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 &phi;C31 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 diagno