202564 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Effects of various test media on the activities of 21 antimicrobial agents against Haemophilus influenzae. PG - 3269-76 AB - As considerable variation in the antimicrobial susceptibility of Haemophilus influenzae has been reported, the effects of various test media on the susceptibility of H. influenzae were studied. MICs were determined by three laboratories for 21 antimicrobial agents against a panel of 100 selected isolates. Testing was performed using a reference NCCLS frozen broth microdilution method with Haemophilus test medium (HTM) broth and dried commercial MIC trays rehydrated with the following media: in-house and commercially prepared HTM broth, Mueller-Hinton broth with 2% lysed horse blood and NAD, IsoSensitest broth with 2% lysed horse blood and NAD, and IsoSensitest broth-based HTM. Overall, all results were very reproducible, with the MIC at which 50% of the isolates tested are inhibited (MIC(50)), MIC(90), and geometric mean MIC being within one doubling dilution by all six methods and at all three testing centers for 15 of the 21 agents tested. Interlaboratory differences were more marked than intralaboratory differences or differences among media. Cefprozil, cefaclor, and trimethoprim-sulfamethoxazole results differed the most, while results for ampicillin, amoxicillin-clavulanic acid, cefdinir, cefixime, ceftriaxone, and clarithromycin were the most reproducible. However, these variations in results caused considerable differences in susceptibility rates for agents for which NCCLS susceptible breakpoints were close to the geometric mean MIC, particularly for cefaclor and cefprozil. This was much less of a problem when pharmacokinetic-pharmacodynamic breakpoints were used. Reproducible susceptibility results were obtained for a wide range of agents against H. influenzae in three laboratories using a variety of media that support the growth of this fastidious species. AD - Department of Pathology, Case Western Reserve University, Cleveland, Ohio, USA. mrj6@po.cwru.edu FAU - Jacobs, Michael R AU - Jacobs MR FAU - Bajaksouzian, Saralee AU - Bajaksouzian S FAU - Windau, Anne AU - Windau A FAU - Appelbaum, Peter C AU - Appelbaum PC FAU - Lin, Gengrong AU - Lin G FAU - Felmingham, David AU - Felmingham D FAU - Dencer, Christine AU - Dencer C FAU - Koeth, Laura AU - Koeth L FAU - Singer, Mendel E AU - Singer ME FAU - Good, Caryn E AU - Good CE LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Anti-Bacterial Agents) RN - 0 (Culture Media) SB - IM MH - Anti-Bacterial Agents/pharmacology MH - Comparative Study MH - Culture Media MH - Haemophilus influenzae/*drug effects/growth & development MH - Humans MH - Microbial Sensitivity Tests/methods/standards MH - Reference Standards MH - Reproducibility of Results MH - Research Support, Non-U.S. Gov't EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3269-76. PMID- 12202565 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Detection of Trichomonas vaginalis on modified Columbia agar in the routine laboratory. PG - 3277-80 AB - Broth culture of Trichomonas vaginalis is considered the "gold standard" for the diagnosis of trichomoniasis. Two studies were carried out to evaluate modified Columbia agar (MCA) for the isolation of T. vaginalis from clinical samples. Study I compared isolation on MCA to that on liquid medium with 889 vaginal samples. Out of 63 samples positive for T. vaginalis (7.1% of total), MCA identified 62 (98.4%) and broth identified 58 (92.1%). In study II, trichomoniasis was diagnosed within the scope of a screening program for a total of 39,585 men and women by culture on MCA and direct microscopy. Culture on MCA detected 199 (98.5%) and Gram staining detected 163 (80.7%) of 202 positive specimens. Wet-mount preparations used for symptomatic patients identified 103 (92.8%) of 111 cases. Culture of T. vaginalis from clinical samples on MCA is highly sensitive and reliable, as well as timesaving, and therefore suitable for screening of symptomatic and asymptomatic individuals. AD - Outpatients' Center for Diagnosis of Infectious Venerodermatological Diseases, A-1210 Vienna, Austria. angelika.stary@univie.ac.at FAU - Stary, Angelika AU - Stary A FAU - Kuchinka-Koch, Angelika AU - Kuchinka-Koch A FAU - Teodorowicz, Lilianna AU - Teodorowicz L LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Culture Media) RN - 0 (Gram's stain) RN - 0 (Phenazines) RN - 548-62-9 (Gentian Violet) RN - 9002-18-0 (Agar) SB - IM MH - *Agar MH - Animals MH - Culture Media MH - Female MH - Gentian Violet MH - Humans MH - Laboratories MH - Laboratory Techniques and Procedures MH - Male MH - Mass Screening MH - Phenazines MH - Sensitivity and Specificity MH - Specimen Handling/methods MH - Trichomonas Vaginitis/*diagnosis/parasitology MH - Trichomonas vaginalis/*growth & development/*isolation & purification MH - Urethra/parasitology MH - Vagina/parasitology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3277-80. PMID- 12202566 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Mycobacterium microti infection (vole tuberculosis) in wild rodent populations. PG - 3281-5 AB - Mycobacterium microti (vole tuberculosis) infections in small wild mammals were first described more than 60 years ago in several populations in Great Britain. Few studies of vole tuberculosis have been undertaken since then, and little is known about the relationship between M. microti isolates originating from different populations or at different times or of the prevalence of this infection in wild rodent populations, despite human cases of M. microti infections being increasingly reported. In this study, field voles (Microtus agrestis), bank voles (Clethrionomys glareolus), and wood mice (Apodemus sylvaticus) were found to be infected, with up to 8% having external tuberculous signs, in wild populations in Northumberland and Cheshire, England. Spoligotyping applied directly to the clinical material simultaneously detected and typed M. microti bacteria in skin lesions, lymph glands, and internal abcesses. IS6110 restriction fragment length polymorphism typing of cultured bacteria was used to compare these isolates with previously isolated strains from both animals and humans. This demonstrated that although the current rodent isolates were distinct from those isolated from voles in the 1930s in Great Britain, they had a high degree of similarity to these strains and were distinct from the M. microti isolates from humans, a pig, and a ferret from The Netherlands. Thus, M. microti infection seems to be widespread in wild rodent populations, but more studies are needed to understand how M. microti might be transmitted from animals to humans and to determine better the zoonotic risk posed. AD - Centre for Comparative Infectious Diseases, University of Liverpool, Liverpool, United Kingdom. rachel@naturebureau.co.uk FAU - Cavanagh, Rachel AU - Cavanagh R FAU - Begon, Michael AU - Begon M FAU - Bennett, Malcolm AU - Bennett M FAU - Ergon, Torbjorn AU - Ergon T FAU - Graham, Isla M AU - Graham IM FAU - De Haas, Petra E W AU - De Haas PE FAU - Hart, C A AU - Hart CA FAU - Koedam, Marianne AU - Koedam M FAU - Kremer, Kristin AU - Kremer K FAU - Lambin, Xavier AU - Lambin X FAU - Roholl, Paul AU - Roholl P FAU - Soolingen Dv, Dick van AU - Soolingen Dv D LA - eng PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Culture Media) RN - 0 (DNA, Bacterial) RN - 0 (Oligonucleotides) SB - IM MH - Animals MH - *Animals, Wild MH - Culture Media MH - DNA, Bacterial/analysis MH - England/epidemiology MH - Microtinae MH - Muridae MH - Mycobacterium/*classification/genetics/isolation & purification MH - Mycobacterium Infections/epidemiology/microbiology/pathology/veterinary MH - Oligonucleotides/analysis MH - Polymorphism, Restriction Fragment Length MH - Research Support, Non-U.S. Gov't MH - Rodent Diseases/*epidemiology/*microbiology/pathology MH - Tuberculosis/epidemiology/microbiology/pathology/*veterinary EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3281-5. PMID- 12202567 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Simultaneous detection of Anaplasma marginale and a new Ehrlichia species closely related to Ehrlichia chaffeensis by sequence analyses of 16S ribosomal DNA in Boophilus microplus ticks from Tibet. PG - 3286-90 AB - To identify ehrlichial agents in Boophilus microplus ticks, DNA samples of B. microplus collected from the Tibet Autonomous Region and Sichuan Province of China were screened by a nested PCR. Sixteen of 43 (37%) DNA samples of B. microplus from Tibet were positive in nested PCR analysis. All 27 samples from Sichuan were negative. The screen identified two ehrlichial agents based on different 16S rRNA genes that were found after amplifying and sequencing the 5'-end fragments of the 16S rRNA genes. One sequence was identical to that of the gene of Anaplasma marginale, an etiological agent of animal anaplasmosis. The other sequence was most similar to that of the gene of Ehrlichia chaffeensis, an etiological agent of human monocytic ehrlichiosis. The sequence of 1,501 bases from the novel ehrlichial agent was obtained and showed the greatest levels of sequence similarity (97 to 98%) to 16S rRNA gene sequences of the members of the E. canis group of the genus EHRLICHIA: Sequence comparison of the 16S rRNA gene with the members of the genus Ehrlichia reveals that the novel ehrlichial agent detected in B. microplus ticks is a new species of the genus Ehrlichia and is most closely related to E. chaffeensis. AD - Department of Microbiology, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China. bohaiwen@sohu.com FAU - Wen, Bohai AU - Wen B FAU - Jian, Rui AU - Jian R FAU - Zhang, Youzhi AU - Zhang Y FAU - Chen, Rong AU - Chen R LA - eng SI - GENBANK/AF414399 PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (DNA, Ribosomal) RN - 0 (RNA, Ribosomal, 16S) SB - IM MH - Anaplasma/classification/genetics/*isolation & purification MH - Animals MH - Base Sequence MH - DNA, Ribosomal/analysis MH - Ehrlichia/*classification/genetics/*isolation & purification MH - Ehrlichia chaffeensis/*classification/genetics MH - Molecular Sequence Data MH - Phylogeny MH - Polymerase Chain Reaction/methods MH - RNA, Ribosomal, 16S/*genetics MH - Research Support, Non-U.S. Gov't MH - Sequence Analysis, DNA MH - Tibet MH - Ticks/*microbiology EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3286-90. PMID- 12202568 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - Rapid detection of methicillin resistance in coagulase-negative Staphylococci with the VITEK 2 system. PG - 3291-5 AB - The aim of the present study was to evaluate the accuracy of the new VITEK 2 system (bioMerieux, Marcy l' Etoile, France) for the detection of methicillin resistance in coagulase-negative staphylococci (CoNS) by using AST-P515 and AST-P523 test cards. Analyses of the VITEK 2 oxacillin MIC determination evaluated according to the actual breakpoint (>/=0.5 micro g/ml) of the National Committee for Clinical Laboratory Standards resulted in a high sensitivity of 99.2% but a moderate specificity of 80%. The newly included oxacillin resistance (OR) test of the VITEK 2 system displayed a high sensitivity and a high specificity of 97.5 and 98.7%, respectively. Concordance between the results of the mecA PCR and the VITEK 2 oxacillin MIC was observed for almost all Staphylococcus epidermidis strains, but the reduced specificity was attributable to higher oxacillin MICs for mecA-negative non-S. epidermidis strains, especially S. saprophyticus, S. lugdunensis, and S. cohnii. Evaluation of alternative oxacillin MIC breakpoints of 1, 2, or 4 micro g/ml resulted in improved degrees of specificity of 84, 90.7, and 97.3%, respectively. Only minor changes occurred in the corresponding sensitivity values, which were 98.4, 97.5, and 97.5%, respectively. Methicillin resistance in CoNS was detected after 7 and 8 h in 91.1 and 93.5% of the mecA-positive strains, respectively, by the VITEK 2 OR test and in 86.3 and 89.5% of the mecA-positive strains, respectively, by VITEK 2 oxacillin MIC determination. After 7 and 8 h the VITEK 2 OR test classified 59.2 and 78.9% of the mecA-negative strains, respectively, as susceptible to oxacillin, whereas comparable values were obtained 2 h later by VITEK 2 oxacillin MIC determination. The results of our study encourage the use of the VITEK 2 system, which proved to be a highly reliable and rapid phenotypic method for the detection of methicillin resistance in CoNS. AD - Institut fur Medizinische Mikrobiologie und Immunologie, Universitatsklinikum Hamburg-Eppendorf, D-20246 Hamburg, Germany. horstko@uke.uni-hamburg.de FAU - Horstkotte, Matthias A AU - Horstkotte MA FAU - Knobloch, Johannes K-M AU - Knobloch JK FAU - Rohde, Holger AU - Rohde H FAU - Dobinsky, Sabine AU - Dobinsky S FAU - Mack, Dietrich AU - Mack D LA - eng PT - Evaluation Studies PT - Journal Article PL - United States TA - J Clin Microbiol JID - 7505564 RN - 0 (Bacterial Proteins) RN - 0 (Carrier Proteins) RN - 0 (Coagulase) RN - 0 (Penicillin-Binding Proteins) RN - 0 (Penicillins) RN - 0 (Reagent Kits, Diagnostic) RN - 66-79-5 (Oxacillin) RN - EC 2.3.2.12 (Peptidyl Transferases) RN - EC 2.4.1.- (Hexosyltransferases) RN - EC 3.4.17.8 (Muramoylpentapeptide Carboxypeptidase) SB - IM MH - *Bacterial Proteins MH - Carrier Proteins MH - Coagulase/*metabolism MH - *Hexosyltransferases MH - Humans MH - *Methicillin Resistance MH - Microbial Sensitivity Tests/methods/standards MH - Muramoylpentapeptide Carboxypeptidase MH - Oxacillin/pharmacology MH - Penicillin-Binding Proteins MH - Penicillins/pharmacology MH - *Peptidyl Transferases MH - *Reagent Kits, Diagnostic MH - Research Support, Non-U.S. Gov't MH - Sensitivity and Specificity MH - Staphylococcal Infections MH - Staphylococcus/*drug effects MH - Time Factors EDAT- 2002/08/31 10:00 MHDA- 2002/11/26 04:00 PST - ppublish SO - J Clin Microbiol 2002 Sep;40(9):3291-5. PMID- 12202569 OWN - NLM STAT- MEDLINE DA - 20020830 DCOM- 20021114 LR - 20041117 PUBM- Print IS - 0095-1137 VI - 40 IP - 9 DP - 2002 Sep TI - New variants of Vibrio cholerae O1 biotype El Tor with attributes of the classical biotype from hospitalized patients with acute diarrhea in Bangladesh. PG - 3296-9 AB - The sixth pandemic of cholera and, presumably, the earlier pandemics were caused by the classical biotype of Vibrio cholerae O1, which was progressively replaced by the El Tor biotype representing the seventh cholera pandemic. Although the classical biotype of V. cholerae O1 is extinct, even