Microbiological

Biotech Histochem, 2000 Sep, 75(5), 203 - 42
A review of the potential and versatility of colloidal gold cytochemical labeling for molecular morphology; Bendayan M; In the present article we review several postembedding cytochemical techniques using the colloidal gold marker . Owing to the high atomic number of gold, the colloidal gold particles are electron dense . They are spherical in shape and can be prepared in sizes from 1 to 25 nm, which renders this marker among the best for electron microscopy . In addition, because it can be bound to several molecules, this marker has the advantage of being extremely versatile . Combined to immunoglobulins or immunoglobulin-binding proteins (protein A), it has been applied successfully in immunocytochemistry . Colloidal gold particles 5-15 nm in size are excellent for postembedding cytochemistry . Particles of smaller size, such as 1 nm, must be silver enhanced to be visualized by transmission electron microscopy . We have elected to review the superiority of indirect immunocytochemical approaches using IgG-gold or protein A-gold (protein G-gold and protein AG-gold) . Lectins or enzymes can be tagged with colloidal gold particles, and the corresponding lectin-gold and enzyme-gold techniques have specific advantages and great potential . Using an indirect digoxigenin-tagged nucleotide and an antidigoxigenin probe, colloidal gold technology can also be used for in situ hybridization at the electron microscope level . Affinity characteristics lie behind all cytochemical techniques and several molecules displaying high affinity properties can also be beneficial for colloidal gold electron microscopy cytochemistry . All of these techniques can be combined in various ways to produce multiple labelings of several binding sites on the same tissue section . Colloidal gold is particulate and can easily be counted; thus the cytochemical signal can be evaluated quantitatively, introducing further advantages to the use of the colloidal gold marker . Finally, several combinations and multiple step procedures have been designed to amplify the final signal which renders the techniques more sensitive . The approaches reviewed here have been applied successfully in different fields of cell and molecular biology, cell pathology, plant biology and pathology, microbiology and virology . The potential of the approaches is emphasized in addition to different ways to assess specificity, sensitivity and accuracy of results.

Acta Cytol, 2000 Sep-Oct, 44(5), 752 - 9
Interpreting microbiopsies in cervical smears . A cytohistologic approach; Mravunac M et al.; OBJECTIVE: To assess interobserver variation in the diagnosis of thick tissue specimens (microbiopsies) in cytology smears and histologic sections taken from them, to evaluate the applicability of MIB-1 in histologic sections from microbiopsies and to evaluate whether processing microbiopsies in inconclusive smears has additional diagnostic value . STUDY DESIGN: Cytologic smears were selected in which there were diagnostic disagreements between pathologists and cytologists and microbiopsies were present . Interobserver variation among three pathologists and three cytologists in the diagnosis of these microbiopsies was investigated . The smears were processed for histologic sections, and interobserver variation between pathologist diagnoses were analyzed . An additional histologic slide stained for MIB-1 was used for consensus diagnosis . The consensus diagnosis was compared with available follow-up and its sensitivity and specificity determined . The value of applying the microbiopsy technique in slides diagnosed as inadequate or atypical squamous cells of undetermined significance (ASCUS) was analysed . RESULTS: From a series of 62,334 cervical smears, 49 with microbiopsies were selected . It was possible to derive histologic slides from 38 cases . Interobserver variability in the diagnosis of microbiopsies and histologic sections from them was moderate--kappa = .44 (SE = .06) and kappa = .44 (SE = .09), respectively . In the consensus meeting for all cases, a conclusive diagnosis was reached . The Pearson correlation coefficient between the consensus diagnosis and MIB-1 staining was r = .62 . The sensitivity of the consensus diagnosis for the follow-up diagnosis was 71% and the specificity 60% . Diagnosis on approximately 50% of slides diagnosed as inadequate or ASCUS could be made . CONCLUSION: The histotechnical workup of microbiopsies is not difficult; however, their diagnosis can be a problem . Adequate diagnostic criteria are not available . Aided by MIB-1 staining, histologic sections from microbiopsies can be diagnosed, and the diagnoses correlated with follow-up in most cases . Processing of microbiopsies in smears with an inconclusive cytologic diagnosis or a diagnosis of ASCUS allowed correct diagnosis in 50% of cases in this study.

J Clin Microbiol, 2000 Oct, 38(10), 3892 - 5
Granulomatous amebic encephalitis in a patient with AIDS: isolation of acanthamoeba sp . Group II from brain tissue and successful treatment with sulfadiazine and fluconazole; Seijo Martinez M et al.; A patient with AIDS, treated with highly active antiretroviral therapy and trimethoprim-sulfamethoxazole, presented with confusion, a hemifield defect, and a mass lesion in the right occipital lobe . A brain biopsy confirmed granulomatous amebic encephalitis (GAE) due to Acanthamoeba castellanii . The patient was treated with fluconazole and sulfadiazine, and the lesion was surgically excised . This is the first case of AIDS-associated GAE responding favorably to therapy . The existence of a solitary brain lesion, absence of other sites of infection, and intense cellular response in spite of a very low CD4 count conditioned the favorable outcome . We review and discuss the diagnostic microbiologic options for the laboratory diagnosis of infections due to free-living amebae.

J Clin Microbiol, 2000 Oct, 38(10), 3837 - 9
Distribution of a specific 500-base-pair fragment in mycobacterium bovis isolates from Sardinian cattle; Sechi LA et al.; Amplification of a specific, 500-bp fragment from Mycobacterium bovis isolates and use of the fragment to differentiate between Mycobacterium tuberculosis and M . bovis was previously reported (J . G . Rodriguez, G . A . Meja, P . Del Portillo, M . E . Patarroyo, and L . A . Murillo, Microbiology 141:2131-2138, 1995) . In the present study, 30 M . bovis isolates from Sardinian cattle were examined for the presence of this 500-bp fragment; 4 of the 30 isolates lacked the fragment . This result indicates that identification of M . bovis strains by amplification of the 500-bp sequence may lead to false-negative results.

J Clin Microbiol, 2000 Oct, 38(10), 3722 - 8
Helicobacter aurati sp . nov., a urease-positive Helicobacter species cultured from gastrointestinal tissues of Syrian hamsters; Patterson MM et al.; A novel helicobacter with the proposed name Helicobacter aurati (type strain MIT 97-5075c) has been isolated from the inflamed stomachs and ceca of adult Syrian hamsters . The new species is fusiform with multiple bipolar sheathed flagella and periplasmic fibers; it contains urease and gamma-glutamyl transpeptidase . By 16S rRNA sequencing and repetitive element PCR-based DNA fingerprinting, it was found that H . aurati represents a distinct taxon and clusters with Helicobacter muridarum, Helicobacter hepaticus, and Helicobacter sp . MIT 94-022 . H . aurati was recovered from hamsters housed in various research and vendor facilities . Further studies are necessary to define its association with disease and other microbiota in hamsters, as well as its impact on research projects involving hamsters . H . aurati (GenBank accession number AF297868) can be used in animal experiments to define the factors that are important for gastric helicobacter pathogenesis.

Commun Dis Public Health, 2000 Sep, 3(3), 158 - 62
Serological diagnosis of syphilis . PHLS Syphilis Serology Working Group; Egglestone SI et al.; The availability of an increasing number of enzyme immunoassays (EIAs) for detecting syphilis antibodies makes it appropriate to review approaches to syphilis serology and to assess the role of syphilis EIAs in routine diagnostic microbiology laboratories . This paper summarises the principles and practice of syphilis serology and provides recommendations on the use of laboratory tests for syphilis in UK diagnostic microbiology laboratories . The main recommendations are summarised in a testing algorithm . Treponemal EIAs are an appropriate alternative to the use of combined Venereal Disease Research Laboratories/rapid plasma reagin and Treponema pallidum haemagglutination assay (TPHA) tests for screening for syphilis . If a treponemal EIA is used for screening an alternative treponemal test, such as TPHA, should be used for confirmatory testing . The fluorescent treponemal antibody-absorbed test is probably best reserved for specimens giving discrepant results . Such specimens may be referred to the PHLS laboratories that provide confirmatory treponemal testing for reference testing and to facilitate collection of surveillance data on what remains an important public health problem.

JPEN J Parenter Enteral Nutr, 2000 Sep-Oct, 24(5), 296 - 303
Hazard analysis and critical control point system approach in the evaluation of environmental and procedural sources of contamination of enteral feedings in three hospitals; Lucia Rocha Carvalho M et al.; BACKGROUND: The administration of contaminated diets may contribute to severe infections, mainly in immunosuppressed patients . To determine the microbiologic quality of enteral feedings and the critical control points involved in the processing of the formulas, a study was carried out in three hospitals in Sao Paulo, Brazil . METHODS: One hundred sixteen diets were evaluated according to the system known as hazard analysis and critical control points (HACCP) . The techniques included the monitoring of the cleaning and disinfection of utensils, surfaces, and equipment; time and temperature controls; and microbiologic analyses that comprised the enumeration of facultative aerobic mesophilic bacteria, yeasts and molds, coliforms, and Escherichia coli . The diets were assessed based on the British Dietetic Association and the Food and Drug Administration standards . RESULTS: The hospital-formulated diets and the commercially made powdered feeds presented statistically significant higher counts of mesophilic bacteria and percentages of no compliance with the standards for coliforms when compared with the commercially prepared, ready-to-feed enteral formula supplied in cans . According to the British standards, 77% and 38% of the diets were inadequate for mesophilic and coliform bacteria, respectively, just after preparation . After 24-hour storage in refrigerators, the percentages rose to 83% and 45%, respectively; the mesophilic and coliforms counts were also significantly higher . The following critical control points (CCP) were identified: cleaning and disinfecting of surfaces, utensils, and equipment; the time spent in preparation; the water used for reconstitution; the final temperature of the diet; the exposure to room temperatures; the lack of a chilling step; the refrigeration temperature; the inadequate disinfecting of the handlers' hands; and the lack of external cleaning and disinfecting of the cans before opening . CONCLUSIONS: The results of this study are worrisome and show the need for implementation of good practices of hygiene and handling of the diets, and the regular monitoring of their preparation . The microbiologic quality of the enteral diets was compromised according to international standards, representing a potential risk of infection to the patients who require enteral nutrition.

Appl Environ Microbiol, 2000 Oct, 66(10), 4440 - 8
Identification and analysis of the polyhydroxyalkanoate-specific beta-ketothiolase and acetoacetyl coenzyme A reductase genes in the cyanobacterium Synechocystis sp . strain PCC6803; Taroncher-Oldenburg G et al.; Synechocystis sp . strain PCC6803 possesses a polyhydroxyalkanoate (PHA)-specific beta-ketothiolase encoded by phaA(Syn) and an acetoacetyl-coenzyme A (CoA) reductase encoded by phaB(Syn) . A similarity search of the entire Synechocystis genome sequence identified a cluster of two putative open reading frames (ORFs) for these genes, slr1993 and slr1994 . Sequence analysis showed that the ORFs encode proteins having 409 and 240 amino acids, respectively . The two ORFs are colinear and most probably coexpressed, as revealed by sequence analysis of the promoter regions . Heterologous transformation of Escherichia coli with the two genes and the PHA synthase of Synechocystis resulted in accumulation of PHAs that accounted for up to 12.3% of the cell dry weight under high-glucose growth conditions . Targeted disruption of the above gene cluster in Synechocystis eliminated the accumulation of PHAs . ORFs slr1993 and slr1994 thus encode the PHA-specific beta-ketothiolase and acetoacetyl-CoA reductase of Synechocystis and, together with the recently characterized PHA synthase genes in this organism (S . Hein, H . Tran, and A . Steinbuchel, Arch . Microbiol . 170:162-170, 1998), form the first complete PHA biosynthesis pathway known in cyanobacteria . Sequence alignment of all known short-chain-length PHA-specific acetoacetyl-CoA reductases also suggests an extended signature sequence, VTGXXXGIG, for this group of proteins . Phylogenetic analysis further places the origin of phaA(Syn) and phaB(Syn) in the gamma subdivision of the division Proteobacteria.

Pharmacoeconomics, 2000 Jul, 18(1), 63 - 72
Health-related quality of life in asymptomatic patients with HIV . Evaluation of the SF-36 health survey in Italian patients; Arpinelli F et al.; OBJECTIVE: To investigate the psychometric performance and clinical validity of the 36-Item Short Form (SF-36) health survey when completed by asymptomatic HIV-positive Italian patients and to compare their health profile with a representative sample of 2031 Italian citizens (the Italian norm) . PATIENTS AND METHODS: This was an observational, multicentre, cross-sectional survey . Microbiologists throughout Italy recruited asymptomatic HIV-positive individuals who were aged at least 18 years and aware of their infection . Investigators collected demographic, social, clinical and treatment data . Patients, classified into 2 clinical categories (A1 and A2) according to explicit pre-defined criteria, completed the SF-36 health survey in the context of a medical visit . RESULTS: Between April and July 1996, 46 microbiologists recruited 214 patients (201 evaluable) . No inconsistent responses were observed in 96% of the sample . The usually recommended psychometric standards were satisfied, and the internal consistency reliability indices were always greater than 0.70 . Weak to moderate associations were found between SF-36 health survey scores and physicians' estimates of patients' physical performance, while no significant associations were found with CD4+ counts . On average, HIV-positive patients reported lower scores than the Italian norm, and patients in category A2 showed lower scores than patients in A1 . These differences were more relevant in scales describing role limitations, general health perception, and psychological well-being . CONCLUSION: Our study showed that the SF-36 health survey maintained its psychometric properties in a sample of Italian asymptomatic HIV-positive patients and produced data that showed its validity and robustness in such a setting.

Microbes Infect, 2000 Aug, 2(10), 1265 - 76
Overview of the epidemiology, microbiology, and pathogenesis of Leptospira spp . in humans; Plank R et al.; Leptospirosis is probably the world's most widespread zoonosis . It remains underdiagnosed largely due to the broad spectrum of signs and symptoms attributable to this spirochetal pathogen . Leptospira spp . cause a diversity of diseases from flu-like illness to Weil's syndrome with multi-organ failure . Recent epidemics may herald a change in virulence or an alteration in the balance between humans and their interactions with other host species and the environment.

J R Coll Physicians Lond, 2000 Jul-Aug, 34(4), 363 - 5
A new joint training programme in infectious diseases and medical microbiology; Cohen J et al.; The increasing overlap between the disciplines of medical microbiology and infectious diseases prompted the Joint Royal Colleges Committee on Infection and Tropical Medicine to set up a working party to examine how trainees could obtain certification in both subjects . Following widespread consultations, a scheme was developed that entails six years of training and leads to the award of CCSTs in both microbiology and infectious diseases . Both Royal Colleges and the Specialist Training Authority have approved the scheme . Joint training will be demanding and will not be suitable for everyone; it represents an alternative approach to training in the infection disciplines that will run alongside the existing monospecialty training programmes.

Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi, 2000 Aug, 11(1), 19 - 26
{Comparative evaluation of BACTEC MGIT 960 system with MB/BacT and egg-based media for recovery of mycobacteria}; Saitoh H et al.; Fully automated, nonradiometric mycobacteria culture systems, BACTEC MGIT 960 (Becton Dickinson Microbiology Systems, Sparks, Md, U.S.A.) and MB/BacT (Organon Teknika, Durham, NC, U.S.A.) were evaluated in comparison with three different eggbased media (3% Ogawa, Ogawa K, and Vite) for the ability to detect mycobacteria in clinical sputum specimens . Sputum specimens were processed by semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) for the automated systems, and by cetylpyridium chloride-succinic acid-NaCl for the egg-based media . A total of 954 sputum specimens were processed, and the recovery of mycobacteria by the BACTEC MGIT 960 was performed in a commercial laboratory . Overall, the frequency of breakthrough contamination was <1% for the three egg-based media, ranging from 0 . 42% to 0.63% . Whereas, the frequency of false positives due to breakthrough contamination was 1.89% for MB/BacT and 20.1% for BACTEC MGIT 960 . A total of 237 isolates of Mycobacterium tuberculosis complex and 167 isolates of nontuberculous mycobacteria (NTM) were recovered . The highest recovery ratio was obtained by MB/BacT (95.8%), followed by the egg-based media, Vite (74.3%), Ogawa K (65.8%), and 3% Ogawa (58.9%) . The recovery ratio by BACTEC MGIT 960 was the lowest, and estimated at 43.1%, mainly due to a high frequency of breakthrough contamination . However, even after omission of these false positives reported by BACTEC MGIT 960, the recovery ratio by this system was comparable to that of 3% Ogawa media . The time to detection of 50% of positive cultures of M . tuberculosis complex by BACTEC MGIT 960 and MB/BacT was 20 days and 17 days, respectively.>From these results, it may be concluded that MB/BacT is superior to BACTEC MGIT 960 and egg-based media for the recovery of mycobacteria from sputum specimens . Furthermore, based on the outcome of this study, we think that considerable improvements are necessary for the clinical application of BACTEC MGIT 960 . These improvements should particularly be focused on reducing the false positive ratio caused by contamination, and culture media, which effectively support mycobacterial growth.

JAMA, 2000 Sep 27, 284(12), 1546 - 8
Clinical features and treatment of peristomal pyoderma gangrenosum; Hughes AP et al.; CONTEXT: Peristomal pyoderma gangrenosum (PPG), an unusual variant of pyoderma gangrenosum, has been reported almost exclusively in patients with inflammatory bowel disease (IBD) and is frequently misdiagnosed . OBJECTIVE: To better characterize the clinical manifestations, diagnosis, and management of PPG . DESIGN, SETTING, AND PATIENTS: Retrospective analysis of 7 patients with PPG observed in a university-affiliated community setting between 1988 and December 1999 . MAIN OUTCOME MEASURES: Clinical and histopathologic features, associated disorders, and microbiologic findings . RESULTS: Two patients had Crohn disease, 2 had ulcerative colitis, and 3 had abdominal cancer . Five patients had at least 1 relapse of PPG after initial healing . Although 3 of 4 patients with IBD had active bowel disease, a parallel course with PPG occurred in only 1 patient . Both patients whose stoma was relocated developed an ulcer at the new site . Effective therapies included topical superpotent corticosteroids; intralesional injection of triamcinolone acetonide at the ulcer margin; topical cromolyn sodium; oral dapsone, prednisone, cyclosporine, mycophenolate mofetil; and intravenous infliximab . CONCLUSION: Our experiences demonstrate that although PPG has been most often reported in patients with IBD, it may occur in the absence of IBD . Biopsy of the skin lesion is not diagnostic but excludes other causes . Relocation of the stoma may be associated with a new ulceration and should be avoided . Trauma to the skin of a predisposed patient may elicit the pustules or ulcerations associated with pathergy . JAMA . 2000;284:1546-1548.

Eur Arch Otorhinolaryngol, 2000, 257(6), 300 - 3
The pathology and clinical features of "glue ear": a review; Rinaldo A et al.; The clinical entity known as "glue ear" is synonymous with mucoid otitis media and represents the most common cause of hearing loss in children . This review considers the current state of our knowledge of mucoid otitis media, discussing definition and terminology, pathology, etiology and microbiology, clinical features, differential diagnosis and treatment.

Rev Med Liege, 2000 Jun, 55(6), 510 - 5
{Malaria: from diagnosis to curative treatment}; Delanaye P et al.; Malaria is a parasitic disease, with variable severity, provoked by the Plasmodium . It is present in tropical zones . The diagnosis is evoked by a fever occurring in a subject coming from a zone at risk, and is confirmed by microbiology . Considering the high prevalence of resistance to chloroquine, the treatment rests on quinine (or its derivatives) associated (or not) with an antibiotic . The severe forms of malaria, due to Plasmodium falciparum, are responsible for a high mortality rate . It requires urgent hospital management is required . Criteria defining this form deserve to be perfectly known.

Am J Forensic Med Pathol, 2000 Sep, 21(3), 264 - 9
Persian leopard (Panthera pardus) attack in Oklahoma: case report; Vogel JS et al.; The authors report a fatal case of a Persian leopard (Panthera pardus) attack in an animal sanctuary in Oklahoma . The victim was a 53-year-old Costa Rican woman who was attempting to feed the animal when she was attacked and killed . Autopsy, radiography, fingerprint analysis, microbiologic cultures, and dental impressions were used to evaluate the case . These simple techniques can be applied to similar cases involving wild and domestic animal attacks.

Acta Ophthalmol Scand, 2000 Aug, 78(4), 411 - 5
Microbiopsy in healthy rabbit corneas . A long-term study; Kompa S et al.; PURPOSE: Corneal biopsies are important in diagnosing multiple corneal diseases . They were previously performed by way of keratectomy, a method that causes corneal opacity and topographic changes due to scarring . Microbiopsy is a new way to perform corneal biopsies . Before microbiopsy may be performed on human corneas, the safety of this procedure has to be proved concerning clinical development, histological changes and topographic changes after multiple biopsies . METHODS: The healthy right cornea of 24 rabbits was punctured . 12 microtrephinations in 4 different symmetric patterns were performed . The clinical development of the bioptic sites as well as the topographic changes were observed over 5 months . After enucleation, serial sections of the corneas were analysed histologically . RESULTS: Out of 294 performed biopsies, 291 samples could be collected . 4 perforations occurred . The initial epithelial defect closed within 3 days . A pale stromal scar remained . The histological analysis of these scars showed a facette underlined by a dense hypocellular fibrous layer and a typical star-shaped figure consisting of a loose hypercellular stromal tissue . Only dioptric power of corneas with circle-pattern showed a statistically significant decrease . CONCLUSION: Micropuncture is a safe and efficient bioptic procedure . Even 12 micropunctures do not lead to significant changes of dioptric power in most patterns . Further studies are necessary to evaluate the reproducibility of refractive changes by circle patterns and corrections of astigmatisms.

J Am Med Inform Assoc, 2000 Sep-Oct, 7(5), 475 - 87
Exploring performance issues for a clinical database organized using an entity-attribute-value representation; Chen RS et al.; 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.

J Cell Sci, 2000 Oct, 113 (Pt 19), 3355 - 6
Cellular microbiology: an integrated approach to understanding pathogenesis of infection
Mitchell TJ.
Cellular Microbiology by P . Cossart, P . Boquet, S . Normark and R . Rappuoli ASM Press (2000) pp . 392 . ISBN 1-55581-157-4 $75.95 The term 'cellular microbiology' was coined by the editors of this book in 1996 . Since then several volumes and journals addressing this subject have been produced and Cellular Microbiology is a valuable contribution to an exciting field . In studying the pathogenesis of infectious diseases it is clear that an understanding of the interaction between the host and the pathogen requires a knowledge of both and here we have an excellent source of information for the integrated study of microbiology and cell biology . The first two chapters are a useful general introduction to the two subject areas and will be particularly useful for scientists of the 'opposite' discipline i.e . microbiologists will find the cell biology section particularly useful and vice versa . The chapters then progress through the encounter of the pathogen with the eukaryotic cell, from the interactions occurring at the cell surface, the mechanisms of attachment and the interaction with the cell cytoskeleton, to the action of bacterial toxins and the consequences of these interactions . Although the book relates mainly to the interaction of bacteria with the host there are also many interesting discussions on parasites and some fungi . Later chapters describe the interaction of pathogens with the immune system, covering the use of bacterial products as tools in cell biology and describing some of the underlying methodology involved in cellular microbiology; there are also interesting chapters on Type III and Type IV secretion systems . As with any textbook in a rapidly expanding area there is a danger that the information contained will rapidly become dated . This may apply especially to the chapters on secretion systems which are the subject of intense research effort . The chapters are all relatively short and can be read as stand alone 'mini-reviews' of the area . This will be very useful for students studying these areas . Cellular Microbiology is very well written and readable . The contributors are all experts in their respective fields and manage to communicate a sense of excitement that will be particularly valuable for readers relatively new to the area . The appearance of the book is enhanced by high quality diagrams and colour plates and the inclusion of some nice electron micrographs . Summary boxes provide useful background information relevant to the area being discussed; selected readings at the end of each chapter provide a good starting point for those wishing to find further information; however, there is no extensive referencing in the text, which may be a problem for the more specialised reader . Overall, this is an excellent textbook . I am sure it will serve both as an essential teaching aid for undergraduate cellular microbiology courses (which are becoming increasingly common) as well as for graduate students and other researchers in this expanding area . The editors and contributors are to be congratulated on producing such a wide-ranging and readable book . I am sure it will both consolidate interest in the area as well as recruit new members to the group of people who are fascinated by the study of cellular microbiology.

Med Parazitol (Mosk), 2000 Jul-Sep, (3), 9 - 16
{The indirect immunofluorescence reaction in the laboratory diagnosis of ixodid tick-borne borreliosis}; Korenberg EI et al.; Serological verification of ixodid tick-borne borrelioses (TBB) is carried out in Russia by using IFA with the standard antigen prepared from the strain Ip-21 B . afzelii by the Laboratory of Infection Vectors, Gamaleya Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences . This study is under way at approximately 50 research and practical institutions that are situated in 30 large administrative territories of the country . The nIFA data on 371 sera from 214 patients with TBB and 229 sera from controls who had other infections have been analyzed . Changes in the proportion antibody sera, mean geometrical and cut-off antibody titers in patients with TBB were compared with those in the controls . The values of seroconversion were analyzed in relation to the time lasted after the onset, the first serum collection, and an interval of reexamination . The specific results of tested of the coded sera were obtained during the international interlaboratory blind experiment . It is concluded that nIFA yields results that are suitable for TBB verification.

Exp Gerontol, 2000 Aug, 35(5), 513 - 9
Summary: experimental gerontology in Austria; Wick G; Austria has a long and successful tradition in geriatric medicine, nursing and sociology . In contrast, research in experimental gerontology has only come into focus more recently . Gerontological research is performed in various laboratories and clinics of the Medical Schools of the Universities of Vienna, Graz and Innsbruck, respectively, as well as in the School of Veterinary Medicine in Vienna, the Institute for Applied Microbiology of the Vienna University for Agricultural Sciences, the Faculty of Natural Sciences of the University of Salzburg, and in extra-universitary institutions such as the Ludwig-Boltzmann-Institute for Aging Research in Vienna and the Institute for Biomedical Aging Research of the Austrian Academy of Sciences in Innsbruck . Therefore, the following brief reflections will be organized in a topographic fashion.

Diagn Microbiol Infect Dis, 2000 Aug, 37(4), 231 - 5
Intra and inter technologist variability in the quality assessment of respiratory tract specimens; Cooper GM et al.; The majority of microbiology laboratories have implemented quality improvement procedures such as a Q scoring system to assess the nature of clinical specimens . Our study reviewed the sources and the amount of variation when Q scoring of lower respiratory secretions was performed . In total, 450 slides representing lower respiratory tract secretions were Q scored by three experienced technologists . Total agreement regarding the number of neutrophils, squamous epithelial cells and Q scores was 76%, 57% and 57% respectively . The major factor influencing Q score values was the enumeration of epithelial cells . From our findings, we expect that there is greater variability in Q scoring then is generally acknowledged and there is a substantial degree of subjectivity on part of individual technologists reading gram stains.

Biochem Biophys Res Commun, 2000 Aug 28, 275(2), 374 - 81
Molecular characterization of key diphtheria toxin:receptor interactions; Brooke JS et al.; The major amino acids necessary for diphtheria toxin (DT) binding to its receptor have been identified previously . Studies by W . H . Shen et al . (J . Biol . Chem . 269, 29077-29084, 1994) and by J . H . Cha et al . (Mol . Microbiol . 29 (5), 1275-1284, 1998) suggested that the positively charged nature of the single amino acid residue, (516)Lys of DT, is crucial for binding to the DT receptor, whereas the negatively charged (141)Glu of the DT receptor is the most important residue for toxin binding . Here, we hypothesize that key interactions occur between these two oppositely charged amino acid residues . Reciprocal substitution of the residues at these positions between the toxin and the receptor was performed, which resulted in a partial reconstitution of the toxin:receptor interaction . This study provides the first biological data that characterizes the specific interaction of these two key residues with each other and also the additional interactions between other positively charged residues of DT and (141)Glu of the DT receptor .

Cesk Fysiol, 2000 Feb, 49(1), 30 - 3
{Ethics and legal aspects of use of animals in experimental studies}; Vaculin S; Animals have been used in research since ancient times already . Originally they were used for anatomical and physiological demonstrations . Later they started to be used by another fields--microbiology, toxicology and pharmacology, surgery . Animal use is frequently implicated in relation to ethical problems . Actual view is based on biocentric opinion, which part is formed by "three R's" concept--reduction, replacement, refinement in animal use . Laws applied to animal use in Czech Republic are anchored in Law on Animal Protection and Decree of Ministry of Agriculture on Experimental Animal Use and Breeding . Owing to militant behaviour of organisations involved in animal rights movement there is no adequate propagation of experiments, which are ethically and juridically clear and which particularly help mankind in a struggle against diseases.

Biosci Biotechnol Biochem, 2000 Jul, 64(7), 1352 - 8
The characteristics and applications of recombinant cholesterol dehydrogenase; Kishi K et al.; Mass production of an r-CDH derived from Nocardia species was made possible by gene technology . (Horinouchi et al., Applied and Environmental Microbiology, 57, 1386-1393 (1991)) . However, the characteristics of the r-CDH have not been studied in detail and have not been improved enough for industrial use . We accordingly characterized both the native-CDH and the r-CDH prepared from Streptomyces lividans . Both CDHs were monomers with molecular masses of 37 kDa . The Km of r-CDH was 2.50 x 10(-3) M for cholesterol and 2.33 x 10(-4) M for NAD . The activators of CDHs were TritonX-100 and cholate . TritonX-405, Ag+, and Zn2+ inhibited both enzymes . The residual activity of native CDH after heat treatment was 32% (37 degrees C, 60 min), while the r-CDH showed a residual activity of 87% (37 degrees C, 60 min) . The r-CDH is an enzyme with high substrate specificity for cholesterol as well as native CDH and higher thermal stability than native CDH . We have developed a novel serum cholesterol assay using the r-CDH, which permits the direct measurement of cholesterol by measuring NADH reaction products . We conclude that this r-CDH enzyme is useful and can be used to measure cholesterol in a clinical chemistry setting.

Rev Esp Cardiol, 2000 Jul, 53(7), 940 - 6
{Coxiella burnetii endocarditis: long-term clinical course in 20 patients}; Sanchez-Recalde A et al.; INTRODUCTION AND OBJECTIVES: Coxiella burnetii is a causative agent of increasingly frequent subacute infective endocarditis, and is associated with elevated morbimortality . Our aim in the present study was to assess the clinical, serological and therapeutic long-term evolution of 20 patients with Coxiella burnetii endocarditis . METHODS: Twenty patients (13 male and 7 female, age 42 +/- 10 years) admitted between 1982 and 1996 were retrospectively studied . All of them fulfilled the Duke criteria modified by Raoult for Q fever endocarditis . RESULTS: Endocarditis involved prosthetic and native valves in 14 and 6 patients, respectively . All patients except one received antibiotic treatment . Patients treated with doxycycline in monotherapy showed worse evolution than those treated with doxycycline in combination with other antibiotics . Valve replacement was performed in 15 patients, due to prosthetic dysfunction in most of them . The overall mortality was 40% (8 patients) . At follow-up of 74 months (range 19-156) (mean 74 +/- 47) all patients showed persistent high levels of phase I antibodies . At follow-up of 15 to 65 months (32 +/- 30) antibiotic treatment was suspended in five patients because they were asymptomatic and without microbiologic findings of valvular endocarditis . CONCLUSIONS: Q fever endocarditis was associated with severe complications, which often required valve replacement . All patients showed persistent high serological titers of Coxiella burnetii endocarditis without other signs of active infection . This finding raises the issue of suspending antibiotic treatment in patients with negative microbiologic findings and questions the persistence of abnormal serology as a monitor of treatment efficacy.

Pol Merkuriusz Lek, 2000 May, 8(47), 322 - 4
{Pneumonia in HIV-infected patients: clinical observations}; Wnuk A et al.; OBJECTIVES: To determine the occurrence of pneumonias in HIV-infected patients in our hospital during 1990-1999; to evaluate the clinical significance of pneumonias in HIV-seropositive patients; to estimate the ethiology of pulmonary infection . MATERIALS AND METHODS: One hundred and two HIV-infected patients, 17 (16.6%) female and 85 (83.3%) male with mean age of 29 +/- 4.5 yrs, were retrospectively analysed . All patients had a physical examination particularly concerning the clinical symptoms of pulmonary infection, X-ray exam and tuberculin skin test (PPD) . The stage of HIV infection according to the 1993 CDC classification was determined . All patients had the microbiology test of sputum (Pc, TB, fungi, other pathogen) . In some cases the bronchofiberoskopy with bronchoalveolar lavage (BAL) was performed . RESULTS: One hundred and two HIV-positive patients had 129 episodes of pneumonia . We determine the bacterial ethiology in 94/129 (72.9%) cases--TB in 11/129 (8.5%) cases . Fourteen patients had 23 episodes of Pneumocystis carinii pneumonia (PcP) . Three patients had CMV--pneumonitis, detected post mortem . In seven cases the ethiology of pulmonary infection was unknown . In summary the ethiology of pneumonia was determined in 58/129 (44.9%) cases . Thirty three patients were died . The pulmonary infections were main cause of death in 23 (67.7%) persons . CONCLUSIONS: In the era of high active antiretroviral therapy (HAART) the pulmonary infections in HIV-positive patients are the main cause of death as before . The PPD test is useless in HIV-positive patients . We make a note the increase cases of TB in HIV-infected patients during the time of observations.

Cent Eur J Public Health, 2000 Jul, 8 Suppl, 97 - 9
Utilization of microbiotests to assess the contamination of water-bases; Torokne A et al.; In the present study 4 hazardous wastes originated from 4 different typical industrial activities (leachates of waste oil, waste stockpiles of aluminium factory, sewage of leather industry and of the chemical industry) were examined with several Toxkit microbiotests: the Algaltoxkit, the Protoxkit, the Daphtoxkit and the Thamnotoxkit . The data obtained with the microbiotests were compared to those obtained with the standard conventional acute toxicity tests . The goal of the study was to try out the reproducibility of the tests on the same samples, prepared in the same laboratory and provided to all the participants of the interlaboratory exercise . The first reproducibility evaluation of the tests was made with the reference toxicant potassium dichromate (K2Cr2O7) . The outcome was that the lowest coefficients of variation were found at the Thamnotoxkit and highest with Algaltoxkit . The same findings were observed with the industrial waste samples . Different responses were obtained depending on the preparation and the character of the waste . For example, whereas a strong (and highest) toxic effect was found with the Thamnotoxkit, the algal cells in the Algaltoxkit test grew better than in the controls, when exposed to the same waste sample . The sensitivity of the Protoxkit ciliates was the lowest for all the wastes examined . In conclusion of this study, the experience gained and data obtained indicate that the Toxkit microbiotests are suited for routine monitoring after modification of some technical aspects.

Arch Pediatr, 2000 Jun, 7 Suppl 3, 531s - 535s
{Viral co-infections in immunocompetent infants with bronchiolitis: prospective epidemiologic study}; Brouard J et al.; The nature of viral infection was prospectively investigated in 202 immunocompetent infants with bronchiolitis . Nasal aspirates were evaluated by immunofluorescence assay, viral isolation technique and polymerase-chain-reaction-hybridization assay . In 55 infants (27%) more than one respiratory virus were detected . A Rotavirus was found in 40 infants (20%), without any relationship with the respiratory viral status, respiratory syncytial virus being the main virus (46/55), and the association of respiratory syncytial virus and adenovirus being the most frequent (21/55) . No difference was found between monoviral infections on the one hand and simultaneous viral infections on the other hand according to age, weight, neonatal disease, past history of personal or familial atopy, central temperature, Silverman's index, oxygen dependency, length of hospitalization, microbiology data . There was no indication that simultaneous virus infections were associated with an increased severity of the bronchiolitis in immunocompetent infants.

Respiration, 2000, 67(4), 402 - 7
Clinical features associated with a delayed diagnosis of cystic fibrosis; McCloskey M et al.; BACKGROUND: The majority of patients with cystic fibrosis (CF) are diagnosed in the first decade of life . In a small number of patients, the diagnosis is not made until later . OBJECTIVE: In this study, the clinical and genetic features of patients diagnosed after the age of 10 were examined . METHODS: All living patients in Northern Ireland diagnosed prior to 1983, when neonatal screening was introduced, were studied . A total of 103 patients were identified of whom 18 were diagnosed after the age of 10 . The relationships between late diagnosis and clinical presentation, sputum microbiology, pancreatic sufficiency, nutritional status, genotype and distance from the regional CF centres was determined by multiple regression analysis . RESULTS: All 18 late-diagnosed patients had a sweat (chloride >70 mmol/l) . Late diagnosis was significantly related to carriage of the R117H mutation (r(2) = 0.45) and pancreatic sufficiency (r(2) = 0.37) . There was a weak relationship with pulmonary function (r(2) = 0.09) . CONCLUSIONS: In Northern Ireland, late diagnosis in mainly associated with pancreatic function and carriage of the R117H mutation .

Angew Chem Int Ed Engl, 2000 Jul 17, 39(14), 2396 - 2407
Microchips as Controlled Drug-Delivery Devices; Santini Jr JT et al.; Controlled-release systems are common in a number of product areas, including foods, cosmetics, pesticides, and paper . Microencapsulated systems, for example, are used for the release of flavors and vitamins in foods, fragrances in perfumes, and inks in carbonless copy paper . Controlled-release systems for drug delivery first appeared in the 1960s and 1970s . In the past three decades, the number and variety of controlled release systems for drug-delivery applications has increased dramatically . Many of these use polymers having particular physical or chemical characteristics such as biodegradability, biocompatibility, or responsiveness to pH or temperature changes . However, recent advances in the field of microfabrication have created the possibility of a new class of controlled-release systems for drug delivery, namely, that of small, programmable devices . Their small size, potential for integration with microelectronics, and ability to store and release chemicals on demand could make controlled-release microchips useful in a number of areas, including medical diagnostics, analytical chemistry, chemical detection, industrial process monitoring and control, combinatorial chemistry, microbiology, and fragrance delivery . More importantly, drug-delivery microchips resulting from this convergence of controlled release and microfabrication technologies may provide new treatment options to clinicians in their fight against disease.

Int J Syst Evol Microbiol, 2000 Jul, 50 Pt 4, 1695 - 6
The status of the genus Pelczaria (Poston 1994) and the species Pelczaria aurantia (Poston 1994) . Request for an Opinion; Tindall BJ et al.; Based upon the results of another publication {P . Schumann et al . (2000) . Int J Syst Evol Microbiol 50, 1421-1424) it is concluded that the culture Pelczaria aurantia ATCC 49321T (= DSM 12801T) currently being distributed does not conform to the description of the type strain of Pelczaria aurantia (Poston 1994) and the type species of the genus Pelczaria (Poston 1994) . It is proposed that the Judicial Commission consider (1) that the organism currently deposited as ATCC 49321T and DSM 12801T be recognized as a member of the species Kocuria rosea; (2) that the organism deposited as ATCC 49321T and DSM 12801T as the type strain of the species does not represent a strain of the species Pelczaria aurantia; (3) to place the name Pelczaria aurantia (Poston 1994) on the list of rejected names if a suitable replacement strain, or a neotype, cannot be found within 2 years of publication of this Request (Rule 18c); (4) to place the genus name Pelczaria (Poston 1994) on the list of rejected names {c.f . Recommendation 20d (3)} if a suitable replacement type strain or a neotype for the type species of the genus Pelczaria (Poston 1994) cannot be found as outlined in (3).

Pharmacotherapy, 2000 Aug, 20(8), 931 - 40
A new ciprofloxacin stepdown program in the treatment of high-risk febrile neutropenia: a clinical and economic analysis; Marra CA et al.; STUDY OBJECTIVE: To determine treatment outcomes and economic impact of a ciprofloxacin stepdown program for high-risk febrile neutropenic adults from the hospital's perspective . DESIGN: Unblinded, two-phase, single-center study . SETTING: Adult leukemia and stem cell transplant unit . PATIENTS: High-risk adults with febrile neutropenia . INTERVENTION: Two conditions were analyzed: a multidisciplinary ciprofloxacin stepdown program involving a reduction in parenteral ciprofloxacin dose from 400 to 200 mg (i.v.-i.v.) and conversion to oral ciprofloxacin (i.v.-p.o.) when criteria were met; and no i.v.-i.v . stepdown program . MEASUREMENTS AND MAIN RESULTS: Forty-six sequential treatment courses were compared with 42 treatment course from 6-month periods in preintervention (P1) and postintervention (P2) phases . Assessed parameters were clinical and microbiologic outcomes, adverse drug reactions (ADRs), and direct medical resource use and costs (1998 $Canadian) for the episode of febrile neutropenia . A decision analytic model was used to map probabilities and costs and to conduct sensitivity analyses . To supplement standard statistical testing, 1,000 bootstrap samples were created, and the mean cost difference was calculated between phases for each sample . Patient demographics, percentage i.v.-p.o . stepdown, and duration of therapy were similar between phases . Clinical success (83% P1, 81% P2), microbiologic eradication (15% P1, 24% P2), and possible ADRs (6% P1, 9% P2) did not differ . Intravenous-to-intravenous dose stepdown occurred in 33% of P2 and no P1 treatment courses (p<0.001) . Resource use and costs were similar between phases, although a reduction was seen in the drug's mean total cost/day ($58 P1, $52 P2, p=0.04) . There was also a trend toward a decrease in mean total treatment costs ($4,843 P1, $3,493 P2, p=0.08) . In 1,000 bootstrap samples, 99.8% showed a cost advantage for P2 . The model was robust to sensitivity analyses . CONCLUSION: This intervention influenced administration of ciprofloxacin without apparent compromise of patient outcomes and resulted in a reduction in total costs of treating febrile neutropenia.

New Microbiol, 2000 Jul, 23(3), 339 - 46
Search for malaria parasites by PCR and Southern blot in patients with imported malaria in Italy; Manca N et al.; The present study evaluates the sensitivity, specificity and usefulness of a PCR method with Southern blot hybridization to detect malaria parasites in blood samples from subjects with a suspect clinical diagnosis of malaria imported to Italy . Plasmodia were detected by PCR using a genus-specific primer-set corresponding to the sequences common to P . falciparum, P . vivax, P . malariae and P . ovale, as described by Arai (Arai et al., Nucleosides Nucleotides, 1994, 13, 1363-1364) and Kimura (Kimura et al., Journal of Clinical Microbiology, 1995, 33, 2342-2346) . In addition, four distinct tandemly repetitive species-specific probes, described by Kawai (Kawai et al., Analytical Biochimestry, 1993, 209, 63-69), were synthesized to specifically detect the four malaria parasites species by Southern blot hybridization . Fifteen blood samples from 12 patients (7 with malaria) were tested and the genus-specific PCR method showed a sensitivity of 100% and a specificity of 100%, when compared to microscopy, in detecting malaria parasites in the tested blood samples . Fourteen samples (nine were positive and five negative by PCR) were confirmed by Southern blot, whereas only one P . vivax positive sample was not hybridized with the species-specific probes . We conclude that this PCR method with Southern blot hybridization may be useful in detecting malaria parasites in patients with malaria imported to Italy.

Acta Cytol, 2000 Jul-Aug, 44(4), 515 - 23
Accuracy of cytology vs . microbiopsy for the diagnosis of well-differentiated hepatocellular carcinoma and macroregenerative nodule . Definition of standardized criteria from a study of 100 cases; Longchampt E et al.; OBJECTIVE: To determine the accuracy of ultrasound (US)-guided fine needle aspiration (FNA) for the diagnosis of well-differentiated hepatocellular carcinoma (wd HCC) and macroregenerative nodule (MRN) and to identify the most useful cytologic and histologic criteria to distinguish between those two diagnoses . STUDY DESIGN: Cytologic and histologic specimens of 50 wd HCC and 50 MRN were reviewed blindly and the diagnosis compared to the final clinical diagnosis . Twenty-eight cytologic and 25 histologic criteria were examined and subjected to statistical analysis . RESULTS: Among 100 cases studied, the final diagnosis was available for 43 . In those 43 cases, combining analysis of cytologic and histologic specimens, the sensitivity of US-guided FNA was of 75% and the specificity 100% . Cytologic analysis was better than isolated histologic analysis, with a sensitivity of 75% vs . 68%, respectively . Sensitivity of cytologic diagnosis was lower for smaller nodules and for those located in poorly accessible hepatic segments . With the use of stepwise logistic regression analysis, four cytologic features (increased nuclear/cytoplasmic ratio, cellular monomorphism, nuclear crowding, loss of bile duct cells) and four histologic features (increased nuclear/cytoplasmic ratio, decreased Kupffer cells, cellular monomorphism, increased trabeculae thickness) were identified as predictive of HCC.

Pediatr Transplant, 2000 Aug, 4(3), 182 - 5
A comparison of microbiologic flora of the sinuses and airway among cystic fibrosis patients with maxillary antrostomies; Dosanjh A et al.; The placement of maxillary antrostomies among cystic fibrosis (CF) patients has been used as a treatment to allow localized antibiotic lavage of infected sinus passages . This procedure is increasingly recommended by lung transplantation centers as a prerequisite prior to accepting a CF patient as a candidate for transplantation . Our study attempts to define the degree of identity between sinus, endotracheal and sputum cultures from 35 patients . The samples (n = 137) were collected within two weeks of each other . An analysis of the microbiologic type, strain, and antibiotic resistance patterns was undertaken . Randomization analysis was performed and a p-value of < 0.05 was considered significant . The results indicated a high degree of correlation between sinus-sputum pairs (n = 55) and endotracheal samples (p < 0.008) . This study provides evidence that there is a potential for cross-infection between sinus passages and the lower airway . The localized irrigation of CF sinus cavities post-transplantation may be warranted in an attempt to reduce bacterial counts and potential direct infection of the allograft . However, it is unlikely that this will eliminate this risk because bacterial colonization continues and the CF trachea is another source of infection.

Ann Biol Clin (Paris), 2000 Jul-Aug, 58(4), 431 - 7
{Standards and recommendations for hospital environmental hygiene}; Brocard-Lemort C; The hospital laboratory has important responsibilities in the survey, control and prevention of nosocomial infections . Contamination of environment (surfaces, air, liquids) has been implicated in nosocomial infections . It is essential to develop a monitoring system for biocontamination control in the controlled environment areas . Classification of zones at risk (low, medium, high and very high risk) should be performed according to relevant guidelines or regulations where these exist . According to the classification of the zone at risk appropriate levels of biocontamination will be assigned . Standards for acceptable levels of microbiologic contamination will be suggested because no uniform agreement have been achieved because of the lack of correlation between these levels and occurrence of clinical infection.

Southeast Asian J Trop Med Public Health, 1999, 30 Suppl 3, 32 - 8
Twenty-years of experiences in external quality assurance in Korea; Cho HI; The Korean External Quality Assurance Scheme (KEQAS) was set up by a voluntary organization, the Korean Society of Quality Control in Clinical Pathology (KSQC) in 1976 . The KSQC was reorganized as the Korean Association of Quality Assurance for Clinical Laboratories (KAQACL) in 1991 . The KAQACL is a government authorized organization . With the growing popularity of the KEQAS, its coverage has expanded to include ten disciplines, and the number of participants has grown to 470 clinical laboratories by the end of 1998 . The ten programs are: clinical chemistry, clinical microbiology, diagnostic hematology, diagnostic immunology . blood bank, urinalysis, immunoassay, TDM (therapeutic drug monitoring), inborn errors of metabolism and diagnostic genetics . KAQACL membership accreditation is based on the QAP return rate, of which the minimum criterion for acceptability is 50% . Membership in KAQACL is highly regarded by the Inspection and Accreditation Program of the Korean Hospital Association and is an essential requirement for outreach services of commercial laboratories . While monitoring the quality of laboratory performances during the last two decades, the KEQAS has provided information on reliability of laboratory performances regarding methods, manufacturers and laboratory groups . The inter-laboratory coefficient variations (CVs) and the variance index score (VIS) have gradually decreased over the years, particularly during the 1990s . The participants who achieved good performances (less than 100 VIS) in clinical chemistry tests have gradually increased in number up to 83.2% of all participants.

Rev Neurol, 2000 May 16-31, 30(10), 964 - 7
{Genetic and environmental factors in multiple sclerosis}; Fernandez O; INTRODUCTION: Multiple sclerosis is expressed in genetically susceptible persons randomly affected by an unknown environmental factor, probably a virus, which sets off an abnormal immune process . DEVELOPMENT AND CONCLUSION: Race is an important predictive risk factor for multiple sclerosis, but genetics alone do not explain the occurrence of this disease . The genetic predisposition is complex, since several loci are involved in the susceptibility . Probably the immune process seen in multiple sclerosis is a consequence not a cause . The cause is possibly unique and due to infection . The causative agent may be rare or on the contrary may be frequent, but has different biological effects on susceptible persons . Epidemiology alone is not sufficient to find the cause . This requires collaboration from the basic sciences (genetics, microbiology, etc.) . The results of epidemiological methods would be much improved if a biological marker for the disease was available.

Semin Arthritis Rheum, 2000 Jun, 29(6), 373 - 8
Cytopenia and past human parvovirus B19 infection in patients with primary Sjögren's syndrome; Ramos-Casals M et al.; OBJECTIVES: To determine the clinical significance of human parvovirus B19 infection in patients with primary Sjogren's syndrome (SS) and to investigate the immunologic and hematologic features related to B19 infection . METHODS: We included 80 consecutive patients with primary SS (74 women and 6 men), with a mean age of 62 years (range, 24 to 87 years) that were seen in our Unit . All patients fulfilled the European Community criteria for SS . As controls, we included 140 consecutive sera samples analyzed for B19 antibodies in our Microbiology Department and obtained from adult inpatients and outpatients of our Hospital . Serum from all patients and controls was tested for antibodies to B19 by enzyme-linked immunosorbent assay (ELISA) . Additionally, the presence of B19 DNA in serum and in circulating leukocytes was investigated by nested polymerase chain reaction (PCR) . RESULTS: Serological evidence of past B19 infection (positive IgG antibodies without IgM antibodies) was present in 28 (35%) patients with primary SS . None of these patients showed evidence for B19 viremia, and B19 virus DNA was not detected in the circulating leukocytes of IgG-B19(+) patients . Positivity for IgM antibodies to B19 was not detected in any patient . When compared with patients without evidence of past B19 infection, those with primary SS and past B19 infection showed a higher prevalence of cytopenia (57% v 15%; P < .001), and, specifically, of leukopenia (36% v 4%; P < .001) . Additionally, when compared with controls positive for IgG-B19, SS patients with these antibodies had a higher prevalence of cytopenia (57% v 13%; P < .001), leukopenia (36% v 3%; P < .001) and thrombocytopenia (21% v 0%; P = .003) . CONCLUSIONS: Serological evidence of past B19 infection is associated with the presence of cytopenia in our patients with primary SS . A possible relationship between B19 infection and the presence of cytopenia in primary SS may occur in some patients immunologically or genetically predisposed.

J Clin Microbiol, 2000 Aug, 38(8), 2853 - 7
vacA genotypes in Helicobacter pylori strains isolated from children with and without duodenal ulcer in Brazil; De Gusmao VR et al.; Data concerning the association between vacA genotypes and disease in children in both developed and developing countries are scarce, especially because of the small number of children with a duodenal ulcer studied . The vacA genotypes of Helicobacter pylori strains obtained from 65 children (24 with a duodenal ulcer and 41 without a duodenal ulcer; 33 girls; mean age, 10.2 years; age range, 1 to 17 years) were investigated as described by J . C . Atherton et al . (J . Clin . Microbiol . 37:2979-2982, 1999) . Ten (15.4%) children were infected with more than one H . pylori strain . None of these patients were included in our analysis of the relationship between gastric disorders and specific vacA genotypes . The s1 allele was detected in all H . pylori strains isolated from patients with a duodenal ulcer and from 21 (58.3%) patients without a duodenal ulcer (P = 0.003) . Strains with the s2 allele were found only in patients without ulcer (n = 15; 41.7%) . Most s1 strains had the s1b allele (97.5%), a result similar to that reported for adults from the Iberian peninsula, which could reflect the Brazilian population origin . One untypeable s1 strain was isolated . The m1 allele was also more frequently found in strains obtained from duodenal ulcer patients (P = 0.028) . The m2 allele was found in strains obtained from 20 (36 . 4%) children, 3 (15.8%) with an ulcer and 17 (47.2%) without an ulcer . Only one m hybrid strain (m1 and m2 hybrid) was detected . It was demonstrated for the first time that the frequencies of colonization with strains with the s1 allele (14.3% in children up to 8 years of age and 85.7% in older patients; P = 0.012) and of strains with the m1 allele (11.1% in patients up to the age 8 years and 88.9% in older children; P = 0.013) increase with age.

Rapid Commun Mass Spectrom, 2000, 14(15), 1337 - 44
Compound-specific isotope analysis . Application to archaeology, biomedical sciences, biosynthesis, environment, extraterrestrial chemistry, food science, forensic science, humic substances, microbiology, organic geochemistry, soil science and sport; Lichtfouse E; The isotopic composition, for example, (14)C/(12)C, (13)C/(12)C, (2)H/(1)H, (15)N/(14)N and (18)O/(16)O, of the elements of matter is heterogeneous . It is ruled by physical, chemical and biological mechanisms . Isotopes can be employed to follow the fate of mineral and organic compounds during biogeochemical transformations . The determination of the isotopic composition of organic substances occurring at trace level in very complex mixtures such as sediments, soils and blood, has been made possible during the last 20 years due to the rapid development of molecular level isotopic techniques . After a brief glance at pioneering studies revealing isotopic breakthroughs at the molecular and intramolecular levels, this paper reviews selected applications of compound-specific isotope analysis in various scientific fields .

Adolesc Med, 2000 Jun, 11(2), 279 - 92
Acute and chronic viral hepatitis; O'Connor JA; Viral hepatitis is the most common cause of acute and chronic hepatitis . The term viral hepatitis generally refers to infections resulting from one of the hepatotrophic viruses: hepatitis A, B, C, D, and E . The last 10 years have brought many important advances in understanding the epidemiology, pathogenesis, molecular biology, and immunoprophylaxis of infections caused by hepatotrophic viruses . Development of sensitive and specific immunoassays has enabled detection of specific agents . This has allowed for identification of infected patients and monitoring response to therapy . Additionally, serologic markers have allowed for isolation of contaminated blood products and a reduction in the spread of disease . The remaining challenge is the application of this knowledge to the treatment and prevention of viral hepatitis . This article explores the risk factors, epidemiology, microbiology, clinical and laboratory diagnosis, treatment, and prevention of the hepatotrophic viral infections.

J Clin Periodontol, 2000 Jul, 27(7), 473 - 80
Serum IgG reactivity to subgingival bacteria in initial periodontitis, gingivitis and healthy subjects; Tanner AC et al.; BACKGROUND/AIMS: Established periodontal diseases may be associated with antibody responses to periodontal pathogens, but it is not known at which stage of disease this antibody response is initiated . This study aimed to characterize the host systemic response in initial periodontitis, gingivitis, and periodontal health, to evaluate whether elevated serum antibodies to subgingival species could be detected in initial periodontitis . METHOD: Human systemic immune response were evaluated to 40 subgingival bacterial species in 16 healthy, 21 gingivitis, 11 initial periodontitis and 5 progressing recession adults . Subjects had minimal periodontal attachment level (AL) loss at baseline . Disease categories were determined after 12 months monitoring at three-month intervals . Increased AL loss > or = 1.5 mm (disease activity) at interproximal sites defined initial periodontitis, recession was characterized by AL loss at buccal sites . Serum IgG antibodies were evaluated semi-quantitatively by immunoblot from blood taken at baseline, active and final visits . RESULTS: No antibody was detected from 55% of reactions . When detected, levels were below those reported for advanced periodontitis subjects . There were no major differences in serum antibody levels between healthy, gingivitis and initial periodontitis subjects, despite differences in the subgingival microbiota . Serum antibodies for more species were detected in recession subjects, compared with the other study subjects . No changes in antibody levels were detected between baseline, active, and final visits . No systematic association between species colonization and presence of systemic antibody was observed . CONCLUSIONS: This study did not detect differential elevation of mean serum antibody levels in initial periodontitis subjects, suggesting that serum antibody levels are not sensitive risk markers for initial periodontitis.

Aust Vet J, 2000 May, 78(5), 334 - 8
Towards an understanding of equine pleuropneumonia: factors relevant for control; Racklyeft DJ et al.; OBJECTIVE: To review relevant literature on factors associated with the development of equine pleuropneumonia . DESIGN: A review of the literature using a range of databases including Current Contents, Medline, ChemAbstracts, Biological Abstracts and CAB and a comprehensive search strategy which involved use of keywords, author and subject category searches . Additional sources included review of articles cited by key accumulated references . RESULTS: Since the early years of this century, many of the "gaps" in our knowledge of the pathogenesis of this disease have been filled . We now know that equine pleuropneumonia results from contamination of the lower respiratory tract with bacteria similar to the normal oropharyngeal microbiota of the horse and that transportation of any mode, especially over long distances (and consequently with no or short rest periods), is the single most important predisposing factor for this disease . This is associated with restraint of horses such that they are unable to lower their heads, which leads to increased opportunity for lower respiratory tract contamination and a reduced opportunity for clearance . Strenuous exercise also results in lower respiratory tract contamination and exercise subsequent to transportation exerts additive detrimental effects on the defenses of the lower respiratory tract . CLINICAL IMPLICATIONS: While modern veterinary medicine and surgery have significantly reduced the death rate from pleuropneumonia, horses that develop the disease have a high probability of not returning to their prior use . This underscores the importance of developing the most effective strategies for its prevention.

Diagn Microbiol Infect Dis, 2000 Jul, 37(3), 165 - 8
Evaluation of a water-extracted pharmamedia agar medium for the culture of fungi; Slifkin M; The purpose of this investigation was to determine the value of a water-extracted Pharmamedia agar medium for the culture of fungi . The growth of the fungi on this medium was compared to that on Sabouraud brain heart infusion agar . The Pharmamedia extract agar supported the growth of the 42 filamentous and yeast or yeast-like fungi that were examined . The medium was relatively clear to easily permit observation of the diagnostic microscopic and macroscopic architecture of the organisms . The Pharmamedia extract agar also permitted the conversion of the mold-to-yeast phase with Blastomyces dermatitides and Penicillium marneffei.This investigation supports the use of Pharmamedia as a culture medium for the clinical microbiology laboratory . Its relative simplicity of preparation and effective use as a culture medium, as well as a conversion medium, emphasizes its useful application for clinical mycology.

Diagn Microbiol Infect Dis, 2000 Jul, 37(3), 151 - 5
Evaluation of the BIOMIC video reader system for routine use in the clinical microbiology laboratory; Geiss HK et al.; The BIOMIC Video Reader System (Giles Scientific, New York, NY, USA) is a semi-automated AST method that combines disk diffusion testing with automated reading and data interpretation . We evaluated this system with 497 strains comprising a total of 5821 drug organism combinations (DOC) from our routine diagnostic laboratory . Additionally, we compared the time required of the manual and the automated method . The overall agreement of interpretative categories of all DOC was 96.1% . However, comparing complete tests the agreement was only 70.8% . The average time required of the BIOMIC system to complete a test was more than twice as long as that of the manual method . Our data suggest that the tested version of the BIOMIC system cannot be recommended for routine use in diagnostic laboratories.

Keio J Med, 2000 Jun, 49(2), 80 - 3
Seeing through the stratum corneum; Marks R; The stratum corneum (SC) provides a vital barrier membrane between the external environment and the vulnerable internal tissues of the skin . It impedes the flow of water, the penetration of xenobiotics, and invasion of pathogenic micro-organisms . It also has protective capacity against ultraviolet radiation and thermal injury . As routine histopathology provides a misleading picture of a disorganized and shadowy SC, we would recommend the skin surface biopsy technique . This painless technique is easy and reliable in obtaining information from the SC . It demonstrates the geometric patterns of the surface, the openings of the eccrine ducts and hair follicles . The skin surface biopsy technique is also ideal for the investigation of the in situ microbiology of skin . Staining with periodic acid Schiff reagent makes it possible to see ringworm fungi, pityriasis versicolor, candida species, or erythrasma micro-organisms . Scanning electron microscopy can be employed when the higher magnification is needed . Histochemical applications include silver staining for melanin particle, potassium ferricyanide staining for blood pigments and lipid staining with Sudan red, for sebum . The rate of movement of topically applied drugs into the skin can be measured using the skin surface biopsy technique . The concentration of radiolabelled drugs can be counted and compared . Comedogenicity and DNA analysis are other applications of this non-invasive technique.

Micron, 2001 Jan, 32(1), 5 - 6
Microbiology: impact on research in life sciences; Arber W; In this essay with a flavor of science history, the influence of imaging techniques, as compared to other research strategies, on microbiologic investigations is discussed . Using a few selected examples, to what degree microbiology became a leading science during the last 50 years in gaining knowledge in life sciences, particularly with regard to molecular genetics and more recently molecular evolution is also discussed.

Arch Ophthalmol, 2000 Jul, 118(7), 996 - 1000
Indwelling temporary retrobulbar catheter for long-lasting titratable local anesthesia; Jonas JB et al.; OBJECTIVE: To evaluate an indwelling temporary retrobulbar catheter for repeatable injections of local anesthetics for long-lasting and titratable retrobulbar anesthesia in intraocular surgery . PARTICIPANTS: The prospective clinic-based study included 153 patients who underwent vitreoretinal surgery (n=111) or buckling procedures with cryocoagulation (n=34) . The mean duration of surgery was 84.7 +/- 49.5 minutes (range, 25-310 minutes) . Using commercially available retrobulbar needles with a diameter of 0.60 or 0.80 mm and a length of 38 mm, 5 mL of 2% mepivacaine hydrochloride was injected . Through the same needle, a 28-gauge commercially available flexible catheter was introduced into the retrobulbar space . The needle was withdrawn and the catheter was fixed . When the patients started to feel pain during surgery, 2 mL of mepivacaine hydrochloride was reinjected through the catheter . RESULTS: Ten to 240 minutes after the start of the operation, 96 patients needed an intraoperative reinjection of mepivacaine after which they felt comfortable again . Forty-two patients needed a second reinjection of mepivacaine 30 to 270 minutes after the start of the operation, and 13 patients needed a third reinjection 45 to 145 minutes after the start of surgery . Removal of the catheter after surgery was unremarkable . No infections were observed . Microbiologic examination results of the catheter tip were negative for organisms . Diplopia or other motility problems were not detected . Introduction and fixation of the catheter took less than 5 minutes in all patients . CONCLUSIONS: An indwelling temporary retrobulbar catheter for repeatable intraoperative injections of local anesthetics is simple, effective, and useful, and in comparison with general anesthesia, it is a time-saver for long-lasting and titratable local anesthesia in intraocular surgery . Arch Ophthalmol . 2000;118:996-1000

J Gen Virol, 2000 Aug, 81(Pt 8), 2067 - 75
Distribution of chicken anaemia virus in the reproductive tissues of specific-pathogen-free chickens; Cardona CJ et al.; The specific-pathogen-free (SPF) flocks of chickens maintained by the Department of Microbiology and Immunology at Cornell University became infected, inadvertently, with chicken anaemia virus (CAV), as demonstrated by seroconversion . Chickens from five flocks representing three different strains were examined for the presence of CAV using nested PCR . Virus was detected in ovaries, infundibula, vas deferentia, testes and spleens . Ovaries were positive in 38 to 72% of the hens in four flocks with 13 to 56 birds examined per flock . Interestingly, the ovaries were often the only positive tissues, while a few hens had only positive spleens . In roosters, the vas deferens was positive in 30 to 79% of the birds with 5 to 19 birds examined per flock; the vas deferens was the only positive tissue in 20 to 37% . Individual cells in the theca externa and rare epithelial cells in the infundibular epithelium were positive for CAV by in situ PCR . Positive cells were not detected in testes or vas deferentia . The SH-1 strain of CAV was isolated from these tissues and partially sequenced . Only minor sequence differences were found compared to CIA-1 and Cux-1 . Embryos from matings between persistently infected dams and sire had CAV-positive cells in mesenchyme near the developing vertebral column . The data show that CAV persists in the reproductive tissues far longer than previously thought, and that it can be vertically transmitted from persistently infected birds.

Oral Oncol, 2000 Jul, 36(4), 373 - 81
Defining mechanisms of action of interleukin-11 on the progression of radiation-induced oral mucositis in hamsters; Sonis ST et al.; Oral ulcerative mucositis is a common toxicity associated with drug and radiation therapy for cancer . It impacts on quality of life and economic outcomes, as well as morbidity and mortality . Mucositis is often associated with dose limitations for chemotherapy or is a cause for dose interruption for radiation . The complexity of mucositis as a biological process has only been recently appreciated . It has been suggested that the condition represents a sequential interaction of oral mucosal cells and tissues, pro-inflammatory cytokines and local factors such as saliva and the oral microbiota . The recognition that the pathophysiology of mucositis is a multifactorial process was partially suggested by the observation that interleukin-11 (IL-11), a pleotropic cytokine, favorably altered the course of chemotherapy-induced mucositis in an animal model . In the current study, we evaluated a series of biologic and morphologic outcomes to determine their roles and sequence in the development of experimental radiation-induced mucositis and to evaluate the effects of IL-11 in attenuating them . Our results suggest that IL-11 favorably modulates acute radiation-induced mucositis by attenuating pro-inflammatory cytokine expression . Data are also presented which help define the pathobiological sequence of mucositis.

Rev Belge Med Dent, 1999, 54(3), 190 - 4
{The sampling of plaque specimens in oral microbiology}; Bollen C; Sampling is the first and most important step in the bacterial analysis of periodontal or endodontal infections . Sterile paperpoints are the ultimate tool to perform the sampling . Sampling is preferably done under continuous CO2-flushing, to preserve an anaerobic environment for the bacteria . For the dilution and the transport of the bacteria, Reduced Transport Fluid (RTF) is the most reliable fluid, when culturing is indicated . In all other cases, sterile physiologic water is sufficient.

Ann R Australas Coll Dent Surg, 1998 Oct, 14, 62 - 5
The use of systemic antibiotics in dental practice; Bischof WH; Clinical dental practice is immersed in the primary treatment and corrective management of infectious bacterial diseases of the oral and peri-oral region . Systemic antibiotics play a key role in achieving either therapeutic and/or prophylactic treatment goals . There are a number of theoretical considerations associated with the use of systemic antibiotics in dental practice . These are related to the principles of infection management, microbiology and host response, and the pharmacology of the particular agent . In the clinical setting, these principles are modulated by a number of factors . These factors need to be understood to ensure appropriate prescribing of antibiotics . Minimizing the occurrence of antibiotic misuse and abuse has global implications for the containment of resistant bacterial strains . This paper discusses some of the current understanding of prescribing practices by dentists . Also presented is an outline of the factors that may be considered in the use of antibiotics.

Acta Otorhinolaryngol Ital, 2000 Feb, 20(1), 47 - 53
{Difficulties in the clinical, radiological and therapeutic evaluation of the initial stage of mucormycosis of the rhinosinus}; Galli J et al.; The initial diagnosis and subsequent treatment of rhino-orbital mucormycoses is quite difficult, particularly because the patient may find it difficult to accept aggressive therapeutic protocols, even when free of any endocranial involvement . The authors draw inspiration from a clinical case of rhino-orbital mucormycosis in a patient suffering from decompensated type I diabetes mellitus to discuss the main clinical-diagnostic and therapeutic aspects of this disorder . Timely medical-surgical treatment proves extremely important for prognosis, preventing the intracranial extension of the lesion which is the cause of death in 80% of such cases . As regards the diagnostic protocol, careful clinical, radiological monitoring with CT and NMR--in strict interdisciplinary cooperation between otorhinolaryngologist, radiologist, ophthalmologist, microbiologist and histopathologist--is especially important . Radical surgery, at times demolition, associated with correction of the metabolic decompensation, systemic therapy with Amphotericin B and localbi-weekly medication for three months constitutes the best therapeutic protocol for treatment of this disorder.

Appl Environ Microbiol, 2000 Jul, 66(7), 2732 - 42
Archaeal population dynamics during sequential reduction processes in rice field soil; Lueders T et al.; The population dynamics of Archaea after flooding of an Italian rice field soil were studied over 17 days . Anoxically incubated rice field soil slurries exhibited a typical sequence of reduction processes characterized by reduction of nitrate, Fe(3+), and sulfate prior to the initiation of methane production . Archaeal population dynamics were followed using a dual approach involving molecular sequence retrieval and fingerprinting of small-subunit (SSU) rRNA genes . We retrieved archaeal sequences from four clone libraries (30 each) constructed for different time points (days 0, 1, 8, and 17) after flooding of the soil . The clones could be assigned to known methanogens (i.e., Methanosarcinaceae, Methanosaetaceae, Methanomicrobiaceae, and Methanobacteriaceae) and to novel euryarchaeotal (rice clusters I, II, and III) and crenarchaeotal (rice clusters IV and VI) lineages previously detected in anoxic rice field soil and on rice roots (R . Grosskopf, S . Stubner, and W . Liesack, Appl . Environ . Microbiol . 64:4983-4989, 1998) . During the initiation of methanogenesis (days 0 to 17), we detected significant changes in the frequency of individual clones, especially of those affiliated with the Methanosaetaceae and Methanobacteriaceae . However, these findings could not be confirmed by terminal restriction fragment length polymorphism (T-RFLP) analysis of SSU rDNA amplicons . Most likely, the fluctuations in sequence composition of clone libraries resulted from cloning bias . Clonal SSU rRNA gene sequences were used to define operational taxonomic units (OTUs) for T-RFLP analysis, which were distinguished by group-specific TaqI restriction sites . Sequence analysis showed a high degree of conservation of TaqI restriction sites within the different archaeal lineages present in Italian rice field soil . Direct T-RFLP analysis of archaeal populations in rice field soil slurries revealed the presence of all archaeal lineages detected by cloning with a predominance of terminal restriction fragments characteristic of rice cluster I (389 bp), Methanosaetaceae (280 bp), and Methanosarcinaceae/rice cluster VI (182 bp) . In general, the relative gene frequency of most detected OTUs remained rather constant over time during the first 17 days after flooding of the soil . Most minor OTUs (e.g., Methanomicrobiaceae and rice cluster III) and Methanosaetaceae did not change in relative frequency . Rice cluster I (37 to 30%) and to a lesser extent rice cluster IV as well as Methanobacteriaceae decreased over time . Only the relative abundance of Methanosarcinaceae (182 bp) increased, roughly doubling from 15 to 29% of total archaeal gene frequency within the first 11 days, which was positively correlated to the dynamics of acetate and formate concentrations . Our results indicate that a functionally dynamic ecosystem, a rice field soil after flooding, was linked to a relatively stable archaeal community structure.

Antibiot Chemother, 2000, 50, 10 - 9
Serum and plasma parameters in clinical evaluation of neutropenic fever; Sudhoff T et al.; Clinicians are searching for a marker which may add to exclusion or diagnosis of relevant infection underlying neutropenic fever . The rise of such a parameter should ideally precede the date of significant microbiologic findings or justify additional intensive search for a focus of infection even in patients without pyrexia . However, the literature concerning the significance of CRP, proinflammatory cytokines and soluble adhesion molecules in the clinical evaluation of neutropenic fever is surprisingly small . In the case of procalcitonin, available data look very preliminary . Furthermore, in case of CRP, it appears that the widespread view that its determination may add substantially to the clinical evaluation of neutropenic fever is not well founded by most clinical trials listed here . Most of the studies available demonstrate several limitations such as poor design and small size of the study population . Additionally, studies were heterogeneous with respect to patients recruited (children and adults, patients with leukemia and patients with solid tumors) and compared different categories of febrile episodes . None of the investigators analyzed cost-effectiveness or impact of serial measurements of these parameters on patients' outcome . To our knowledge no single multicenter trial has been published addressing this issue . Although the group of proinflammatory cytokines and known acute-phase reactants will surely grow, more data on relevance of the available parameters in the diagnosis of neutropenic fever are needed.

Am J Surg, 2000 Feb 1, 179(2 Suppl 1), 36 - 39
Pneumonia in the surgical patient: duration of therapy and does the organism matter?
Spain DA.
Pneumonia is a frequent complication in the critically ill surgical or trauma patient . Despite its common occurrence and increased attention recently, many basic issues, such as how long to treat and whether the exact causative organism even matters, remain unanswered . Currently, cessation of treatment is based on clinical response, although some data suggest that both the microbiologic and immunologic response within the lung are also important . A thorough knowledge of the likely causative organisms for both early and late pneumonia should allow safe and efficacious empiric antibiotic selection.

Am J Surg, 2000 Feb 1, 179(2 Suppl 1), 31 - 35
Multifactorial analyses in the diagnosis of pneumonia arising in the surgical intensive care unit; Polk HC et al.; The diagnosis of ventilator-associated pneumonia in the surgical intensive care unit continues to be problematic . The majority of intensive care units use clinical criteria based on chest x-ray; fever; leukocytosis; alterations in the pulse oximeter observations; the need to alter modes and amounts of ventilatory support; and more specific microbiologic studies, such as appropriate sputum, Gram stain, and culture to identify pneumonia . Diagnosing pneumonia based on clinical criteria alone is often difficult and inaccurate, which may lead to inappropriate use and choice of antibiotics . Invasive diagnostic techniques, such as protected specimen brush and bronchoalveolar lavage, provide an important microbiologic diagnosis . However, the cost and inconvenience limit broad usage . Furthermore, those results that return positive are often too late to dictate the need for, or direction of, therapy . Our use of a "pneumonia grid" may help identify patients likely to have a poor outcome . Until a readily available and cost-effective diagnostic study for pneumonia is developed, clinical criteria remain vital in routine practice.

Am J Surg, 2000 Feb 1, 179(2 Suppl 1), 26 - 29
Diagnosis of acute respiratory distress syndrome and differentiation from ventilator-associated pneumonia; Croce MA; Acute respiratory distress syndrome (ARDS) remains a significant cause of morbidity and mortality in surgical intensive care unit patients . A relatively predictable sequence of pathophysiologic events occur in the lung, which involve inflammatory mediators and neutrophils . The characteristic radiographic findings of ARDS may mimic ventilator-associated pneumonia (VAP), making the diagnosis of VAP difficult . The standard clinical criteria of fever, leukocytosis, purulent sputum, and infiltrate on chest radiograph are not specific for pneumonia in the surgical patient . The use of bronchoscopy with bronchoalveolar lavage and quantitative cultures can differentiate local and systemic inflammatory response to injury or blood loss from invasive bacterial pneumonia . Basing antibiotic therapy solely on the results of quantitative cultures is safe, because quantitative cultures identify VAP in less than half the patients with clinical evidence of pneumonia . Empiric therapy should be based on the microbiology of the intensive care unit rather than the results of the Gram stain.

Sports Med, 2000 Jun, 29(6), 439 - 47
Tinea gladiatorum: wrestling's emerging foe; Kohl TD et al.; Tinea corporis, or ringworm, has become a common nuisance in competitive wrestling . Although it is a fairly benign infectious skin disease, it has significant effects on the ability of a wrestler to compete because of infection control issues . Very little has been published in the medical literature describing this problem . The majority of the literature has described outbreaks in an isolated group of wrestlers . One must examine ringworm infections in wrestlers as an entity distinct from tinea corporis infections typically seen in the paediatric population, thus the term 'tinea gladiatorum' . Tinea gladiatorum outbreaks have been caused by the dermatophyte, Trichophyton tonsurans . The epidemiology and microbiology point to person-to-person contact as the main source of transmission in wrestlers . The clinical features of tinea gladiatorum may or may not be consistent with those found in the general population . Ancillary tests, including potassium hydroxide preparations and fungal cultures may have to be done to confirm the diagnosis . Treatment guidelines for tinea corporis have failed to produce the desired goals in this particular population . More research studying different treatment regimens in the wrestling environment is needed to define the optimal treatment to return wrestlers to competition quickly without putting other wrestlers at risk for infection . Intuitive hygiene practices have been suggested to prevent spread of the infection, but they have not been substantiated . Anecdotal reports suggest that hygiene practices fall short of producing adequate primary or secondary prevention . Pharmaceutical prophylaxis has been effective, but universal drug prophylaxis carries risks including drug adverse effects and potential drug resistance . The role of potential asymptomatic carriers of dermatophytes has yet to be elucidated in the origin and/or perpetuation of tinea gladiatorum outbreaks . There are many unanswered questions about tinea gladiatorum . Sports medicine professionals must work to define this entity more completely before making recommendations about treatment, prevention and infection control . The ultimate goal is the eradication of tinea infections from the wrestling world . Energy should be focused on primary and secondary prevention, as well as treatment . Without a thorough knowledge of tinea gladiatorum as a distinct disease entity, wrestling has been losing its battle with this formidable opponent.

J Bacteriol, 2000 Jul, 182(14), 4087 - 95
Expansion of the clavulanic acid gene cluster: identification and in vivo functional analysis of three new genes required for biosynthesis of clavulanic acid by Streptomyces clavuligerus; Li R et al.; Clavulanic acid is a potent inhibitor of beta-lactamase enzymes and is of demonstrated value in the treatment of infections by beta-lactam-resistant bacteria . Previously, it was thought that eight contiguous genes within the genome of the producing strain Streptomyces clavuligerus were sufficient for clavulanic acid biosynthesis, because they allowed production of the antibiotic in a heterologous host (K . A . Aidoo, A . S . Paradkar, D . C . Alexander, and S . E . Jensen, p . 219-236, In V . P . Gullo et al., ed., Development in industrial microbiology series, 1993) . In contrast, we report the identification of three new genes, orf10 (cyp), orf11 (fd), and orf12, that are required for clavulanic acid biosynthesis as indicated by gene replacement and trans-complementation analysis in S . clavuligerus . These genes are contained within a 3.4-kb DNA fragment located directly downstream of orf9 (cad) in the clavulanic acid cluster . While the orf10 (cyp) and orf11 (fd) proteins show homologies to other known CYP-150 cytochrome P-450 and {3Fe-4S} ferredoxin enzymes and may be responsible for an oxidative reaction late in the pathway, the protein encoded by orf12 shows no significant similarity to any known protein . The results of this study extend the biosynthetic gene cluster for clavulanic acid and attest to the importance of analyzing biosynthetic genes in the context of their natural host . Potential functional roles for these proteins are proposed.

Ann N Y Acad Sci, 2000, 899, 136 - 47
Free radicals and antioxidants in the year 2000 . A historical look to the future; Gutteridge JM et al.; In the late 1950's free radicals and antioxidants were almost unheard of in the clinical and biological sciences but chemists had known about them for years in the context of radiation, polymer and combustion technology . Daniel Gilbert, Rebeca Gerschman and their colleagues related the toxic effects of elevated oxygen levels on aerobes to those of ionizing radiation, and proposed that oxygen toxicity is due to free radical formation, in a pioneering paper in 1956 . Biochemistry owes much of its early expansion to the development and application of chromatographic and electrophoretic techniques, especially as applied to the study of proteins . Thus, superoxide dismutase (SOD) enzymes (MnSOD, CuZnSOD, FeSOD) were quickly identified . By the 1980's Molecular Biology had evolved from within biochemistry and microbiology to become a dominant new discipline, with DNA sequencing, recombinant DNA technology, cloning, and the development of PCR representing milestones in its advance . As a biological tool to explore reaction mechanisms, SOD was a unique and valuable asset . Its ability to inhibit radical reactions leading to oxidative damage in vitro often turned out to be due to its ability to prevent reduction of iron ions by superoxide . Nitric oxide (NO.) provided the next clue as to how SOD might be playing a critical biological role . Although NO . is sluggish in its reactions with most biomolecules it is astoundingly reactive with free radicals, including superoxide . Overall, this high reactivity of NO . with radicals may be beneficial in vivo, e.g . by scavenging peroxyl radicals and inhibiting lipid peroxidation . If reactive oxygen species are intimately involved with the redox regulation of cell functions, as seems likely from current evidence, it may be easier to understand why attempts to change antioxidant balance in aging experiments have failed . The cell will adapt to maintain its redox balance . Indeed, transgenic animals over-expressing antioxidants show some abnormalities of function . There must therefore be a highly complex interrelationship between dietary, constitutive, and inducible antioxidants with the body, under genetic control . The challenge for the new century is to be able to understand these relationships, and how to manipulate them to our advantage to prevent and treat disease.

Gene, 2000 Jun 13, 251(1), 81 - 90
The FK520 gene cluster of Streptomyces hygroscopicus var . ascomyceticus (ATCC 14891) contains genes for biosynthesis of unusual polyketide extender units; Wu K et al.; FK520 (ascomycin) is a macrolide produced by Streptomyces hygroscopicus var . ascomyceticus (ATCC 14891) that has immunosuppressive, neurotrophic and antifungal activities . To further elucidate the biosynthesis of this and related macrolides, we cloned and sequenced an 80kb region encompassing the FK520 gene cluster . Genes encoding the three polyketide synthase (PKS) subunits (fkbB, fkbC and fkbA), the peptide synthetase (fkbP), the 31-O-methyltransferase (fkbM), the C-9 hydroxylase (fkbD) and the 9-hydroxyl oxidase (fkbO) had the same organization as the genes reported in the FK506 gene cluster of Streptomyces sp . MA6548 (Motamedi, H., Shafiee, A., 1998 . The biosynthetic gene cluster for the macrolactone ring of the immunosuppressant FK506 . Eur . J . Biochem . 256, 528-534) . Disruption of a PKS gene in the cluster using the φC31 phage vector, KC515, led to antibiotic non-producing strains, proving the identity of the cluster . Previous labeling data have indicated that FK520 biosynthesis uses novel polyketide extender units (Byrne, K.M., Shafiee, A., Nielson, J., Arison, B., Monaghan, R.L., Kaplan, L., 1993 . The biosynthesis and enzymology of an immunosuppressant, immunomycin, produced by Streptomyces hygroscopicus var, ascomyceticus . Dev . Ind . Microbiol . 32, 29-45) . Genes in the flanking regions of the FK520 cluster were identified that appear to be involved in synthesis of these extender units . All but two of these genes were homologous to genes with known function . In addition to a crotonyl-CoA reductase gene (fkbS), at least two other genes are proposed to be involved in biosynthesis of the atypical PKS extender unit ethylmalonyl-CoA, which accounts for the ethyl side chain on C-21 of FK520 . A set of five contiguous genes (fkbGHIJK) is proposed to be involved in biosynthesis of an unusual PKS extender unit bearing an oxygen on the alpha-carbon, and leading to the 13- and 15-methoxy side chains . These putative precursor synthesis genes in the flanking regions of the FK520 cluster are not found in the flanking regions of the rapamycin cluster (Molnar, I., Aparicio, J.F., Haydock, S.F., Khaw, L.E., Schwecke, T., Konig, A., Staunton, J., Leadlay, P.F., 1996 . Organisation of the biosynthetic gene cluster for rapamycin in Streptomyces hygroscopicus: analysis of genes flanking the polyketide synthase . Gene 169, 1-7), consistent with labeling data showing that rapamycin biosynthesis uses only malonyl and methylmalonyl extender units.

J Forensic Sci, 2000 May, 45(3), 624 - 32
Beta-hydroxybutyric acid--an indicator for an alcoholic ketoacidosis as cause of death in deceased alcohol abusers; Iten PX et al.; We analyzed the postmortem blood of a total of 100 fatal cases for beta-hydroxybutyric acid (BHBA) . In 25 cases of sudden and unexpected death of alcoholics we found pathologically increased levels of BHBA of 1260 to 47200 (median 8000) micromol/L . This led us to the diagnosis of an alcoholic ketoacidosis (AKA) as cause of death in these cases . The control group of 69 postmortem cases revealed that BHBA concentrations below 500 can be regarded as normal, and values up to 2500 micromol/L as elevated . Our study shows that BHBA values over 2500 micromol/L could lead to death, if no medical attention is sought . During storage we did not find any indication of postmortem formation or decomposition of BHBA in blood in vitro or in the corpses . In our opinion, BHBA should be considered the diagnostic marker of choice for the postmortem determination of alcoholic ketoacidosis (AKA) as the cause of death . The classical indications of such deaths are: unexpected death of a chronic alcoholic; none or only traces of ethanol in the blood; increased acetone blood concentration; and neither autopsy, histology, microbiology, nor toxicology reveal the cause of death . In six further cases a diabetic ketoacidosis (DKA) was diagnosed as the cause of death.

J Food Prot, 2000 Jun, 63(6), 827 - 31
Epidemiology, microbiology, and risk assessment of waterborne pathogens including Cryptosporidium; Haas CN; Cryptosporidium is one of a suite of relatively recently emerging pathogens of concern in drinking water . Based on human dose-response tests, guidelines for exposure yielding defined levels of endemic risk have been developed . This risk assessment procedure is grounded in the process used for chemical risk assessment . From outbreak data, critical concentrations in water that may lead to epidemic levels have been postulated . Development of these levels will be discussed . Validation of the information using outbreak reports from the 1993 Milwaukee incident can be made . Use of this approach must be tempered by the existence of substantial waterborne cases in the absence of detectable oocyst levels as in the Las Vegas outbreak, and (apparent) high levels of oocysts without (apparent) significant health effects as in the case of the (at the time of this writing) ongoing incident in Sydney, Australia.

Zh Mikrobiol Epidemiol Immunobiol, 1999 Sep-Oct, (5), 22 - 6
{The current status and outlook for molecular genetic methods in solving the tasks of medical microbiology}; Gintsburg AL et al.; The article deals with modern methods, viz . PCR, molecular display and genotherapy, which permit the new approach to the solution of problems connected with the identification of infective agents, the study of the mechanisms of the pathogenesis of infectious diseases and their treatment . In this article concrete examples, clearly demonstrating how each of the above-mentioned technologies makes it possible to broaden the circle of problems solved in infectious pathology of man, are presented.

Ir J Med Sci, 2000 Jan-Mar, 169(1), 60 - 2
The effects of blood on rapid urease testing for Helicobacter pylori in mucosal biopsies from the gastric antrum; Lee JM et al.; BACKGROUND: While the eradication of Helicobacter pylori in patients with bleeding peptic ulcer disease (PUD) decreases the rate of ulcer re-bleeding, the sensitivity of the rapid urease test (RUT) for H . pylori diagnosis is lower in this setting . The aim of this study was therefore to determine if exposing a gastric biopsy specimen to blood before its use in the RUT (CLOtest) could account for these findings . METHODS: In patients undergoing endoscopy for the evaluation of dyspepsia gastric mucosal biopsies were obtained for H . pylori diagnosis (RUT, microbiology, and histology) . Mucosal biopsies from each patient were also exposed to blood for 15, 30 and 45 minutes before use in the RUT . RESULTS: Using a combination of diagnostic tests (histology, microbiology and routine CLOtest) as the 'gold standard', the sensitivity, specificity, positive predictive value and negative predictive value of the CLOtest remained above 90% despite prior exposure of the gastric biopsy specimen to blood, and these values were not significantly different from the performance characteristics of the CLOtest processed in a routine manner . CONCLUSION: The exposure of gastric mucosal biopsy specimens to blood alone is not the explanation for the reduced sensitivity of the RUT in patients with bleeding peptic ulcers.

Clin Chim Acta, 2000 Jul, 297(1-2), 305 - 11
Towards European urinalysis guidelines . Introduction of a project under European Confederation of Laboratory Medicine; Kouri TT et al.; Improved standardized performance is needed because urinalysis continues to be one of the most frequently requested laboratory tests . Since 1997, the European Confederation of Laboratory Medicine (ECLM) has been supporting an interdisciplinary project aiming to produce European urinalysis guidelines . More than seventy clinical chemists, microbiologists and ward-based clinicians, as well as representatives of manufacturers are taking part . These guidelines aim to improve the quality and consistency of chemical urinalysis, particle counting and bacterial culture by suggesting optimal investigative processes that could be applied in Europe . The approach is based on medical needs for urinalysis . The importance of the pre-analytical stage for total quality is stressed by detailed illustrative advice for specimen collection . Attention is also given to emerging automated technology . For cost containment reasons, both optimum (ideal) procedures and minimum analytical approaches are suggested . Since urinalysis mostly lacks genuine reference methods (primary reference measurement procedures; Level 4), a novel classification of the methods is proposed: comparison measurement procedures (Level 3), quantitative routine procedures (Level 2), and ordinal scale examinations (Level 1) . Stepwise strategies are suggested to save costs, applying different rules for general and specific patient populations . New analytical quality specifications have been created . After a consultation period, the final written text will be published in full as a separate document.

J Infect, 2000 Mar, 40(2), 184 - 204
Efficacy of long-term antibiotic suppressive therapy in proven or suspected infected abdominal aortic grafts; Roy D et al.; We have reviewed our experience of long-term antibiotic suppressive therapy in patients who underwent repair of an abdominal aortic aneurysm (AAA) and developed proven or strongly suspected infection of a graft . Five patients with abdominal aortic repair complicated by proven or suspected graft infections were treated with continuing antibiotic suppressive therapy based on microbiology culture reports . Two patients developed infection of an established graft, two patients had a graft inserted into an infected area and one patient was thought to be at high risk of developing infection of a recently placed graft . All patients had severe co-existent medical problems and were considered too ill to tolerate further definitive surgery . Response to therapy was monitored by absence of symptoms, fever, inflammatory markers and survival . All patients are alive with a median survival of 32 months, the longest having survived for 6 years . In selected patients with abdominal aortic graft infections, indefinite antibiotic suppressive therapy may be an acceptable alternative to further surgery.

J Infect Dis, 2000 Jun, 181 Suppl 3, S452 - 4
Polymerase chain reaction detection of Chlamydia pneumoniae in circulating white blood cells; Boman J et al.; Several recently published studies suggest that Chlamydia pneumoniae may represent a risk factor for atherosclerosis or its complications . In order to establish whether C . pneumoniae is causally linked to atherosclerotic diseases, clinical intervention trials may be needed . However, to identify eligible subjects with a persistent C . pneumoniae infection and to monitor the effect of antibiotic therapy, there is a need for a reliable diagnostic marker . Blood-based polymerase chain reaction assays may be of value for identifying patients persistently infected with C . pneumoniae and for assessing the microbiologic efficacy of antichlamydial therapy in clinical intervention trials.

Epidemiol Mikrobiol Imunol, 2000 Apr, 49(2), 83 - 6
{History of the J.E . Purkynĕ Czech Medical Society}; Tichacek B; The author presents some basic data from the time of the foundation of the Czech Physicians Society (1862), the predecessor of the present Czech J . E . Purkyne Medical Association . Its beginnings date back to the year 1949 . The author mentions also data associated with the foundation of the present Society of Epidemiology and Microbiology of the Czech J . E . Purkyne Medical Association.

Acta Haematol, 2000, 103(2), 90 - 5
Evidence of lymphocyte alveolitis by bronchoalveolar lavage in thalassemic patients with pulmonary dysfunction; Filosa A et al.; Pulmonary dysfunction represents one of the least studied complications in thalassemic patients . Probably, it is due to the absence of pulmonary symptoms . There are few works in the literature, and contradictory results have been published . The aim of this study was to define the spirometric pattern and the possible causes of lung impairment by testing bronchoalveolar lavage (BAL) with pathological pulmonary function tests (PFTs) . Furthermore, diffusion capacity tests for carbon monoxide corrected for hemoglobin value (Dco*) were performed . We studied 48 thalassemic patients (27 F and 21 M), with an age range from 8 to 23 years, divided into two groups on the basis of PFTs results . Thus, group A was formed by 16 patients with restrictive spirometric patterns of whom 14 had also reduced Dco* values and group B consisted of 32 patients with normal PFTs and Dco* values . Patients of group A underwent chest high-resolution computing tomography (CHRCT) and BAL whose fluid was analyzed by microbiologic and cytological assays . A pathological CHRCT picture was present in 8 patients . Nine out of 16 patients who accepted to undergo BAL had a chronological age greater than 17 years with a mean bone age of 13.9 years . BAL results showed lymphocyte alveolitis in 6 patients and a normal cytogram in 3, while alveolar iron-laden macrophages were present in 4 out of 6 patients with alveolitis and 2 out of 3 patients with normal cytogram . Moreover, all examined BAL fluids showed a normal CD4/CD8 ratio, while only 2 patients showed an altered serum CD4/CD8 ratio . We demonstrated the presence of (1) lung-restrictive syndrome in 16 of the oldest thalassemic patients; (2) lymphocyte alveolitis in 6 patients, and (3) a picture of interstitial fibrosis by CHRCT in 8 of them . All these data are suggestive of a diagnosis of interstitial lung disease secondary to thalassemia . BAL helped to identify the presence of alveolar iron-laden macrophages that represented a local defense mechanism against free iron . This latter finding therefore might be the primary cause of the lung impairment promoting an oxidative damage . Further studies are needed to investigate this hypothesis and therapeutical potentials .

Prim . Care Update Ob Gyns . 1998 Jul 1;5(4):150.
Safety and efficacy comparison of two dosing regimens of MetroGel-Vaginal(R) in the treatment of bacterial vaginosis: a multicenter, randomized, single-blind, parallel comparison; Soper DE et al.; Objective: To compare the safety and efficacy of 0.75% metronidazole vaginal gel (MGV) administered QD for 5 days to MGV administered BID for 5 days for the treatment of bacterial vaginosis (BV).Methods: Nonpregnant women 18 years or older with a diagnosis of BV were enrolled into this trial . Patients were determined to have BV if their vaginal discharge had >/=20% clue cells and if they had at least 2 of the following criteria: homogeneous discharge, positive KOH amine odor test, or vaginal pH >/=4.7 . In addition, microbiologic confirmation of BV at baseline was required to be evaluable . Patients were evaluated at baseline and at 7-12 days after completion of therapy . If patients remained free of BV after therapy, they were evaluated at a follow-up visit 28-35 days after therapy.Results: A total of 514 patients were enrolled in the trial; 252 in the QD treatment group and 262 in the BID treatment group . Clinical cure rates based on patients considered evaluable at first return visits were 77% and 80% for QD and BID dosing regimens, respectively . Clinical cure rates based on patients considered evaluable at the final visit were 58% and 61% for QD and BID dosing regimens, respectively . Clinical cure rates at the final return visit based on intent-to-treat analysis were 53% and 57% for the QD and BID regimens, respectively . Two adverse events in two patients were reported as serious . Both adverse events resolved and neither event was judged to be related to therapy.Conclusion: MetroGel-Vaginal administered QD for 5 days was found to be as effective as MetroGel-Vaginal administered BID for 5 days in the treatment of bacterial vaginosis 28-35 days after therapy . The number of patients reporting adverse events judged to be related to therapy was also statistically comparable in both treatment groups.

Prim . Care Update Ob Gyns . 1998 Jul 1;5(4):150.
The association of asymptomatic bacterial vaginosis with upper genital tract infection; Bennett BB et al.; Objective: To determine whether asymptomatic bacterial vaginosis (BV) is associated with upper genital tract inflammation or bacterial colonization.Methods: Fifty nonpregnant women with intact uteri who planned to undergo gynecologic surgery and had no recent symptoms of vaginal infection were enrolled . We obtained a vaginal swab for Gram stain, endocervical swab for chlamydia and gonorrhea DNA probe testing, and Pipelle endometrial biopsy for aerobic and anaerobic cultures and histology . We correlated surgical findings and histology of available surgical specimens with the microbiologic results . The diagnosis of BV was made according to Speigel's criteria . Bacteria isolated from the uterus were classified as high virulence versus low virulence . Contingency tables were analyzed using the chi-square or Fisher Exact tests.Results: Twenty-one of 50 patients had BV on Gram stain, 3 had intermediate BV, and 3 had unreadable slides . Eleven patients had histologic endometritis, including 7 of the 21 with BV . The association between BV and endometritis was not statistically significant . Thirty-eight patients had a positive endometrial culture; 18 of these also had BV and 3 had intermediate BV . Of patients with a positive endometrial culture, isolates from those who also had BV or intermediate BV were more likely to be highly virulent pathogens compared to those who tested negative for BV (P <.05) . No patients had positive gonorrhea or chlamydia tests . Of the 37 patients who underwent abdominal surgery, 20 had visible adhesions . The association between adhesions and endometritis, BV, or positive endometrial cultures was not significant.Conclusion: Patients with asymptomatic BV are more likely than patients without BV to have highly virulent bacteria isolated from the endometrium . BV may predispose to upper genital tract infection.

Arch Pathol Lab Med, 2000 Jun, 124(6), 883 - 7
Identification of the asexual state of Rhizopus species on histologic tissue sections in a patient with rhinocerebral mucormycosis; Ciesla MC et al.; Mucormycosis is an infection caused by a group of fungi in the order Mucorales in the phylum Zygomycota . The most well-known form of this disease is rhinocerebral mucormycosis, which usually develops in diabetic or immunocompromised patients . The fungal hyphal elements are easily detected in biopsy specimens by direct or histologic examination . However, the confirmatory identification of the genus or species requires culture of the specimen . This article presents a case of rhinocerebral mucormycosis in which presumptive identification of the genus was made without microbiologic cultures and was based on the extraordinarily rare appearance of fungal sporangia and sporangiospores in histologic tissue sections . Identification of these structures allowed an early and accurate diagnosis of rhinocerebral invasive mucormycosis.

J Clin Microbiol, 2000 Jun, 38(6), 2065 - 75
Monoclonal antibodies directed against conserved epitopes on the nucleocapsid protein and the major envelope glycoprotein of equine arteritis virus; Weiland E et al.; We recently developed a highly effective immunization procedure for the generation of monoclonal antibodies (MAbs) directed against the porcine reproductive and respiratory syndrome virus (E . Weiland, M . Wieczorek-Krohmer, D . Kohl, K . K . Conzelmann, and F . Weiland, Vet . Microbiol . 66:171-186, 1999) . The same method was used to produce a panel of 16 MAbs specific for the equine arteritis virus (EAV) . Ten MAbs were directed against the EAV nucleocapsid (N) protein, and five MAbs recognized the major viral envelope glycoprotein (G(L)) . Two of the EAV G(L)-specific MAbs and one antibody of unknown specificity neutralized virus infectivity . A comparison of the reactivities of the MAbs with 1 U.S . and 22 newly obtained European field isolates of EAV demonstrated that all N-specific MAbs, the three nonneutralizing anti-G(L) MAbs, and the weakest neutralizing MAb (MAb E7/d15-c9) recognized conserved epitopes . In contrast, the two MAbs with the highest neutralization titers bound to 17 of 23 (MAb E6/A3) and 10 of 23 (MAb E7/d15-c1) of the field isolates . Ten of the virus isolates reacted with only one of these two MAbs, indicating that they recognized different epitopes . The G(L)-specific MAbs and the strongly neutralizing MAb of unknown specificity (MAb E6/A3) were used for the selection of neutralization-resistant (NR) virus variants . The observation that the E6/A3-specific NR virus variants were neutralized by MAb E7/d15-c1 and that MAb E6/A3 blocked the infectivity of the E7/d15-c1-specific NR escape mutant confirmed that these antibodies reacted with distinct antigenic sites . Immunoelectron microscopy revealed for the first time that the antigenic determinants recognized by the anti-G(L) MAbs were localized on the virion surface . Surprisingly, although the immunofluorescence signal obtained with the neutralizing antibodies was relatively weak, they mediated binding of about three times as much gold granules to the viral envelope than the nonneutralizing anti-G(L) MAbs.

J Theor Biol, 2000 Jun 21, 204(4), 533 - 41
Length distribution of the peptidoglycan chains in the sacculus of Escherichia coli; Koch AL; The stress-bearing fabric of bacteria is made of peptidoglycan . This crosslinked fabric is formed from disaccharide pentapeptide units that are transported through the cytoplasmic membrane and then polymerized in two directions: (i) to form oligoglycan chains; and (ii) to cross link these chains by tail-to-tail bonds from the muropeptides to the protruding peptides of other chains . The distribution of the glycan chain lengths is reminiscent of the "most probable distribution of polymer chemistry . Of course, the process is more complex than solely the random addition of units to growing chains . The complexity precludes mathematical analysis, but computer modeling of the Monte Carlo type is capable of including a range of possibilities . At each time point a specified number of disaccharides are singly added to the muramic acid residue ends of existing chains chosen at random . The transfer is in exchange for the cleavage of pyrophosphate bactoprenol that transported the disaccharide pentapeptide through the membrane . The progam then selects, again at random, which chain to cleave and between which two disaccharides of the chain the cleavage event is to occur . The cleavage generates an N -acetyl 1,6 anhydro-muramic acid end and a non-reducing N -acetyl glucosamine end . The simulation can be modified so that the program does not cleave off a disaccharide next to either end of the chain . Comparisons are shown with the experimental results of Obermann & H}oltje (1994 . Microbiology140, 79-87.) They obtained their data by taking the results with normal growing cells and subtracting the similar data from minicells to estimate the chain length distribution in the cylinder part of the cell . In its most basic form the computer simulation has only one fitted parameter, K, which is the number of disaccharides added to the murein for every internal cleavage event . In this form the fitting to the experimental results is poor . One possible reason for this is that the tension on the chains, and therefore the probability of being cleaved by autolysins varies with orientation of the chain on the cylinder surface . It is well known that the tension in the cylindrical wall is twice as large in the circumferential direction as in the axial one, so one class would consist of those chains aligned longitudinally, subject to lower stress, and would have a higher energy of activation for autolysis than chains aligned circumferentially . A good fit is obtained on the assumption that there are only two classes of chains; one more likely to be cleaved than the other . The key point is that only two processes: adding of disaccharide pentapeptides at random to glycan chains and cleavage between the disaccharides at random, together with the assumption that the wall is less easily hydrolysed in the axial direction is sufficient to account for the experimental distribution .

Am Heart J, 2000 Jun, 139(6), 945 - 51
Clinical information determines the impact of transesophageal echocardiography on the diagnosis of infective endocarditis by the duke criteria; Roe MT et al.; BACKGROUND: Although transesophageal echocardiography (TEE) is more sensitive than transthoracic echocardiography (TTE) in detecting echocardiographic evidence of infective endocarditis (IE), the impact of TEE on the clinical diagnosis of IE has not been clearly delineated . METHODS AND RESULTS: We studied 112 patients with 114 suspected episodes of IE over a 6-year period who underwent both TTE and TEE during their diagnostic evaluation . Using the results of these studies along with clinical and microbiologic data, we attempted to determine the incremental value of TEE to the Duke Endocarditis Diagnostic Criteria . Patients were initially classified into a diagnostic category of the Duke criteria with TTE data, and then the diagnostic classification was reconsidered with TEE data . A diagnostic category reassignment occurred in 25 of 114 episodes of IE evaluated when TEE results were incorporated into the evaluation with the Duke criteria (22 patients were reclassified from possible IE to definite IE whereas 3 patients were reclassified from rejected to possible IE) . Diagnostic reclassification occurred in 9 (11%) of the 80 episodes of suspected IE with native cardiac valves and 13 (34%) of 34 episodes with prosthetic cardiac valves . Most patients reclassified from possible IE to definite IE with TEE data (19 of 22) had an intermediate clinical likelihood of IE, whereas 92% of patients had negative TTE results . Pathologic examination of valvular tissue in 22 of the 114 episodes of suspected IE revealed that the positive predictive value of the Duke criteria with TEE data for diagnosis of IE was 85% in patients with native valves and 89% in patients with prosthetic valves . CONCLUSIONS: When clinical evidence of IE is present, TEE improves the sensitivity of the Duke criteria to diagnose definite IE . TEE data appears to be especially useful for the diagnostic evaluation of patients with suspected IE who have prosthetic valves.

Dis Colon Rectum, 2000 May, 43(5), 701 - 6; discussion 706-7
Simple and effective surgical treatment of pilonidal sinus: asymmetric excision and primary closure using suction drain and subcuticular skin closure; Akinci OF et al.; PURPOSE: This study was planned to evaluate prospectively the results of 112 pilonidal sinus cases treated surgically by using asymmetric excision and primary closure with suction drain and subcuticular skin closure . It is aimed at elimination of the causative factors of pilonidal sinus . METHOD: The patient's age, profession, weight and height, symptoms and signs, duration of symptoms, previous treatments, operation time and cost, hospital stay, return to normal activity, complications, pathologic and microbiologic examinations, and recurrences were noted . All pilonidal sinus cases except pilonidal abscess and extensive gluteal involvement were treated surgically . The procedure consists of an eccentric, elliptical excision of the affected tissue, mobilization of the flap to the sacrococcygeal fascia and the suturing of its edge to the lateral one . Penrose drains were placed in the first eight (7.14 percent) cases, but suction drains were placed in others . The cases were followed up for a mean of 2.4 years . RESULTS: Twenty-eight (25 percent) cases had undergone previous operative procedures . Of 112 patients 106 (94.6 percent) were male . Mean age was 22.1 years . Mean history of disease was 4.2 years . The overall complication rate was 7.14 percent . Two (1.8 percent) wound infections, two wound breakdowns, three (2.7 percent) collections, and one (0.9 percent) recurrence were recorded . The collections were reduced to zero after first eight cases by using a suction drain . Sixty-eight of the patients (60.7 percent) had body weight over 90 kg, and the mean body mass index was 24.8 . The mean hospital stay was 2.6 days, and the mean time off work was 12.4 days . The average healing time was 13.2 days . There were no anesthetic or surgical deaths . CONCLUSION: The natal cleft is flattened and the incision scar and the incision line is transferred from the midline to the lateral side by performing the asymmetric excision and primary closure, and thus the essential cause of pilonidal sinus is eliminated . The procedure is simple, the complications and recurrences are very low, and it is seen to be an excellent procedure in the surgical treatment of uncomplicated pilonidal sinus disease.

Med Confl Surviv, 2000 Jan-Mar, 16(1), 42 - 59
Preventing deliberate disease; Pearson GS; The danger from deliberate disease as a weapon of war--biological weapons--is examined and it is concluded that biological weapons currently pose the greatest danger of all weapons of mass destruction . The ongoing work of the Ad Hoc Group negotiating a Protocol to strengthen the effectiveness and improve the implementation of the Biological and Toxins Weapons Convention is analysed . The Protocol is nearing completion and contains provisions for declarations of the most relevant facilities, for declaration follow up procedures including infrequent randomly-selected visits to declared sites, for investigations of non-compliance concerns as well as measures to promote international co-operation for peaceful purposes in microbiology and biotechnology . It is concluded that an efficient and effective Protocol is achievable within the coming year.

Ginecol Obstet Mex, 2000 Apr, 68, 154 - 9
{Effectiveness and safety of ciclopirox olamine 1% vaginal cream versus terconazole 0.8% vaginal cream in the treatment of genital candidiasis}; Garcia Figueroa RG et al.; A multicenter randomized study was performed . One hundred and seventy patients were selected . The patients were 18 years and older . They presented signs and symptoms of genital candidiasis and had positive smear culture for Candida . Eighty five patients were assigned to receive Cicloprox olamine 1%, and eighty five patients were assigned to receive Terconazol 0.8% . The treatment lasted six days with the objective to compare the clinical and antifungal efficiency and safety with both treatments . The result of mixed efficiency (clinical and microbiology) for Cicloprox olamine was 48 cases (62.3%) at the end of the treatment were cured--day 7-; and at the continuation--day 21-42 of them had the same result (55.3%); improvement was seen at the end in 25 cases (32.5%) and at the continuation 21 cases had the same result (27.6%) . For Terconazol the result was 45 cases (61.6%) were cured at the end of the treatment, and at the continuation 39 had the same result (57.4%); improvement was seen at the end in 23 cases (31.5%) and at the continuation 22 cases had the same result (32.4%) . We conclude that both treatments are effective and well tolerated for genital candidiasis treatment.

J Infect Dis, 2000 May, 181(5), 1643 - 51 Epub 2000 May 15.
Persistent diarrhea signals a critical period of increased diarrhea burdens and nutritional shortfalls: a prospective cohort study among children in northeastern Brazil; Lima AA et al.; Persistent diarrhea (PD; duration >/=14 days) is a growing part of the global burden of diarrheal diseases . A 45-month prospective cohort study (with illness, nutritional, and microbiologic surveillance) was conducted in a shantytown in northeastern Brazil, to elucidate the epidemiology, nutritional impact, and causes of PD in early childhood (0-3 years of age) . A nested case-control design was used to examine children's diarrhea burden and nutritional status before and after a first PD illness . PD illnesses accounted for 8% of episodes and 34% of days of diarrhea . First PD illnesses were preceded by a doubling of acute diarrhea burdens, were followed by further 2.6-3.5-fold increased diarrhea burdens for 18 months, and were associated with acute weight shortfalls . Exclusively breast-fed children had 8-fold lower diarrhea rates than did weaned children . PD-associated etiologic agents included Cryptosporidium, Giardia, enteric adenoviruses, and enterotoxigenic Escherichia coli . PD signals growth shortfalls and increased diarrhea burdens; children with PD merit extended support, and the illness warrants further study to elucidate its prevention, treatment, and impact.

J R Coll Surg Edinb, 2000 Apr, 45(2), 135 - 7
Mycobacterium tuberculosis presenting as sternal osteomyelitis; Stewart KJ et al.; A 54-year-old female presented with a presternal abscess and developed axillary lymphadenopathy . Imaging confirmed the presence of sternal osteomyelitis . The osteomyelitis was cured by resection and muscle flap reconstruction . Although tuberculosis was suspected, the organism was only cultured after the fourth surgical procedure . Surgeons should be aware that negative microbiology does not exclude a diagnosis of Mycobacterium tuberculosis.

J Infect Dis, 2000 May, 181(5), 1713 - 9 Epub 2000 May 15.
Early detection of aspergillus infection after allogeneic stem cell transplantation by polymerase chain reaction screening; Hebart H et al.; Invasive aspergillosis (IA) has become a major cause of mortality in patients after allogeneic stem cell transplantation . To assess the potential of prospective polymerase chain reaction (PCR) screening for early diagnosis of IA, 84 recipients of an allogeneic stem cell transplant were analyzed with the investigators blinded to clinical and microbiologic data . Of 1193 blood samples analyzed, 169 (14.2%) were positive by PCR . In patients with newly diagnosed IA (n=7), PCR positivity preceded the first clinical signs by a median of 2 days (range, 1-23 days) and preceded clinical diagnosis of IA by a median of 9 days (range, 2-34 days) . Pretransplantation IA (relative risk {RR}, 2.37), acute graft-versus-host disease (RR, 2.75), and corticosteroid treatment (RR, 6.5) were associated with PCR positivity . The PCR assay revealed a sensitivity of 100% (95% confidence interval {CI}, 48%-100%) and a specificity of 65% (95% CI, 53%-75%) . None of the PCR-negative patients developed IA during the study period . Thus, prospective PCR screening allows for identification of patients at high risk for subsequent onset of IA.

Nature, 2000 May 11, 405(6783), 175 - 8
Detection and classification of atmospheric methane oxidizing bacteria in soil; Bull ID et al.; Well-drained non-agricultural soils mediate the oxidation of methane directly from the atmosphere, contributing 5 to 10% towards the global methane sink . Studies of methane oxidation kinetics in soil infer the activity of two methanotrophic populations: one that is only active at high methane concentrations (low affinity) and another that tolerates atmospheric levels of methane (high affinity) . The activity of the latter has not been demonstrated by cultured laboratory strains of methanotrophs, leaving the microbiology of methane oxidation at atmospheric concentrations unclear . Here we describe a new pulse-chase experiment using long-term enrichment with 12CH4 followed by short-term exposure to 13CH4 to isotopically label methanotrophs in a soil from a temperate forest . Analysis of labelled phospholipid fatty acids (PLFAs) provided unambiguous evidence of methane assimilation at true atmospheric concentrations (1.8-3.6 p.p.m.v.) . High proportions of 13C-labelled C18 fatty acids and the co-occurrence of a labelled, branched C17 fatty acid indicated that a new methanotroph, similar at the PLFA level to known type II methanotrophs, was the predominant soil micro-organism responsible for atmospheric methane oxidation.

Int J Periodontics Restorative Dent, 1999 Dec, 19(6), 555 - 67
The short-term effect of apically repositioned flap surgery on the composition of the subgingival microbiota; Levy RM et al.; The purpose of this investigation was to examine the short-term effect of apically repositioned flap surgery on clinical and microbiologic parameters in patients with adult periodontitis . A total of 11 patients with moderate to advanced periodontitis received apically repositioned flap surgery . Subjects were monitored during a 3-month pretreatment phase, the baseline surgical phase, and for 3 months post-surgery . Clinical assessments including plaque accumulation, gingival redness, suppuration, bleeding on probing, pocket depth, and attachment level were made at 6 sites per tooth . Subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 29 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization . The mean levels and percentage of sites colonized by each species (prevalence) were computed for each subject at each visit . After surgery, there was a significant decrease in mean pocket depth and percentage of sites exhibiting gingival redness . Significant decreases were seen in the percentage of sites that had attachment levels < 4 mm, with a significant increase in the percentage of sites with attachment levels of 4 to 6 mm after therapy . The mean total DNA probe count for all bacterial species was significantly decreased by both scaling and root planing and surgical therapy . P gingivalis and B forsythus, 2 bacteria previously shown to be susceptible to mechanical therapy, exhibited statistically significant decreases in mean total DNA probe count . Because surgical therapy decreased levels of the suspected periodontal pathogens C rectus, P nigrescens, and C gracilis, it may be speculated that there was a potential added beneficial effect of surgery on the periodontal microbiota.

Am J Gastroenterol, 2000 May, 95(5), 1166 - 70
Rapid urease tests lack sensitivity in Helicobacter pylori diagnosis when peptic ulcer disease presents with bleeding; Lee JM et al.; OBJECTIVE: The eradication of Helicobacter pylori (H . pylori) in patients with bleeding peptic ulcer disease (PUD) decreases the rate of ulcer rebleeding . Although all methods for H . pylori diagnosis have been extensively evaluated in uncomplicated PUD the efficacy of the commonly used rapid urease test (RUT) has not been established in patients with bleeding peptic ulcer disease . The aim of this study was to evaluate the efficacy of the RUT (CLOtest) in patients with bleeding duodenal ulcers (DUs) . METHODS: Consecutive patients with symptoms of upper GI tract hemorrhage and a DU at the time of endoscopy were evaluated . The presence of H . pylori infection was determined by RUT, microbiology, and histology . Consecutive patients with uncomplicated DUs were similarly evaluated . The prevalence of H . pylori as determined by the RUT alone was compared to that determined by a combination of all tests in both patient groups . RESULTS: Fifty-five patients with bleeding DUs and 69 with nonbleeding DUs were evaluated . The prevalence of H . pylori in patients presenting with bleeding was 72.7% (95% confidence interval {CI} 61.0-84.5%) and lower than the prevalence rate of 92.8% (95% CI 86.6-98.8%) in patients with uncomplicated PUD (p < 0.05) . The prevalence of H . pylori in the bleeding DU group as determined by RUT alone (54.5%) was less than that determined by a combination of all tests (73%) with a false-negative rate of 10 of 40 (25%; 95% CI 11.6-38.4%) (p < 0.05) . This false-negative rate was significantly greater than that observed in the group presenting with dyspepsia (1 of 64 {1.6%; 95% CI 0-4.6%}) (p < 0.01) . CONCLUSIONS: The prevalence of H . pylori is lower in patients with bleeding DUs when compared to patients with uncomplicated DUs . In addition, the sensitivity and negative predictive value of the RUT is lower in patients presenting with bleeding, and other methods of H . pylori diagnosis should be used in this patient group.

J Nucl Med, 2000 May, 41(5), 896 - 902
Scintigraphic evaluation of experimental chronic osteomyelitis; Dams ET et al.; Assessment of disease activity and disease extent in chronic osteomyelitis remains a difficult diagnostic problem . Radiography is not particularly sensitive . Scintigraphic techniques can be more helpful, but the routinely available agents lack specificity (99mTc-methylene diphosphonate {MDP}, 67Ga-citrate) or are laborious to prepare (111In-leukocytes) . We evaluated the performance of 2 new radiopharmaceuticals, 99mTc-polyethyleneglycol (PEG) liposomes and 99mTc-hydrazinonicotinamide (HYNIC)-immunoglobulin G (IgG), in an experimental model of chronic osteomyelitis . Methods: Chronic osteomyelitis was induced in rabbits by inserting S . aureus into the right reamed and washed femoral canal . The canal was closed with cement . A sham operation was performed on the left femur . Routine radiographs were obtained immediately after surgery and before scintigraphy . Four weeks after surgery, each rabbit was injected with 37 MBq 99mTc-PEG liposomes, 99mTc-HYNIC-IgG, and 99mTc-MDP on 3 consecutive days and imaged up to 4 (MDP) or 22 (liposomes and IgG) h after injection . On day 4, rabbits received either 18 MBq 111In-granulocytes or 67Ga-citrate and were imaged up to 44 h after injection . Uptake in the infected femur was determined by drawing regions of interest . Ratios of infected-to-sham-operated femur were calculated . After the last image, the rabbits were killed, and the left and right femur were scored for microbiologic and histopathologic evidence of osteomyelitis . RESULTS: 99mTc-PEG liposomes and 99mTc-HYNIC-IgG correctly identified all 6 rabbits with osteomyelitis . 11In-granulocytes and 67Ga-citrate gave equivocal results in 1 infected rabbit . 99mTc-MDP missed 1 case of osteomyelitis . The uptake in the affected region did not differ significantly between the agents, although 99mTc-MDP tended to have higher values (MDP, 4.75 +/- 1.23 percentage injected dose per gram {%ID/g}; 67Ga, 2.05 +/- 0.54 %ID/g; granulocytes, 1.56 +/- 0.83 %ID/g; liposomes, 1.75 +/- 0.76 %ID/g, and IgG, 1.96 +/- 0.27 %ID/g) . The ratios of infected-to-normal femur were also not significantly different for the respective radiopharmaceuticals . Radiography visualized only severe osteomyelitis . CONCLUSION: In this rabbit model, 99mTc-PEG liposomes and 99mTc-HYNIC-IgG performed at least as well as 111In-granulocytes and 67Ga-citrate in the localization of chronic osteomyelitis . The ease of preparation, the better image quality, and the lower radiation dose suggest that 99mTc-PEG liposomes and 99mTc-HYNIC-IgG might be suitable alternatives for 67Ga-citrate and 111In-granulocytes in the scintigraphic evaluation of osteomyelitis.

Forensic Sci Int, 2000 May 15, 110(2), 139 - 44
Significance of various analytical methods with reference to the causes and manners of death in alcoholics; Thomsen JL; It was the aim of the present investigation to apply a broad spectrum of analyses to forensic autopsies of alcoholics in order to estimate the significance of the various analytical methods with reference to the cause and manner of death . The analyses were performed on a consecutive series of 73 medico-legal autopsies in alcoholics . Both extensive histology as well as toxicology and microbiology were used . The microbiology did not contribute substantially to the determination of the cause of death, while histology was decisive in six cases . Toxicology analyses were necessary for determining the cause of death in 37 cases . The results of the investigation may help in the selection of analytical priorities.

Hunan Yi Ke Da Xue Xue Bao, 1998, 23(6), 566 - 8
{Detection of interleukin-8 and nitric oxide in chronic prostatitis}; Hu F et al.; We detected levels of interleukin-8(IL-8) and nitric oxide(NO) in 62 prostatic fluids . The results were that microbiologic exam, IL-8 content, and NO level in control group were all negative; there was not significant difference on the microbiologic exam and NO level between the chronic prostatitis group and non-prostatitis group; there was significant difference between two groups of IL-8 positive ratio . These results suggest that the IL-8 detected in the prostatic fluid of patients with prostatitis may be a diagnostic indicator of prostatic infection.

Infect Dis Obstet Gynecol, 2000, 8(2), 83 - 7
Association of lower genital tract inflammation with objective evidence of endometritis; Peipert JF et al.; The purpose of this report is to evaluate the association between lower genital tract inflammation and objectively diagnosed endometritis . We analyzed the first 157 patients enrolled in the PEACH study, a multicenter randomized clinical trial designed to compare the effectiveness of outpatient and inpatient therapy for PID . Women less than 38 years of age, who presented with a history of pelvic discomfort for 30 days or less and who were found to have pelvic organ tenderness (uterine or adnexal tenderness) on bimanual examination, were initially invited to participate . After recruitment of the first 58 patients (group 1) we added the presence of leukorrhea, mucopurulent cervicitis, or untreated positive test for N . gonorrhoeae or C . trachomatis to the inclusion criteria (group 2, N = 99) . We compared rates of endometritis in the two groups and calculated the sensitivity, specificity, and predicted values of the presence of white blood cells in the vaginal wet preparation . The rate of upper genital tract infection in group 1 was 46.5% (27/58) compared to 49.5% (49/99) in group 2 . Microbiologic evidence of either N . gonorrhoeae or C . trachomatis increased from 22.4% in group 1 to 38.3% in group 2 . The presence of vaginal white blood cells or mucopus has a high sensitivity (88.9%), but a low specificity (19.4%) for the diagnosis of upper genital-tract infection . Assessment of the lower genital tract for evidence of infection or inflammation is a valuable component of the diagnostic evaluation of pelvic inflammatory disease . The presence of either mucopus or vaginal white blood cells is a highly sensitive test for endometritis in patients with pelvic pain and tenderness.

Am J Surg, 2000 Feb, 179(2A Suppl), 36S - 39S; discussion 39S-40S
Pneumonia in the surgical patient: duration of therapy and does the organism matter?
Spain DA.
Pneumonia is a frequent complication in the critically ill surgical or trauma patient . Despite its common occurrence and increased attention recently, many basic issues, such as how long to treat and whether the exact causative organism even matters, remain unanswered . Currently, cessation of treatment is based on clinical response, although some data suggest that both the microbiologic and immunologic response within the lung are also important . A thorough knowledge of the likely causative organisms for both early and late pneumonia should allow safe and efficacious empiric antibiotic selection.

Am J Surg, 2000 Feb, 179(2A Suppl), 31S - 35S
Multifactorial analyses in the diagnosis of pneumonia arising in the surgical intensive care unit; Polk HC et al.; The diagnosis of ventilator-associated pneumonia in the surgical intensive care unit continues to be problematic . The majority of intensive care units use clinical criteria based on chest x-ray; fever; leukocytosis; alterations in the pulse oximeter observations; the need to alter modes and amounts of ventilatory support; and more specific microbiologic studies, such as appropriate sputum, Gram stain, and culture to identify pneumonia . Diagnosing pneumonia based on clinical criteria alone is often difficult and inaccurate, which may lead to inappropriate use and choice of antibiotics . Invasive diagnostic techniques, such as protected specimen brush and bronchoalveolar lavage, provide an important microbiologic diagnosis . However, the cost and inconvenience limit broad usage . Furthermore, those results that return positive are often too late to dictate the need for, or direction of, therapy . Our use of a "pneumonia grid" may help identify patients likely to have a poor outcome . Until a readily available and cost-effective diagnostic study for pneumonia is developed, clinical criteria remain vital in routine practice.

Am J Surg, 2000 Feb, 179(2A Suppl), 26S - 29S; discussion 30S
Diagnosis of acute respiratory distress syndrome and differentiation from ventilator-associated pneumonia; Croce MA; Acute respiratory distress syndrome (ARDS) remains a significant cause of morbidity and mortality in surgical intensive care unit patients . A relatively predictable sequence of pathophysiologic events occur in the lung, which involve inflammatory mediators and neutrophils . The characteristic radiographic findings of ARDS may mimic ventilator-associated pneumonia (VAP), making the diagnosis of VAP difficult . The standard clinical criteria of fever, leukocytosis, purulent sputum, and infiltrate on chest radiograph are not specific for pneumonia in the surgical patient . The use of bronchoscopy with bronchoalveolar lavage and quantitative cultures can differentiate local and systemic inflammatory response to injury or blood loss from invasive bacterial pneumonia . Basing antibiotic therapy solely on the results of quantitative cultures is safe, because quantitative cultures identify VAP in less than half the patients with clinical evidence of pneumonia . Empiric therapy should be based on the microbiology of the intensive care unit rather than the results of the Gram stain.

Br Dent J, 2000 Mar 25, 188(6), 308 - 12
The British Nutrition Foundation Oral Task Force report--issues relevant to dental health professionals; Moynihan P; A recent report on diet and oral health by the British Nutrition Foundation reviews the anatomy, microbiology and pathology of a number of oral diseases including dental caries, tooth-wear, oral cancer, periodontal disease and enamel defects . The role of nutritional factors in the aetiology and prevention of these oral diseases is discussed . The report states that improvements in the levels of caries in the UK are halting and remain unacceptably high in some 'at risk groups'--including the socially deprived . It states that a two-pronged attack, i.e . reduced frequency of consumption of sugary foods and use of fluoride is necessary to address the problem . To address the increased prevalence of oral cancer avoiding tobacco, limiting alcohol and increasing fruit and vegetable consumption are recommended . Recommendations for approaches to improve oral health are made for health authorities, industry, schools and health professionals and it is recommended that the Government make adequate resources available to improve oral health preventive strategies.

J Biol Chem, 2000 May 12, 275(19), 14537 - 40
Characterization of a P-type Na+-ATPase of a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum; Ueno S et al.; A facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum, possesses a P-type Na(+)-stimulated ATPase in the membrane (Koyama, N . (1999) Curr . Microbiol . 39, 27-30) . In this study, we attempted to purify and characterize the enzyme . The ATPase appears to consist of a single polypeptide with an apparent molecular mass of 100 kDa . The enzyme exhibited an optimum pH for activity at around 9 . The enzyme was strongly inhibited by vanadate (50% inhibition observed at 3 microm) and forms an acylphosphate intermediate, suggesting a P-type ATPase . The enzyme, when reconstituted into soybean phospholipid vesicles, exhibited ATP-dependent (22)Na(+) uptake, which was completely inhibited by gramicidin . The reconstituted vesicles exhibited a generation of membrane potential (positive, inside) . The enzyme is likely to be involved in an electrogenic transport of Na(+).

Indian J Pediatr, 1999 Nov-Dec, 66(6), 923 - 8
Diagnosis of cystic fibrosis: Indian perspective; Kabra SK et al.; Cystic fibrosis (CF) is one of the common life limiting inherited diseases in Caucasian population . Recent reports suggest that the diagnosis of cystic fibrosis in Indian children is missed or delayed due to low index of suspicion . The diagnosis of cystic fibrosis is suspected by the typical clinical features and should be confirmed by doing sweat chloride estimation . If sweat test is not available, ancillary tests including blood electrolyte and acid base balance, airway microbiology, tests to identify pancreatic insufficiency and semen analysis for obstructive azoospermia in post pubertal boys should be carried out . Positive results of these tests make the suspicion very strong . A strongly suspected case should be treated as cystic fibrosis, but for giving a diagnosis of CF, sweat test should be done from the nearest centre where it is available . In the presence of typical clinical features with borderline sweat chloride values sweat test should be repeated 2-3 times and the child should be investigated for alternative diagnosis . In the absence of alternative diagnosis with consistently high or borderline sweat chloride values an attempt should be made to get tests for mutations.

Cochrane Database Syst Rev . 2000;(2):CD000026.
Routine versus selective antifungal administration for control of fungal infections in patients with cancer; Gotzsche PC et al.; BACKGROUND: Systemic fungal infection is considered to be an important cause of morbidity and mortality in cancer patients, particularly those with neutropenia . Antifungal drugs are often given prophylactically, or to patients with persistent fever . OBJECTIVES: The objective of this review was to assess the effect of antifungal drugs in cancer patients with neutropenia . SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (October 1997), MEDLINE (to September 1997) and the reference lists of articles . We searched the proceedings of the ICAAC, General Meeting of the ASM (from 1990 to 1995), and the 7th European Congress of Clinical Microbiology and Infectious Diseases (1995) and contacted researchers in the field . SELECTION CRITERIA: Randomised trials of amphotericin B, AmBisome, fluconazole, ketoconazole, miconazole, or itraconazole compared with placebo or no treatment in cancer patients with neutropenia . DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility, methodological quality and abstracted data . MAIN RESULTS: Twenty-five trials involving 2912 patients were included . In 19 trials, prophylactic or later treatment with antifungal drugs had no effect on mortality (odds ratio 0.92, 95% confidence interval 0.75 to 1.14) . Only amphotericin showed a significant benefit (odds ratio 0.58, 95% confidence interval 0.37 to 0.93) based on seven trials, but the studies were small and the difference in number of deaths was only 15 . Overall it would be necessary to treat 59 patients (95% confidence interval 37 to 131) with an antifungal drug to prevent one case of fungal invasion in surviving patients, although only amphotericin and fluconazole showed a clear beneficial effect . Antifungal treatment decreased fungal colonisation and the need for additional antifungal therapy, but there was heterogeneity across the trials . REVIEWER'S CONCLUSIONS: Routine prophylactic or later therapy with antifungal drugs in cancer patients with neutropenia does not appear to have a beneficial effect on mortality and only a modest effect on fungal invasion.

Cochrane Database Syst Rev . 2000;(2):CD000405.
Enteral antibiotics for preventing necrotising enterocolitis in low birthweight or preterm infants; Bury RG et al.; BACKGROUND: Necrotising enterocolitis continues to be a problem, particularly in preterm neonates . There have been reports published suggesting that the use of enteral antibiotics may be effective as prophylaxis . This systematic review was undertaken to clarify the issue . OBJECTIVES: To evaluate the benefits and harms of enteral antibiotic prophylaxis for necrotising enterocolitis in low birth weight and preterm infants . SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal trials, MEDLINE (search terms: necrotizing enterocolitis, antibiotics; Limits: newborn infant), previous reviews with cross references, abstracts, conference and symposia proceedings, expert informants and journal handsearching in the fields of Neonatal Pediatrics and Microbiology . SELECTION CRITERIA: All randomized or quasi-randomized controlled trials where enteral antibiotics were used as prophylaxis against NEC in LBW (<2500g) and/or preterm (<37 weeks gestation) infants . DATA COLLECTION AND ANALYSIS: The standard method of the Cochrane Collaboration and its Neonatal Review Group was used . The methodological quality of each trial was reviewed by the second author who was blinded to the trial authors and institutions . Each author extracted data separately before comparison and resolution of differences . Relative risk (RR), risk difference (RD), and number needed to treat were used in the analysis . MAIN RESULTS: The administration of prophylactic enteral antibiotics resulted in a statistically significant reduction in NEC {RR 0.47 (0.23, 0.98); RD -0.072 (-0.136, -0.008); NNT 13.9 (7.4, 125)} . There was a reduction in NEC-related deaths which was of borderline statistical significance {RR 0.16 (0.02, 1.26); RD -0.097 (-0.183, -0.010); NNT 10.3 (5.46, 100)} . There were no significant differences in all deaths (one trial only) or in NEC-like enteropathies (one trial only) . There was a statistically significant increase in the incidence of colonisation with resistant bacteria {RR 1.73 (1.00, 2 . 97); RD 0.123 (0.008, 0.238); NNT 8.1 (4.2, 125)} . REVIEWER'S CONCLUSIONS: There is insufficient evidence to support the use of enteral antibiotic prophylaxis for NEC in clinical practice . To address this question further, a large trial would be required with a sample size sufficient to examine all the important benefits and harms . Adverse outcomes associated with infection, particularly with resistant bacteria, should be evaluated.

Genetics, 2000 May, 155(1), 7 - 16
Negligible genetic diversity of mycobacterium tuberculosis host immune system protein targets: evidence of limited selective pressure; Musser JM et al.; A common theme in medical microbiology is that the amount of amino acid sequence variation in proteins that are targets of the host immune system greatly exceeds that found in metabolic enzymes or other housekeeping proteins . Twenty-four Mycobacterium tuberculosis genes coding for targets of the host immune system were sequenced in 16 strains representing the breadth of genomic diversity in the species . Of the 24 genes, 19 were invariant and only six polymorphic nucleotide sites were identified in the 5 genes that did have variation . The results document the highly unusual circumstance that prominent M . tuberculosis antigenic proteins have negligible structural variation worldwide . The data are best explained by a combination of three factors: (i) evolutionarily recent global dissemination in humans, (ii) lengthy intracellular quiescence, and (iii) active replication in relatively few fully immunocompetent hosts . The very low level of amino acid diversity in antigenic proteins may be cause for optimism in the difficult fight to control global tuberculosis.

Pediatrics, 2000 May, 105(5), 1041 - 5
Systemic candidiasis in extremely low birth weight infants receiving topical petrolatum ointment for skin care: a case-control study; Campbell JR et al.; BACKGROUND AND OBJECTIVE: An increase in the incidence of systemic candidiasis (SC) followed a change in skin care for extremely low birth weight (ELBW) infants in our neonatal intensive care unit (NICU) . We sought to determine whether the use of topical petrolatum ointment (TPO) for skin care of ELBW infants was associated with risk for SC . STUDY DESIGN: Case-control study . SETTING: A 48-bed NICU in a private hospital in Houston, Texas . PATIENTS: Ten ELBW infants with and 30 without SC admitted to the NICU from December 1, 1997 through July 31, 1998 . METHODS: ELBW infants with SC were identified using hospital microbiology and infectious disease consultation databases . A case was defined as an infant weighing </=1000 g at birth with Candida spp isolated from a normally sterile body site . Three infants without SC were matched to each case by birth weight, admission date, and survival to the age of SC onset for the case . Data were collected by retrospective medical record review . Molecular analysis of Candida isolates was performed by karyotyping and restriction fragment length polymorphism using pulsed-field gel electrophoresis . RESULTS: Case infants had a mean (+/- standard deviation) age of onset of 21.5 +/- 24 days . Infants with SC and controls did not differ in birth weight, gestational age, or duration of therapy with steroids, antibiotics, insulin, or total parenteral nutrition . Although cases were more likely to be born vaginally and had a longer duration endotracheal intubation than controls, these differences were not significant . The odds ratio for skin care with TPO in case infants versus control infants was 11 (95% confidence interval: 1.9-63) . Skin care with TPO was discontinued and the incidence of SC decreased to baseline . Several Candida spp and genetic profiles were identified, suggesting that there was not a common source outbreak . CONCLUSIONS: We conclude that the use of TPO promoted an increase in the incidence of SC in ELBW infants . Additional investigation of potential infectious risks for ELBW infants receiving TPO skin care is warranted.

J Clin Microbiol, 2000 May, 38(5), 2005 - 6
Brucella abortus infection acquired in microbiology laboratories; Fiori PL et al.; We report an outbreak of laboratory-acquired Brucella abortus infection originating in the accidental breakage of a centrifuge tube . A total of 12 laboratory workers were infected (attack rate of 31%), with an incubation time ranging from 6 weeks to 5 months . Antibody titers were evaluated weekly in all personnel exposed, allowing the diagnosis of the infection in most cases before the onset of clinical symptoms, so that specific therapy could be administrated.

JAMA, 2000 Apr 19, 283(15), 2003 - 7
The case against anergy testing as a routine adjunct to tuberculin skin testing; Slovis BS et al.; Although anergy testing is commonly used to help interpret negative tuberculin skin test results, the validity of this approach has not been demonstrated . Specific issues include lack of a standardized protocol for antigen selection, number needed to reliably evaluate inability to respond, and uniform criteria for defining cutaneous reactivity, as well as regional variation in skin test reactivity . Tuberculin skin testing is used to screen for latent infection and to evaluate the need for isoniazid prophylaxis . The presence or absence of reactivity to control antigens does not affect this decision . The results of anergy testing also do not predict the risk for progression to active disease in either HIV-negative or HIV-positive patients . In HIV-negative patients with active tuberculosis, 10% to 20% have negative tuberculin test results, and 5% to 10% have a negative tuberculin result but have a positive reaction to another antigen . A negative tuberculin skin test result does not exclude either latent infection or active disease, even in the presence of a reaction to other antigens . Neither anergy testing nor tuberculin testing obviates the need for microbiologic evaluation when there is suspicion for active tuberculosis infection . Therefore, anergy testing is not useful in screening for asymptomatic tuberculous infection or for diagnosing active tuberculosis.

Phytochemistry, 2000 Apr, 53(7), 747 - 57
Fungicidal activity of natural and synthetic sesquiterpene lactone analogs; Wedge DE et al.; Fungicidal activity of 36 natural and synthetic sesquiterpene lactones with guaianolide, trans, trans-germacranolide, cis, cis-germacranolide, melampolide, and eudesmanolide carbon skeletons was evaluated against the phytopathogenic fungi Colletotrichum acutatum, C . fragariae, C . gloeosporioides, Fusarium oxysporum, Botrytis cinerea, and Phomopsis sp . Dose-response data for the active compounds dehydrozaluzanin C, dehydrocostuslactone, 5alpha-hydroxydehydrocostuslacone, costunolide, and zaluzanin C are presented . A new 96-well microbioassay procedure for fast and easy evaluation of antifungal activity was used to compare these compounds with commercial fungicide standards . Some structure-activity conclusions are also presented.

Plasmid, 2000 May, 43(3), 214 - 22
Identification of a potential membrane-targeting region of the replication initiator protein (TrfA) of broad-host-range plasmid RK2; Kim PD et al.; Plasmid RK2 codes for two species of the replication initiator protein TrfA (33 and 44 kDa) . Both polypeptides are strongly associated with membrane fractions of Escherichia coli host cells (W . Firshein and P . Kim, Mol . Microbiol . 23, 1-10, 1997) . We investigated the role of a 12-amino-acid hydrophobic region (HR) in the membrane association of TrfA . Epitope-tagged polypeptide fragments of TrfA that contained HR were expressed and found to be associated with membrane fractions . Site-directed mutagenesis of trfA revealed that changes of specific amino acids in HR can affect both TrfA association with the membrane and its ability to support replication of an RK2 oriV plasmid in vivo . These results are consistent with the hypothesis that membrane association of TrfA is functionally relevant and that the HR region of TrfA is involved in membrane association and DNA replication in vivo .

J Assoc Physicians India, 1999 Sep, 47(9), 866 - 8
Seroprevalence of IgG antibodies to Helicobacter pylori among gastrointestinal endoscopists, radiologists and paramedical personnel; Kamat AH et al.; BACKGROUND: The route of transmission of Helicobacter pylori is unknown . Since the organism has been isolated from saliva, gastric juice and stool, medical personnel could be at high risk for acquiring the infection during procedures like gastrointestinal endoscopy . AIMS: To study whether endoscopy is a professional hazard for acquisition of H . pylori . METHODS: We studied the prevalence of IgG antibodies to H . pylori in endoscopists (n = 17), radiologists (n = 17) and personnel from paraclinical branches (n = 35); microbiology (n = 21), pathology (n = 7) and forensic medicine (n = 7); among the paraclinical personnel five were at high risk because they worked with cultures of H . pylori . Subjects answered a questionnaire regarding upper gastrointestinal symptoms, and precautions taken at the work place against infection . The serum was tested for IgG antibodies to H . pylori using a microwell ELISA and a rapid card test . RESULTS: H . pylori antibodies were present in five (29.4%) endoscopists, three (17.6%) radiologists and seven (20%) paraclinical personnel; only one of the 5 high risk para medical personnel was positive . There was no correlation between the duration of performing endoscopies and the H . pylori IgG status . CONCLUSION: Endoscopy is not a risk factor for acquiring H . pylori infection.

Nippon Ishinkin Gakkai Zasshi, 2000, 41(2), 109 - 14
Are giant cells conidia in Sporothrix schenckii? -Freeze-fracture electron microscopic observation-; Maeda M et al.; Sporothrix schenckii is a well-known pathogenic dimorphic fungus . In this study, we focused on the plasma membrane ultrastructures of giant cells of S . schenckii seen mainly on Sabouraud's dextroseagar slant medium . In the organisms grown for 1, 2, 3, 4, 6 and 8 weeks at 27 and 37 degrees C on brain heart infusion and Sabouraud's dextrose agar slant media, the number of conidia, hyphae, brownish and non-brownish giant cells were counted in ten separated areas under light microscope (x100) to determine the culture conditions under which giant cells were generated . The results showed that brownish giant cells were predominantly seen after longer cultivation periods . Using freeze-fracture electron microscopy, larger oval- or round shaped cells can be identified as conidia by their plasma membrane ultrastructure characteristics, i.e, trench-like invaginations seen in ordinary mature conidia (Maeda M et al.; Can J Microbiol 33: 40, 1987) . From these structural features seen by freeze-fracture electron microscopy, giant cells appeared possibly be conidia and were suggested to be starved because of their predominant existence under longer cultivation conditions.

Med Microbiol Immunol (Berl), 1999 Dec, 188(3), 139 - 44
An improved recombinant IgG immunoblot for serodiagnosis of Lyme borreliosis; Wilske B et al.; We have previously described the use of the following recombinant antigens for serodiagnostic immunoblots: p83/100, p39, OspC and p41 (flagellin) internal fragment {Wilske et al . (1993) Med Microbiol Immunol 182:255-270; Rossler et al . (1997) J Clin Microbiol 35:2752-2758} . In our currently used immunoblot p83/100 is derived from strain PKo (Borrelia afzelii), p39 (BmpA) and OspC from strains PKa2 (B . hurgdorferi sensu stricto), PKo and PBi (B . garinii), respectively; the p41 (flagellin) internal fragments were cloned from strains PKo and PBi . In this study we describe the use of two additional recombinantly expressed highly immunogenic proteins Osp 7 (derived from PKo) and p58 (derived from PBi) . A clinically well-defined panel of sera from 147 Lyme borreliosis patients and 139 controls previously tested by a standardized whole cell lysate immunoblot {Hauser et al . (1997) J Clin Microbiol 35:1433-1444} was investigated in the recombinant immunoblot without (old recombinant immunoblot) and with Ospl7 and p58 (new recombinant immunoblot) for IgG antibodies . The sensitivity of the recombinant IgG immunoblot for diagnosis of stage II and stage III could be significantly improved by addition of Osp17 and p58 without loss of specificity . With the exception of sera from patients with erythema migrans the diagnostic sensitivity is comparable to the whole cell lysate IgG immunoblot . The main advantage of the recombinant immunoblot is the easy identification of diagnostic bands, whereas the identification of bands in the whole cell lysate immunoblot is difficult . The recombinant immunoblot is especially suitable where large series of sera need to be investigated.

Indian J Pediatr, 1998 Jan-Feb, 65(1), 99 - 105
Study of gene probes in childhood leprosy; Dayal R et al.; The objective was to detect nucleic acids of M . leprae in skin lesions of leprosy patients and study the effect of treatment on these nucleic acids, using r-RNA gene probes, using a cross sectional study . The study was carried out at Department of Paediatrics, S.N . Medical College, Agra and Department of Microbiology, Central JALMA Institute for leprosy, Agra . The study included 32 cases of leprosy less than 16 years of age, divided into 3 groups viz . without treatment (12 cases), in middle of treatment (11 cases) and at the end of treatment (9 cases) . All cases were subjected to a detailed history and thorough clinical examination . All of them had smear examination and lepromin test done and their skin biopsies were subjected to gene detection . Nucleic acids were isolated from skin biopsies of all cases by standard procedure . After dot blotting of these nucleic acids, they were hybridised with radioactive (p32) r-RNA probes . The results were interpreted after getting the X-ray films processed with background signals from controls . Majority of cases were between 13-16 years of age . As age advanced, the disease moved from tuberculoid end of spectrum towards lepromatous end (p < 0.05) . Majority of paucibacillary (PB) cases were lepromin positive while majority of multibacillary (MB) cases were lepromin negative (p < 0.05) . In specimens of untreated cases, 50% of PB specimens and 87.5% of MB specimens were positive for r-RNA probes . In multibacillary type 100% smear positive specimens and 67% smear negative specimens were positive for r-RNA probes . In patients during the middle of treatment positivity for r-RNA decreased and 20% of PB specimens and 16.6% MB specimens were positive . At the end of treatment (1 year for PB cases and 2 years for MB cases), the results of r-RNA were negative, which indicated that the treatment regimens used in the study were effective . This study supports the usefulness of r-RNA probes as a diagnostic and therapeutic tool in childhood leprosy.

Indian J Pediatr, 1997 Nov-Dec, 64(6), 747 - 53
Inflammatory diseases of the ear; Cheng AT et al.; The inflammatory diseases of external and middle ear are one of the commonest conditions encountered by the pediatric physician . Inner ear inflammations are less common and need special and urgent attention . Special management in each case requires detailed history, examination, necessary investigations and appropriate referral to otolaryngologist when necessary . The article is aimed to help formulate a plan in managing the inflammatory conditions of ear . Otalgia constitutes the most prominent of the symptoms in external and middle ear inflammations whereas vertigo, tinnitus and sensory hearing loss form the symptom complex for inner ear infections . It is necessary to understand the basic pathophysiology of the inflammatory condition to be able to institute a targetted treatment . The audiometry impedance studies, microbiology of discharge and occasionally ABR and CT scan from the mainstay of investigative workup . The treatment is specific and based on the precise diagnosis . It often requires the help of an otolaryngologist . Decisions may have to be made with regards to the need for any surgical intervention particularly in acute otitis media, an external canal abscess or an acute mastoiditis . A case of chronic otitis media with facial palsy or vertigo (labyrinthitis being a possibility) needs urgent intervention.

Crit Rev Biotechnol, 2000, 20(1), 17 - 48
Growth of filamentous fungi in submerged culture: problems and possible solutions; Gibbs PA et al.; Filamentous fungi are important organisms industrially and continue to attract research interest as microbiologists attempt to overcome the problems associated with their behavior in submerged culture . This review critically examines the literature describing these problems and where available suggests possible solutions to them . The influence of the chemical and physical environment on culture morphology, the process engineering challenges presented by different fungal morphologies, and the relationship between fungal morphology and metabolite production are all discussed.

Antonie Van Leeuwenhoek, 2000 Feb, 77(2), 103 - 16
Microbiology of petroleum reservoirs; Magot M et al.; Although the importance of bacterial activities in oil reservoirs was recognized a long time ago, our knowledge of the nature and diversity of bacteria growing in these ecosystems is still poor, and their metabolic activities in situ largely ignored . This paper reviews our current knowledge about these bacteria and emphasises the importance of the petrochemical and geochemical characteristics in understanding their presence in such environments.

FEBS Lett, 2000 Apr 14, 471(2-3), 251 - 5
Evidence that the glucoamylases and alpha-amylase secreted by Aspergillus niger are proteolytically processed products of a precursor enzyme; Dubey AK et al.; A 125-kDa starch hydrolysing enzyme of Aspergillus niger characterised by its ability to dextrinise and saccharify starch {Suresh et al . (1999) Appl . Microbiol . Biotechnol . 51, 673-675} was also found to possess activity towards raw starch . Segregation of these activities in the 71-kDa glucoamylase and a 53-kDa alpha-amylase-like enzyme supported by antibody cross-reactivity studies and the isolation of mutants based on assay screens for the secretion of particular enzyme forms revealed the 125-kDa starch hydrolysing enzyme as their precursor . N-terminal sequence analysis further revealed that the 71-kDa glucoamylase was the N-terminal product of the precursor enzyme . Immunological cross reactivity of the 53-kDa amylase with antibodies raised against the precursor enzyme but not with the 71- and 61-kDa glucoamylase antibodies suggested that this enzyme activity is represented by the C-terminal fragment of the precursor . The N-terminal sequence of the 53-kDa protein showed similarity to the reported Taka amylase of Aspergillus oryzae . Antibody cross-reactivity to a 10-kDa non-enzymic peptide and a 61-kDa glucoamylase described these proteins as products of the 71-kDa glucoamylase . Identification of only the precursor starch hydrolysing enzyme in the protein extracts of fungal protoplasts suggested proteolytic processing in the cellular periplasmic space as the cause for the secretion of multiple forms of amylases by A . niger.

J Biol Chem, 2000 Apr 21, 275(16), 12164 - 74
Multiple O-glycoforms on the spore coat protein SP96 in Dictyostelium discoideum . Fuc(alpha1-3)GlcNAc-alpha-1-P-Ser is the major modification; Mreyen M et al.; A decreased level of fucosylation on certain spore coat proteins of Dictyostelium discoideum alters the permeability of the spore coat . Here the post-translational modifications of a major spore coat protein, SP96, are studied in a wild type strain (X22) and a fucosylation-defective mutant (HU2470) . A novel phosphoglycan structure on SP96 of the wild type strain, consisting of Fuc(alpha1-3)GlcNAc-alpha-1-P-Ser(,) was identified by electrospray ionization mass spectrometry and NMR . It was shown using monosaccharide and gas chromatography mass spectrometry analysis that SP96 in the mutant HU2470 contained approximately 20% of wild type levels of fucose, as a result of a missing terminal fucose on the novel glycan structure . The results support previous predictions, based on inhibition studies on different fucose-deficient strains, about the nature of monoclonal antibody epitopes identified by monoclonal antibodies MUD62 and MUD166, which are known to identify O-linked glycans (Champion, A., Griffiths, K., Gooley, A . A., Gonzalez, B . Y., Gritzali, M., West, C . M., and Williams, K . L . (1995) Microbiology 141, 785-797) . Quantitative studies on wild type SP96 indicated that there were approximately 60 sites with phosphodiester-linked N-acetylglucosamine-fucose disaccharide units and a further approximately 20 sites with fucose directly linked to the protein . Over 70% of the serine sites are modified, with less than 1% of these sites as phosphoserine . Threonine and tyrosine residues were not found to be modified.

Dent Update, 1999 Jun, 26(5), 191 - 7
The microbiology of periodontal disease; Jenkinson HF et al.; Periodontal disease result from mixed bacterial infections, in which both host resistance barriers and bacterial interactions are important . Approximately ten bacterial species are strongly implicated with various forms of periodontal disease, although species that cannot yet be cultivated are likely also to be relevant . New technologies have shown that pathogenic bacterial species are present in defined complexes within subgingival plaque, thus identifying specific targets for therapeutic intervention . In light of increasing antibiotic resistance amongst oral bacteria, new strategies for control of periodontal bacterial complexes must be developed that inhibit the bacterial factors necessary for colonization and destruction of host tissues.

Am J Infect Control, 2000 Apr, 28(2), 133 - 7
Improved tuberculosis infection control practices in Maryland acute care hospitals; Fuss EP et al.; BACKGROUND: In 1992 and 1993, the Maryland Hospital Association and the Maryland Department of Health and Mental Hygiene conducted 2 surveys of tuberculosis prevention practices in Maryland hospitals that showed poor compliance with the 1990 Centers for Disease Control and Prevention's guidelines for preventing transmission of tuberculosis in health care facilities . OBJECTIVE: The objective of this study was to assess compliance in 1997 with the Centers for Disease Control and Prevention's guidelines in Maryland acute care hospitals . METHODS: A written questionnaire with 3 components-Infection Control, Employee Health, and Microbiology Laboratory-was mailed to 56 Maryland acute care hospitals . RESULTS: Seventy-three percent of the surveys were returned . One hundred percent of responding hospitals with tuberculosis isolation rooms reported negative pressure ventilation, 6 air exchanges per hour, and air exhausted to the outside or through high-efficiency particulate air filters . One hundred percent of the responding hospitals reported providing National Institute for Occupational Safety and Health-approved respiratory protection for health care workers; this compared with 24% in 1992 (P <.01) . One hundred percent of the responding hospitals reported performing at least annual tuberculin skin testing; this compared with 50% in 1992 (P < . 01) . CONCLUSIONS: The survey results demonstrate excellent compliance with the 1994 Centers for Disease Control and Prevention recommendations for tuberculosis control in Maryland acute care hospitals, even in those facilities determined to be at minimal to low risk for tuberculosis exposure . The proposed Occupational Safety and Health Administration regulations are unlikely to further reduce the risk of tuberculosis exposure to health care workers in Maryland acute care hospitals.

Dermatol Surg, 2000 Mar, 26(3), 235 - 9
Principles of CO2/erbium laser safety; Fader DJ et al.; BACKGROUND: There are a variety of potential hazards with laser technology . METHODS: A review of the literature . OBJECTIVE: To summarize the potential hazards of CO2 and erbium laser technologies and the safety guidelines and equipment developed to minimize them . RESULTS: Laser hazards can be divided into the following categories: mechanical, environmental, macrobiologic, microbiologic, and iatrogenic . CONCLUSION: At the conclusion of this learning activity, the reader should be able to discuss the mechanical, environmental, macrobiologic, microbiologic, and iatrogenic hazards of resurfacing laser technology, the literature cited to support current safety guidelines, and the equipment developed to promote laser safety.

Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 501 - 3
Leptospirillum gen . nov . (ex Markosyan 1972), nom . rev., including Leptospirillum ferrooxidans sp . nov . (ex Markosyan 1972), nom . rev . and Leptospirillum thermoferrooxidans sp . nov . (Golovacheva et al . 1992); Hippe H; The name Leptospirillum ferrooxidans is not in the Approved Lists of Bacterial Names (1980), nor has it been subsequently validly published . In accordance with the rules of the International Code of Nomenclature of Bacteria, the name Leptospirillum for the genus (gen . nov., nom . rev.) and Leptospirillum ferrooxidans for the species (sp . nov., nom . rev.) is revived here . The type species is Leptospirillum ferrooxidans strain L15T (= DSM 2705T) . The second species in the genus is Leptospirillum thermoferrooxidans (Golovacheva et al . 1992) (type strain L-88T; Institute of Microbiology, INMI, Moscow, Russia).

Spine, 2000 Mar 15, 25(6), 745 - 8
Aggressive thoracic actinomycosis complicated by vertebral osteomyelitis and epidural abscess leading to spinal cord compression; Yung BC et al.; STUDY DESIGN: Report of a successfully diagnosed and treated case of spinal cord compression due to epidural actinomycosis . OBJECTIVE: To illustrate that proper use of imaging strategy can greatly facilitate diagnosis and management of this rare condition . SUMMARY OF BACKGROUND DATA: Spinal actinomycosis causing epidural abscess and significant spinal cord compression is an uncommon condition . Although diagnosis is difficult, favorable results are widely reported when specific therapy is instituted . METHODS: A 32-year-old Chinese man had extensive dorsal thoracic soft tissue swelling and lower limb weakness . Collapse of the T5 vertebral body was found on plain radiographs with mediastinal infiltrates on chest radiograph . It took magnetic resonance imaging (MRI) to fully delineate the epidural abscess and dorsal muscular abscesses, which were not depicted by computed tomographic (CT) scan . Diagnosis was made by examination of CT-guided aspirate and tissue recovered during surgery by a microbiologist . The patient received high-dose intravenous penicillin and prompt spinal decompression once diagnosis of actinomycosis was confirmed . RESULTS: The dorsal muscular abscesses and upper thoracic epidural abscess resolved rapidly after intravenous antibiotics and surgical drainage . This was well documented by follow-up MRI and the full recovery of motor power and lower limb sensation in the patient . CONCLUSIONS: High clinical suspicion and proper use of imaging data led to timely diagnosis of this rare case of mediastinal, epidural, and intramuscular thoracic actinomycosis . Specific antibiotic therapy and timely, well-targeted surgical intervention greatly improve the outcome of this condition.

J Basic Microbiol, 2000, 40(1), 25 - 32
Evaluation of two reverse transcription polymerase reaction assays (GEN ETI-K HGV RNA and LCx GBV-C assay) for the detection of GB virus C/hepatitis G virus RNA in clinical samples; Garcia F Jr et al.; Our study evaluates the analytical performance of two amplification methods in the detection of GB Virus C/Hepatitis G Virus, GEN ETI-K HGV RNA (GEN) and the LCx GBV-C Assay (LCx) . GB Virus C RNA was detected by at least one test in 58/315 samples (18.41%) . Fifty-five samples (17.46%) were positive by the GEN method and 51 samples (16.19%) by the LCx method . The same rate of detection was found for 71 haemodialysis patients and 18 non-A non-E hepatitis . Method based differences in prevalence were observed for patient samples from the general population, 8/106 (7.55%) positive by GEN vs 7/106 (6.60%) by LCx; and HIV infected patients, 26/98 (26.53%) vs 23/98 (23.46%) . For chronic type C hepatitis 10/22 (45.5%) were positive by both methods, with two samples discordant . Overall, discordance was observed for ten samples, with seven positive only by the GEN ETI-K HGV RNA, and three positive only by the LCx GBV-C Assay . An additional evaluation of serial samples, from chronic type C hepatitis patients under interferon treatment, revealed three samples which were positive only by the GEN method . Results were 100% concordant for patients under haemodialysis and for non-A non-E hepatitis, 95.9% in the HIV positive group, 90.9% in the chronic type C hepatitis group, and 97.1% in the general population group . Overall, a 97.2% of concordance was found between methods . Both tests have a similar diagnostic performance, though in our opinion, LCx GBV-C Assay better suits the requirements of a clinical microbiology diagnostic laboratory.

Diagn Microbiol Infect Dis, 2000 Jan, 36(1), 7 - 11
Longitudinal 10-year prospective survey of fungaemia in Slovak Republic: trends in etiology in 310 episodes . Slovak Fungaemia study group; Krcmery V Jr et al.; A 10-year prospective survey of fungaemia in the Slovak Republic, involving 31 microbiology laboratories and 71 hospitals, was conducted from 1989-1998 (10 years): 310 fungaemias were analyzed for etiology, clinical characteristics, therapy, and outcome . C . albicans was responsible for 191 (61.6%) fungaemias, non-albicans Candida spp . (NAC) for 97 (31.3%), non-Candida yeasts for 18 (5.8%) and moulds (Fulsarium spp.) for four fungaemias . The most frequent NAC isolated from blood cultures were C . parapsilosis--30 (9.7%), C . krusei--18 (5.8%), C . tropicalis--14 (4.5%), and C . glabrata--10 (3.2%) . Secular trends in etiology showed a sustaining decrease of C . albicans (from 100% in 1989 to 50.7% in 1998) and increase of NAC (from 0% in 1989-1990 to 46.3% in 1998) . Non-Candida yeasts and moulds showed a stable proportion during the investigated period . There were statistically significant differences in etiology of fungaemia various subgroups of patients: non-albicans Candida spp . was significantly more frequent observed among subgroups of patients with pancreatitis and coma (53.3% vs . 31.3%, p < or = 0.02) and less frequently in the subgroup of neonates (15.0% vs . 31.3%, p < or = 0.006) . Vice versa, C . albicans appeared more frequently in neonates (85%).

Commun Dis Public Health, 2000 Mar, 3(1), 46 - 9
Cyclospora infections in England and Wales: 1993 to 1998; Cann KJ et al.; The coccidian protozoon Cyclospora cayetanensis is a treatable cause of prolonged, watery diarrhoea in humans . Microbiology laboratories in England and Wales often restrict testing to those who have recently travelled abroad . Only 44 to 66 laboratory reports of C . cayetanensis are made in England and Wales each year and a large proportion are found to have visited developing countries . Large foodborne outbreaks of infection have arisen in North America among people who have not travelled abroad but no such outbreaks have been identified in the United Kingdom . Public health laboratories in England and Wales were surveyed in 1998 to investigate their procedures for identifying C . cayetanensis . Sixty-eight per cent actively looked for the protozoon, but only half used a recommended method of direct microscopy of formol ether concentrates . National external quality assurance results for all participating UK laboratories were reviewed to assess laboratory proficiency in identification . C . cayetanensis was correctly identified in a wet preparation by 58% of laboratories, the lowest rate for specimens containing a single parasite species . Cyclosporiasis could be acquired in the UK from imported food, but current laboratory procedures might fail to identify it . Ascertainment must improve and awareness needs to be raised among food handlers, public and environmental health workers, laboratory staff, and general practitioners . We recommend that laboratories test all patients with watery diarrhoea for > 1 week for cyclospora, use formol ether concentration and microscopy with a calibrated eyepiece graticule, and confirm diagnoses with the help of a reference laboratory.

Appl Environ Microbiol, 2000 Apr, 66(4), 1410 - 5
The complete amino acid substitutions at position 131 that are positively involved in cold adaptation of subtilisin BPN'; Taguchi S et al.; To ascertain whether position 131 of a mesophilic protease, subtilisin BPN', is a potential critical site for cold adaptation as screened by evolutionary engineering (S . Taguchi, A . Ozaki, and H . Momose, Appl . Environ . Microbiol . 64:492-495, 1998), a full set of subtilisin BPN' mutants with mutations at position 131 was constructed by site-saturation mutagenesis . All mutated enzymes were measured for specific activity at 10 degrees C by the quantitative titer microplate assay system using polyclonal antibody against subtilisin BPN' and a synthetic chromogenic substrate . All the mutants exhibited proteolytic activities almost the same as or higher than that of the wild-type enzyme, suggesting that position 131 may be important for cold adaptation . In comparison with the wild type, purified mutants G131F, G131R, G131M, and G131W were found to acquire proteolytic activities (k(cat)/K(m)) at 10 degrees C that were 150, 94, 84, and 50% higher, respectively . In particular, for the G131F mutant, temperature dependency in enzyme activity was shown by an increase in k(cat) and a decrease in K(m) . All of these amino acid substitution mutants, G131F, G131R, G131M, and G131W, acquired increased proteolytic activities at 10 degrees C for three different synthetic peptide substrates but no increase in caseinolytic activity . Furthermore, they all conferred thermolability on the enzyme to differing extents in terms of the half-life of enzyme inactivation at 60 degrees C . No significant correlation was found between the amino acids preferred for cold adaptation surveyed here and those present at position 131 of subtilisin of psychrophilic cells naturally occurring in cold environments . Based on these findings, position 131 is a contributor in artificial evolution for acquiring a cold-active character and may not be related to physiological requirements for subtilisin-producing cells living in cold environments . Therefore, saturation mutagenesis would be effective in achieving rapid improvement in protein properties via evolutionary engineering.

J Gene Med, 1999 Jan-Feb, 1(1), 13 - 21
Maxillary sinusitis as a surrogate model for CF gene therapy clinical trials in patients with antrostomies; Wagner JA et al.; BACKGROUND: Assessing the biological activity and clinical efficacy of gene therapy is critically important in cystic fibrosis (CF) . It is widely accepted that clinical testing using surrogate markers including pulmonary function will be useful in assessing clinical efficacy . One problem with pulmonary surrogate markers of CF disease is the large number of patients and length of time required to demonstrate clinical efficacy . An alternative to pulmonary testing of new CF treatments is use of the maxillary sinuses as a surrogate model of CF lung disease . Using CF sinusitis as a surrogate model for testing clinical efficacy of new treatments is attractive because CF upper respiratory disease is similar to the lower respiratory disease with respect to electrophysiology and microbiology . METHODS: Sinusitis recurrence in untreated sinuses was analyzed during a prospective, randomized, unblinded, dose-escalation, within-subjects, phase I clinical trial of the adeno-associated virus mediated cystic fibrosis transmembrane conductance regulator (AAV-CFTR) gene transfer . RESULTS: Clinical symptoms combined with sinus endoscopy proved useful in the diagnosis of unilateral and bilateral sinusitis recurrence . Sinusitis recurred at a rate of 45% during one month of follow-up . IL-8 concentration rose in sinus fluids from affected sinuses . Bacterial cultures and increased sinus leukocytes corroborated recurrent sinusitis . Sinus CT scans were also useful in diagnosing recurrent sinusitis in this surrogate model of CF infectious exacerbations . CONCLUSIONS: CF sinusitis as a surrogate for lung disease is particularly well-suited for phase II clinical trials of gene transfer agents, with the potential for measuring clinical efficacy in relatively small numbers of patients over relatively short periods of time.

Lett Appl Microbiol, 2000 Feb, 30(2), 95 - 8
Quantal microbiology; Bridson EY et al.; Quantal microbiology describes a similarity between physics and microbiology . In both sciences there is an apparent dichotomy between the certainty and stability of the macro-subject and the uncertainty/complexity of the individual atom/cell . Classical physics is to quantum mechanics as classical microbiology is to quantal microbiology.

J Vet Diagn Invest, 2000 Mar, 12(2), 118 - 25
A statistical model for assessing sample size for bacterial colony selection: a case study of Escherichia coli and avian cellulitis; Singer RS et al.; A general problem for microbiologists is determining the number of phenotypically similar colonies growing on an agar plate that must be analyzed in order to be confident of identifying all of the different strains present in the sample . If a specified number of colonies is picked from a plate on which the number of unique strains of bacteria is unknown, assigning a probability of correctly identifying all of the strains present on the plate is not a simple task . With Escherichia coli of avian cellulitis origin as a case study, a statistical model was designed that would delineate sample sizes for efficient and consistent identification of all the strains of phenotypically similar bacteria in a clinical sample . This model enables the microbiologist to calculate the probability that all of the strains contained within the sample are correctly identified and to generate probability-based sample sizes for colony identification . The probability of cellulitis lesions containing a single strain of E . coli was 95.4% . If one E . coli strain is observed out of three colonies randomly selected from a future agar plate, the probability is 98.8% that only one strain is on the plate . These results are specific for this cellulitis E . coli scenario . For systems in which the number of bacterial strains per sample is variable, this model provides a quantitative means by which sample sizes can be determined.

Arthritis Rheum, 2000 Mar, 43(3), 593 - 8
Presence of bacterial DNA and bacterial peptidoglycans in joints of patients with rheumatoid arthritis and other arthritides; van der Heijden IM et al.; OBJECTIVE: The continuous presence of bacteria or their degraded antigens in the synovium may be involved in the pathogenesis of rheumatoid arthritis (RA) . The aim of this study was to determine the presence of bacterial nucleic acids and bacterial cell wall constituents in the joints of patients with RA and other forms of arthritis . METHODS: Joint samples were obtained from patients with RA (n = 26), septic arthritis (n = 2), inflammatory osteoarthritis (n = 5), and gout (n = 6), and joint trauma (n = 1) . Universal 16S-ribosomal RNA primers were used to detect the presence of bacterial DNA in these samples, using stringent regimens for sample collection and molecular microbiologic analysis . Automated sequencing and comparative data analysis were performed to identify the species . The presence of bacterial peptidoglycan-polysaccharide complexes in synovial tissue was detected by immunohistologic analysis with a specific antibody . RESULTS: The bacterial species cultured from the synovium could be identified in both of the patients with septic arthritis . DNA amplicons were also detected in the synovial fluid and/or tissue samples from 5 patients with RA and 2 patients with crystal-induced arthritis; these originated from multiple bacterial species . Staining for peptidoglycan-polysaccharide complexes was positive in the synovial tissue of both patients with septic arthritis, 16 with RA, 4 with inflammatory osteoarthritis, 4 with crystal-induced arthropathy, and 1 with joint trauma . The staining was mainly found in cells in the synovial sublining, including macrophages . CONCLUSION: The results indicate that bacterial DNA and bacterial cell wall constituents are retained in the joints of some patients with arthritis, where they might enhance synovial inflammation.

Akush Ginekol (Sofiia), 1999, 38(4), 20 - 2
{Gynecological interventions and cesarean section without suturing of the peritoneum (visceral and parietal)}; Lazarov L et al.; The authors reveal advantages of sewing off technics of parietal and visceral peritoneum in gynaecologic operations and Caesarean sections compared with common routine technics of sewing up . This study is prospective, controlled and includes two years period . 170 cases of gynaecologic operations and 45 Caesarean sections are investigated . It is confirmed the role of preoperative microbiologic investigations of cervical secret and vaginal secret . On the base of results similar to world experience in this field authors recommend described technics to gynaecologists and obstetricians.

Scand J Gastroenterol, 2000 Feb, 35(2), 184 - 9
Local short-chain fatty acids supplementation without beneficial effect on inflammation in excluded rectum; Schauber J et al.; BACKGROUND: Rectal instillation of short-chain fatty acids (SCFA), important nutrients for the colorectal mucosa, has been suggested to be of therapeutic value in distal intestinal inflammation . METHODS: In this study nine patients with Hartmann-closed rectum after colectomy for acute colitis were investigated . In a double-blind crossover trial an enema containing SCFA or a placebo solution was administered twice daily for 3 weeks . Before entry into the protocol, after each treatment period, and 6 weeks after the study period the patients' symptoms were evaluated, rectal endoscopy was performed, histologic samples were scored, and microbiologic analyses were carried out . RESULTS: No significant differences in symptoms, in mucosal inflammation, in histologic scoring, or in microbiologic studies were found between SCFA and placebo periods . Unexpectedly, all but one patient entirely lacked coliform bacteria in the rectum . CONCLUSIONS: In this study SCFA enemas had no beneficial effect on inflammation in excluded rectum in patients earlier submitted to colectomy for colitis . However, a different rectal flora was detected in these patients.

Int J Antimicrob Agents, 2000 Feb, 14(1), 13 - 9
The impact of laboratory reporting practice on antibiotic utilisation; Cunney RJ et al.; Microbiology reports are often misinterpreted by clinicians, which may lead to inappropriate antibiotic prescribing . Restricted release of susceptibilities combined with interpretative comments, can have a positive impact on the level of appropriate antibiotic use . Such a system requires two-way communication between the laboratory and the clinician and the laboratory's reporting practices should encourage such communication . The production and transmission of clinically relevant microbiology reports should be an integral part of infectious disease management programmes in hospitals.

Biochem, Educ. . 2000 Jan 1, 28(1), 20 - 23
The making of a model of Escherichia coli, magnified two million times - the 'Millennium Bug'
Tribe HT.
The author is making for the Millennium a model of Escherichia coli at 2x10(6) with emphasis on its interior composition . At this magnification molecules come into view and metabolic pathways are demonstrable as tangible systems of metabolites and enzymes interacting with one another . The model is intended to have value both for public understanding of science and in teaching of microbiology and biochemistry.

Vet Pathol, 2000 Mar, 37(2), 129 - 36
Expression of viral proteins in feline leukemia virus-associated enteritis; Kipar A et al.; Fourteen cases of feline leukemia virus (FeLV)-associated enteritis were immunohistologically examined for the expression of FeLV proteins gp70, p27, and p15E in the jejunum, mesenteric lymph nodes, spleen, and bone marrow . Results were compared with those of FeLV-infected cats without intestinal alterations . Other viral infections and specific bacterial, fungal, and parasitic infections were excluded by standard microbiologic methods, histopathology, immunohistology, and in situ hybridization . In FeLV-associated enteritis, FeLV gp70 and p15E were strongly expressed in intestinal crypt epithelial cells . In contrast, FeLV-positive cats without intestinal alterations showed only faint staining for gp70 and p15E and comparatively strong p27 expression in these cells . Findings suggest a direct relation between FeLV infection and alterations in intestinal crypt epithelial cells that may be attributed to the envelope proteins gp70 and p15E and/or their precursor protein . Distinct similarities to the intestinal changes in the experimentally induced FeLV-feline AIDS syndrome are obvious, suggesting that naturally occurring feline AIDS variants may be responsible for FeLV-associated enteritis.

Ophthalmology, 2000 Mar, 107(3), 574 - 80
Keratomycosis: clinical and microbiologic experience with dematiaceous fungi; Garg P et al.; OBJECTIVE: To assess the significance of dematiaceous fungi in the causation of keratomycosis . DESIGN: Retrospective, noncomparative case series . PARTICIPANTS: Eighty-eight cases of dematiaceous fungal keratitis seen at the L . V . Prasad Eye Institute, Hyderabad, India from January 1991 through December 1996 . INTERVENTION: Only culture-proven cases were analyzed . MAIN OUTCOME MEASURES: Predisposing factors, clinical characteristics, microbiology, treatment methods, and outcome . RESULTS: Of 557 cases of fungal keratitis seen during the study period, dematiaceous fungi were the etiologic agents in 88 (15.7%), after Fusarium in 210 (37.6%) and Aspergillus species in 170 cases (30.4%), respectively . Trauma was the most common predisposing factor (47.7%) . Fifty-three eyes (61.3%) had the classical clinical picture of yellow-white, dry raised infiltrate with feathery hyphate edges at initial examination . The characteristic macroscopic pigmentation was seen in only 24 eyes (27.27%) . Septate branching fungal filaments were identified in 78 smears (88.63%) on light microscopy, of which 5 (5.7%) also showed the presence of bacteria . Curvularia species dominated the spectrum (22.7%) . Treatment was started in 48 eyes with topical antifungal agents, whereas 37 received both oral and topical antifungal agents . Outcome data were available for 68 cases . Forty-nine (72%) responded to medical therapy, whereas 13 eyes required therapeutic penetrating keratoplasty and 6 eyes had to be eviscerated . CONCLUSIONS: This is the largest series of keratitis caused by dematiaceous fungi reported to date . It clearly brings out the clinical importance of this group of corneal infections.

J Am Vet Med Assoc, 2000 Mar 1, 216(5), 708 - 12, 684-5
Disseminated infection with Phialemonium obovatum in a German shepherd dog; Smith AN et al.; A 4-year-old spayed female German Shepherd Dog was evaluated because of left forelimb lameness . A fungal granuloma on the distal portion of the radius was determined to be the cause of the lameness; the infecting organism was identified as Phialemonium obovatum . Despite aggressive treatment with amphotericin B, itraconazole, and ketoconazole and curettage of the local area, the dog developed systemic disease and was euthanatized 5 months after initial evaluation . Immune dysfunction may have played a role in development of disseminated disease, because although serum concentrations of total IgG, IgA, and IgM were within or greater than reference ranges, results of lymphocyte proliferation assays were abnormal, which indicated cellular immune dysfunction . Infection with Phialemonium obovatum should be considered as a differential diagnosis when branching fungal organisms are detected during histologic, cytologic, or microbiologic evaluation of tissue specimens.

Tuber Lung Dis, 1999, 79(5), 299 - 308
Characterization of a Mycobacterium tuberculosis homologue of the Streptomyces coelicolor whiB gene; Mulder NJ et al.; SETTING: Molecular Research Laboratory, Department of Medical Microbiology, University of Cape Town and Groote Schuur Hospital . OBJECTIVE: Characterize Mycobacterium tuberculosis homologue of the Streptomyces coelicolor, sporulation specific, whiB regulatory gene . DESIGN: The M . tuberculosis whiB3 gene was isolated by enriched cloning of a 2.8 kb BamHl fragment to which the S . coelicolor whiB gene hybridized . Expression of the gene was analysed by S1 nuclease analysis and promoter studies . RESULTS: An open reading frame within the 2.8 kb BamHl fragment was identified as the M . tuberculosis whiB3 gene, one of four whiB homologues in the M . tuberculosis genome . The deduced amino acid sequence has a 92% identity with a M . leprae protein, and 32% identity with the S . coelicolor WhiB protein . S1 nuclease analysis showed that the M . tuberculosis whiB3 gene is constitutively expressed by the cells in liquid culture . Primer extension analysis revealed three transcriptional start sites . Expression from the three potential promoters is growth phase-dependent . CONCLUSION: The M . tuberculosis whiB3 gene is expressed throughout growth, but expression from the individual promoters is growth phase dependent.

Arch Pathol Lab Med, 2000 Mar, 124(3), 357 - 61
Effects of restructuring on the performance of microbiology laboratories in Alberta; Church DL et al.; OBJECTIVE: To evaluate the error rates of organism identification and antibiotic susceptibility proficiency testing challenges before, during, and after microbiology laboratory restructuring in Alberta . METHODS: Alberta Health substantially reduced and redistributed laboratory funds to the regional health authorities in 1995, forcing a dramatic restructure of services . Many rural hospitals expanded their microbiology test menus, and urban centers consolidated microbiology testing into a centralized high-volume laboratory . The Laboratory Proficiency Testing Program of the College of Physicians and Surgeons of Alberta mailed regular test profile surveys to microbiology laboratories during the restructure period to determine the type and extent of changes in services . Based on the types of tests and the extent of analysis being done, most rural B-level and some C-level laboratories were reclassified to the A level . The Laboratory Proficiency Testing Program reviewed the error rates of proficiency challenges based on the performance of different levels of laboratories before and after the period of restructure . RESULTS: Overall performance has improved according to the number of errors documented on identification and susceptibility challenges for laboratories that remained at the same classification (ie, A or C) . The number of major identification errors for laboratories that were reclassified increased, but the rate of major susceptibility errors decreased . More reclassified laboratories do not have dedicated registered technologist(s) who perform microbiology testing and are not supervised by an on-site pathologist and/or medical microbiologist compared with laboratories that remained at the same classification . CONCLUSIONS: Microbiology laboratory restructuring will have adverse effects on the quality of complex testing if experienced technologists are not retained and services are not medically supervised.

Respiration, 2000, 67(1), 24 - 9
Limited additive value of pleural fluid carcinoembryonic antigen level in malignant pleural effusion; Riantawan P et al.; OBJECTIVE: To assess the additive value of pleural fluid carcinoembryonic antigen (CEA.PF) level in the diagnosis of malignant pleural effusion . METHODS: Thoracentesis and closed pleural biopsy were performed in consecutive patients with pleural effusions . CEA.PF, cell analysis, and biochemical, cytopathologic and microbiologic studies were carried out . Further diagnostic interventions were undertaken if initial tests were inconclusive . RESULTS: A total of 176 patients were evaluated . The effusions proved malignant in 78 patients (44%) . Benign etiologies were diagnosed in 89 cases, comprising 51 tuberculous pleurisies, 12 empyemas, 26 others . The cause was unknown in 9 patients . Median (range) in ng/ml of CEA.PF were 233 (1-12,500) in malignant vs . 2.5 (0.3-9) in tuberculosis, 1.4 (0.1-2) in transudates, 19.4 (0.6-312) in empyemas, p < 0.001 . Receiver operating characteristic curve identified 10 ng/ml as the best cut-off for CEA.PF, yielding a sensitivity of 0.77, a specificity of 0.94, a positive and negative predictive value of 0.92 and 0.82, respectively . Among the 78 patients with malignant effusions, CEA.PF was elevated but initial cytopathologic study was nondiagnostic in 14 patients (18%) . Prompted by the raised CEA.PF, further diagnostic interventions were undertaken and secured the diagnosis of malignancy in all of these 14 patients . CONCLUSIONS: CEA.PF level adds limited value on cytopathologic study in the diagnosis of malignant pleural effusions . It potentially identifies 18% of patients with malignant effusions who require further investigations despite negative initial cytopathologic study .

Biochim Biophys Acta, 2000 Mar 15, 1464(1), 27 - 34
Location of tryptophan residues in free and membrane bound Escherichia coli alpha-hemolysin and their role on the lytic membrane properties; Verza G et al.; alpha-hemolysin (HlyA) is an extracellular protein toxin secreted by Escherichia coli that acts at the level of plasma cell membranes of target eukaryotic cells . Previous studies showed that toxin binding to the bilayers occurs in at least two ways, a reversible adsorption and an irreversible insertion . Studies of HlyA insertion into bilayers formed from phosphatidylcholine show that insertion is accompanied by an increase in the protein intrinsic fluorescence . In order to better define structural parameters of the membrane-bound form, the location of tryptophan residues was studied by means of quenchers of their intrinsic fluorescence located at 7, 12 and 16 positions of the acyl chain of phosphatidylcholine . The quenching was progressively weaker suggesting an interfacial location of the Trp . In parallel, HlyA was subjected to oxidation with N-bromosuccinimide to study the role of Trp residues exposed to aqueous media in its structure-function relationship . In the folded toxin molecule, a single residue was susceptible to oxidation with NBS, whereas incubation with LUV of the toxin prior modification prevents its oxidation, suggesting that Trp residue(s) are directly involved in toxin binding and insertion . Finally, the modification of residues exposed to solvent resulted in a complete impairment of the lytic activity . It was concluded that the modification-sensitive Trp residues are essential for the structure and function of native HlyA . These results are consistent with the model proposed by Soloaga et al . (Mol . Microbiol . 31 (1999) 1013-1024) according to which HlyA is bound to a single monolayer through a number of amphipathic instead of inserted transmembrane helices.

Mediators Inflamm, 1999, 8(1), 25 - 9
Regulation of inflammatory responses to Bordetella pertussis by N(G)-monomethyl-L-arginine in mice intranasally infected; Torre D et al.; To investigate effect of MMLA, an inhibitor of nitric oxide (NO) production, on regulation of inflammatory responses to Bordetella pertussis infection, mice were infected intranasally, and treated with various concentrations of MMLA . Ten days after infection, mice treated with MMLA at dosage of 100 mg/kg, given intraperitoneally in a single dose or for 5 consecutive days, showed at histopathologic examination, a significant decrease of intensity of inflammation (scores, 0.6 +/- 0.2 and 0.9 +/- 0.5 respectively) . A decrease of cellular accumulation of neutrophils and lymphocytes in the bronchoalveolar lavage (BAL) fluid was observed in infected mice treated with MMLA, especially at dosage of 10 mg/kg, given in a single dose intraperitoneally . In addition, BP-infected mice treated with MMLA (100 mg/kg, intraperitoneally) for 5 consecutive days showed higher mortality rate than untreated mice infected with B . pertussis, and the number of B . pertussis in lungs of mice treated with MMLA was significantly increased . However, MMLA treatment of infected mice had some effect on levels of IFN-gamma and nitrite/nitrate (end-stable products of NO) in the BAL fluid . This study indicates that NO may play a role either as microbiocidal agent or as a modulator of immune regulation, inasmuch as it may upregulate tissue inflammatory response to B . pertussis.

Otolaryngol Head Neck Surg, 2000 Mar, 122(3), 370 - 3
Endoscopically guided sinonasal cultures: a direct comparison with maxillary sinus aspirate cultures; Vogan JC et al.; Sinusitis is a common medical problem that can at times be challenging to treat . Although most cases respond to empiric therapy, success is not achieved universally . If empiric therapy fails, it is important to identify the causative bacterial pathogen . Antral puncture is the traditional diagnostic method to recover and identify pathogens in sinusitis; however, it remains a painful, invasive test with potential complications . In contrast, rigid sinonasal endoscopy permits recovery of mucopus emanating from the sinus ostia with little pain and few possible complications . Endoscopy also affords important visual information that can confirm or refute a historical/clinical diagnosis of sinusitis . Although previous studies have shown poor correlation between nasal cavity swab cultures and maxillary sinus aspiration cultures, few investigations have compared endoscopically guided middle meatal cultures with cultures obtained from maxillary sinus aspiration . Thirteen patients with maxillary sinusitis in one or both sinuses underwent endoscopically guided culture of the middle meatus and maxillary sinus puncture with aspiration and culture (16 total study samples) . Results from the microbiologic analysis were compared . Endoscopically guided middle meatal cultures accurately identified the predominant bacterial pathogen and correlated with the cultures from maxillary sinus aspiration in more than 90% of infections . These preliminary results suggest that endoscopically guided sinonasal cultures hold promise as a viable alternative to maxillary sinus aspiration . Endoscopically guided cultures appear to be an effective, noninvasive diagnostic tool for otolaryngologists managing sinusitis.

Prog Neurobiol, 2000 Apr, 60(5), 471 - 99
Unraveling of important neurobiological mechanisms by the use of pure, fully differentiated neurons obtained from adult animals; Hyden H et al.; An important, and often overlooked, problem in the neurochemical approach to neurobiological problems is that analysis of tissue involves almost always a heterogeneous population of cells (neurons, glia and other types of tissue cells) . The use of cell cultures has obvious limitations such as that they derive from embryonic or immediately postnatal animals; in addition, the cell culture conditions most certainly are quite different from the real tissue environment for the nerve cells . We underline here an alternative strategy, which is not new, but which, in our view, has already given formidable contributions to neurobiological studies and still is giving results of great importance . This is the technique proposed and used since the late fifties and early sixties by the senior author (H . Hyden) . The method involves the isolation of the big vestibular neurons from the adult rabbit vestibular nucleus . The neurons, fully differentiated and performing a precisely defined function, are obtained rapidly and completely free from surrounding glial cells . The separate microbiochemical study of these cells and their surrounding glia has yielded already in 1962, the information that modifications in gene expression are associated with plastic modifications of the function of the relevant neurons, which take place in the behavioral event of learning . Another important concept was formulated in the same time period following determination of the activities of energy metabolism related enzymes separately in vestibular neurons and their glia under vestibular stimulation . This is the concept that, under increased functional activity glia increases its anaerobic metabolism and passes then on the resulting metabolites to the neurons for aerobic metabolism . Both these concepts (RNA and memory; metabolic cooperation between glia and neurons) are nowadays widely accepted . In addition, this approach with pure big nerve cells has allowed in recent years the discovery of a novel mechanism for chloride extrusion in these cells . This mechanism utilizes structures similar to GABA activated chloride channels in cyclic modifications resulting in the final extrusion of chloride ions . The energy for the process is provided by a protein phosphorylation step . Future approaches are warranted such as the possibility of recognizing by RT-PCR specific neuronal mRNAs and their modification in expression in relation to function and plastic modifications (learning) . Another possible interesting application appears to be the recognition of the mRNAs for GABA(A) receptor subunits expressed here in these neurons in relation to the physiological and pharmacological characteristics of these native neuronal GABA(A) receptors.

Br J Biomed Sci, 1999, 56(2), 105 - 8
Enteric pathogens and their isolation at a cancer hospital in Pakistan; Bhatti AR et al.; Diarrhoeal diseases remain a major cause of mortality and morbidity in developing countries . However, due to lack of funds, supply problems and some inexperience, some laboratories have difficulty identifying a causative agent in stool samples . In the year following the opening of the Shaukat Khanum Memorial Cancer Hospital and Research Centre in Lahore, Pakistan, the microbiology department had not isolated a single enteric pathogen . From January 1996, new culture techniques were introduced, with a resulting increase (10%) in identification of these pathogens . In addition, the introduction of formol-ether concentration made a significant contribution to the number of intestinal parasites seen . This report demonstrates how simple microbiology methods made a difference to the running of the department and, ultimately, to the patients.

Pediatr Infect Dis J, 2000 Feb, 19(2), 118 - 21
Immunoblotting and serology for diagnosis of Helicobacter pylori infection in children; Raymond J et al.; BACKGROUND: The easiest way to identify the presence of current or past Helicobacter pylori infection is to test for antibodies . The aim of this study was to compare an enzyme-linked immunosorbent assay (ELISA) technique based on the detection of IgG antibodies directed against a global antigenic preparation with immunoblotting based on the analysis of IgG antibody reactivity to separate proteins . METHODS: Sera were collected from 80 children (mean age, 9.9 +/- 4.3 years) . The reference tests were microbiologic and histologic examination of gastric biopsies obtained at upper endoscopy . RESULTS: The immunoblotting was more sensitive (100%) and specific (88%) than ELISA (96 and 79%, respectively) in the evaluation of H . pylori infection in children . Its positive predictive value was 92%, and its negative predictive value was 100% . The best performance index of immunoreactive bands to detect antibodies was obtained with the 26-kDa (88.7%), 30-kDa (77.5%) and 19.5-kDa (70%) antigens . Antibodies by immunoblot technique against the CagA antigen were present in 43.1% of children . CONCLUSION: Immunoblotting is highly sensitive and more specific than ELISA in children and provides additional information about the full serologic profile . Immunoblotting may therefore be a useful complement to serology, particularly in cases with doubtful ELISA results.

J Bacteriol, 2000 Mar, 182(6), 1632 - 40
Global regulatory mutations in csrA and rpoS cause severe central carbon stress in Escherichia coli in the presence of acetate; Wei B et al.; The csrA gene encodes a small RNA-binding protein, which acts as a global regulator in Escherichia coli and other bacteria (T . Romeo, Mol . Microbiol . 29:1321-1330, 1998) . Its key regulatory role in central carbon metabolism, both as an activator of glycolysis and as a potent repressor of glycogen biosynthesis and gluconeogenesis, prompted us to examine the involvement of csrA in acetate metabolism and the tricarboxylic acid (TCA) cycle . We found that growth of csrA rpoS mutant strains was very poor on acetate as a sole carbon source . Surprisingly, growth also was inhibited specifically by the addition of modest amounts of acetate to rich media (e.g., tryptone broth) . Cultures grown in the presence of >/=25 mM acetate consisted substantially of glycogen biosynthesis (glg) mutants, which were no longer inhibited by acetate . Several classes of glg mutations were mapped to known and novel loci . Several hypotheses were examined to provide further insight into the effects of acetate on growth and metabolism in these strains . We determined that csrA positively regulates acs (acetyl-coenzyme A synthetase; Acs) expression and isocitrate lyase activity without affecting key TCA cycle enzymes or phosphotransacetylase . TCA cycle intermediates or pyruvate, but not glucose, galactose, or glycerol, restored growth and prevented the glg mutations in the presence of acetate . Furthermore, amino acid uptake was inhibited by acetate specifically in the csrA rpoS strain . We conclude that central carbon flux imbalance, inhibition of amino acid uptake, and a deficiency in acetate metabolism apparently are combined to cause metabolic stress by depleting the TCA cycle.

Int J Infect Dis, 2000, 4(1), 26 - 33
International study comparing cefdinir and cefuroxime axetil in the treatment of patients with acute exacerbation of chronic bronchitis; Van Herwaarden CL et al.; OBJECTIVES: To assess the efficacy and tolerability of three antibiotic regimens in patients with acute exacerbation of chronic bronchitis . METHODS: In this double-blind, randomized, multicentered, parallel-group study, patients received once-daily cefdinir 600 mg, twice-daily cefdinir 300 mg, or twice-daily cefuroxime axetil 250 mg for 10 days . Primary efficacy measures were microbiologic eradication rate, by pathogen and by patient, and clinical response rate, by patient . RESULTS: Of 1045 patients, 589 were evaluable for efficacy . At baseline, most patients had moderate or severe cough and sputum production as well as rhonchi, wheezing, and dyspnea . The microbiologic eradication rates by pathogen were 90% with once-daily cefdinir, 85% with twice-daily cefdinir, and 88% with twice-daily cefuroxime . The corresponding values for microbiologic eradication rate by patient were 90% (once-daily cefdinir), 85% (twice-daily cefdinir), and 86% (twice-daily cefuroxime) . The respective clinical response rates by patient were 81%, 74%, and 80% . There were no significant differences in the incidence of drug-related adverse events or discontinuations due to adverse events . Diarrhea was the most frequent complaint . CONCLUSIONS: The results indicate that the efficacy and tolerability of cefdinir, once or twice daily, and cefuroxime were comparable with no significant differences between the regimens used.

Acta Clin Belg, 1999 Dec, 54(6), 321 - 7
Prevention of pneumococcal disease: an update on the Belgian Consensus Report; Peleman RA et al.; An ad hoc working party on pneumococcal vaccine with representatives of the Belgian Society for Infectiology and Clinical Microbiology, the Belgian Society of Pulmonology and Scientific Societies of General Practitioners reviewed new data on the epidemiology of S . pneumoniae infections in Belgium, on the efficacy and the cost-effectiveness of the 23-valent capsular polysaccharide vaccine . We discuss recent data on vaccination with a new conjugate pneumococcal vaccine, shown to be highly effective in children . The Working Group of the Belgian Scientific Societies endorses the recommendations issued by the Hoge Gezondheidsraad in 1993 and described in a consensus report in 1996.

Adv Perit Dial, 1999, 15, 201 - 4
Exit-site and catheter care: review of important issues; Swartz RD; This brief review addresses the impact that several important aspects of catheter technology and exit-site care have on catheter-related infections and catheter longevity . The discussion includes exit-site microbiology, catheter configuration, exit-site care, and catheter salvage, following which a summary of recommendations is presented.

Biochem Biophys Res Commun, 2000 Feb 24, 268(3), 688 - 91
Expression of a Desulfovibrio tetraheme cytochrome c in Escherichia coli; da Costa PN et al.; A tetraheme cytochrome c was successfully overexpressed for the first time in Escherichia coli . Desulfovibrio desulfuricans ATCC 27774 tetraheme cytochrome c(3) was expressed in aerobically grown Escherichia coli cotransformed with Escherichia coli ccm gene cluster (Arslan et al . (1998) Bioch . Biophys . Res . Commun . 251, 744-747) . The analysis of the produced cytochrome showed that the signal peptide was correctly cleaved, the four heme groups were inserted and the electronic structure around the heme irons was conserved, i.e., the recombinant tetraheme cytochrome was identical to that isolated from the native source . Contradicting previous results which indicated that Escherichia coli was only capable of producing apocytochrome c(3) (Pollock et al . (1989) J . Gen . Microbiol . 135, 2319-2328), the present work proves unequivocally that the holoform can also be obtained .

Mt Sinai J Med, 2000 Jan, 67(1), 76 - 83
The history of liver disease at The Mount Sinai Hospital; Schaffner F; Diseases of the liver and biliary tract interested the physicians of The Mount Sinai Hospital from the time the hospital started until the present . Indeed, the institution has become a well-recognized center for the study of the liver and its diseases . During the first 75 years of the hospital, there were many admissions for hepatobiliary diseases, resulting in many case reports . The evolution of the hospital into a teaching hospital brought with it a more systematic method of studying diseases, not only in Pathology under Paul Klemperer, but in clinical chemistry and microbiology as well . Liver biopsy was also attempted . With the arrival of Hans Popper in 1957, the emphasis shifted to coordinated studies of structure and function under normal circumstances and in diseases as they progressed . Soon, Liver Diseases (Hepatology) were split from Gastroenterology, with Fenton Schaffner as the first chief . Over the next 30 years, more than 1000 papers, chapters and books were published . The main areas of research were fibrosis, cholestasis (especially morphology and bile salt metabolism), toxic liver injury, metabolic transformations and carcinogenesis . Primary biliary cirrhosis and viral hepatitis were and continue to be special interests . Fellows from all over the world were trained and many moved on to leadership positions . Although he was active in the development of the liver transplant program, Popper did not live to see its start . A new generation of hepatologists maintains the interest and position of The Mount Sinai Hospital in this important field of medicine.

Gene, 2000 Jan 11, 241(2), 247 - 54
Two genes encoding a putative multidrug efflux pump of the RND/MFP family are cotranscribed with an rpoH gene in Bradyrhizobium japonicum; Krummenacher P et al.; The rpoH3 gene of Bradyrhizobium japonicum codes for one of three sigma32-type transcription factors in this organism and is flanked by rag (rpoH3-associated) genes comprising the chromosomal arrangement ragABrpoH3ragCD . The first genes in this cluster code for a classical two-component regulatory system with an unknown function (Narberhaus et al., 1997 . Mol . Microbiol . 24, 93-104) . The deduced proteins of the last two genes display a high sequence similarity to heavy metal or multidrug efflux pumps of the RND (Resistance/Nodulation/cell Division)-MFP (Membrane Fusion Protein) family . Reverse transcription and polymerase chain reaction (RT-PCR) analysis demonstrated that ragC is cotranscribed with rpoH3 . Mutant strains carrying disrupted rag genes or an extented deletion of the rag locus exhibited neither an apparent growth defect nor a deficiency in the symbiotic interaction with soybean roots . The minimal inhibitory concentrations (MICs) of various metals and organic compounds were identical for the wild-type and mutant strains . Moreover, translational lacZ fusions to each of the first four genes of the rag cluster showed a very low expression under all conditions tested; hence, the substrate for the putative efflux pump has not been uncovered.

J Clin Pathol, 1999 Oct, 52(10), 773 - 5
Review of the clinical activity of medical microbiologists in a teaching hospital; Wooster SL et al.; BACKGROUND: The clinical interactive role of medical microbiologists has been underestimated and the discipline is perceived as being confined to the laboratory . Previous studies have shown that most microbiology interaction takes place over the telephone . AIM: To determine the proportion of clinical ward based and laboratory based telephone interactions and specialties using a microbiology service . METHODS: Clinical microbiology activity that took place during November 1996 was prospectively analysed to determine the distribution of interactions and specialties using the service . RESULTS: In all, 1177 interactions were recorded, of which nearly one third (29%) took place at the bedside and 23% took place on call . Interactions involving the intensive treatment unit, general ward visits, and communication of positive blood cultures and antibiotic assays were the main areas of activity identified . There were 147 visits to 86 patients on the general wards during the study, with the number of visits to each individual varying from one to eight . The need for repeated visits reflected the severity of the underlying condition of the patients . Ward visits were regarded as essential to obtain missing clinical information, to assess response to treatment, and to make an appropriate entry in a patient's notes . CONCLUSIONS: Ward visits comprise a significant proportion of clinical microbiology interactions and have potential benefits for patient management, service utilisation, and education.

J Bacteriol, 2000 Mar, 182(5), 1321 - 7
Novel phospholipase A activity secreted by Legionella species; Flieger A et al.; Bacterial phospholipases are regarded as a major virulence factor in infection . In bacteria associated with pneumonia, destruction of lung surfactant and host cell membranes by bacterial phospholipases secreted during infection is thought to contribute to the disease . Phospholipase C (PLC) activity has been described in several Legionella species (W . B . Baine, J . Gen . Microbiol . 134:489-498, 1988; W . B . Baine, J . Gen . Microbiol . 131:1383-1391, 1985) . By using detection methods such as thin-layer chromatography and mass spectrometry, PLC activity could not be detected in several strains of Legionella pneumophila . Instead, phospholipid degradation was identified to be caused by a novel PLA activity . We could demonstrate that PLA secretion starts at the mid-exponential-growth phase when bacteria were grown in liquid culture . Several Legionella species secreted different amounts of PLA . Legionella PLA may act as a powerful agent in the mediation of pathogenicity due to destruction of lung surfactant and epithelial cells.

J Bacteriol, 2000 Mar, 182(5), 1286 - 95
WhiD and WhiB, homologous proteins required for different stages of sporulation in Streptomyces coelicolor A3(2); Molle V et al.; The whiD locus, which is required for the differentiation of Streptomyces coelicolor aerial hyphae into mature spore chains, was localized by map-based cloning to the overlap between cosmids 6G4 and D63 of the minimal ordered library of Redenbach et al . (M . Redenbach et al., Mol . Microbiol . 21:77-96, 1996) . Subcloning and sequencing showed that whiD encodes a homologue of WhiB, a protein required for the initiation of sporulation septation in S . coelicolor . WhiD and WhiB belong to a growing family of small (76- to 112-residue) proteins of unknown biochemical function in which four cysteines are absolutely conserved; all known members of this family are found in the actinomycetes . A constructed whiD null mutant showed reduced levels of sporulation, and those spores that did form were heat sensitive, lysed extensively, and were highly irregular in size, arising at least in part from irregularity in septum placement . The whiD null mutant showed extreme variation in spore cell wall deposition; most spores had uniformly thin (20- to 30-nm) walls, but spore chains were frequently observed in which there was irregular but very pronounced (up to 170 nm) cell wall thickening at the junctions between spores . whiD null mutant spores were frequently partitioned into irregular smaller units through the deposition of additional septa, which were often laid down in several different planes, very close to the spore poles . These "minicompartments" appeared to be devoid of chromosomal DNA . Two whiD promoters, whiDp1 and whiDp2, were identified, and their activities were analyzed during development of wild-type S . coelicolor on solid medium . Both promoters were developmentally regulated; whiDp1 and whiDp2 transcripts were detected transiently, approximately at the time when sporulation septa were observed in the aerial hyphae.

Vet Hum Toxicol, 2000 Feb, 42(1), 30 - 5
Ivermectin: an assessment of its pharmacology, microbiology and safety; Burkhart CN; Ivermectin (IVM) is the drug of choice for a variety of parasitic diseases due to its broad spectrum of activity and wide margin of safety . More than 18 million people are treated with IVM each year . Delivery modes include oral, topical, and s.c . injections . Its anti-parasitic activity depends upon species and developmental stages . Although IVM is believed to act mainly through interactions with invertebrate glutamate-gated chloride (GluCl) channel, other targets such as spleen cells and aminobutyric acid receptors may play important roles in the anti-parasitic activity of IVM . As several organisms have evolved resistance to IVM through mutations in p-glycoproteins and/or the GluCl channel itself, research continues on improvement of IVM either through mode of administration or the feasibility of alternative macrolides . An understanding of IVM's pharmacology is essential before improved therapeutics are created.

Curr Opin Ophthalmol, 1999 Dec, 10(6), 452 - 7
Ocular manifestations of systemic infections; Uchio E et al.; Ocular involvement, mainly uveitis or retinochoroiditis, is common in various systemic diseases, such as endogenous endophthalmitis, Lyme disease, human T-cell lymphotropic virus type I infection, toxoplasmosis, and toxocariasis . Recent progress, especially in laboratory microbiologic testing, has enabled us to reliably diagnose many formerly idiopathic intraocular inflammatory diseases . A group of systemic infectious diseases, including those discussed here, are implicated as a body of emerging or re-emerging diseases that have appeared in the past two decades and are thought to have a close relation with global socioenvironmental changes . This paper discusses recent clinical and experimental studies of the most important systemic infectious diseases that affect the eye.

J Hosp Infect, 1999 Dec, 43 Suppl, S285 - 91
Interaction between the microbiology laboratory and clinician: what the microbiologist can provide; Kolmos HJ; The work of the clinical microbiologist comprises three major areas: diagnostic work in the laboratory, advice to clinicians about treatment of infected patients, and infection control . By clinical alertness, either from work in the laboratory or from clinical contacts, the microbiologist may contribute to the recognition of hospital outbreaks . The microbiologist plays a key role in implementing a restrictive antibiotic policy in hospital . Experience shows that a close personal contact with clinicians in the daily treatment of patients is the most efficient way to ensure a rational use of antibiotics and keep the consumption low . Other important measures include the elaboration of antibiotic guidelines and performance of audits . On basis of periodic summaries of laboratory data and data on antibiotic consumption, the microbiologist can keep the clinicians informed about antibiotic resistance and compliance with the antibiotic guidelines . In addition to informal contacts, the microbiologist also interacts with clinicians through participation in infection control and drug and therapeutic committees.

Verh K Acad Geneeskd Belg, 1999, 61(6), 649 - 59
{Report of the first congress of the "European Society for Emerging Infections" . (Budapest, 13-16 September 1998)}; Huygelen C; The first congress of the recently founded European Society for Emerging Infections was held in Budapest from 13 to 16 September 1998 . About 200 physicians, veterinarians, biologists and microbiologists attended this meeting . The euphoria of the 1970's with respect to infectious diseases is now gone . During the last twenty years about thirty new infections agents have been identified and re-emergence of old diseases which had disappeared to a large extent, has been reported in many countries . Most newly emerging diseases in man are of zoonotic origin or are closely related to disease in animal (wild or domestic) showing a parallel pathology . The nature of the etiologic agents varies widely: prions, viruses, chlamydia, rickettsiae, bacteria, protozoa etc . Several factors play a role in the emergence: mutations of the agents themselves; changing habits of man as the host: travel, sexual habits, etc.; modifications of the climate or environment can influence the expansion of vectors . The subjects discussed at the congress covered a wide field of diseases and agents: plague, retroviruses, antibiotic-resistant bacteria, influenza, lyme borreliosis, tick-borne encephalitis, hantaviruses, rickettsioses and ehrlichiose, transmissible spongiform encephalopathies, Borna, lyssaviruses, E . coli, protozoa, chlamydia, etc.

J Clin Microbiol, 2000 Feb, 38(2), 547 - 51
Comparison of sorbitol MacConkey agar and a two-step method which utilizes enzyme-linked immunosorbent assay toxin testing and a chromogenic agar to detect and isolate enterohemorrhagic Escherichia coli; Novicki TJ et al.; Enterohemorrhagic Escherichia coli (EHEC) and specifically serotype O157:H7 are a significant cause of hemorrhagic gastrointestinal disease and the hemolytic uremic syndrome . Methods currently used in clinical microbiology labs, such as sorbitol-MacConkey (SMAC) agar, reliably detect only O157:H7 . We have evaluated a two-step method that has the potential to identify and isolate all EHEC serotypes, including serotype O157:H7 . This method utilizes a chromogenic selective-differential medium for the isolation of E . coli together with an enzyme-linked immunosorbent assay (ELISA) that detects the Shiga-like toxins Stx1 and Stx2 . Both are commercially available and usable in a wide range of clinical microbiology laboratories . Compared to a Vero cell cytotoxic assay, SMAC had sensitivities of 23.5% for the identification of all EHEC serotypes and of 50.0% for the identification of O157:H7 alone . The two-step method had sensitivities of 76.5 and 100%, respectively . The ELISA alone had a sensitivity of 82.4% in the detection of Stx1 and Stx2 . The specificity was 100% in all cases . Overall, 14 EHEC isolates were obtained: 8 (58%) O157:H7, 2 (14%) O26, 2 (14%) O111:NM, 1 (7%) O103:H2, and 1 (7%) O121:H19 . All but one were isolated during the months of May to September . The two-step method was found to be considerably more expensive than SMAC for both positive and negative samples.

Appl Environ Microbiol, 2000 Feb, 66(2), 643 - 50
Endosymbiotic microbiota of the bamboo pseudococcid Antonina crawii (Insecta, Homoptera); Fukatsu T et al.; We characterized the intracellular symbiotic microbiota of the bamboo pseudococcid Antonina crawii by performing a molecular phylogenetic analysis in combination with in situ hybridization . Almost the entire length of the bacterial 16S rRNA gene was amplified and cloned from A . crawii whole DNA . Restriction fragment length polymorphism analysis revealed that the clones obtained included three distinct types of sequences . Nucleotide sequences of the three types were determined and subjected to a molecular phylogenetic analysis . The first sequence was a member of the gamma subdivision of the division Proteobacteria (gamma-Proteobacteria) to which no sequences in the database were closely related, although the sequences of endosymbionts of other homopterans, such as psyllids and aphids, were distantly related . The second sequence was a beta-Proteobacteria sequence and formed a monophyletic group with the sequences of endosymbionts from other pseudococcids . The third sequence exhibited a high level of similarity to sequences of Spiroplasma spp . from ladybird beetles and a tick . Localization of the endosymbionts was determined by using tissue sections of A . crawii and in situ hybridization with specific oligonucleotide probes . The gamma- and beta-Proteobacteria symbionts were packed in the cytoplasm of the same mycetocytes (or bacteriocytes) and formed a large mycetome (or bacteriome) in the abdomen . The spiroplasma symbionts were also present intracellularly in various tissues at a low density . We observed that the anterior poles of developing eggs in the ovaries were infected by the gamma- and beta-Proteobacteria symbionts in a systematic way, which ensured vertical transmission . Five representative pseudococcids were examined by performing diagnostic PCR experiments with specific primers; the beta-Proteobacteria symbiont was detected in all five pseudococcids, the gamma-Proteobacteria symbiont was found in three, and the spiroplasma symbiont was detected only in A . crawii.

Cleve Clin J Med, 2000 Jan, 67(1), 25 - 9, 33-4, 37-8 passim
The dilemma of nosocomial pneumonia: what primary care physicians should know; Khurana PS et al.; Because nosocomial pneumonia is difficult to diagnose and the need to treat it is often urgent, particularly in patients on mechanical ventilation, therapy is often empiric . We review the pathogenesis, risk factors, microbiology, diagnosis, and treatment of this disease.

FEMS Microbiol Rev, 2000 Jan, 24(1), 9 - 20
Vaccines, coming of age after 200 years; Makela PH; An overview on the short, only 200 years, past history and future expectations in the field of vaccines is presented . The focus is on development trends and potential rather than individual vaccines . While the first vaccines were a result of keen observation, the further development has been tightly dependent on the development of microbiology to provide both the knowledge basis and the technology for new vaccines for new purposes . The post-genomic era just starting therefore promises an exponential increase of vaccine research and new vaccines, both improved vaccines with a greater efficacy and less adverse effects to replace old ones and vaccines for prevention of diseases for which no vaccines exist . Furthermore, fully new applications to prevention or treatment of chronic diseases not traditionally associated with infections are expected.

Rinsho Byori, 1999 Dec, 47(12), 1105 - 12
{Department of microbiosensing system using micromachine techniques}; Murakami Y et al.; This paper reviews devices for Point Of Care (POC) testing, especially focusing on current and future biosensors and biosensing fields . POC testing involves on-site diagnosis such as bedside diagnosis and near-patient testing . Biosensors and biosensing devices are a key technology to measure all clinically important parameters . For many reasons such as historical, simplicity, the high stability and activity of the enzyme required, and large demand from diabetics, only glucose sensors are commercially available . Various biosensors are waiting for commercialization . Not only simple biosensors having a biofunctional membrane on a transducer, but more complicated biosensing systems are fabricated on a microchip . Micro-Total Analysis System (mu TAS) is a research field to fabricate miniaturized sensing devices or systems . In these devices or systems, biochemical reactions for detection can be well controlled by running buffers, even though the consumption of reagents and sample is very small . Microarray chip technology, such as the DNA chip, is now receiving much attention in not only genetic research, but other biochemical research and application fields as well . Detection apparatus for the DNA chip is currently bench-top size . According to recent research, however, the size will be reduced as to a portable-size that is suitable for POC testing . Research and development of POC devices are advanced not alternative methods of conventional laboratory test, but as complement technology with advantages such as mobility, speed, and low-cost.

Am J Forensic Med Pathol, 2000 Dec, 21(4), 395 - 400
An analysis of the usefulness of specific stages in the pathologic investigation of sudden infant death; Mitchell E et al.; Retrospective analysis of autopsy findings in 60 infants who had been found unexpectedly dead in their cribs or beds in South Australia from 1994 to 1998 was undertaken to determine the diagnostic usefulness of individual stages in the postmortem investigation . Positive findings occurred in 2 of 43 scene examinations (3%), 2 of 60 external examinations (3%), 2 of 11 radiologic examinations (18%), 8 of 60 internal examinations (13%), 7 of 60 histologic examinations (12%), and 3 of 58 microbiologic examinations (5%) . No positive findings were detected on toxicologic screening . Not every case underwent each diagnostic step . This gave alternative diagnoses to sudden infant death syndrome (SIDS) in 15 cases (25%) . This study demonstrates an increase in the percentage of cases of unexpected infant death due to causes other than SIDS; it also shows the diagnostic yield of individual stages in the postmortem evaluation of such cases . Negative findings were important in giving validity to the diagnosis in the 45 cases that were ultimately designated as SIDS.

Clin Microbiol Rev, 2000 Jan, 13(1), 83 - 121, table of contents
New strategies for prevention and therapy of cytomegalovirus infection and disease in solid-organ transplant recipients; Sia IG et al.; In the past three decades since the inception of human organ transplantation, cytomegalovirus (CMV) has gained increasing clinical import because it is a common pathogen in the immunocompromised transplant recipient . Patients may suffer from severe manifestations of this infection along with the threat of potential fatality . Additionally, the dynamic evolution of immunosuppressive and antiviral agents has brought forth changes in the natural history of CMV infection and disease . Transplant physicians now face the daunting task of recognizing and managing the changing spectrum of CMV infection and its consequences in the organ recipient . For the microbiology laboratory, the emphasis has been geared toward the development of more sophisticated detection assays, including methods to detect emerging antiviral resistance . The discovery of novel antiviral chemotherapy is an important theme of clinical research . Investigations have also focused on preventative measures for CMV disease in the solid-organ transplant population . In all, while much has been achieved in the overall management of CMV infection, the current understanding of CMV pathogenesis and therapy still leaves much to be learned before success can be claimed.

Anal Quant Cytol Histol, 1999 Dec, 21(6), 468 - 72
DNA quantification as a prognostic factor in colorectal adenocarcinoma; Azua J et al.; OBJECTIVE: To determine if DNA quantification, studied in cytologic samples obtained by fiberendoscopy, has predictive value in colorectal adenocarcinoma . STUDY DESIGN: Thirty patients with colorectal cancer diagnosed by cytology, endoscopy and microbiopsy, were selected . The study covered > 10 years . Smears were stained with progressive hematoxylin and processed by computer for DNA evaluation by image cytometry . RESULTS: Four different types of histograms that directly related to malignancy were obtained . These histograms were characterized by the value of entropy, and we established four grades of aggressiveness . Finally, we obtained two large groups: high and low grade of malignancy . We studied the survival times in both groups and used a Kaplan-Meier survival curve for the high grade group . We confirmed the results statistically and found that there was a significant relationship, with P < .05 . CONCLUSION: We strongly advise the use of DNA quantification by image cytometry in daily surgery as a prognostic indicator of survival time in colorectal cancer.

Mol Phylogenet Evol, 1999 Dec, 13(3), 525 - 35
Molecular systematics of marsupials based on the rRNA 12S mitochondrial gene: the phylogeny of didelphimorphia and of the living fossil microbiotheriid Dromiciops gliroides thomas; Palma RE et al.; Nucleotide sequence data from the mitochondrial 12S rRNA gene were used to evaluate the phylogenetic relationships among the major groups of didelphimorph and paucituberculatan marsupials from South America, the microbiotheriid Dromiciops gliroides, and representatives of four orders of Australasian marsupials . Based on approximately 800 bp in 18 genera, we conclude that the didelphids constitute a monophyletic group with large-sized forms differentiated from small opossums, while Caluromys constitutes the sister taxon to didelphids . The peramelid Isoodon was recovered as the sister taxon to the paucituberculatans Caenolestes and Rhyncholestes, although it is in an uncertain phylogenetic position within the marsupial tree . Dromiciops was recovered as a well-differentiated lineage from South American opossums within the Australidelphian radiation of metatherians that include dasyurid, diprotodontian, and notoryctemorph marsupials .

IEEE Trans Biomed Eng, 1999 Dec, 46(12), 1483 - 7
Medium and interface components in impedance microbiology; Felice CJ et al.; Classic impedance microbiology (CIM) is based on the measurement of the impedance components that appear between a pair of electrodes submerged in a cell containing inoculated broth . Either a bipolar or a tetrapolar technique can be applied, requiring about 1 x 10(3) to 3 x 10(7) cells/ml to produce detectable changes in the impedance curves . Theoretical analysis of the electrode-electrolyte interface during bacterial growth is lacking, with no generally accepted measuring standards . Besides, there is considerable disagreement . We separated out the interface and medium components using the frequency variation technique (FVT) and also analyzed the interface reactance-resistance diagram, both before and after bacterial growth . Medium resistance Rm, interface reactance Xi, and interface resistance Ri, were quantified as time functions growth curves, from the complex bipolar impedance seen between two electrodes . We took into account the electrical current density, the temperature and the associated circuitry, also explaining the theoretical and experimental bases that justify the proposed dissecting procedure . It was found that, within the working frequency range, Rm, Ri, and Xi percental growth curves are frequency-independent, i.e., neither Rm(f), nor Xi(f) nor Ri(f) changed their slopes before, during and after bacterial growth . Besides, no alpha-dispersion effect in Rm curves was detected . It is concluded that impedance microbiology could become a fertile area for interdisciplinary knowledge; its development might offer new avenues for basic and applied research.

Enferm Infecc Microbiol Clin, 1999, 17 Suppl 2, 15 - 21
{Systematic reviews on infectious diseases . The Cochrane Collaboration}; Bonfill X et al.; BACKGROUND: Theoretically, doctors update their knowledge from the scientific literature, and bibliographic databases have made it possible to overcome many of the limitations with this information, but not all of them . When faced with a therapeutic issue, the practice of evidence based medicine requires the efficient access to information derived from controlled clinical trials (CCT) . Reviews of the scientific literature are of increasing importance . The Cochrane Collaboration is dedicated to preparing, maintaining and disseminating updated, systematic reviews of CCTs of health care interventions, which are published in the Cochrane Library . OBJECTIVE: To describe the Cochrane Collaboration and to locate, with the greatest exhaustiveness possible, CCTs published in the journals "Enfermedades Infecciosas y Microbiologia Clinica", describe their characteristics and include them in a global database of clinical trials maintained by the Cochrane Collaboration . METHODS: The CCTs were identified by systematic manual searches of all issues of the journal . A descriptive analysis of the CCTs located was made . RESULTS: A total of 24 CCTs were published in 15 years (1.6 per year), 10 were original articles and 14 were communications at congresses . All were done in Spain and the majority were done in hospitals . The most frequently researched question was antibiotic prophylaxis and all evaluated pharmaceutical interventions . The majority of the CCTs published had important information missing (phase of study, randomization, blinding, etc.) . CONCLUSIONS: The identification of CCTs is a prerequisite for the development of systematic reviews . There were few clinical trials published in the 15 years of publication of this journal, and the most common characteristic is, clearly, the absence of basic data . The most common is a lack of on the phase of the clinical trial, the follow-up period, and funding sources . The idea of what experimental clinical investigation in infectious diseases is in our country needs to include the identification of CCTs published in other Spanish and international journals.

Infect Immun, 2000 Jan, 68(1), 214 - 20
Comparative evaluation of low-molecular-mass proteins from Mycobacterium tuberculosis identifies members of the ESAT-6 family as immunodominant T-cell antigens; Skjot RL et al.; Culture filtrate from Mycobacterium tuberculosis contains protective antigens of relevance for the generation of a new antituberculosis vaccine . We have identified two previously uncharacterized M . tuberculosis proteins (TB7.3 and TB10.4) from the highly active low-mass fraction of culture filtrate . The molecules were characterized, mapped in a two-dimensional electrophoresis reference map of short-term culture filtrate, and compared with another recently identified low-mass protein, CFP10 (F . X . Berthet, P . B . Rasmussen, I . Rosenkrands, P . Andersen, and B . Gicquel . Microbiology 144:3195-3203, 1998), and the well-described ESAT-6 antigen . Genetic analyses demonstrated that TB10.4 as well as CFP10 belongs to the ESAT-6 family of low-mass proteins, whereas TB7.3 is a low-molecular-mass protein outside this family . The proteins were expressed in Escherichia coli, and their immunogenicity was tested in cultures of peripheral blood mononuclear cells from human tuberculosis (TB) patients, Mycobacterium bovis BCG-vaccinated donors, and nonvaccinated donors . The two ESAT-6 family members, TB10.4 and CFP10, were very strongly recognized and induced gamma interferon release at the same level (CFP10) as or at an even higher level (TB10.4) than ESAT-6 . The non-ESAT-6 family member, TB7.3, for comparison, was recognized at a much lower level . CFP10 was found to distinguish TB patients from BCG-vaccinated donors and is, together with ESAT-6, an interesting candidate for the diagnosis of TB . The striking immunodominance of antigens within the ESAT-6 family is discussed, and hypotheses are presented to explain this targeting of the immune response during TB infection.

Emerg Infect Dis, 1999 Nov-Dec, 5(6), 779 - 87
Supplementing tuberculosis surveillance with automated data from health maintenance organizations; Yokoe DS et al.; Data collected by health maintenance organizations (HMOs), which provide care for an increasing number of persons with tuberculosis (TB), may be used to complement traditional TB surveillance . We evaluated the ability of HMO-based surveillance to contribute to overall TB reporting through the use of routinely collected automated data for approximately 350,000 HMO members . During approximately 1.5 million person-years, 45 incident cases were identified in either HMO or public health department records . Eight (18%) confirmed cases had not been identified by the public health department . The most useful screening criterion (sensitivity of 89% and predictive value positive of 30%) was dispensing of two or more TB drugs . Pharmacy dispensing information routinely collected by many HMOs appears to be a useful adjunct to traditional TB surveillance, particularly for identifying cases without positive microbiologic results that may be missed by traditional public health surveillance methods.

Pediatr Nephrol, 1999 Nov, 13(9), 771 - 2
Penicillium peritonitis in an adolescent receiving chronic peritoneal dialysis; Equils O et al.; A 19-year-old female on chronic peritoneal dialysis developed acute peritonitis; multiple peritoneal fluid and catheter tip cultures yielded Penicillium species . She promptly responded to catheter removal and intravenous amphotericin B, followed by oral fluconazole, without further recurrences 1 year later . This is the first reported case of Penicillium peritonitis in the pediatric population . We review the microbiology and clinical spectrum of this disease, as well as the few previous reported cases in adults.

Pharmacotherapy, 1999 Dec, 19(12), 1363 - 8
Malaria epidemiology and economics in a pharmacist-managed international travel clinic; McNicholl IR et al.; With high rates of travel and low adherence to malaria prophylaxis, targeting educational efforts to high-risk travelers is vital . We assessed risk factors for acquiring malaria, and resource use and outcomes of these patients in a managed care environment . Patients were identified from January 1, 1994, to December 31, 1997, using microbiology and pharmacy databases, chart reviews, and interviews . Sixteen patients acquired malaria during the study; although only 50% contacted the travel clinic . Only 31% (5) of them had documented adherence . Fifty percent were hospitalized at a cost of $3881/patient . Travelers at greatest risk for nonadherence appear to be expatriates and those visiting Africa . Providers should target these groups with more intensive counseling in an effort to improve therapy adherence and reduce the risk for malaria.

Acta Derm Venereol, 1999 Nov, 79(6), 473 - 4
Direct or referral microscopy of vaginal wet smear for bacterial vaginosis: experience from an STD clinic; Petersen CS et al.; A new wet smear diagnostic criterion for bacterial vaginosis was applied to 124 consecutive female patients attending an STD clinic located in the centre of Copenhagen . Bacterial vaginosis was detected in 54 (44%) women, making bacterial vaginosis the most prevalent pathological condition encountered . A total of 47 (87%) of the women were symptomatic . Concomitant genital infections were found in 13 (24%) of these women, most often as vulvo-vaginal candidiasis . A correct microscopic diagnosis could also be obtained by sending the vaginal smear to the local microbiologist for rehydration and phase contrast microscopy . It is suggested that the previously described vaginal wet smear criteria are used in place of Amsel's criteria for routine diagnosis of bacterial vaginosis.

Commun Dis Public Health, 1999 Dec, 2(4), 301 - 2
Document control and document management; Djemal KK; Most schemes for the accreditation (e.g . United Kingdom Accreditation Service) and certification (e.g . BS EN ISO 9002) of laboratories include a requirement to establish and maintain procedures for the management and control of documents generated internally . Such documents include policy statements, procedures, specifications, and some notices and memoranda . Organisations benefit from using agreed and approved information and from knowing that staff are using agreed and approved methods in their operating procedures . Document control systems are likely to become compulsory as accreditation schemes, such as Clinical Pathology Accreditation (CPA) for clinical microbiology laboratories, align with international standards . The Technical Services Division (TSD) in PHLS Headquarters has been developing a control system for various documents that it issues to the PHLS and control of documentation that forms the TSD quality system . The document control system has recently developed into a document management system that provides a mechanism for managing all documents generated or received by the division . TSD's approach is described here to provide laboratories and other organisations with ideas for how they could set up or develop their own document management system to improve accessibility to information.

Rofo, 1999 Oct, 171(4), 294 - 301
{Importance of digital thoracic radiography in the diagnosis of pulmonary infiltrates in patients with bone marrow transplantation during aplasia}; Weber C et al.; PURPOSE: Evaluation of digitized chest x-ray for the detection of pulmonary infiltrations in bone marrow transplant patients during aplasia . METHODS: Digitized chest x-rays of 40 patients (21 female, 19 male) with "Fever of unknown origin" (FUO) were evaluated concerning radiological signs of pulmonary infiltrations and correlated to clinical findings, blood chemistry, microbiology and bronchoscopy . Additionally, an individual risk profile was established . RESULTS: In 11/40 patients pulmonary infiltrations were detected in digitized chest x-rays (group 1) . 10/11 developed an infectious pulmonary infiltration . 29/40 patients developed no pulmonary infiltration (group 2) . When fever increased for the first time (initial chest x-ray) a sensitivity, specificity, positive and negative predictive value of 46%, 86%, 56%, 81% and for the chest x-rays in progress of 61%, 79% 68% and 73% was found . C-reactive protein and temperature increase occurred statistically significantly earlier (p < 0.05) in group 1 compared to group 2 . The average latency of digital chest x-rays in comparison to c-reactive protein and temperature increase was 6 days . The incidence of risk factors was significantly higher in group 1 in comparison to group 2 (p < 0.05) . CONCLUSION: Digitized chest x-rays are not a reliable method for primary detection of pulmonary infiltrations after bone marrow transplantation . Individual risk factors have to be taken into consideration to indicate further diagnostic methods such as computed tomography at an earlier time.

J Hand Surg {Br}, 1999 Oct, 24(5), 625 - 7
Histoplasmosis of the wrist; Schasfoort RA et al.; We present a case of synovitis of the wrist due to histoplasmosis, diagnosed only after extensive surgery and culturing . Treatment with amphotericin B in combination with radical surgery was effective in curing the disease . This manifestation was probably an exacerbation of a latent chronic infection with Histoplasma capsulatum, although it was unclear why the exacerbation occurred . Synovitis resistant to treatment should be assessed with great care, especially in view of the growing number of immunocompromised patients . Close collaboration between surgeon, rheumatologist, pathologist and microbiologist is paramount in such cases.

Arch Surg, 1999 Dec, 134(12), 1300 - 7; discussion 1307-8
Waiting for microbiologic data to direct therapy against nosocomial infections in febrile surgical patients: are outcomes worsened?
Pelletier SJ, Crabtree TD, Gleason TG, Banas LE, Patel SR, Pruett TL, Sawyer RG.
HYPOTHESIS: Allowing adequate time for laboratory and culture results before initial treatment may be associated with a worse outcome in nosocomial infections . DESIGN: Cohort study of all episodes of nosocomial infection from December 10, 1996, to October 28, 1998 . SETTING: Surgical services at a university hospital . PATIENTS AND METHODS: In surgical patients presenting with fever, 372 episodes of nosocomial infection were evaluated . Nosocomial infections were divided by time from fever to intervention (< or =12, 13-24, and >24 hours) . These groups were subdivided by Acute Physiology and Chronic Health Evaluation II (APACHE II) scores into low (< or =10 {n = 114}), moderate (11-20 {n = 169}), and high severity of illness (>20 {n = 89}) . Pneumonia and bloodstream infections were divided by APACHE II scores into low (< or =15 {n = 55 and n = 56, respectively}) or high severity of illness (>15 {n = 84 and n = 77, respectively}) . MAIN OUTCOME MEASURES: Mortality, length of stay . RESULTS: No difference in outcome was seen between different time intervals from fever to intervention for nosocomial infections in patients with APACHE II scores of no more than 10 . Patients treated more than 24 hours after fever were significantly younger than those treated at no more than 12 and 13 to 24 hours with APACHE II scores of 11 to 20 (P<.05) and more than 20 (P<.05) . Mortality and length of stay for patients treated at later time intervals were comparable with those of patients treated earlier with similar APACHE II scores . There was no difference in outcome for patients with pneumonia or bloodstream infection . CONCLUSIONS: Episodes of infection in which treatment was withheld until initial microbiologic data were available (24 hours) did not have worse outcomes compared with those treated earlier . Waiting for laboratory and culture results to direct antibiotic therapy for nosocomial infections does not appear harmful and may be potentially beneficial.

Vet Microbiol, 1999 Oct, 70(1-2), 1 - 6
Examination of adenovirus-types in intestinal vascular endothelial inclusions in fatal cases of enteric disease in cattle, by in situ hybridisation; Smyth JA et al.; There have been occasional reports in the literature of a severe naturally occurring enteric disease of cattle in which adenoviral inclusions were found in intestinal vascular endothelium . Bovine adenovirus type 10 (BAV-10) was identified by in situ hybridisation (ISH) in the inclusions of all 13 such cattle detected in Northern Ireland {Smyth, J.A., Benko, M., Moffett, D.A., Harrach, B., 1996 . J . Clin . Microbiol . 34, 1270-1274} . The present paper describes ISH examination of the vascular endothelial inclusion bodies in a further seventeen cattle with enteric disease, from Canada, Great Britain, Italy and The Netherlands . BAV-10 was identified in the inclusions of ten cases, and Subgroup 2 BAVs in six cases . BAV-10 was identified in cattle from Canada, The Netherlands and Great Britain . This is the first recognition of BAV-10 infection outside Northern Ireland and New Zealand . The results also show that at least two adenovirus serotypes may induce inclusion bodies in intestinal vascular endothelium of cattle . It will, therefore, be difficult in the short term to develop a simple test to allow diagnosis of this form of adenoviral infection in living animals, and thus to determine the relationship between it and clinical disease.

Rinsho Byori, 1999 Nov, 47(11), 997 - 1004
{Genetic examination in clinical laboratory}; Tozuka M et al.; Genetic technology is finding active application today in the field of clinical laboratory medicine . Genetic examinations are divided into following three main classes: 1) examination for infectious disease according to the detection of the gene derived from bacteria or viruses, 2) examination for inherited disease according to molecular analysis of the genetic variation, 3) examination for oncogene according to molecular analysis of genetic abnormalities . At present, the main genetic examination in a large number of laboratories is for infectious disease because of its relatively simplified technique and high demand . The division of genetics is not a new independent section of clinical laboratory, but rather an ultramodern and powerful tool for existing divisions, such as biochemistry, serology, hematology, microbiology, and pathology . Genetic technology quickly provides results with high sensitivity and reliability, and plays a role at the core of the clinical laboratory . We should remember that the genetic technology is a great present given to clinical laboratories, however, it will eventually change into only one of the routine examinations according to the method of used . Examinations utilized in the clinical laboratory must be well established and standardized . Genetic examinations are no exception to that rule . These tests require a remarkably high precision since the results have an extraordinarily important meaning . There are more than 8,000 inherited diseases for instance . It is difficult to cover all examinations for those 8,000 in one laboratory . We need a network of laboratories that possess a genetic division, so that the examinations for as many inherited disease as possible can be comprehensively offered.

J Periodontol, 1999 Nov, 70(11), 1322 - 9
Longitudinal study of dental implants in a periodontally compromised population; Sbordone L et al.; BACKGROUND: The purpose of this longitudinal study was to determine the clinical status and the composition of the subgingival microbiota of dental implants and natural teeth in patients with a history of periodontitis . METHODS: Twenty-five partially edentulous patients treated for moderate to advanced adult periodontitis and having a total of 42 implants participated in this 3-year study . The assessment of clinical status was done 1, 2, and 3 years after prosthetic loading (T1, T2, and T3, respectively) . Clinical parameters evaluated included probing depth (PD), clinical attachment level (CAL), gingival index (GI), and plaque index (PI) . The subgingival microbiota at peri-implant and periodontal sites were analyzed at T1 and T2 . RESULTS: No significant difference in clinical parameters between implants and teeth and within the 2 groups between different time points was observed through the study . PD and CAL measurements of sampled periodontal and peri-implant sites did not show any statistically significant difference through the study and between the 2 groups . PI of sampled periodontal sites showed a statistically significant improvement during the study . From the morphological observation of the subgingival microbiota, a significant difference in the composition of motile rods between implants and teeth was found at T1 . There were no differences detected in the subgingival microbiota, culturally identified at peri-implant and periodontal sites for the duration of the study . CONCLUSIONS: In conclusion, implants were colonized by the indigenous periodontal microbiota and were well maintained in patients with a history of periodontitis . No significant association between progressing or non-progressing periodontal or peri-implant sampled sites in terms of loss of attachment and infection with at least one of the searched periodontal pathogens was found, suggesting that the presence of putative periodontopathogens at peri-implant and periodontal sites may not be associated with future attachment loss or implant failure.

Head Neck, 2000 Jan, 22(1), 87 - 9
Malignant otitis externa caused by Malassezia sympodialis; Chai FC et al.; BACKGROUND: Malignant otitis externa caused by fungal infections is rare . A review of the literature showed only 9 cases, and the causative fungus in all cases was Aspergillus . This article reports an unusual case caused by Malassezia sympodialis . METHODS: A 53-year-old man with non-insulin dependent diabetes presented with malignant otitis externa . He deteriorated despite treatment with intravenous antipseudomonal therapy and surgical debridement . Microbiologic tests revealed M . sympodialis . He responded rapidly to intravenous amphotericin . RESULTS: Systemic human infections caused by M . sympodialis have not been reported . M . furfur systemic infection is rare and has been associated lipid hyperalimentation by means of a central catheter . Only 1 other case of M . fungemia without these associated risk factors has been reported . CONCLUSIONS: The first case of malignant otitis externa caused by M . sympodialis is presented . It highlights the difficulty of initial biologic diagnosis and the need for lipid-enriched media to grow this fastidious organism .

Immunol Rev, 1999 Oct, 171, 45 - 87
The B-cell system of human mucosae and exocrine glands; Brandtzaeg P et al.; The mucosae and exocrine glands harbour the largest activated B-cell system of the body, amounting to some 80-90% of all immunoglobulin (Ig)-producing cells . The major product of these immunocytes is polymeric (p)IgA (mainly dimers) with associated J chain . Both pIgA and pentameric IgM contain a binding site for the polymeric Ig receptor (pIgR), or secretory component (SC), which is a requirement for their active external transport through secretory epithelia . The pIgR/SC binding site depends on covalent incorporation of the J chain into the quaternary structure of the polymers when they are produced by the local immunocytes . This important differentiation characteristic appears to be sufficient functional justification for the J chain to be expressed also by most B cells terminating at secretory effector sites with IgD or IgG production; they probably represent a "spin-off" from sequential downstream CH switching on its way to pIgA expression, thus apparently reflecting a maturational stage of effector B-cell clones compatible with homing to these sites . Observations in IgA-deficient individuals suggest that the magnitude of this homing is fairly well maintained even when the differentiation pathway to IgA is blocked . Certain microenvironmental elements such as specific cytokines and dendritic cells appear to be required for induction of IgA synthesis, but it remains virtually unknown why this isotype normally is such a dominating product of local immunocytes and why they have such a high level of J chain expression . Also, despite the recent identification of some important requirements in terms of adhesion molecules (e.g . integrin alpha 4 beta 7 and MAdCAM-1) that explain the "gut-seeking" properties of enterically induced B cells, the origin of regionalized homing of B cells to secretory effector sites outside the gut remains elusive . Moreover, little is known about immune regulation underlying the striking disparity of both the class (IgD, IgM) and subclass (IgA1, IgA2, IgG1, IgG2) production patterns shown by local immunocytes in various regions of the body, although the topical microbiota and other environmental stimuli might be important . Rational design of local vaccines will depend on better knowledge of both inductive and migratory properties of human mucosal B cells.

Appl Environ Microbiol, 1999 Dec, 65(12), 5207 - 11
Cloning, overexpression, and mutagenesis of the Sporobolomyces salmonicolor AKU4429 gene encoding a new aldehyde reductase, which catalyzes the stereoselective reduction of ethyl 4-chloro-3-oxobutanoate to ethyl (S)-4-chloro-3-hydroxybutanoate; Kita K et al.; We cloned and sequenced the gene encoding an NADPH-dependent aldehyde reductase (ARII) in Sporobolomyces salmonicolor AKU4429, which reduces ethyl 4-chloro-3-oxobutanoate (4-COBE) to ethyl (S)-4-chloro-3-hydroxybutanoate . The ARII gene is 1,032 bp long, is interrupted by four introns, and encodes a 37,315-Da polypeptide . The deduced amino acid sequence exhibited significant levels of similarity to the amino acid sequences of members of the mammalian 3beta-hydroxysteroid dehydrogenase-plant dihydroflavonol 4-reductase superfamily but not to the amino acid sequences of members of the aldo-keto reductase superfamily or to the amino acid sequence of an aldehyde reductase previously isolated from the same organism (K . Kita, K . Matsuzaki, T . Hashimoto, H . Yanase, N . Kato, M . C.-M . Chung, M . Kataoka, and S . Shimizu, Appl . Environ . Microbiol . 62:2303-2310, 1996) . The ARII protein was overproduced in Escherichia coli about 2, 000-fold compared to the production in the original yeast cells . The enzyme expressed in E . coli was purified to homogeneity and had the same catalytic properties as ARII purified from S . salmonicolor . To examine the contribution of the dinucleotide-binding motif G(19)-X-X-G(22)-X-X-A(25), which is located in the N-terminal region, during ARII catalysis, we replaced three amino acid residues in the motif and purified the resulting mutant enzymes . Substrate inhibition of the G(19)-->A and G(22)-->A mutant enzymes by 4-COBE did not occur . The A(25)-->G mutant enzyme could reduce 4-COBE when NADPH was replaced by an equimolar concentration of NADH.

Virus Res, 1999 Dec 15, 65(2), 121 - 9
Genetic diversity of GBV-C/HGV strains among HIV infected-IVDU and blood donors from Buenos Aires, Argentina; Oubina JR et al.; GBV-C/HGV RNA was investigated in serum samples from 70 HIV(+) intravenous drug users (IVDU), as well as from 200 blood donors from Buenos Aires, Argentina . Viral RNA was demonstrated in 21 IVDU by reverse transcription-nested PCR of the 5' UTR . c-DNA amplified products were analyzed and their sequences compared with those downloaded from GenBank . A phylogenetic tree based on 171 sequences demonstrated the presence of three major genogroups, including two subgroups, within local samples, i.e . group 1 (n=1), 2a (n=11), 2b (n=4) and 3 (n=5) . These results agreed entirely with those obtained by a novel RFLP (J . Clin . Microbiol . 37, 1340-1347, 1999) of the same 5' UTR amplicons . As expected, GBV-C/HGV RNA prevalence was significantly higher among IVDU than among blood donors (P<0.0001), although within the latter group an unexpectedly high rate was also detected, since 11 of 200 sera (5.5%) proved positive . These viral isolates were ascribed either to subgroup 2a (n=5), subgroup 2b (n=5) or genogroup 3 (n=1) . Briefly, this partial view of GBV-C/HGV molecular epidemiology in Argentina shows: (i) different rates of GBV-C/HGV infection within both IVDU and blood donors; (ii) a high prevalence of viral RNA among blood donors; and (iii) a predominant circulation of genogroup 2, with minor contribution of groups 3 and 1.

Rev Enferm, 1999 Sep, 22(9), 571 - 8
{Nursing and the prevention of biological risks: from education to professional activity}; Plitt Gomez C et al.; OBJECTIVE: To discover the training nursing students receive regarding prevention and control of infection inside hospital settings and the means of self-protection as well as making people aware of the need to improve this training if deemed necessary . METHODS: This is a transversal descriptive study which took place in the nursing schools in Spain . Data was gathered by means of a questionnaire each respondent filled in by themselves . The lack of a response was also studied . A followup meeting was held with those who participated in this study to seek consensus in the search for solutions to those necessary changes detected . RESULTS: 86% responded to the questionnaire . During the course of career preparation, these were the average number of hours dedicated to various areas of study: 34.5 to Biological Statistics, 40 to Epidemiology, 28 to Infectious Diseases, 29 to Microbiology, 9 to Infection Control, 11 to Concepts of Aspesis and Antisepsis, and 36 to Methods of Research and Investigation . More than 50% of the professors teaching these classes are licensed nurses . Student followup during their clinical practice was carried out basically by nurses in university hospitals . 39% of nursing schools teach the concept and content for universal precautions prior to clinical practice . In the majority of public nursing schools, the followup of a student after a biological accident is carried out by the preventive medicine service, whereas this is done by the school insurance company in private nursing schools . Therefore, significant differences exist . COMMENTS: The theoretical knowledge taught is adequate; the number of hours dedicated to each subject varies among the schools; it is necessary to come to an agreement on the minimum levels of training required before engaging in clinical practice in a safe manner . Universal precautions should be taught in all schools before clinical practice occurs.

Am J Ophthalmol, 1999 Oct, 128(4), 511 - 2
Identification of bacterial pathogens in patients with endophthalmitis by 16S ribosomal DNA typing; Knox CM et al.; PURPOSE: To determine whether sequence analysis of 16S ribosomal DNA (rDNA) can be used to detect bacterial pathogens in patients with postoperative endophthalmitis . METHODS: In 10 eyes of 10 patients, vitreous specimens were collected for culture and rDNA typing . Variable segments of each ribosomal DNA specimen were amplified by polymerase chain reaction (PCR), sequenced, and aligned by BLAST, a computer alignment program, against sequences in GenBank at the National Institutes of Health . RESULTS: Specimens were available from five eyes with bacterial endophthalmitis diagnosed by Gram stain or culture . Amplified 16s rDNA sequences from the eyes of three patients were identical to microbiologic results . Polymerase chain reaction results were negative in two cases in which unusual organisms were detected . All five control specimens from patients with nonbacterial endophthalmitis or uveitis were PCR negative . Approximately 48 to 72 hours are required under ideal conditions for final species identification with this ribosomal typing technique . CONCLUSIONS: 16S rDNA typing shows potential as a relatively rapid technique for identifying bacteria in vitreous samples.

Biochimie, 1999 Aug-Sep, 81(8-9), 921 - 9
Division-associated changes in membrane viscosity of Escherichia coli; Binenbaum Z et al.; Septum formation is initiated by the FtsZ ring assembly in the middle of rod-shape bacteria . The mechanism which determines the division site in the membrane and makes it recognizable by FtsZ is still unknown . We have recently demonstrated that the putative division membrane domains can be visualized by a fluorescent membrane probe (Fishov and Woldring, Mol . Microbiol., 1999) and that these domains can be dissipated by interrupting the process of coupled transcription and translation of proteins (Binenbaum et al., Mol . Microbiol., 1999) . Here, we examined the membrane dynamics of Escherichia coli during division and after a reversible division arrest . Anisotropy of DPH fluorescence, used as an indicator of membrane dynamics (viscosity), correlated with the rate of division in synchronous cells . It decreased during filamentation caused by drugs or by temperature, but not in the ftsZ mutant and when DNA replication was blocked by nalidixic acid . Based on previous data, we incline to interpret these results as reflecting formation and dissipation of putative membrane domains marking the division sites; domains are formed by partitioning nucleoids and dissipate while used for constriction or after the nucleoids have been segregated too far in a filament.

Biochimie, 1999 Aug-Sep, 81(8-9), 897 - 900
Visualizing multiple constrictions in spheroidal Escherichia coli cells; Zaritsky A et al.; An Escherichia coli cell grows by elongation and divides in a perpendicular plane . Alternating planes of successive divisions in three dimensions can only be ascertained when multiple constrictions exist simultaneously in large, spheroidal cells (with extended constriction process), if the division signals are enhanced . Large, spheroidal cells are obtained by a brief mecillinam treatment, and more frequent divisions are achieved by manipulating the rate of chromosome replication without affecting cell mass growth rate . Such a procedure has recently been performed by thymine-limitation of E . coli K12 strain CR34 (Zaritsky et al., Microbiology 145 (1999), 1052-1022) . Enhancing the replication rate in cells with multi-forked replicating chromosomes (by addition of deoxyguanosine) shortens the intervals between successive terminations and thus triggers divisions more frequently . Monoclonal antibodies against FtsZ were used to visualize the rings of secondary constrictions, but apparent shortage of FtsZ to complete rings over wide cells allowed assembly of arcs only . The arcs observed were not parallel nor perpendicular; the tilted constriction planes are consistent with our 3-D 'nucleoid segregation'model for division under conditions which relieve the cylindrical constraint for nucleoid segregation by the bacillari peptidoglycan sacculus (Woldringh et al . , J . Bacteriol . 176 (1994) 6030-6038) . The shortage in FtsZ may explain the longer time required to complete the division process in wide cells with long circumferences, observed during thymine step-up . Overexpression of fusion protein FtsZ-GFP on a multi-copy plasmid should circumvent the shortage.

FEBS Lett, 1999 Oct 22, 460(1), 139 - 44
Sequencing, expression and biochemical characterization of the Porphyromonas gingivalis pepO gene encoding a protein homologous to human endothelin-converting enzyme; Awano S et al.; We have determined the nucleotide sequence of the clone pAL2 obtained from Porphyromonas gingivalis 381 in the previous study {Ansai et al . (1995) Microbiology 141, 2047-20521 . The DNA sequence analysis of this fragment revealed one complete ORF and one incomplete ORF . The ORF encoded a protein (PgPepO) of 690 amino acids with a calculated molecular weight of 78796 . The deduced amino acid sequence exhibited a significant homology with human endothelin-converting enzyme (ECE)-1 . Recombinant PgPepO was purified to homogeneity and characterized . The purified enzyme was strongly inhibited by phosphoramidon, and converted big endothelin-1 to endothelin-1 . Furthermore, the purified PgPepO strongly cross-reacted with a monoclonal antibody against rat ECE-1 . These results indicate that PgPepO has striking similarity to mammalian ECE in structure and function.

Appl Microbiol Biotechnol, 1999 Oct, 52(4), 464 - 73
Bacterial magnetosomes: microbiology, biomineralization and biotechnological applications; Schuler D et al.; Magnetotactic bacteria orient and migrate along geomagnetic field lines . This ability is based on intracellular magnetic structures, the magnetosomes, which comprise nanometer-sized, membrane-bound crystals of the magnetic iron minerals magnetite (Fe3O4) or greigite (Fe3S4) . Magnetosome formation is achieved by a mineralization process with biological control over the accumulation of iron and the deposition of the mineral particle with specific size and orientation within a membrane vesicle at specific locations in the cell . This review focuses on the current knowledge about magnetotactic bacteria and will outline aspects of the physiology and molecular biology of the biomineralization process . Potential biotechnological applications of magnetotactic bacteria and their magnetosomes as well as perspectives for further research are discussed.

J Agric Food Chem, 1999 Apr, 47(4), 1295 - 319
Chemistry, biochemistry, nutrition, and microbiology of lysinoalanine, lanthionine, and histidinoalanine in food and other proteins; Friedman M; Heat and alkali treatments of foods, widely used in food processing, result in the formation of dehydro and cross-linked amino acids such as dehydroalanine, methyldehydroalanine, beta-aminoalanine, lysinoalanine (LAL), ornithinoalanine, histidinoalanine (HAL), phenylethylaminoalanine, lanthionine (LAN), and methyl-lanthionine present in proteins and are frequently accompanied by concurrent racemization of L-amino acid isomers to D-analogues . The mechanism of LAL formation is a two-step process: first, hydroxide ion-catalyzed elimination of H(2)S from cystine and H(2)O, phosphate, and glycosidic moieties from serine residues to yield a dehydroalanine intermediate; second, reaction of the double bond of dehydroalanine with the epsilon-NH(2) group of lysine to form LAL . Analogous elimination-addition reactions are postulated to produce the other unusual amino acids . Processing conditions that favor these transformations include high pH, temperature, and exposure time . Factors that minimize LAL formation include the presence of SH-containing amino acids, sodium sulfite, ammonia, biogenic amines, ascorbic acid, citric acid, malic acid, and glucose; dephosphorylation of O-phosphoryl esters; and acylation of epsilon-NH(2) groups of lysine . The presence of LAL residues along a protein chain decreases digestibility and nutritional quality in rodents and primates but enhances nutritional quality in ruminants . LAL has a strong affinity for copper and other metal ions and is reported to induce enlargement of nuclei of rats and mice but not of primate kidney cells . LAL, LAN, and HAL also occur naturally in certain peptide and protein antibiotics (cinnamycin, duramycin, epidermin, nisin, and subtilin) and in body organs and tissues (aorta, bone, collagen, dentin, and eye cataracts), where their formation may be a function of the aging process . These findings are not only of theoretical interest but also have practical implications for nutrition, food safety, and health . Further research needs are suggested for each of these categories . These overlapping aspects are discussed in terms of general concepts for a better understanding of the impact of LAL and related compounds in the diet . Such an understanding can lead to improvement in food quality and safety, nutrition, microbiology, and human health.

Ceska Gynekol, 1999 Jul, 64(4), 238 - 41
{Evaluation of the role of the HPV test in routine clinical practice: prospective study}; Holub Z et al.; OBJECTIVE: To determine the incidence of papillomavirus and to asses the role a HPV DNA in screening of cervical intraepithelial neoplasia . DESIGN: Czech National Collaborative Trial . SETTING: Department of Gynaecology and Obstetrics, Kladno Hospital, Department of Microbiology, KLINLAB, Prague, outpatient clinics (Kladno, Bruntal, Unhost') . METHODS: The study group includes of 3,264 women, their cervical scrapings were sent to KLINLAB during November 1996 to June 1998, because of abnormal results of primary screening . Patients with ASCUS, AGNUS, LSIL, HSIL and/or abnormal colposcopic findings were eligible for this study . Only completely filled in records of a subgroup of 1,158 patients were assessed for sensitivity and specificity for CIN in screening methods . RESULTS: The overall sensitivity of cytology and basic colposcopy were 84.7% and 64.9% respectively . The overall sensitivity of HPV DNA testing was 66% . The sensitivity of cytology of women with minor cytological abnormities for major cervical disease (CIN 2/3 and microinvasive cancer) in this group of patients was only 35% . Combined cytology with HPV DNA test gave an increased sensitivity for major cervical disease of 83% . CONCLUSION: 153 (66.4%) of the cervical intraepithelial neoplasias grade 2/3 displayed negative and borderline or mild cytological changes . The combined use of cytology with HPV testing increased the sensitivity with a reduction of specificity . However, it is evident from these results that HPV DNA testing alone cannot be used as a diagnostic tool but could usefully improve the level of screening CIN in common clinical practice.

J Gynecol Obstet Biol Reprod (Paris), 1999 Sep, 28(5), 425 - 32
{Contribution of ultrasonography-guided microbiopsy in breast diseases}; Escolano E et al.; OBJECTIVE: To assess the usefulness of core biopsy under ultrasonography (CBUS) . MATERIALS AND METHODS: 165 US guided breast lesions biopsies were reviewed . Forty-eight underwent surgery and the 117 remaining cases were followed up for at least one year . Forty-four lesions were malignant and 89 were no palpable . Lesion size ranged from 3 to 35 mm (mean 14.8 mm) . Core biopsy was performed with an automatic device with a 18 Gauge needle using a long-throw (2.2 cm excursion) . Needle length was 10 cm . At least two passes were performed . RESULTS: Five false negatives were obtained which were all explained . All the infiltrative carcinomas without microcalcifications as mammographic sign were correctly diagnosed . A resolutive hematoma was the only complication to occur . None of the lesions with follow up showed any evolution . With CBUS, sensitivity for malignancy was 91% and specificity 100% . This is in agreement with previous studies . CONCLUSION: Methodology and indications of such core biopsies are peculiar . Actually, the lesion must be seen with ultrasonography and should not be superficial, be correlated with microcalcifications, or be a cyst even with a thick content . This method is safe and comfortable . It is not expensive when compared with surgery or stereotaxy . It should avoid useless surgical open biopsies as it allows histologic diagnosis with Scarff and Bloom grading and hormonal receptor status . So, US guided core-biopsy should contribute to increase the predictive positive value of surgical biopsy in senologic screening.

J Bacteriol, 1999 Nov, 181(22), 7115 - 25
Two new early bacteriophage T4 genes, repEA and repEB, that are important for DNA replication initiated from origin E; Vaiskunaite R et al.; Two new, small, early bacteriophage T4 genes, repEA and repEB, located within the origin E (oriE) region of T4 DNA replication, affect functioning of this origin . An important and unusual property of the oriE region is that it is transcribed at early and late periods after infection, but in opposite directions (from complementary DNA strands) . The early transcripts are mRNAs for RepEA and RepEB proteins, and they can serve as primers for leading-strand DNA synthesis . The late transcripts, which are genuine antisense RNAs for the early transcripts, direct synthesis of virion components . Because the T4 genome contains several origins, and because recombination can bypass a primase requirement for retrograde synthesis, neither defects in a single origin nor primase deficiencies are lethal in T4 (Mosig et al., FEMS Microbiol . Rev . 17:83-98, 1995) . Therefore, repEA and repEB were expected and found to be important for T4 DNA replication only when activities of other origins were reduced . To investigate the in vivo roles of the two repE genes, we constructed nonsense mutations in each of them and combined them with the motA mutation sip1 that greatly reduces initiation from other origins . As expected, T4 DNA synthesis and progeny production were severely reduced in the double mutants as compared with the single motA mutant, but early transcription of oriE was reduced neither in the motA nor in the repE mutants . Moreover, residual DNA replication and growth of the double mutants were different at different temperatures, suggesting different functions for repEA and repEB . We surmise that the different structures and protein requirements for functioning of the different origins enhance the flexibility of T4 to adapt to varied growth conditions, and we expect that different origins in other organisms with multiorigin chromosomes might differ in structure and function for similar reasons.

Biotech Histochem, 1999 Jul, 74(4), 194 - 212
Ruthenium red and the bacterial glycocalyx; Fassel TA et al.; Ruthenium red, a promising cationic reagent for electron microscopy (EM), has long been an important tool in histology . The reagent was initially used by botanists as a semispecific stain for pectic substances, but it has gradually been embraced by investigators in microbiology and the animal sciences as a stain for anionic glycosylated polymeric substances . Luft developed a reliable method and demonstrated that ruthenium red was a useful reagent for visualizing ultrastructural detail . Many investigators, using modifications of Luft's approach, have identified numerous applications for this important reagent . Ruthenium red has been used to show the ultrastructural detail of bacterial glycocalyces . Strong, sharp and consistent observations of this ultrastructural component of the bacterial cell have given a better understanding its fibrous anionic matrix . Any variations in staining owing to artifactual alteration of the fine delicate ultrastructural features have been overcome by incorporation of diamine lysine into ruthenium red methods, thus providing flexible processing times under less than ideal laboratory sampling conditions . Ruthenium red has broad utility in the biological sciences, and in combination with lysine, it is an excellent EM stain for enhanced visualization of bacterial glycocalyx from culture or from clinical specimens.

Pharmacotherapy, 1999 Nov, 19(11), 1252 - 60
Pharmacodynamic characterization of nephrotoxicity associated with once-daily aminoglycoside; Murry KR et al.; STUDY OBJECTIVE: To characterize nephrotoxicity associated with an individualized serum concentration target-specific, once-daily aminoglycoside (ODA) program . DESIGN: Concurrent and retrospective study . SETTING: University-affiliated trauma hospital . PATIENTS: Two hundred patients treated with ODA and 100 treated with individualized traditional dosing (TDA) . INTERVENTIONS: Empiric dosing for both groups was based on patient-specific pharmacokinetics and severity of infection . Regimens were modified according to predetermined target maximum and minimum serum concentrations for both groups . MEASUREMENTS AND MAIN RESULTS: Nephrotoxicity occurred in 7.5% patients treated with ODA and 14.7% receiving TDA (p=0.05) . Minimum serum concentrations, length of aminoglycoside therapy, and cumulative area under the curve (AUC) were all dependently related to nephrotoxicity, and concomitant vancomycin and other nephrotoxic drugs were independently related to the disorder . The cumulative AUC was greatest in patients receiving TDA (p=0.03), and the modeled probability of becoming toxic at any given cumulative AUC was significantly greater with TDA than with ODA (p<0.01) . Clinical and microbiologic outcomes were similar between groups . Maximum concentration:minimum inhibitory concentration ratios were higher (p<0.01) and number of days to organism eradication was shorter in the ODA group (p=0.04) . CONCLUSION: The trend was toward decreased nephrotoxicity in patients treated with ODA compared with TDA, and at any given cumulative AUC, the risk of toxicity was lower for ODA.

Scand J Prim Health Care, 1999 Sep, 17(3), 185 - 90
Long-lasting improvement in general practitioners' prescribing of antibiotics by means of medical audit; Munck AP et al.; OBJECTIVE: To evaluate the usefulness of medical audit of GPs' antibiotic prescription habits . DESIGN: Medical audit according to the APO method . Registration of antibiotic prescriptions for respiratory tract infections during a 3 year period . Intervention with courses, visits to the laboratory, and distribution of recommendations concerning diagnosis and treatment of respiratory infections . SETTING: 24 Danish GPs in cooperation with Audit Project Odense (APO) and Department of Clinical Microbiology, Odense University Hospital . 207 GPs acted as controls . MAIN OUTCOME MEASURES: Changes in the number of antibiotic prescriptions and in the penicillin/broad-spectrum antibiotic ratio . RESULTS: The proportion of antibiotic prescriptions was reduced during the investigation period, but a similar reduction was found in the control groups . Only for acute sinusitis was a lasting decrease not found in the control groups recorded . The penicillin/broad-spectrum antibiotic ratio increased in the intervention group (1.33 in 1992, 1.94 in 1993 and 2.70 in 1995) . This increase was significantly higher than in the control groups . The change was seen for acute sinusitis, bronchitis, and pneumonia, but not for acute otitis media or acute tonsillitis . The changes induced from 1992 to 1993 were maintained or increased from 1993 to 1995 although the educational measures had stopped . CONCLUSION: Medical audit according to the APO method is a useful tool for inducing and maintaining desirable changes in GPs' prescription habits.

Mol Biotechnol, 1999 Aug, 12(1), 75 - 99
Application of nucleic acid amplification in clinical microbiology; Lisby G; The use of nucleic acid amplification methods in routine clinical microbiology laboratories is becoming increasingly widespread . The theory of polymerase chain reaction is described, including discussion of suitable microbal targets, extraction of nucleic acid from clinical samples, choice of primers, optimization of the process, laboratory design, contamination, and other problems as well as quality control . Other nucleic acid amplification methods such as ligase chain reaction, self-sustained sequence replication, strand displacement amplification, and branched DNA signal amplification are described and the choice of technology is discussed.

Oral Microbiol Immunol, 1999 Aug, 14(4), 257 - 65
Identification of oral Actinomyces species using DNA probes; Ximenez-Fyvie LA et al.; Oral Actinomyces comprise a major segment of both the supra- and subgingival microbiota; however, little is known about the distribution of individual species in different sites or clinical conditions . The purpose of the present investigation was to develop DNA probes for suggested species and genotypes of oral Actinomyces . Whole genomic DNA probes to 12 human oral species and/or serotypes were labeled with digoxigenin and used to seek cross-reactions among the taxa using the checkerboard DNA-DNA hybridization assay . The Actinomyces formed three distinct groups: 1) Actinomyces georgiae, Actinomyces meyeri and Actinomyces odontolyticus serotypes I and II; 2) Actinomyces viscosus and Actinomyces naeslundii serotypes I, II, III and WVA 963; and 3) Actinomyces gerencseriae and Actinomyces israelii . Cross-reactions among taxa were detected and minimized by increasing the temperature of the post-hybridization high-stringency wash to 80 degrees C . Despite the elevation in high stringency wash temperature, cross-reactions among strains of the A . naeslundii/A . viscosus group persisted . Probes for two of the three currently recognized genospecies in this group were prepared by removing the DNA in common between cross-reacting species using subtraction hybridization and polymerase chain reaction . Nine species and genospecies could be clearly separated by a combination of whole genomic and subtraction hybridization probes and by increasing the high-stringency wash temperature . A total of 195 fresh isolates of Actinomyces were grouped in a blind study using DNA probes and separately by SDS-PAGE protein profiles . Concordance between the two methods was 97.3% . The probes and hybridization conditions were tested for their ability to detect the Actinomyces species and genospecies in samples of supragingival and subgingival plaque from periodontitis subjects using checkerboard DNA-DNA hybridization . The probes detected the species in samples of supragingival and subgingival plaque . We concluded that whole genomic and subtraction hybridization DNA probes facilitate the detection and enumeration of species and genospecies of Actinomyces in plaque samples.

Neurosurgery, 1999 Nov, 45(5), 1025 - 92
A legacy of microneurosurgery: memoirs, lessons, and axioms; Yasargil MG; This article is written at the request of the editor . It contains my autobiographical sketch, professional memories, lessons, axioms, and reflections on the present problems in neurodiagnosis and neurotherapy . The combination of microsurgical techniques, the bipolar coagulation technique, the concept of arachnoidal exploration, and the concept of segmental and compartmental occurrence of vascular and neoplastic lesions of the central nervous system, with their predilection sites, allowed microneurosurgery to gradually unfold and proceed within the last 30 years as a continuation of conventional neurosurgical principles established by the founder generation . Today, the lesions in each region of the central nervous system can be accessed without using computer-assisted targeting and navigation technology and can be selectively eliminated ("pure lesionectomy") with acceptable outcomes; the mortality and morbidity rates have been reduced remarkably . Further scientific and technological advances will promote the ongoing evolution in neurodiagnosis and neurotherapy . Competitive neurospecialties are welcomed in the interest of patients, medical sciences, and surgical advances . The younger generation of neurosurgeons will have spent more time in laboratory training, deepening their knowledge of neuroanatomy and gaining experience in surgical techniques . The achievements, limits, and problems of neurosurgery in relation to technology, medical and surgical standards, and controversial treatment options have been presented thoroughly in numerous professional publications . However, the relationship of neurosurgery to the evolution of integral neurophysiology and biochemistry has hitherto been inadequately evaluated . The advances in microbiology, anesthesiology, and topographic neurology have been viewed as essential components of neurosurgery's foundations . A critical analysis proves that this is only partially true . The turning point in the development from craniospinal surgery to physiological neurosurgery began with the research of Th . Kocher, V . Horsley, H . Cushing, and W . Dandy concerning the importance of the cerebrospinal fluid system . This was the first step in a trend toward integral neurophysiology, which initiated neurosurgical procedures on a routine basis . The intensive research on the hypothalamus by R.W . Hess and associates led to intensified studies on the autoregulated integral functional units of the central nervous system ("dynamic homeostasis," in the words of W.B . Cannon) . This slowly developing but exciting history of neurophysiology requires patient study to seek out solutions for the present difficulties in neurodiagnosis and neurotherapy, which constitute a similar situation to that encountered by the pioneer surgeons at the end of the last century . In pertinent sections, my personal opinions relating to observations and experiences with a large number of operated patients with vascular and neoplastic lesions are presented . The predilection sites of brain tumors in the neopallial and paleopallial (limbic-paralimbic) areas and brainstem, and their expansive but usually not infiltrative growth, are discussed and documented . The current hypothesis of infiltrative growth of gliomas is opposed . The microsurgical technique for the treatment of various types of lesions is summarized . The principal microsurgical instruments and apparatus are presented with some remarks relating to their conception and manufacture.

J Hosp Infect, 1999 Oct, 43(2), 123 - 9
A study of infection in elderly nursing/residential home and community-based residents; Yates M et al.; It is commonly believed that patients admitted to hospital from nursing homes/residential homes (NHRH) with infections are less likely to respond to treatment and have a higher fatality rate than counterparts admitted from their own homes ('the Community') . It is also believed that NHRH's harbour a reservoir of unusual and resistant organisms . These preconceptions may influence how these patients are managed . A database of 10593 sequential admissions to a Geriatric Medical unit over a three-year period was used to identify NHRH and community populations with a principal diagnosis of infection . They were investigated using the Department of Microbiology's database . The admission rate in the NHRH group was twice that of the community group . There were no significant differences in length of stay (LOS) {16 +/- 2 vs 17 +/- 2 days (s.e.m.)}, or mean survival time (ST)(61 days (37-84) vs 48 days (25-72): 95% confidence intervals) between the two groups . Subgroups of the NHRH group did have significantly different survival times . Fatality rate was not significantly different between the NHRH (40%) or Community (35%) groups . Both the NHRH and community group underwent very similar levels of investigation (189 vs 200 investigations performed) . The types and frequencies of pathogen seen in the two groups were very similar.

J Clin Virol, 1999 Sep, 14(1), 17 - 23
Problems with biotin-labelled virions as probes in poliovirus-specific mu-capture-IgM assays; Valtanen S et al.; BACKGROUND: We have previously developed a mu-capture-based radioimmunoassay (RIA) for detecting virus-specific IgM for the diagnosis of poliomyelitis . To probe captured IgM we used radiolabelled, purified preparations of representatives of each poliovirus serotype (Roivainen M, Agboatwalla M, Stenvik M, Rysa T, Akram DS, Hovi T . J Clin Microbiol 1993;31:2427-32) . However, this assay is not directly applicable for wider use because preparation and handling of radioactive reagents is cumbersome and potentially hazardous . OBJECTIVES: To develop a non-radioactive modification of the assay retaining the number of steps and reagents to a minimum . STUDY DESIGN: Replacement of radioactive labelling by in vitro biotinylation of purified virions, and detection of bound virions with horseradish peroxidase-conjugated streptavidin . To study sensitivity and poliovirus serotype-specificity, 129 sera and 115 CSF specimens from children with acute poliomyelitis were used in comparative tests with the in-house RIA . In addition, sera from 40 healthy adults and 11 paired sera from patients with non-polio enterovirus infection were used to assess specificity . RESULTS: While results with the new test on specimens from clinically confirmed polio patients revealed some correlation with those obtained in the in-house RIA, studies on sera from healthy adults indicated, that non-specific binding of biotinylated virions is difficult to control . Moreover, examination of sera from patients with non-polio enterovirus infection suggested frequently occurring cross-reactivity between immune responses induced by polio- and other enterovirus infections . The latter were also seen in the RIA . CONCLUSION: Cross-reactive epitopes between poliovirus serotypes and between polioviruses and other enteroviruses may compromise the use of an assay for virus-specific IgM for poliovirus diagnosis . Biotinylation of the virions seemed to aggravate these problems.

AIDS, 1999 Oct 22, 13(15), 2143 - 9
Frequent detection of HIV-1 in the gastric aspirates of neonates born to HIV-infected mothers; Mandelbrot L et al.; OBJECTIVE: To evaluate the frequency and correlates of oral route exposure of infants born to HIV-1-infected women . METHODS: A multicenter study was performed within the prospective French Perinatal Cohort Study of mother-to-child HIV transmission . Oropharyngeal and gastric aspirates from 122 neonates were studied by reverse transcriptase (RT) polymerase chain reaction (PCR) for the presence of HIV-1, as well as for standard microbiology (Gram staining and culture) . RESULTS: Aspirates from 101 neonates were analyzed by RT-PCR; 28% of these were positive for HIV RNA . Another 21 aspirates could not be tested because of PCR inhibition . The median concentration of HIV RNA in the positive aspirates was 126 copies/ml (range: 8-1270) . Detection of HIV-1 in the aspirate was significantly related to high maternal plasma-viral load, presence of blood in the aspirate, positive Gram stain or culture, episiotomy or perineal lesions, and sexually transmitted infections during the pregnancy . Most of the mothers received zidovudine prophylaxis during pregnancy and delivery . Among the six infants who were infected with HIV, three had positive aspirates . Of the three assumed to have acquired the infection intrapartum, only one had an HIV RNA-positive aspirate . CONCLUSION: Exposure of the fetus to HIV via the oral route occurs frequently, even in the presence of zidovudine prophylaxis, and is likely to be one of the mechanisms of intrapartum transmission, but not the only one.

Appl Environ Microbiol, 1999 Nov, 65(11), 5186 - 91
Epitope identification for a panel of anti-Sinorhizobium meliloti monoclonal antibodies and application to the analysis of K antigens and lipopolysaccharides from bacteroids; Reuhs BL et al.; In two published reports using monoclonal antibodies (MAbs) generated against whole cells, Olsen et al . showed that strain-specific antigens on the surface of cultured cells of Sinorhizobium meliloti were diminished or absent in the endophytic cells (bacteroids) recovered from alfalfa nodules, whereas two common antigens were not affected by bacterial differentiation (P . Olsen, M . Collins, and W . Rice, Can . J . Microbiol . 38:506-509, 1992; P . Olsen, S . Wright, M . Collins, and W . Rice, Appl . Environ . Microbiol . 60:654-661, 1994) . The nature of the antigens (i.e., the MAb epitopes), however, were not determined in those studies . For this report, the epitopes for five of the anti-S . meliloti MAbs were identified by polyacrylamide gel electrophoresis-immunoblot analyses of the polysaccharides extracted from S . meliloti and Sinorhizobium fredii . This showed that the strain-specific MAbs recognized K antigens, whereas the strain-cross-reactive MAbs recognized the lipopolysaccharide (LPS) core . The MAbs were then used in the analysis of the LPS and K antigens extracted from S . meliloti bacteroids, which had been recovered from the root nodules of alfalfa, and the results supported the findings of Olsen et al . The size range of the K antigens from bacteroids of S . meliloti NRG247 on polyacrylamide gels was altered, and the epitope was greatly diminished in abundance compared to those from the cultured cells, and no K antigens were detected in the S . meliloti NRG185 bacteroid extract . In contrast to the K antigens, the LPS core appeared to be similar in both cultured cells and bacteroids, although a higher proportion of the LPS fractionated into the organic phase during the phenol-water extraction of the bacteroid polysaccharides . Importantly, immunoblot analysis with an anti-LPS MAb showed that smooth LPS production was modified in the bacteroids.

Microbiology, 1999 Oct, 145 ( Pt 10), 2769 - 75
The relationship between critical pressure and width of gas vesicles in isolates of Planktothrix rubescens from Lake Zürich; Bright DI et al.; The mean critical collapse pressure (p(c)) of gas vesicles in 81 strains of the cyanobacterium Planktothrix rubescens from Lake Zurich, Switzerland, was bimodally distributed between a minimum of 0.86 MPa and a maximum of 1.17 MPa . Measurements were made of the cylinder diameter (d) of gas vesicles isolated from seven of the strains . The mean diameter, which varied from 48 to 61 nm, was inversely related to p(c), in keeping with the theory of strength of thin-walled rigid cylinders . These measurements extended the range of p(c)-width relationship of gas vesicles, which can be described by the expression p(c) = 461(d/nm)(-1.53) MPa . p(c) was correlated with gas vesicle genotype (see the accompanying paper by S . J . Beard, B . A . Handley, P . K . Hayes & A . E . Walsby, Microbiology 145, 2757-2768): of the 81 strains investigated, all those with the gas vesicle genotype GV2 produced gas vesicles with a mean p(c) of less than 1.0 MPa, whereas those of GV3 had a mean p(c) of greater than 1.0 MPa . It is suggested that gas vesicles of the GV3 strains, which are narrower and stronger than any previously recorded in freshwater cyanobacteria, have evolved to withstand the high hydrostatic pressures during deep winter mixing in Lake Zurich.

Microbiology, 1999 Oct, 145 ( Pt 10), 2757 - 68
The diversity of gas vesicle genes in Planktothrix rubescens from Lake Zürich; Beard SJ et al.; Part of the gas vesicle gene cluster was amplified by PCR from three strains of Planktothrix rubescens isolated from Lake Zurich, Switzerland . Each contains multiple alternating copies of gvpA and gvpC . All of the gvpA sequences in the different strains are identical . There are two types of gvpC: gvpC20, of length 516 bp, encodes a 20 kDa protein of 172 amino acid residues (whose N-terminal amino acid sequence is homologous with the sequence of GvpC in Planktothrix {Oscillatoria} agardhii); gvpC16, of length 417 bp, encodes a 16 kDa protein of 139 amino acid residues that differs in lacking an internal 33-residue section . An untranslated 72 bp fragment from the 3' end of gvpC, designated omegaC, is also present in some strains . The two types of gvpC and presence of omegaC could be distinguished by the different lengths of PCR amplification products obtained using pairs of oligonucleotide primers homologous to internal sequences in gvpC and gvpA . Three genotype classes were found: GV1, containing only gvpC20; GV2, containing gvpC20 and omegaC; and GV3, containing gvpC16, gvpC20 and omegaC . Subclasses of GV2 and GV3 contained either one or two copies of omegaC . The accompanying paper by D . I . Bright & A . E . Walsby (Microbiology 145, 2769-2775) shows that strains of the GV3 genotype produce gas vesicles with a higher critical pressure than those of GV1 and GV2 . A PCR survey of 185 clonal cultures of P . rubescens isolated from Lake Zurich revealed that 3 isolates were of genotype GV1, 73 were of GV2 and 109 were of GV3 . The PCR technique was used to distinguish the gas vesicle genotype, and thence the associated critical-pressure phenotype, of single filaments selected from lakewater samples . Sequence analysis of the 16S rDNA and of regions within the operons encoding phycoerythrin, phycocyanin and Rubisco confirmed that these strains of Planktothrix form a tight phylogenetic group.

Zentralbl Veterinarmed B, 1999 Sep, 46(7), 457 - 66
Serological cross-sectional study of paratuberculosis in cattle in Austria; Gasteiner J et al.; From 1995 to 1997, the prevalence of serum antibodies against Mycobacterium avium subspecies paratuberculosis (M . av . ssp . ptbc.)--the causal agent of paratuberculosis (Johne's Disease)--was examined in 11,028 Austrian cattle . Samples from the four oldest cattle on 2757 farms were collected according to a specific sampling schedule for this epidemiological study . District, age and breed of animals were included as variables in this study . For antibody screening against M . avium subspecies paratuberculosis, a modified, commercially available ELISA (ALLIED Monitors, Fayette, USA) was employed . A total of 2253 samples that were found to be positive or questionable were subjected to further testing with a more specific ELISA (Institute of Microbiology and Infectious Animal Diseases) . Results of this study were used for statistical analysis . The average prevalence of antibodies to M . avium subspecies paratuberculosis was 1.99% in Austria . The highest prevalence was seen in 6-year-old cattle (2.84%) and Holstein Frisian cattle (3.51%) . Sero-positive animals were found on 6.96% of farms tested, and the prevalence was highest in Vorarlberg, followed by Salzburg, the Tyrol, Styria and Carinthia . This study is unique in Europe in the use of an adequate random sampling plan for an investigation of this magnitude.

Curr Opin Obstet Gynecol, 1999 Oct, 11(5), 435 - 9
Recurrent pregnancy loss: an update; Kutteh WH; This review highlights recent studies that investigate causes and treatments for recurrent pregnancy loss . Generally the causes of recurrent pregnancy loss are classified as genetic, endocrinologic, anatomic, immunologic, microbiologic, and environmental . The majority of recent work has focused on potential autoimmune and alloimmune causes; however, controversy still exists over appropriate testing and treatment . Reports have investigated the potential associations between autoimmune factors (antithyroid antibodies and antiphospholipid antibodies) and alloimmune factors (natural killer cells, cytotoxic T cells, and embryotoxic factors) and recurrent pregnancy loss . Increasingly, clinical reports are suggesting intravenous immunoglobulin as a potential treatment for these immunologic problems . Several lines of investigation have suggested certain hypercoagulable states as causative of recurrent pregnancy loss . New studies relating recurrent pregnancy loss to endocrinologic aberrations (hyperprolactinemia and hyperandrogenism) as well as social/environmental factors (stress, caffeine use, tobacco use, human immunodeficiency virus, and history of induced abortion) have been made . A summary of proposed evaluation and treatment options is presented.

Clin Infect Dis, 1999 Nov, 29(5), 1210 - 9
Aspergillosis in children with cancer: A 34-year experience; Abbasi S et al.; A retrospective review of medical records, microbiology and pathology laboratory records, and nosocomial infection surveillance data was undertaken to describe the experience with culture-documented aspergillus infection in pediatric cancer patients at our facility . Sixty-six patients were identified from a 34-year period . The most common underlying diagnosis was leukemia . Risk factors included neutropenia, immunosuppression, and prior antibiotic therapy . On the basis of clinical presentation, 23 patients were believed to have disseminated disease and 43 to have localized disease . The lung was the most frequently affected organ . Despite aggressive medical and surgical management, overall mortality was 85% within the first year after diagnosis . Patients who presented with disease in sites other than the lungs fared better than patients with initial pulmonary involvement (P=.0014) . Aspergillosis continues to be associated with poor outcome . Development of improved medical and adjuvant therapies, including surgery, is warranted.

Gene, 1999 Sep 3, 237(1), 123 - 33
Sequence, overproduction and purification of the family 11 endo-beta-1,4-xylanase encoded by the xyl1 gene of Streptomyces sp . S38; Georis J et al.; The xyl1 gene encoding the Xyl1 xylanase of Streptomyces sp . strain S38 was cloned by screening an enriched DNA library with a specific DNA probe and sequenced . Three short 5 bp -CGAAA- sequences are located upstream of the Streptomyces sp . S38 xyl1 gene 105, 115 and 250 bp before the start codon . These sequences, named boxes 1, 2 and 3, are conserved upstream of the Actinomycetales xylanase genes and are specifically recognized by a DNA-binding protein (Giannotta et al., 1994 . FEMS Microbiol . Lett . 142, 91-97) and could be probably involved in the regulation of xylanase production . The Xyl1 ORF encodes a 228 residue polypeptide and the Xyl1 preprotein contains a 38 residue signal peptide whose cleavage yields a 190 residue mature protein of calculated M(r) = 20,585 and basic pI value of 9.12 . The molecular mass of the produced and purified mature protein determined by mass spectrometry (20,586 +/- 1 Da) and its pI (9.8) agree with these calculated values . Its N-terminal amino-acid sequence confirmed the proposed cleavage site between the signal peptide and the mature protein . Comparisons between Xyl1 and the 62 other xylanases belonging to family 11 allowed the construction of a phylogenetic tree and revealed its close relationship with Actinomycetales enzymes . Moreover, nine residues were found to be strictly conserved among the 63 xylanases.

J Clin Microbiol, 1999 Nov, 37(11), 3676 - 80
Phenotypic and genotypic heterogeneity among cultivable pathogen-related oral spirochetes and Treponema vincentii; Riviere GR et al.; Recent findings challenge the assumption that pathogen-related oral spirochetes (PROS) are related to Treponema pallidum . Treponema vincentii, grown in OMIZ-Pat media, cross-reacted with monoclonal antibody H9-2 against T . pallidum, and cultivable PROS had 16S rRNA gene sequences similar to those of T . vincentii (C.-B . Choi, C . Wyss, and U . B . Gobel . J . Clin . Microbiol . 34:1922-1925, 1996) . Aims of the present study were to determine whether antigen phenotypes of oral treponemas were influenced by growth conditions and to evaluate the genetic relatedness of cultivable PROS to T . pallidum and T . vincentii . Results show that three T . pallidum monoclonal antibodies (H9-1, H9-2, and F5) cross-reacted with whole cells from four Treponema species grown in modified OMIZ-Pat medium, but not with treponemas grown in NOS medium . Only H9-2 reacted in immunoblots with reduced proteins from cultivable PROS and T . vincentii . Three of five PROS isolates were amplified by T . vincentii-specific PCR, and one was amplified by Treponema medium-specific PCR . None were amplified by T . pallidum-specific PCR . Three of five PROS isolates had 16S ribosomal DNA restriction fragment length polymorphism patterns identical to that of T . vincentii, and the patterns of two isolates resembled that of T . medium . Arbitrarily primed-PCR profiles from whole genomic DNA were distinct among five PROS isolates and two T . vincentii strains . Thus, PROS isolates represent a heterogeneous group of treponemas that share some 16S rRNA gene sequences with T . vincentii and T . medium, but not with T . pallidum . It is proposed that the PROS nomenclature be dropped.

J Clin Microbiol, 1999 Nov, 37(11), 3448 - 51
Prevalence of Borrelia burgdorferi and granulocytic and monocytic ehrlichiae in Ixodes ricinus ticks from southern Germany; Baumgarten BU et al.; A total of 287 adult Ixodes ricinus ticks, collected in two regions of southern Germany (Frankonia and Baden-Wurttemberg) where Borrelia burgdorferi infections are known to be endemic, were examined for the presence of 16S ribosomal DNA specific for the Ehrlichia phagocytophila genogroup, E . chaffeensis, E . canis, and B . burgdorferi by nested PCR . Totals of 2.2% (6 of 275) and 21.8% (65 of 275) of the ticks were positive for the E . phagocytophila genogroup and B . burgdorferi, respectively . Two ticks (0.7%) were coinfected with both bacteria . Of 12 engorged I . ricinus ticks collected from two deer, 8 (67%) were positive for the E . phagocytophila genogroup and one (8%) was positive for B . burgdorferi . There was no evidence of infection with E . canis or E . chaffeensis in the investigated tick population . The nucleotide sequences of the 546-bp Ehrlichia PCR products differed at one or two positions from the original sequence of the human granulocytic ehrlichiosis (HGE) agent (S.-M . Chen, J . S . Dumler, J . S . Bakken, and D . H . Walker, J . Clin . Microbiol . 32:589-595, 1994) . Three groups of sequence variants were detected; two of these were known to occur in other areas in Europe or the United States, whereas one has not been reported before . Thus, in the German I . ricinus tick population closely related granulocytic ehrlichiae are prevalent, which might represent variants of E . phagocytophila or the HGE agent.

J Microbiol Methods, 1999 Oct, 38(1-2), 155 - 67
Standardisation of restriction fragment length polymorphism analysis for Mycobacterium avium subspecies paratuberculosis; Pavlik I et al.; DNA from 1008 strains of Mycobacterium avium subspecies paratuberculosis, digested by restriction endonucleases PstI and BstEII, was hybridised with a standard IS900 probe prepared by PCR and labelled non-radioactively by ECL . DNA fingerprints were scanned by CCD camera and analysed using the software Gel Compar (Applied Maths, Kortrijk, Belgium) . Thirteen restriction fragment length polymorphism (RFLP) (PstI) types were detected, which where designated as A, B, C, D, E, F, G, H, I, J, K, L and M in accordance with the study of Pavlik et al . (1995) {Pavlik, I., Bejckova, L., Pavlas, M., Rozsypalova, V., Koskova, S., 1995 . Characterization by restriction endonuclease analysis and DNA hybridization using IS900 of bovine, ovine, caprine and human dependent strains of Mycobacterium paratuberculosis isolated in various localities . Vet . Microbiol . 45, 311-318} . Twenty RFLP (BstEII) types were detected and designated as C1-3, C5, C7-20, S1 and I1 in accordance with the study by Collins et al . 1990 {Collins, D.M., Gabric, D.M., de Lisle, G.W., 1990 . Identification of two groups of Mycobacterium paratuberculosis strains by restriction endonuclease analysis and DNA hybridization . J . Clin . Microbiol . 28, 1591-1596} . A combination of both RFLP (PstI) and RFLP (BstEII) results revealed a total of 28 different RFLP types . All the RFLP types and detailed protocols are available at Intemet web site WWW...: /www.vri.cz/wwwrflptext.htm.

Vet Microbiol, 1999 Sep 1, 69(1-2), 111 - 3
Treatment of feline leukemia virus (FeLV) infection; Hartmann K et al.; FeLV infection is still considered to account for most disease-related deaths in pet cats . Different treatment attempts with various drugs were performed in the past but none resulted in healing or complete virus elimination . Therefore, it caused a sensation when Horber and Mayr {Horber, D., Mayr, B., 1991 . Prax . 19, 311-314; Horber, D., Schnabl, W., Mayr, B., 1992 . Tierarztl . Umschau 47, 556-560; Mayr, B., Horber, D., 1992 . Kleintierprax . 37, 515-518} published that they were able to cure 80 to 100% FeLV-infected cats from viremia by using an immunomodulating compound . Articles in cat breeder and cat owner journals appeared assuming that obviously there is a rescue for FeLV-infected cats suffering from this deadly infection . The immunomodulator {Buttner, M., 1993 . Comp . Immun . Microbiol . Infect . Dis . 18, 1-10} used in those studies was the so-called 'paramunity inducer' PIND-ORF (Baypamun, Bayer, Leverkusen, Germany) consisting of inactivated parapox ovis virus . Since that time, Baypamun is the most commonly used drug for treatment of FeLV infection in Germany and other European countries . Four placebo-controlled double-blind trials were performed to determine the therapeutic efficacy of Baypamun and other compounds in naturally FeLV-infected cats under controlled conditions.

Appl Environ Microbiol, 1999 Oct, 65(10), 4497 - 505
Localization and in situ activities of homoacetogenic bacteria in the highly compartmentalized hindgut of soil-feeding higher termites (Cubitermes spp.); Tholen A et al.; Methanogenesis and homoacetogenesis occur simultaneously in the hindguts of almost all termites, but the reasons for the apparent predominance of methanogenesis over homoacetogenesis in the hindgut of the humivorous species is not known . We found that in gut homogenates of soil-feeding Cubitermes spp., methanogens outcompete homoacetogens for endogenous reductant . The rates of methanogenesis were always significantly higher than those of reductive acetogenesis, whereas the stimulation of acetogenesis by the addition of exogenous H(2) or formate was more pronounced than that of methanogenesis . In a companion paper, we reported that the anterior gut regions of Cubitermes spp . accumulated hydrogen to high partial pressures, whereas H(2) was always below the detection limit (<100 Pa) in the posterior hindgut, and that all hindgut compartments turned into efficient H(2) sinks when external H(2) was provided (D . Schmitt-Wagner and A . Brune, Appl . Environ . Microbiol . 65:4490-4496, 1999) . Using a microinjection technique, we found that only the posterior gut sections P3/4a and P4b, which harbored methanogenic activities, formed labeled acetate from H(14)CO(3)(-) . Enumeration of methanogenic and homoacetogenic populations in the different gut sections confirmed the coexistence of both metabolic groups in the same compartments . However, the in situ rates of acetogenesis were strongly hydrogen limited; in the P4b section, no activity was detected unless external H(2) was added . Endogenous rates of reductive acetogenesis in isolated guts were about 10-fold lower than the in vivo rates of methanogenesis, but were almost equal when exogenous H(2) was supplied . We conclude that the homoacetogenic populations in the posterior hindgut are supported by either substrates other than H(2) or by a cross-epithelial H(2) transfer from the anterior gut regions, which may create microniches favorable for H(2)-dependent acetogenesis.

J Biochem (Tokyo), 1999 Oct, 126(4), 762 - 8
Characterization and application to hot start PCR of neutralizing monoclonal antibodies against KOD DNA polymerase; Mizuguchi H et al.; DNA polymerase from Pyrococcus kodakaraensis KOD1 (KOD DNA polymerase) is one of the most efficient thermostable PCR enzymes exhibiting higher accuracy and elongation velocity than any other commercially available DNA polymerase {M . Takagi et al . (1997) Appl . Environ . Microbiol . 63, 4504-4510} . However, even when KOD DNA polymerase was used for PCR, troubles with nonspecific DNA amplification and primer dimer formation still remain because of undesirable DNA polymerase activity during the first denaturing step of PCR . In order to inhibit this undesirable DNA polymerase activity (hot start PCR), two neutralizing monoclonal antibodies (mAbs), 3G8 and betaG1, to KOD DNA polymerase were obtained . Both of these antibodies belong to subclass IgG(1), <font face="k">k . K(d) values were 7.3 x 10(-8) for 3G8 and 1.1 x 10(-6) for betaG1 . Nucleotide sequencing of cDNAs of these monoclonal antibodies revealed their sequences to differ in their CDRs (complementarity determining region) . Exonuclease activity measurement and epitope mapping revealed that the epitope for 3G8 is located in conserved regions among alpha-like (family B) DNA polymerases (Region II), and the epitope for betaG1 is located in the 3'-5' exonuclease domain . When hot start PCR with each of these mAbs was performed, the specificity of target gene amplification became much higher than in reactions without monoclonal antibody . Furthermore, this method can easily be applied to long distance PCR (>17.5 kbp).

Gastrointest Endosc, 1999 Oct, 50(4), 461 - 7
Disposable-sheath, flexible gastroscope system versus standard gastroscopes: a prospective, randomized trial; Mayinger B et al.; BACKGROUND: Endoscopically transmitted infections due to inadequate disinfection are rare but well-recognized complications . A new prototype of a flexible, fiberoptic endoscope has been developed that uses a disposable sheath to cover and protect all working surfaces of the endoscope from contamination . The present study investigated the function, reprocessing, and hygienic status of this endoscope system in comparison with standard systems . METHODS: In a prospective, randomized trial, 100 upper endoscopic procedures (50 standard, 50 sheathed) were performed . Analog rating scales were used to evaluate endoscopic performance and reprocessing . Reprocessing time, procedure duration, insertion depth, total instrument downtime, and problems occurring during the procedure were recorded . Microbiologic swabs were obtained from each endoscope . RESULTS: Mean procedure duration was slightly longer with the sheathed system than with standard endoscope (9.9 vs . 8.4 min) . Set up and reprocessing times, however, were significantly shorter with the Endosheath (8.9 vs . 48.4 min with the standard endoscope) . The disposable-sheath endoscope system permits the rate of performance of procedures to be increased by a factor of up to 3.0 . Both endoscopists and reprocessing personnel preferred the standard endoscope . No post-procedure sheath leakage or rupture was seen . CONCLUSION: Our results suggest that the "disposable endoscope" may have important advantages in terms of decreased instrument turn-around time and potentially improved safety.

Biosci Biotechnol Biochem, 1999 Aug, 63(8), 1321 - 8
Effects of high pressure on lipids and biomembranes for understanding high-pressure-induced biological phenomena; Kato M et al.; This review covers high pressure effects on lipids, lipid bilayers, and biochemical observations recently found in the field of high-pressure bioscience and biotechnology including deep-sea microbiology and food science . To explain these phenomena in a unified model, recent studies of physical and chemical properties of artificial membranes and natural membranes are summarized . On the basis of this newly described knowledge, high pressure effects on biochemical events are considered at the molecular level and concluded that high pressure induces decreases in biomembrane fluidity and phase transitions that result in breakage of the membrane, and finally, leads to the destruction of bilayer membrane accompanied by denaturation of membrane-associated proteins.

Pediatr Pulmonol, 1999 Oct, 28(4), 231 - 41
Epidemiologic study of cystic fibrosis: design and implementation of a prospective, multicenter, observational study of patients with cystic fibrosis in the U.S . and Canada; Morgan WJ et al.; Cystic fibrosis (CF) is a complex illness characterized by chronic lung infection leading to deterioration in function and respiratory failure in over 85% of patients . An understanding of the risk factors for that progression and the interaction of these factors with current therapeutic strategies should materially improve the prevention of this progressive lung disease . The Epidemiologic Study of Cystic Fibrosis (ESCF) was therefore designed as a multicenter, longitudinal, observational study to prospectively collect detailed clinical, therapeutic, microbiologic, and lung function data from a large number of CF treatment sites in the U.S . and Canada . The ESCF also serves an important role as a phase-IV study of dornase alfa . To be eligible for enrollment, subjects must have the diagnosis of CF and receive the majority of their care at an ESCF site . In this paper, the authors present the ESCF study design in detail . Further, enrollment data collected at 194 study sites in 18,411 subjects enrolled from December 1, 1993 to December 31, 1995 are presented in summary form . This comprehensive study is unique in the detail of clinical data collected regarding patient monitoring and therapeutic practices in CF care . Two companion articles present data regarding practice patterns in cystic fibrosis care, including data on resource utilization and prescribing practices .

J Am Dent Assoc, 1999 Sep, 130(9), 1291 - 8
Dental infection control at the year 2000: accomplishment recognized; Molinari JA; OVERVIEW: Health care providers have strived to address the many elements necessary to achieve effective infection control . Historical progress in microbiology and hospital asepsis provided the foundation for advances in dental preventive practices . Evidence supporting the application of current practices includes a long history of scientific and clinical investigations, technological advances in equipment and materials, and periodic publication of updated recommendations by professional health care organizations . CLINICAL IMPLICATIONS: The routine application of precautions such as multiple aseptic procedures, latex gloves, masks, protective eyewear, clinic coats, automated instrument decontamination devices, time-efficient heat sterilization modalities, chemical disinfectants, waste management procedures and single-use disposable items have created a safer environment for dental personnel and patients alike.

Cornea, 1999 Sep, 18(5), 616 - 20
Chrysosporium parvum keratomycosis; Wagoner MD et al.; PURPOSE: To report a case of corneal infection with Chrysosporium parvum, a filamentous fungus usually associated with pulmonary infections . METHODS: A 43-year-old Saudi man had a corneal stromal infiltrate and perforation of his left eye . He was treated with a therapeutic penetrating keratoplasty and topical and systemic antifungal therapy . Corneal scrapings, microbiologic evaluation, and histopathologic examination of the surgical specimen were performed to establish the diagnosis . After the development of recurrent stromal keratitis at the graft-host junction, similar diagnostic and therapeutic maneuvers were performed . RESULTS: Corneal scrapings and histopathologic examination were positive for numerous septate hyphae with endospores, consistent with a diagnosis of filamentous keratomycosis . Microbiologic isolation confirmed the diagnosis of Chrysosporium parvum . Similar diagnostic maneuvers for recurrent keratitis produced identical results . CONCLUSION: To our knowledge, this is the first case of Chrysosporium parvum keratomycosis.

Vestn Ross Akad Med Nauk, 1999, (8), 15 - 20
{Luminescent ultramicroanalysis for the simultaneous determination of biological and chemical substances: the methodology and the ways for solution}; Chudinov AV et al.; The next revolution in in-vitro bioaffinity assays will be associated with the minimization of the liquid phase of an experiment and with the extremely rapid detection of a large number of samples . The ultra detection unit is one of the essential elements of such devices . The fluorescence detection systems are most promising . The fluorescence detection limit of europium ions is as low as 10(-15) M in the complex with beta-diketone . The authors' approach to designing new highly effective chelates is to synthesize a stable chelate wherein several molecules of beta-diketones are properly oriented and chemically fixed . The molecular mechanic calculation of three dimensional structure of different Eu chelates should be used in optimizing the strategy of chemical synthesis of new compounds . New chelates can be efficiently used for the development of a new approach to measuring distances equal to over 10 nm in the biological systems . Pesticides also represent challenging targets because the levels of compounds are to be detects in minor concentrations, thus the time-resolution fluorescence assay may have some advantages due to its high sensitivity . DNA-based microbiochips are the most promising area in the practical use of a new chelate . A novel europium chelate is a key compound for the development of new methods for studying biological and chemical objects, as well as new diagnostic systems based on a basically new instrumental base that is characterized by extremely high miniaturization.

Clin Ther, 1999 Aug, 21(8), 1418 - 25
Computer-assisted evaluation of antibiotic regimen coverage and cost; Day D et al.; Evaluation of the cost-effectiveness of antibiotic regimens has become an essential part of drug selection for pharmacists and physicians . However, these evaluations can be complicated, time-consuming, and, if the data used are not based on local conditions, misleading . The computer program Dare to Compare 98 was developed to provide an analysis of empiric antibiotic regimens . The Infectious Disease Challenge portion of the program lists the commonly identified pathogens in specific infectious diseases . The Antibiogram Susceptibility Reports section generates susceptibility reports based on local antibiogram data or data from institutions nationwide that have similar demographic profiles . Susceptibility information is combined with cost data in the Quality/Cost Index section . Comparison of the quality and cost index values allows the user to determine which regimens provide the optimal microbiologic activity and cost values for treatment of a particular infectious disease based on local data . Thus Dare to Compare 98 can help pharmacists and physicians evaluate antibiotic regimens and their suitability for inclusion in formularies and disease-management algorithms.

Arch Immunol Ther Exp (Warsz), 1999, 47(4), 237 - 44
Development and disappearance of tolerance to induction of interferon and tumor necrosis factor response in athletes treated with natural immunostimulant; Inglot AD et al.; The effect of nonspecific immunostimulation was examined in 15 basketball players subjected to extensive physical effort . The Tolpa* Torf Preparation (TTP*), a natural immunostimulating drug, was applied orally, one 5 mg tablet daily, in two 21-day cycles, separated by 2-week hiatus . Blood samples were collected 4 times, after each of two TTP* cycles and after the first and second hiatus . Whole blood assay was used to determine the spontaneous and induced production of interferon (IFN) and tumor necrosis factor (TNF) . The levels of the cytokines were measured by microbioassays . TTP* stimulated synthesis of IFN and TNF in the whole blood cultures . However, after the oral administraton of TTP* for 3 weeks the leukocytes of the athletes developed hyporeactivity to IFN induction by TTP* and to a lesser extent to another "superinducer"--a mixture of phytohemagglutinin and bacterial lipopolysaccharide . The hyporeactivity state disappeared spontaneously within 2 weeks . In contrast, the tolerance to TNF induction did not develop during the TTP* administration . The increase of immunoglobulins, mainly of IgM and IgG classes and an acute phase protein--alpha1-antitrypsin, was observed at the late phase of the treatment . We suggest that the cytokine levels may be early markers for immunoprophylaxis . Furthermore, high production of IFN and TNF may be associated with extensive physical effort.

J Bacteriol, 1999 Sep, 181(18), 5581 - 90
Cloning and molecular characterization of the genes for carbon monoxide dehydrogenase and localization of molybdopterin, flavin adenine dinucleotide, and iron-sulfur centers in the enzyme of Hydrogenophaga pseudoflava; Kang BS et al.; Carbon monoxide dehydrogenases (CO-DH) are the enzymes responsible for the oxidation of CO to carbon dioxide in carboxydobacteria and consist of three nonidentical subunits containing molybdopterin flavin adenine dinucleotide (FAD), and two different iron-sulfur clusters (O . Meyer, K . Frunzke, D . Gadkari, S . Jacobitz, I . Hugendieck, and M . Kraut, FEMS Microbiol . Rev . 87:253-260, 1990) . The three structural genes of CO-DH in Hydrogenophaga pseudoflava were cloned and characterized . The genes were clustered on the chromosome in the transcriptional order cutM-cutS-cutL . The cloned cutM, cutS, and cutL genes had open reading frames of 864, 492, and 2,412 nucleotides, coding for proteins with calculated molecular weights of 30,694, 17,752, and 87,224, respectively . The overall identities in the nucleotide sequence of the genes and the amino acid sequence of the subunits with those of other carboxydobacteria were 64.5 to 74.3% and 62.8 to 72.3%, respectively . Primer extension analysis revealed that the transcriptional start site of the genes was the nucleotide G located 47 bp upstream of the cutM start codon . The deduced amino acid sequences of the three subunits of CO-DH implied the presence of molybdenum cofactor, FAD, and iron-sulfur centers in CutL, CutM, and CutS, respectively . Fluorometric analysis coupled with denaturing polyacrylamide gel electrophoresis of fractions from hydroxyapatite column chromatography in the presence of 8 M urea of active CO-DH and from gel filtration of spontaneously inactivated enzyme revealed that the large and medium subunits of CO-DH in H . pseudoflava bind molybdopterin and FAD cofactors, respectively . Iron-sulfur centers of the enzyme were identified to be present in the small subunit on the basis of the iron content in each subunit eluted from the denaturing polyacrylamide gels.

Med J Aust, 1999 Aug 2, 171(3), 127 - 31
Vancomycin and teicoplanin use in Victorian hospitals . The Victorian Drug Usage Evaluation Group; Robertson MB et al.; OBJECTIVE: To determine patterns of prescribing of glycopeptide antibiotics (vancomycin and teicoplanin) in Victorian hospitals and identify areas for targeted intervention . DESIGN: A concurrent, observational, multisite evaluation of drug use . SETTING: Thirty-five Victorian hospitals, 1-14 September 1997 . STUDY POPULATION: Patients commencing a glycopeptide antibiotic course . MAIN OUTCOME MEASURES: Rate of glycopeptide antibiotic use; indications; duration of use; main hospitals using glycopeptide antibiotics . RESULTS: 293 patients (269 adults and 24 neonates) commenced on 302 glycopeptide antibiotic courses: 296 intravenous (i.v.) vancomycin courses and three each of oral vancomycin and parenteral teicoplanin . The overall rate of use was 10.3 courses per 1000 inpatient separations . Of 271 i.v . vancomycin courses for adults, 176 (65%) were for treatment--120 empirically . The median duration of treatment courses was 4.7 days (interquartile range, 2.0-8.2 days) . A flucloxacillin-resistant organism was confirmed for 44% of treatment courses . Ninety-five i.v . vancomycin courses were for prophylaxis, including for cardiac (54%) and vascular surgery (21%); 82% of prophylactic courses were administered for less than 24 hours . Of all the glycopeptide antibiotic courses, 69% were administered at five major metropolitan hospitals . CONCLUSIONS: Glycopeptide antibiotic use in Victoria is concentrated in the major metropolitan hospitals . Prolonged durations of vancomycin therapy, including for surgical prophylaxis and empirical therapy not subsequently confirmed by microbiology findings, would be suitable targets for interventional strategies.

J Clin Pathol, 1999 Apr, 52(4), 271 - 3
Comparative study of seven commercial yeast identification systems; Verweij PE et al.; AIMS: To compare the performance of seven commercial yeast identification methods with that of a reference method, and to compare the costs of the commercial kits . METHODS: Clinical yeast isolates (n = 52), comprising 19 species, were identified using Vitek, Api ID 32C, Api 20C AUX, Yeast Star, Auxacolor, RapID Yeast Plus system, and Api Candida and compared with a reference method which employed conventional tests . RESULTS: The percentage of correctly identified isolates varied between 59.6% and 80.8% . Overall, the highest performance was obtained with Api Candida (78.8%) and Auxacolor (80.8%) . Among germ tube negative yeast isolates, Auxacolor and Api Candida both identified 93.1% of isolates correctly . All systems failed to identify C norvegensis, C catenulata, C haemulonii, and C dubliniensis . In comparison with Auxacolor, the Api Candida is less expensive and requires less bench time . CONCLUSIONS: Auxacolor and Api Candida appeared to be the most useful systems for identification of germ tube negative yeast isolates in clinical microbiology laboratories, although one should be aware that several germ tube negative Candida species cannot be identified by these systems.

Compr Ther, 1999 Jun-Jul, 25(6-7), 326 - 9
Causes, diagnosis and treatment of pharyngitis; Huovinen P; Pharyngitis is the only respiratory tract infection in which microbiologic diagnosis has immediate influence on the treatment decision . Microbiologic diagnosis should always precede antibiotic treatment of pharyngitis to ensure optimal treatment . Penicillin administered for 10 days is still the drug of choice.

Clin Exp Allergy, 1999 Sep, 29(9), 1260 - 4
Usefulness of early serial measurement of specific and total immunoglobulin E in the diagnosis of gastro-allergic anisakiasis; Daschner A et al.; BACKGROUND: Sensitization to Anisakis simplex (A . simplex) has been documented to produce severe allergic reactions following ingestion of mainly raw or under-cooked parasitized fish . False positive skin prick tests (SPT) or specific IgE against this nematode and cross-reactivity restricts diagnosis . Gastric anisakiasis and gastro-allergic anisakiasis occur if fish is parasitized by live A . simplex OBJECTIVE: To investigate if serial serological analysis could be useful in the diagnosis of acute parasitation by this nematode . METHODS: We included 41 patients who experienced an allergic reaction and/or abdominal symptoms after ingestion of raw or undercooked fish and displayed specific IgE against A . simplex . Total and specific IgE were determined two times: in the 24-h period after onset of clinical symptoms and after 1 month . SPTs were performed against A . simplex and implicated fish . A fibre optic gastroscopy was performed in 22 patients . RESULTS: Median total IgE was 80.0 (Interquartile range {IQR} 41.5-186.5) kU/L in the first evaluation and 247.0 (IQR 96.5-649.5) kU/L after 1 month . Median specific IgE against A . simplex was 11.4 (IQR 7.1-33.5) kU/L in the first 24 h and 36.8 (IQR 19.5-79.5) kU/L after 1 month . A rise of total IgE was observed in 35 of 41 patients (P<0.00001) and a rise in specific IgE against A . simplex in 37 of 41 patients (P<0.00001) . Mean percentage increment was 392% (215-571%; 95% confidence interval {CI}) for total IgE and 339% (177-502%; 95% CI) for specific IgE . In nine of 22 gastroscopic examinations at least one larva, identified as A . simplex, could be detected by our microbiology service . In this group (n = 9) a rise of total and specific IgE was detected in eight patients (89%) (P = 0.02) . CONCLUSIONS: We consider a rise of total and specific IgE in the first month after an allergic reaction as a useful tool in the diagnosis of gastro-allergic anisakiasis (together with patient's history), even if the parasite cannot be seen with fibre optic gastroscopy . The important rise of total and specific IgE against A . simplex can be considered as a reaction induced by the live parasitizing larva in the context of a polyclonal immunological stimulation.

Acta Physiol Scand, 1999 Aug, 166(4), 261 - 77
The impact of biochemical methods for single muscle fibre analysis; Pette D et al.; Biochemical methods for single muscle fibre analysis provide sensitive measures for elucidating muscle fibre heterogeneity . The understanding of the complexity of skeletal muscle fibres, initially based on qualitative histochemistry and immunohistochemistry, has been greatly expanded by quantitative micromethods, such as microphotometry and microbiochemical assays . Assessment of metabolic enzyme activity levels has revealed pronounced scattering within and between different fibre types and has highlighted the use of specific enzyme activity ratios as discriminative measures . With the exception of type I fibres, metabolic properties are loosely coupled with molecular properties of the myofibrillar apparatus . As such, myosin heavy chain (MHC) isoforms appear to be the best choice for fibre type delineation . Among the two available methods for MHC-based fibre type distinction, single fibre electrophoresis appears to be superior to immunohistochemistry . The electrophoretic separation of MHC isoforms in single fibres is quantitative and, as opposed to immunohistochemistry, yields important information on MHC isoform proportions in hybrid fibres . Histochemical staining for myofibrillar ATPase activity can, thus, be correlated in most cases with specific MHC isoform profiles . Single fibre studies have demonstrated a relationship between ATP phosphorylation potential and MHC isoform complement . This relationship corresponds to different tension costs and provides an additional rationale for the MHC-based fibre type diversity and transitions . The combination of reverse transcriptase (RT) with polymerase chain reaction (PCR) has proved to be a highly sensitive tool and has extended single fibre analysis to the level of MHC mRNA isoforms . Application of RT-PCR techniques to single fibre fragments identified by their MHC protein isoform profile, provides insights at two levels of expression and, thus, has extended our knowledge on the plasticity of muscle and the dynamical state of muscle fibres.

Infect Control Hosp Epidemiol, 1999 Aug, 20(8), 543 - 8
A decade of prevalence surveys in a tertiary-care center: trends in nosocomial infection rates, device utilization, and patient acuity; Weinstein JW et al.; OBJECTIVE: To evaluate the usefulness of repeated prevalence surveys to determine trends in the rates of nosocomial infections and to detect changes in risk factors (e.g., use of invasive devices) associated with nosocomial infections . PATIENTS AND METHODS: Ten annual prevalence surveys were conducted by trained infection control practitioners between 1985 and 1995 for acute-care patients on the medical, surgical, pediatric, and obstetric-gynecologic services at a 900-bed, tertiary-care, teaching hospital with 750 acute-care beds . The same methods of chart review and concurrent reporting from nursing, the microbiology and clinical laboratory, and the pharmacy were used each year to collect data on the prevalence of nosocomial infections, invasive-device utilization, and abnormal laboratory indicators . Although data were collected on a single day, a period-prevalence study approach was used, because charts were reviewed for any infection data occurring within the 7 days prior to the survey . RESULTS: The hospital census for acute care patients, as measured by the prevalence surveys, declined sharply over the 10 years, from 673 to 575 patients (P = .02) . However, the medical service census increased from 150 to 188 patients (P = .01) . During the same period, there was a significant decrease in the mean length of stay, from 7.3 to 6.0 days (P = .01), and a concomitant increase in the mean diagnosis related-group case-mix index, from 1.03 to 1.24 (P = .001) . Overall, nosocomial infection rates remained unchanged over the study period (mean of 9.85 infections per 100 patients), but rates of nosocomial bloodstream infection increased from 0.0% in 1985 to 2.3% in 1995 (P = .05) . Nosocomial infection rates were significantly higher on the medical and surgical services than on other services (P<.001) . Utilization rates increased significantly for Foley catheters (9.0% to 16.0%, P = .002) and ventilators (5.0% to 8.0%, P = .05) . CONCLUSIONS: Despite apparent increases in the severity of illness of our patients, overall rates of nosocomial infection remained stable during a decade of study . Rates of nosocomial bloodstream infection increased, in parallel with National Nosocomial Infection Surveillance System data . We found repeated prevalence surveys to be useful in following trends and rates of infection, device utilization, and abnormal laboratory values among patients at our institution . Such methodologies can be valuable and low-cost components of a comprehensive infection surveillance, prevention, and control program and other potential quality-improvement initiatives, because they enable better annual planning of departmental strategies to meet hospital needs.

AIDS, 1999 Aug 20, 13(12), 1511 - 5
Phase 1 trial of nonoxynol-9 film among sex workers in South Africa; Rustomjee R et al.; OBJECTIVES: To assess the acceptability and safety of a vaginal nonoxynol-9 film in a group of sex workers at a truck stop in South Africa . DESIGN: A randomized double-blinded crossover trial was conducted between April 1995 and July 1995 . INTERVENTION: Seventy-two mg nonoxynol-9 film and an identical glycerine placebo film . METHODS: Following informed consent, each study participant was randomly assigned the designated pre-coded film for 1 month . The second month was a film-free washout period and the participants used the alternate film in the third month . Besides measuring behavioural and clinical outcomes, colposcopy examination for genital lesions, serology and microbiology investigations for sexually transmitted diseases and semi-quantitative PCR for vaginal HIV load estimates were performed . RESULTS: Twenty women participated in the study . The women reported, on average, 19 sexual encounters per week . Vaginal intercourse was protected 25% of the time by condoms . On average, 11 vaginal films, either nonoxynol-9 or placebos were inserted per week . There were no statistically significant differences between the two treatment groups for genital lesions (P = 0.29), reported side effects (P = 0.73), and viral load (P = 0.9) . However, the proportions of clinically detected genital lesions (six out of eight versus two out of eight) and self-reported side-effects (five out of eight versus three out of eight) were higher in the nonoxynol-9 group when compared with the placebo group . Incident sexually transmitted diseases occurred more frequently in the placebo group . An increased viral load was associated with the development of a genital lesion (relative risk, 6.0; 95% confidence interval, 0.81-44.4) . CONCLUSIONS: The 72 mg film formulation of nonoxynol-9 was an acceptable product for use in this population of sex workers . Although no statistically significant differences in adverse outcomes were detected, clinically there appeared to be an increase in minor lesions and self-reported side-effects with nonoxynol-9 and less protection against sexually transmitted diseases with the placebo . Furthermore, HIV shedding was correlated with the presence of incident vaginal or cervical lesions . This brings into question the potential narrow margin of safety for this product; additional Phase 2 studies are therefore required.

J Bacteriol, 1999 Sep, 181(17), 5419 - 25
New sporulation loci in Streptomyces coelicolor A3(2); Ryding NJ et al.; Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the grey polyketide spore pigment, and such white (whi) mutants had been used to define eight sporulation loci, whiA, whiB, whiD, whiE, whiG, whiH, whiI, and whiJ (K . F . Chater, J . Gen . Microbiol . 72:9-28, 1972; N . J . Ryding, Ph.D . thesis, University of East Anglia, 1995) . In an attempt to identify new whi loci, we mutagenized S . coelicolor M145 spores with nitrosoguanidine and identified 770 mutants with colonies ranging from white to medium grey . After excluding unstable strains, we examined the isolates by phase-contrast microscopy and chose 115 whi mutants with clear morphological phenotypes for further study . To exclude mutants representing cloned whi genes, self-transmissible SCP2*-derived plasmids carrying whiA, whiB, whiG, whiH, or whiJ (but not whiD, whiE, or whiI) were introduced into each mutant by conjugation, and strains in which the wild-type phenotype was restored either partially or completely by any of these plasmids were excluded from further analysis . In an attempt to complement some of the remaining 31 whi mutants, an SCP2* library of wild-type S . coelicolor chromosomal DNA was introduced into 19 of the mutants by conjugation . Clones restoring the wild-type phenotype to 12 of the 19 strains were isolated and found to represent five distinct loci, designated whiK, whiL, whiM, whiN, and whiO . Each of the five loci was located on the ordered cosmid library: whiL, whiM, whiN, and whiO occupied positions distinct from previously cloned whi genes; whiK was located on the same cosmid overlap as whiD, but the two loci were shown by complementation to be distinct . The phenotypes resulting from mutations at each of these new loci are described.

Anaesthesia, 1999 Sep, 54(9), 868 - 72
Antiseptic-bonded central venous catheters and bacterial colonisation; Hannan M et al.; This study was undertaken to evaluate the impact of chlorhexidine/silver sulphadiazine-bonded catheters on the incidence of colonisation and catheter-related sepsis in critically ill patients . Threehundred and fifty-one catheters were inserted into 228 patients during the study period, 174chlorhexidine/silver sulphadiazine-bonded catheters and 177 standard catheters . Indications for catheter removal were: death, clinical redundancy and clinical evidence of local or systemic infection . All catheter tips were sent to the microbiology laboratory for semiquantitative analysis of bacterial colony count . Seventy-one (40.2%) of the standard catheters and 47 (27.2%) of the antiseptic-bonded catheters were found to be colonised on removal (p < 0.01) . Eight cases (4.7%) of catheter-related sepsis were associated with standard catheters and three cases (1.7%) with antiseptic-bonded catheters, however, this reduction was not statistically significant . Our results indicate that the use of antiseptic-bonded catheters in critically ill patients significantly reduces the incidence of bacterial colonisation.

Infect Immun, 1999 Sep, 67(9), 4819 - 26
beta-Chemokines enhance parasite uptake and promote nitric oxide-dependent microbiostatic activity in murine inflammatory macrophages infected with Trypanosoma cruzi; Aliberti JC et al.; In the present study, we describe the ability of Trypanosoma cruzi trypomastigotes to stimulate the synthesis of beta-chemokines by macrophages . In vivo infection with T . cruzi led to MIP-1alpha, RANTES, and JE/MCP1 mRNA expression by cells from peritoneal inflammatory exudate . In addition, in vitro infection with T . cruzi resulted in expression of beta-chemokine MIP-1alpha, MIP-1beta, RANTES, and JE mRNA by macrophages . The expression of the beta-chemokine MIP-1alpha, MIP-1beta, RANTES, and JE proteins by murine macrophages cultured with trypomastigote forms of T . cruzi was confirmed by immunocytochemistry . Interestingly, macrophage infection with T . cruzi also resulted in NO production, which we found to be mediated mainly by beta-chemokines . Hence, treatment with anti-beta-chemokine-specific neutralizing antibodies partially inhibited NO release by macrophages incubated with T . cruzi parasites . Further, the addition of the exogenous beta-chemokines MIP-1alpha, MIP-1beta, RANTES, and JE/MCP-1 induced an increased T . cruzi uptake, leading to enhanced NO production and control of parasite replication in a dose-dependent manner . L-NMMA, a specific inhibitor of the L-arginine-NO pathway, caused a decrease in NO production and parasite killing when added to cultures of macrophages stimulated with beta-chemokines . Among the beta-chemokines tested, JE was more potent in inhibiting parasite growth, although it was much less efficient than gamma interferon (IFN-gamma) . Nevertheless, JE potentiates parasite killing by macrophages incubated with low doses of IFN-gamma . Together, these results suggest that in addition to their chemotactic activity, murine beta-chemokines may also contribute to enhancing parasite uptake and promoting control of parasite replication in macrophages and may play a role in resistance to T . cruzi infection.

Bone Marrow Transplant, 1999 Aug, 24(3), 325 - 9
Mycobacterial central venous catheter tunnel infection: a difficult problem; Ward MS et al.; We report our experience of non-tuberculous mycobacterial infection associated with the tunnel of Hickman-Broviac central venous catheters in immunosuppressed patients with haematological malignancies undergoing high-dose chemotherapy supported by BMT . The problem is rare and difficult to treat . Our cases are unique in developing tunnel site mycobacterial infection well after the tunnelled catheters were removed . We diagnosed one case of Mycobacterium chelonae, which is a well-documented cause of such infections, and two cases of Mycobacterium haemophilum, which are the first reported cases in this setting . Early wide surgical excision of the infected tunnel site and prolonged antibiotic therapy is necessary . Despite these measures recurrence occurred in two cases . Close liaison with the microbiology laboratory is needed to ensure the appropriate culture media and conditions are used for these fastidious organisms . Empiric antibiotic regimens should be based on the likely organism . Drugs active against M . chelonae and M . haemophilum should be included.

Bone Marrow Transplant, 1999 Jul, 24(2), 139 - 45
Cord blood banking in London: the first 1000 collections; Armitage S et al.; The London Cord Blood Bank was established with the aim of collecting, processing and storing 10000 unrelated stem cell donations for the significant number of children in the UK requiring transplantation, for whom a matched unrelated bone marrow donor cannot be found . Collection is performed at two hospitals by dedicated cord blood bank staff after delivery of the placenta . Mothers are interviewed regarding medical, ethnic and behavioural history by nurse counsellors and sign a detailed consent form . Donations are returned to the bank for processing . Volume reduction is undertaken by a simple, closed, semi-automated blood processing system, with excellent recovery of progenitor cells . Units are cryopreserved and stored in the vapour phase of liquid nitrogen . Blood samples from mothers and cord blood donations are tested for the UK mandatory red cell and microbiology markers for blood donors . Donations are typed for HLA-A, B and DR at medium resolution (antigen split) level using sequence-specific oligonucleotide probing and sequence-specific priming techniques . The selection of collection hospitals on the basis of ethnic mix has proven effective, with 41.5% of donations derived from non-European caucasoid donors . Bacterial contamination of collections has been dramatically reduced by implementation of improved umbilical cord decontamination protocols.

Electrophoresis, 1999 Jul, 20(10), 2013 - 26
Proteomics for genetic and physiological studies in plants; Thiellement H et al.; Proteomics is becoming a necessity in plant biology, as it is in medicine, zoology and microbiology, for deciphering the function and role of the genes that are or will be sequenced . In this review we focus on the various, mainly genetic, applications of the proteomic tools that have been developed in recent years: characterization of individuals or lines, estimation of genetic variability within and between populations, establishment of genetic distances that can be used in phylogenetic studies, characterization of mutants and localization of the genes encoding the revealed proteins . Improvements in specifically devoted software have permitted precise quantification of the variation in amounts of proteins, leading to the concept of "protein quantity loci" which, combined with the "quantitative trait loci" approach, results in testable hypotheses regarding the role of "candidate proteins" in the metabolism or phenotype under study . This new development is exemplified by the reaction of plants to drought, a trait of major agronomic interest . The accumulation of data regarding genomic and cDNA sequencing will be connected to the protein databases currently developed in plants.

Am J Vet Res, 1999 Aug, 60(8), 960 - 4
Efficacy of florfenicol in the treatment of experimentally induced infectious bovine keratoconjunctivitis; Dueger EL et al.; OBJECTIVE: To evaluate efficacy of florfenicol in an induced model of infectious bovine keratoconjunctivitis, using a blinded randomized, controlled trial . ANIMALS: 48 male Holstein calves, 2 to 4 months old . PROCEDURE: Moraxella bovis infection was induced in all calves . When corneal ulcers developed, each calf was assigned randomly to 1 of 3 treatment groups, using a block design determined by corneal ulcer size (day 0) . Calves were treated with florfenicol (20 mg/kg of body weight, IM) on days 0 and 2 (IM group; n = 16) . Calves of a second group received a single dose of florfenicol (40 mg/kg, SC) on day 0 (SC group; n = 16) . The third group of calves was not treated (control group; n = 16) . Corneal ulcers were photographed, and each calf was assessed for 30 days after treatment for 10 clinical signs of infection . Corneal ulcer surface areas were measured, and clinical scores were calculated . Ocular secretions for microbiologic culture were obtained weekly from each eye . RESULTS: A Cox regression model indicated that, after adjustment for initial ulcer size, healing rates were 6.2 and 4.8 times greater in calves of the IM and SC groups, respectively, compared with the control group . Clinical scores and surface area measurements for treatment groups were significantly smaller than those for controls during posttreatment weeks 1 through 4 . From day 8 through day 29, M bovis was isolated from ocular secretions of 14 of 16 control calves and 1 of 32 treated calves . CONCLUSIONS AND CLINICAL RELEVANCE: Parenterally administered florfenicol reduces corneal ulcer healing time, lessens clinical severity, and reduces the amount of bacterial shedding from calves infected with M bovis.

Clin Infect Dis, 1999 Jun, 28(6), 1200 - 5
Use of Sulesomab, a radiolabeled antibody fragment, to detect osteomyelitis in diabetic patients with foot ulcers by leukoscintigraphy; Harwood SJ et al.; Diabetic patients suspected of having osteomyelitis secondary to foot ulcers underwent scintigraphic imaging with Sulesomab, an anti-granulocyte antibody Fab' fragment labeled with technetium-99m . Among 122 patients who had osteomyelitis confirmed or excluded by histopathologic and/or microbiologic techniques, Sulesomab had a 91% sensitivity, a 56% specificity, and an accuracy of 80% . One planar imaging session was usually sufficient for diagnosis, typically requiring 20-30 minutes of camera time 1-2 hours after injection . Compared with ex vivo autologous white blood cell (WBC) scans, Sulesomab performed comparably but with significantly greater sensitivity (92% vs . 79%; P < .05) . Sulesomab results were more sensitive than radiography (90% vs . 62%; P < .05) and more specific than bone scans (50% vs . 21%; P < .05) and would have altered management plans in most patients . No related adverse events occurred, and there was no induction of human anti-mouse antibody . Sulesomab is an effective and rapid imaging agent that is diagnostically comparable or superior to WBC scans in this setting, with significant advantages in safety and ease of use.

Diagn Cytopathol, 1999 Sep, 21(3), 180 - 7
Cytologic aspect of fibrolamellar hepatocellular carcinoma in fine-needle aspirates; Perez-Guillermo M et al.; This is a review of the cytologic and clinicopathological findings seen in a series of six fibrolamellar hepatocellular carcinomas (FL-HCC) studied by means of fine-needle aspiration (FNA) . A comparison of several cellular measurements (cell, nuclear and nucleolar sizes, and N/C ratios) of FL-HCC, ordinary hepatocellular carcinoma (O-HCC), and normal hepatocytes was also carried out in order to find out if these figures could be of help in the cytologic diagnosis . Aspirates were made up of a rather monotonous population of large discohesive cells resembling the morphology of the oncocytes seen in thyroid aspirates; trabecular arrangement of tumor cells was not observed . Cytoplasmic pale bodies and hyaline cytoplasmic bodies were seen in variable quantities . Microbiopsies displaying the fibrolamellar pattern were observed in four cases . FL-HCC individual tumor cells were larger than individual O-HCC tumor cells (P < 0.001), as were nuclear (P < 0.007) and nucleolar sizes (P < 0.001), but N/C ratio of O-HCC was higher than the N/C ratio of FL-HCC (P < 0.005) . Based on the findings, a single cell aspirated from an FL-HCC is three times the size of a normal hepatocyte and 1.60 times the size of a single cell aspirated from a well-differentiated O-HCC . The cytologic findings of FL-HCC are very characteristic and permit a correct diagnosis of this liver malignancy, provided the cytopathologist is aware of the clinical, demographic, CT-image, biochemical, and pathological features of this neoplasm . Diagn . Cytopathol . 21:180-187, 1999 .

Microbiol Immunol, 1999, 43(5), 449 - 59
Association of a cellular 21-kDa transmembrane protein (VAP21) with enveloped viruses; Yamamoto K et al.; We reported previously that the rabies virions contained a 21-kDa cellular transmembrane protein (referred to as VAP21) as a minor component (Sagara, J . et al, Microbiol . Immunol . 41(12): 947-955, 1997) . In this study, we further examined the possible interactions of VAP21 with other enveloped viruses, including the vesicular stomatitis virus (VSV; negative-stranded RNA virus), Sindbis virus (positive-stranded RNA virus) and herpes simplex virus type 1 (HSV-1; double-stranded DNA virus) . An immunoblot analysis demonstrated that all of these enveloped viruses contained VAP21 in the virion as a minor component . Immunoprecipitation studies suggested that VAP21 was associated with certain viral proteins in the cell, such as the matrix (M) protein of VSV, a capsid protein of Sindbis virus, and at least a capsid protein (VP5) of HSV-1 . The association was disrupted by treatment with 0.5% sodium dodecyl sulfate, but resistant to the treatment with 1% NP-40 plus 1% deoxycholate . These results suggest that: 1) VAP21 is not primarily associated with the viral transmembrane glycoprotein but rather with the internal viral protein, and, 2) this association would cause the efficient incorporation of VAP21 into the virion.

J Clin Virol, 1999 Aug, 13(3), 121 - 30
HCV genotyping by three methods: analysis of discordant results based on sequencing; Furione M et al.; BACKGROUND: Correct genotyping of hepatitis C virus (HCV) RNA-positive serum samples may have important clinical and therapeutic implications . OBJECTIVES: Three methods were compared to improve accuracy of HCV genotyping . STUDY DESIGN: A panel of 144 HCV RNA-positive sera prospectively tested by a modified Okamoto's type-specific reverse transcription-nested polymerase chain reaction (RT-nPCR) (Okamoto H, Tokita H, Sakamoto M, Kojima M, Iizuka H, Mishiro S . J Gen Virol 1993; 74: 2385-2390) was retrospectively analyzed by two recently described methods which were reported to identify all HCV types and the majority of HCV subtypes: (i) a restriction fragment length polymorphism (RFLP) analysis of PCR products amplified from the 5' untranslated region (5'UTR) of the viral genome (Pohjanpelto P, Lappalainen M, Widell A, Asikainen K, Paunio M . Clin Diagn Virol 1996; 7: 7-16); and (ii) a type-specific RT-nPCR relevant to the core region (Ohno T, Mizokami M, Wu R, Saleh M, Ohba K, Orito E, Mukaide M, Williams R, Lau J . J Clin Microbiol 1997; 35: 201-207) . The panel (according to results given by the modified Okamoto's method) consisted of: (i) 105 sera belonging to five different HCV subtypes; (ii) 20 specimens containing a mixture of > or = 2 genotypes; and (iii) 19 untypeable clinical samples . RESULTS: There was agreement of the three methods for 78/144 (54.2%) blood samples, whereas discordant results were obtained for the remaining 66 samples, 56 of which could be typed by sequencing . Of these, 51 (91.7%) were correctly typed by RFLP, 37 (66.0%) by Ohno's and 27 (48.2%) by the modified Okamoto's procedure . The overall genotyping sensitivity of each method over the total number of 134 samples whose genotype was ascertained, was 96.2% for RFLP, 85.8% for Ohno's and 78.3% for the modified Okamoto's procedure . CONCLUSIONS: RFLP analysis, notwithstanding some limitations in subtyping efficiency of genotype 1 samples, appears superior to the two RT-nPCR methods because: (i) it is able to type a larger number of samples; (ii) it is more efficient in identifying genotypes 2a/c, which are widespread in Italy; (iii) it is highly sensitive (together with Ohno's method) in recognizing genotypes 3 and 4.

FEMS Immunol Med Microbiol, 1999 Aug 1, 25(1-2), 1 - 6
The role of infection in sudden infant death syndrome; Blackwell CC et al.; Studies on the potential role of infectious agents in sudden infant death syndrome (SIDS) have been published over the years in a variety of journals . The aim of this special issue of FEMS Immunology and Medical Microbiology is to bring together a group of the most recent studies from Europe, Australia and Canada which cover epidemiology and laboratory studies examining hypotheses relating to infection and inflammation in SIDS . The articles in this issue examine evidence for the involvement of specific micro-organisms in SIDS and the problems relating to experimental studies on infection in relation to the underlying pathology of these deaths . There is an update on the evidence for the common bacterial hypothesis proposed in 1987 examining risk factors identified in epidemiological studies, particularly how the prone sleeping position could affect bacterial colonisation or induction of toxins . Evidence for induction of inflammatory responses in SIDS infants is reviewed and the relation of these responses to mechanisms proposed as causes of death assessed . Factors found to be associated with reduction of the risk of SIDS (breast feeding and immunisation) are examined in relation to some of the toxigenic bacteria implicated in these deaths . Finally, the high incidence of SIDS in some ethnic groups is examined as a potential model to investigate the contributions of genetic, environmental and cultural differences to susceptibility of infants not only to SIDS but to serious respiratory tract infections.

Rinsho Byori, 1999 Jul, 47(7), 637 - 41
{Laboratory automation and information technology in clinical microbiology--computerized support system and clinical competence of laboratory physician}; Kumasaka K; Laboratory automation and new information technology have considerable potential to improve care through protocols that reduce errors and guide diagnosis and therapy . Computer-based decision-support systems are now in place in many hospitals in the USA . For example, optimal decisions about the use of antibiotics in critically ill patients require access to a large amount of complex information, therefore a computerized decision-support program linked to computer-based patient records can assist physicians in the use of antiinfective agents and improve the quality of care . The support systems can educate physicians, guide their clinical reasoning, and measure the quality of the care that they provide . The installation of the computer components of the system is relatively easy, but the human components of the system may be much more difficult to transfer from hospital to hospital . The successful operation of the system should require a high level of clinical competence in every staff member of the hospital . The project should encompass a broad range of complex clinical conditions and decision algorithms . It must not be focused solely on the sophistication of laboratory methods . We, laboratory physicians/clinical pathologists must modify our behavior effectively and accept the value and limitations of laboratory automation and information technology . We must work more closely with physicians and other health care professionals as the best clients of our clinical laboratories and establish a good collaborative partnership with them.

J Biol Chem, 1999 Aug 13, 274(33), 22977 - 84
Functional and biochemical characterization of Escherichia coli sugar efflux transporters; Liu JY et al.; A family of bacterial transporters, the SET (sugar efflux transporter) family, has been recently reported (Liu, J . Y., Miller, P . F., Gosink, M., and Olson, E . R . (1999) Mol . Microbiol . 31, 1845-1851) . In this study, the biochemical and cell biological properties of the three Escherichia coli members (SetA, SetB, and SetC) of the family are characterized . We show that both SetA and SetB can transport lactose and glucose . In addition, SetA has broad substrate specificity, with preferences for glucosides or galactosides with alkyl or aryl substituents . Consistent with the observed in vitro substrate specificities, strains that hyperexpress SetA or SetB are desensitized to lactose analogues as measured by induction of the lac operon . In addition, strains that hyperexpress SetA are resistant to the growth inhibitory sugar analogue o-nitrophenyl-beta-D-thiogalactoside . Strains disrupted for any one or all of the set genes are viable and show no defects in lactose utilization nor increased sensitivity to inducers of the lac operon and nonmetabolizable sugar analogues . The data suggest that the set genes are either poorly expressed under normal laboratory growth conditions or are redundant with other cellular gene products.

J Am Optom Assoc, 1999 Jun, 70(6), 355 - 83
Antiretroviral therapy for HIV infection: a review of current medications and therapies; Bright DC; BACKGROUND: Management of HIV infection has been revolutionized in the past 3 years by the introduction of drugs significantly more potent than those previously available . Simultaneously revelations about dynamics of HIV infection and viral replication have altered concepts about HIV disease, as well as the need for therapeutic interventions . METHODS: A literature search was conducted of journals listed in the Index Medicus, as well as abstracts from annual meetings of the American Society for Microbiology, the Infectious Disease Society of America (Conference on Retroviruses and Opportunistic Infections), and the International Conferences on AIDS . RESULTS: Antiretroviral drugs of four differing pharmacologic classes are able to achieve profound suppression of viral replication when combined in varying regimens . Costs of medications remain high, many toxicities are associated with drug therapy, and regimens are both complex and daunting . In spite of many difficulties associated with highly active anti-retroviral therapy (HAART), many patients are able to achieve improved quality of life and longer survival . CONCLUSIONS: Optometrists, as members of the health care team, play a role in diagnosis and management of HIV patients, in providing encouragement and empathy to patients--particularly emphasizing the need for adherence to complex therapy regimens, as well as the importance of regular followup.

Perit Dial Int, 1999 May-Jun, 19(3), 259 - 62
Peritonitis due to Stenotrophomonas maltophilia in patients undergoing chronic peritoneal dialysis; Taylor G et al.; The occurrence of cases of Stenotrophomonas maltophilia peritonitis in chronic peritoneal dialysis (PD) patients prompted a review of our experience with this condition . A search of microbiology records revealed seven episodes of S . maltophilia peritonitis in 7 patients in 1996 - 3.8% of all PD patients - compared to no cases in 1994 and 1995 (p = 0.01) . Patients ranged in age from 16 to 64 years; there were 3 males and 4 females . Six of seven episodes of peritonitis were community acquired and one was hospital acquired . No temporal clustering of cases was seen . Patients were from different urban and rural communities . Patients used the same commercially supplied dialysate fluid, different dialysis techniques, and were taught a no-touch technique for connection . Treatment of peritonitis required removal of the Tenckhoff catheter in 4 of 7 cases . Fingerprinting of six available isolates by polymerase chain reaction using primers derived from the conserved region of the 16/23Sr RNA gene sequence and pulsed field gel electrophoresis revealed all to be unique strains . A case-control study comparing 7 S . maltophilia cases to 21 PD controls showed case patients to be younger and more likely to be on immunosuppressive therapy . We conclude that S . maltophilia has emerged as an important cause of peritonitis in our continuous ambulatory PD population . Evidence to date suggests community acquisition with no evidence of a common source.

Antonie Van Leeuwenhoek, 1999 Apr, 75(3), 201 - 6
Pyrolysis mass spectrometry analysis of free-living and symbiotic cyanobacteria; West NJ et al.; The potential of pyrolysis mass spectrometry to distinguish closely related cyanobacterial strains was assessed by using the technique to compare symbiotic cyanobacteria isolated from the hornwort Phaeoceros laevis and free-living cyanobacterial strains at the same field site . The same strains had previously been compared using polymerase chain reaction-based DNA fingerprinting techniques (West & Adams 1997, Appl . Environ . Microbiol . 63: 4479-4484) . Many of the strains were grouped identically by the two techniques, although there were some differences, possibly resulting from the ability of these cyanobacteria to develop a range of specialised cell types having different chemical compositions to the vegetative cells . Although growth conditions were chosen to suppress cellular differentiation, this may not always have been completely successful . With careful control of growth conditions pyrolysis mass spectrometry has considerable potential as an additional tool for the phenetic comparison of cyanobacterial strains . It has the advantage that analysis is directly derived from whole cells, and hence is simpler and cheaper than DNA-based methods, although it does require the growth of axenic strains . The technique may be particularly useful in the study of some of the more cryptic unicellular and non-heterocystous filamentous cyanobacterial groups, in which the lack of cellular differentiation should minimise any variability in the chemical composition of cells.

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