Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 61 - 3 {The future is molecular (micro)biology}; Perez-Trallero E; Until the end of the decade of the 1980's molecular biology did not obtain a significance worthy of standing out in the field of clinical microbiology . From then on and with oscillating movements ranging from euphoria to deception we arrived to the second half of the decade of the 1990's . Since then the enormous potential of molecular biology for the development of clinical microbiology and of medicine in general became evident . The versatility and high diagnostic capacity that will be able to be obtained by the use of genetics helped by bioinformatics make molecular biology one of the indispensable instruments for its progression . Knowing how to adapt the procedures of the future without losing the human factor that modulates, translates and enriches them will be the definitive challenge of our speciality of Clinical Microbiology. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 54 - 60 {Bacterial identification methods}; Gobernado M et al.; In clinical microbiology the identification of bacteria has been, and continues to be, based on traditional classical taxonomy of their phenotypic characteristics such as morphology, appearance in culture media, physiological and biochemical properties, macromolecular degradation, type of respiratory enzyme, nutritional requirements, chemotaxonomic characteristics, inhibition by various substances and reaction to antibodies . Simple techniques are used for their identification such as microscopic examination before or after staining and cultures, as well as more complex procedures, some of which are automated . The future of microbiology applied to the identification of bacteria causing human infections is directed toward the preservation of clinical data as the basis of the entire process and to maintaining microscopy, cultures and simple techniques, that is, to conserving the best of classical methods while at the same time incorporating new technological knowledge and developing techniques of rapid diagnosis . Microbiologists, as well as clinicians, should prepare themselves for the changes represented by conserving traditional methods and applying new technologies. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 49 - 53 {The present and future of the microbiology laboratory in the control of human immunodeficiency virus (HIV) infection}; Moreno S et al.; Until recently, the role of the Microbiology Laboratory with regard to HIV infection was limited to the serological diagnosis, if the contribution to the management of opportunistic infections that complicate the evolution of the disease is excluded . The follow-up of the outcome and monitoring of the response to antiretroviral therapy has been done through CD4 1 lymphocite counts at the Immunology laboratory . In recent years, other important technics for the clinical care of HIV-infected patients have been incorporated to the laboratory of Microbiology: measurement of plasma HIV RNA levels, which has become the main parameter to monitor the response to therapy and for the taking of decisions regarding the beginning and changes of treatment; resistance testing to antiretroviral drugs, not yet fully implemented in our laboratories; and therapeutic drug monitoring, with a role still to be defined . For the near future, new tools will come that, firstly, will improve the existing ones (greater sensitivity to detect HIV RNA in plasma, greater ease of interpretation of the results of genotypic resistance testing, wide access to phenotypic studies) and, secondly, new tools that will allow a more individualized care of the patients (monitoring of some toxic side effects, pharmacogenomics, immunogenomics). Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 44 - 8 {What can the microbiology laboratory currently offer in the professional training of clinical microbiologists?}; Ausina V; In this chapter it is pointed out the scarce information on clinical mycobacteriology included in the current program of the Clinical Microbiology specialty and the needs of a more prolonged and specific formation in this field . The current Mycobacteriology laboratory is an excellent model to evaluate the impact, value and limitations of the new diagnostic techniques . The laboratory of Mycobacterology can currently offer excellent tools for the diagnosis and control of the infections caused by mycobacterias . The current laboratory is inconceivable without a close collaboration among microbiologists, clinicians and epidemiologists . Only in this collaborative scenario where the technology is open and receptive to resolution of xlinical problems we will be able to educate our spaecialists in the future . Probably there is not any other area in the clinical laboratory where the technical and professional competences of the microbiologists are so well defined and limited. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 24 - 7 {Clinical microbiology and infectious diseases}; Gudiol F; Clinical microbiology and infectious diseases, which in Spain began to trace a common route more than twenty-five years ago, share many aspects from a language and scientific community to healthcare, educational and research objectives and should be viewed as complementary specialties . While maintaining their identities, both in the professional and academic spheres, both specialties should move toward models of integration that would make them stronger and more efficient, thus guaranteeing their future as well-established specialties in our health system. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 17 - 23 {Systems of quality management in clinical laboratories: certification and accreditation}; Gimeno C; The aim of this article is to review the concept of quality and the vigent rules in order to implant a quality system in the Clinical Microbiology laboratories . We describe the different ways of its application (autonomic administrative autorization, certification based in the ISO 9001:2000 and accreditation following the ISO 17025 rule) . We denote the advantages and the disadvantages of each one and also the keys to put them into practice. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 15 - 6 {Management in clinical microbiology: Activity measurement and outsourcing}; Picazo JJ; More than 20 years ago the Spanish health system understood that accurate determination of a set of variables related to the health activity performed (workload, quality assurance, activity costs, etc.) was essential . The creation of an Activities Catalogue is a key requirement in evaluating our work.Outsourcing is only possible if quality of care is maintained, otherwise other hospital services would suffer, irreparably affecting patients . A different matter would be the appropriate concentration of particular resources that could (and should) take place in public institutions. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 12 - 4 {Coordination among clinical microbiology laboratories}; Martin R; Microbiology laboratories are facing, during last years, new organizative models wich sometimes prioritize economical management instead of their assistential, docent and research work . The microbiologist should contribute with our experience to make compatible good microbiology practice with the system health management . We propose to stablish different assistential laboratory levels and the coordination among them. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 9 - 11 {Medical microbiology in the European Union}; Andreu A; Although in the majority of countries on the European Union (EU) the Clinical o Medical Microbiology is a recognized (with the exceptions of Portugal, France and Belgium) and well established specialty (with the exception of Portugal), the training programme as well as the circumstances that conform their practice are quite different . In the construction of the new Europe there is the feeling of the necessity to unify the Medical Microbiology in an only doctrinaire body with the goal to obtain the same certification and to facilitate the free movement of microbiologists throughout the EU . To this purpose, the Commission of Medical Microbiology of the Union of European Medical Specialist (UEMS), with the support of the European Society of Clinical Microbiology and Infectious Diseases (ESMID), has elaborate a training programme to be discussed and approved by the Scientific Societies and the National Specialties Commissions of each country, and then be presented to the appropriated EU organisms . The core training programme, with 5 years period, aims to provide the trainee with both the theoretical foundation and the practical, technical, clinical and managerial skills necessary for the independent specialist practice of Medical Microbiology . This programme contain a high clinical training, meaning that the microbiologist specialist is not only who set protocols and maintain standards within the laboratory but also who goes out as adviser for infectious diseases management, control and prevention of the hospitals acquired infection and control of antibiotic usage, among other responsibilities. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 7 - 8 {The clinical microbiologist of the future}; Ausina V; In this chapter the origin of the crisis and the current situation of the specialty on Microbiology and Parasitology in Spain is analized . The competences of the future Clinical Microbiologists are defined. Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 2 - 6 {Clinical microbiology in the XXI century . A new scenario}; Perea EJ; The centre-stage position recently reached by Microbiology has not spared our professional activity numerous problems.Professional difficulties imposed by social and economic circumstances are discussed . These circumstances induce our work to take place on a not so favourable stage.Financial limitations imposed on our professional activity by the current trend to provide free healthcare for all are considered; along with the alternatives that could be adopted on a socio-economic or individual basis.On a human level, the cost has been even greater due to the reduction in resources . This process has gone through several phases such as the reduction in staff, with the resulting increase in workload for the remaining team members, possibly leading sub-contracting or outsourcing.Finally, a number of initiatives aimed at increasing the awareness and appreciation of the function of Clinical Microbiologists by both management and society are suggested . Two examples are the compilation of reports on the significance and benefits of our profession, and refocusing our activity ensure a better future for Clinical Microbiology. Indian J Pathol Microbiol, 2003 Oct, 46(4), 541 - 50 Nine-banded armadillo and leprosy research; Job CK; In this presentation an attempt has been made to describe the nine-banded armadillo as an animal model, probably the only one in which lepromatous leprosy similar to that found in humans can be experimentally produced . Some unique features of the physiology of the animal are mentioned . The pathology and the microbiology of leprosy in the armadillo are described in detail . The discovery of lepromatous leprosy in the wild armadillos in the southern parts of United States, the transmission of disease among them through trauma and thorn pricks and the pathogenesis of the disease are presented . The impact of leprosy in the wild animals may have on human leprosy is discussed. Asia Pac J Clin Nutr . 2003 Nov;12(Suppl):S50. Sustainably managing food production resources to maximise human nutritional benefit; Beeton R; Nutrition is an integrating discipline that draws on a diverse range of reductionist disciplines . It embraces biochemistry, molecular biology, microbiology, psychology, physiology and epidemiology to name a few . The benefits of the discipline for humans lie in the efficient production of food and its intelligent use by consumers . Food is not simply fuel; it is overlain by culture and identity, is in many ways an index of the status of consumers and is symbolic of their stage of economic and consciousness 'development' . The challenges for the future lie in the social context of food production . The sustainable production of food and its safety for humans reflects self image and a new consciousness . Human nutritional benefit is problematic . The benefit can be an expression of health or it can be an expression of well-being that embraces both health and the environments both actual and vicarious . The latter is the forum for nutritional futures . Future nutritional concerns will see food as a component of the body that is clean and a component of the 'soul' that is green . It is relatively simple to understand the system that renders quality assured safe food with certain nutritional properties . It is far more difficult to understand the lifecycle impacts of food . The problem is that while the nutritional properties of two items of the same food may be identical the whole lifecycle impact on the environment of the two identical items may differ enormously . This in turn reflects the spatial and temporal heterogeneity of food production systems and is further complicated by markets that segment on quality variables (objective and perceptual) rather than source lifecycle variables . The lifecycle approach to sustainable food production is holistic and has to be challenging to a profession that is fragmented in character . The divide is not however between positivism and constructivism but between holism and reductionism . Such a divide is not easily understood save through a paradigm shift to a systems and systematic approach. Diagn Microbiol Infect Dis, 2004 Mar, 48(3), 173 - 4 What happens when automated blood culture instrument detect growth but there are no technologists in the microbiology laboratory? Savinelli T, Parenteau S, Mermel LA. We investigated blood cultures that revealed growth at night when no technologists were in the microbiology laboratory and at other times when the laboratory was staffed . On average, it took 7 h and 26 min and 2 h and 12 min for laboratory personnel to report positive blood culture Gram stain results to physicians when growth was detected by automated instruments during the former and latter time periods, respectively . When positive blood culture results led to a change in therapy, it took an average of 8 h and 21 min and 5 h and 26 min in the former and latter groups, respectively, from detection to when the order was written. Infect Dis Obstet Gynecol, 2003, 11(3), 167 - 9 Tubo-ovarian abscess occurring 16 years after supracervical hysterectomy; Tohya T et al.; BACKGROUND: Supracervical hysterectomy is seldom performed and there are few reports of tubo-ovarian abscess (TOA) after supracervical hysterectomy . CASE: The case of a 49-year-old woman with a right TOA is reported . This patient had received a supracervical hysterectomy 16 years earlier due to rupture of the uterus . At this admission, she presented with complaints of, lower abdominal pain and fever . Bimanual and transvaginal ultrasound examinations demonstrated a tender mass in the right adnexal region . Laparotomy, pathologic examination and microbiologic study confirmed the diagnosis of right TOA . CONCLUSION: After supracervical hysterectomy, patients may develop endocervicitis, parametritis and/or TOA . This series may be a subtype of ascending infections in the female genital tract. Ann Epidemiol, 2004 Feb, 14(2), 137 - 42 Recurrent gastroenteritis among infants in Western Australia: a seven-year hospital-based cohort study; Lee AH et al.; PURPOSE: To investigate factors that affect the frequency of recurrent gastroenteritis among infants in Western Australia (WA) . METHODS: A 7-year retrospective cohort study was undertaken on all infants born in 1995 who were admitted for gastroenteritis during their first year of life (n=514) . Linked hospitalization records of the cohort were retrieved to derive the number of readmissions, microbiologic diagnoses, patient demographics, and co-morbidities at the index episode . A negative binomial regression model adjusting for inter-hospital variations was used to determine the prognostic factors influencing recurrent gastroenteritis . RESULTS: Diarrhea with no specific etiology accounted for 54.7% of the cases presented at index admission and 55.8% of the total 676 admissions for the cohort . Of the 514 infants, 119 (23%) experienced repeated episodes of gastroenteritis . The lowest proportion of recurrences was 15.4% for patients initially admitted for bacterial or viral diarrhea . Over 85% of the recurrences from either bacterial and viral diarrhea or etiology unspecified were readmitted under the same category . Aboriginality and dehydration were significantly associated with the recurrence frequency, the adjusted incidence rate ratio being 2.86 (95% CI, 1.92-4.26) and 0.66 (95% CI, 0.49-0.88), respectively . Aboriginal infants contributed to 58% of those patients in the cohort who sustained repeated episodes of gastroenteritis . The proportion of patients with the recurrent disease was also significantly higher for Aboriginals (39%) than for non-Aboriginals (15%) . The effect of dehydration was evident after accounting for within hospital correlations . CONCLUSIONS: Hospitalizations for recurrent gastroenteritis were more frequent among Aboriginal children than non-Aboriginal children in WA . Readmissions were also related to the presence of dehydration at the index episode . These findings have implications for preventive strategies to reduce the burden of gastroenteritis. MMWR Morb Mortal Wkly Rep, 2004 Mar 12, 53(9), 192 - 4 Mycobacterium chelonae infections associated with face lifts--New Jersey, 2002-2003; Centers for Disease Control and Prevention (CDC); In March 2003, the New Jersey Department of Health and Senior Services (NJDHSS) was notified about three patients who acquired surgical-site infections caused by Mycobacterium chelonae after having face lifts (i.e., rhytidectomies) performed at an outpatient surgical center . NJDHSS learned subsequently of another patient with M . chelonae infection who had a rhytidectomy performed at a second surgical center . The four patients received diagnoses of M . chelonae infection during March 2002-February 2003 . NJDHSS conducted an epidemiologic, environmental, and microbiologic investigation . This report summarizes the results of that investigation, which identified contaminated methylene blue used as a tissue-marking agent as the source of infection . Surgeons should use only sterile, single-use, tissue-marking agents during procedures that require aseptic technique, and clinicians should consider M . chelonae when evaluating surgical-site infections. Water Res, 2004 Mar, 38(6), 1514 - 9 Mechanisms of inactivation of hepatitis A virus in water by chlorine dioxide; Li JW et al.; In this study, to elucidate the mechanisms of inactivation of hepatitis A virus (HAV) by chlorine dioxide, cell culture, enzyme-linked immunosorbent assay (ELISA), and long-overlapping RT-PCR were used to detect the infectivity, antigenicity, and entire genome of HAV before and after disinfection . The results revealed the complete inactivation of infectivity after a 10-min exposure to 7.5mg of chlorine dioxide per liter; and the highest level of sensitivity in the 5'non-translated regions (5'NTR) (the sequence from bp 1 to 671), inactivation of which took as much time as the inactivation of infectivity of HAV by chlorine dioxide; the complete destruction of antigenicity after a 10-min exposure to 7.5mg of chlorine dioxide per liter . It is suggested that the inactivation mechanism of HAV by chlorine dioxide was due to the loss of the 5'NTR and/or destruction of the antigenicity, which is not similar to that of chlorine (Appl Environ Microbiol 68: 4951). Waste Manag, 2004, 24(3), 227 - 40 Modelling the biochemical degradation of solid waste in landfills; White J et al.; This paper describes the concept of a generic spatially distributed numerical model that has been developed to contain and link sub-models of landfill processes in order to simulate solid waste degradation and gas generation in landfills . The model includes the simulation of the transport of leachate and gases, and the consolidation of the solid waste . The structure of the model consists of linked discrete constant volume elements . The paper outlines the theoretical background that provides the framework to contain the numerical procedures that make up the model . Details are also given of the approach to the modelling of the chemistry and microbiology of solid waste degradation. Eur J Clin Microbiol Infect Dis, 2004 Apr, 23(4), 289 - 99 Epub 2004 Mar 10. False-positive results and contamination in nucleic acid amplification assays: suggestions for a prevent and destroy strategy; Borst A et al.; Contamination of samples with DNA is still a major problem in microbiology laboratories, despite the wide acceptance of PCR and other amplification techniques for the detection of frequently low amounts of target DNA . This review focuses on the implications of contamination in the diagnosis and research of infectious diseases, possible sources of contaminants, strategies for prevention and destruction, and quality control . Contamination of samples in diagnostic PCR can have far-reaching consequences for patients, as illustrated by several examples in this review . Furthermore, it appears that the (sometimes very unexpected) sources of contaminants are diverse (including water, reagents, disposables, sample carry over, and amplicon), and contaminants can also be introduced by unrelated activities in neighboring laboratories . Therefore, lack of communication between researchers using the same laboratory space can be considered a risk factor . Only a very limited number of multicenter quality control studies have been published so far, but these showed false-positive rates of 9-57% . The overall conclusion is that although nucleic acid amplification assays are basically useful both in research and in the clinic, their accuracy depends on awareness of risk factors and the proper use of procedures for the prevention of nucleic acid contamination . The discussion of prevention and destruction strategies included in this review may serve as a guide to help improve laboratory practices and reduce the number of false-positive amplification results. Hepatogastroenterology, 2004 Jan-Feb, 51(55), 313 - 5 Improvement of complete blood count in patients with iron deficiency anemia and Helicobacter pylori infection after the eradication of Helicobacter pylori; Hacihanefioglu A et al.; BACKGROUND/AIMS: Recent reports support the possible association between Hp infection and iron deficiency anemia . In the present study, the effects of the eradication therapy on iron deficiency anemia were investigated . METHODOLOGY: Fourteen women with iron deficiency anemia were enrolled (mean age: 36.4 years; range: 20-52 years old) . None of the patients received iron supplementation . Several examinations including upper and lower gastrointestinal endoscopy were performed to reveal any gastrointestinal bleeding sites in all patients . Gastric biopsies during endoscopy were taken from the subjects except one whose serum anti-Hp IgG was positive . After diagnosing the Hp infection by means of microbiology, histology and Gram stain, a combination therapy consisting of lansoprosol, clarithromycin and amoxicillin was administered to each patient . Hematologic examinations and the body iron status were evaluated periodically, following the eradication therapy . RESULTS: Endoscopic findings were as follows: Seven patients with antral gastritis, two patients with pangastritis, whereas five patients were found to be endoscopically normal . None of the subjects were found to have gastrointestinal bleeding of any type . Serum hemoglobin, iron and transferrin saturations of the patients were found to be increased at 20-24 weeks of follow-up after the eradication therapy . Serum ferritin levels were not found to be increased . CONCLUSIONS: Hp infection may be involved in cases of iron deficiency anemia of unknown origin, and the eradication of the infection may improve blood parameters other than serum ferritin levels. In Vivo, 2004 Jan-Feb, 18(1), 33 - 5 The 500-base-pair fragment of the putative gene RvD1-Rv2031c is also present in the genome of Mycobacterium tuberculosis; Metaxa-Mariatou V et al.; BACKGROUND: It has been proposed that differentiation between M . bovis and M . tuberculosis is possible by using a PCR assay for the 500bp fragment present only in the M . bovis genome . MATERIALS AND METHODS: Forty clinical samples and 16 clinical isolates from the Department of Microbiology, as well as 4 clinical isolates obtained from another laboratory, were tested for the purpose of this study . As controls we tested 2 M . bovis (M . bovis BCG Pasteur TMC1011 and M . bovis BCG Copenhagen), 1 H37Rv M . tuberculosis strain, 2 M . avium (ATCC15765 and ATCC1975, respectively) and 1 M . paratuberculosis (ATCC19698) strains . RESULTS: None of the mtp40- negative clinical isolates amplified the 500bp fragment, whereas 4 out of 17 mtp40-positive clinical isolates scored positive for the 500bp fragment . All clinical isolates scored positive for IS6110, mtp40, the pncA and oxyR PCR's . All but one of the clinical isolates amplified the 500bp fragment . Sequence analysis of the pncA and oxyR PCR products revealed the presence of nucleotide C at position 169 and G at position 285 respectively, suggesting M . tuberculosis as the causative agent . CONCLUSION: Our data suggest that the 500bp PCR fragment is present not only in M . bovis but also in M . tuberculosis. Am J Otolaryngol, 2004 Jan-Feb, 25(1), 5 - 10 Chronic rhinosinusitis and adenoid hypertrophy in children; Tuncer U et al.; PURPOSE: The purpose of this study was to investigate the relationship between adenoid microbiology, adenoid size, and maxillary sinus microbiology in children with both chronic rhinosinusitis and adenoid hypertrophy . METHODS: The children with both chronic rhinosinusitis and adenoid hypertrophy were admitted to this prospective study . The study included 30 children . The diagnosis of chronic rhinosinusitis was based on clinical and radiologic examinations . Adenoid hypertrophy was classified as medium and large based on the preoperative flexible fiberoptic endoscopy and nasal endoscopy during surgery . Maxillary sinus aspiration and adenoidectomy was performed in all patients . Sinus aspirate and adenoid tissue specimens were cultured . The correlation of culture results was investigated . The relationship between adenoid size and maxillary sinus culture results was analyzed by using chi-square test . RESULTS: Adenoid sizes were medium in 12 (40%) and large in 18 (60%) cases . Bacterial growth was found on 14 of 30 (47%) sinus aspirate, and all adenoid specimen cultures showed bacterial growth (100%) . There was no statistical correlation between cultures obtained from the adenoid tissue with those from the maxillary sinus . The relationship between adenoid size and maxillary sinus culture results was not found statistically significant (chi(2) = 0.96, P = 1.0) . CONCLUSION: The reason that there was no correlation between cultures obtained from the adenoid tissue with those from the maxillary sinus is that it seems possible that the adenoids act as a barrier causing mechanical obstruction rather than a nidus for chronic sinus infection . However, there is no relationship between adenoid size and maxillary sinus culture positivity . Medium adenoids causing partial obstruction may lead to changes in the microenvironment and may start bacterial growth in children with positive maxillary culture . Further investigation is needed to explain the association between adenoid hypertrophy and rhinosinusitis . Adenoidectomy helps to resolve the symptoms of chronic rhinosinusitis in the children with both chronic rhinosinusitis and adenoid hypertrophy. Syst Parasitol, 2004 Mar, 57(3), 211 - 9 Description of the nymph and larva and redescription of the female of Ixodes neuquenensis Ringuelet, 1947 (Acari: Ixodidae), a parasite of the endangered Neotropical marsupial Dromiciops gliroides Thomas (Microbiotheria: Microbiotheriidae); Guglielmone AA et al.; The female of Ixodes neuquenensis Ringuelet, 1947 (Acari: Ixodidae) is redescribed and the nymph and larva are described from specimens collected from the endangered marsupial Dromiciops gliroides Thomas (Microbiotheria: Microbiotheriidae) in Argentina . At first sight the female of I . neuquenensis resembles a member of the subgenus Ixodes Latreille, 1795 . However, the female of I . neuquenensis is peculiar in having the combination of two spurs on coxae II-IV and a pair of chitinous plaques internal to coxa I . Both the nymph and larva have an anterior and posterior process on palpal article I, characteristics of the subgenus Ixodiopsis Filippova, 1957 and some representatives of the subgenus Pholeoixodes Schulze, 1942 . Analysis of 16S mitochondrial rDNA sequences showed no strong relationship with any known Ixodes subgenus. Cell Microbiol, 2004 Apr, 6(4), 333 - 43 Focusing light on infection in four dimensions; Roux P et al.; The fusion of cell biology with microbiology has bred a new discipline, cellular microbiology, in which the primary aim is to understand host-pathogen interactions at a tissue, cellular and molecular level . In this context, we require techniques allowing us to probe infection in situ and extrapolate quantitative information on its spatiotemporal dynamics . To these ends, fluorescent light-based imaging techniques offer a powerful tool, and the state-of-the-art is defined by paradigms using so-called multidimensional (multi-D) imaging microscopy . Multi-D imaging aims to visualize and quantify biological events through time and space and, more specifically, refers to combinations of: three (3D, volume), four (4D, time) and five (5D, multiwavelength)-dimensional recordings . Successful multi-D imaging depends upon understanding the available technologies and their limitations . This is especially true in the field of microbiology where visualization of infectious/pathogenic activities inside living host systems presents particular technical challenges . Thus, as multi-D imaging rapidly becomes a common bench tool to the cellular microbiologist, this review provides the new user with some of the necessary technical insight required to get the best from these methods. J Mol Evol, 2003, 57 Suppl 1, S3 - 12 Radiation of extant marsupials after the K/T boundary: evidence from complete mitochondrial genomes; Nilsson MA et al.; The complete mitochondrial (mt) genomes of five marsupial species have been sequenced . The species represent all three South American orders (Didelphimorphia, Paucituberculata, and Microbiotheria) . Phylogenetic analysis of this data set indicates that Didelphimorphia is a basal marsupial lineage followed by Paucituberculata . The South American microbiotherid Dromiciops gliroides (monito del monte) groups with Australian marsupials, suggesting a marsupial colonization of Australia on two occasions or, alternatively, a migration of an Australian marsupial lineage to South America . Molecular estimates suggest that the deepest marsupial divergences took place 64-62 million years before present (MYBP), implying that the radiation of recent marsupials took place after the K/T (Cretaceous/Tertiary) boundary . The South American marsupial lineages are all characterized by a putatively non-functional tRNA for lysine, a potential RNA editing of the tRNA for asparagine, and a rearrangement of tRNA genes at the origin of light strand replication. Orthopade, 2004 Mar, 33(3), 273 - 86 {Hematogenous osteomyelitis in adults}; Strecker W et al.; Hematogenous osteomyelitis (HOM) in adults is a very rare event in industrialised countries . However, in tropical regions the morbidity of HOM is more important, primarily due to the impact of sickle cell disease, thalassemia, HIV-infection and tuberculosis . HOM is most commonly caused by pyogenic bacteria and mycobacteria, but infections by fungi, viruses and parasites must also be considered . In spite of modern diagnostic procedures such as nuclear and magnetic resonance imaging, the histopathologic and microbiologic examination of bone remains the gold standard for diagnosing OM . Other diagnoses should also be considered . Nonbacterial osteomyelitic lesions (plasmacellular OM, sclerosing OM, SAPHO syndrome) as well as acute leukemia, malignant bone tumors (i.e., Ewing's sarcoma, osteosarcoma) are conditions with similar presentations . Acute HOM is best managed by appropriate antibiotic therapy . In case of failure and in chronic HOM, surgical debridement is mandatory. Appl Environ Microbiol, 2004 Mar, 70(3), 1830 - 2 Detection of Escherichia coli serogroups O26 and O113 by PCR amplification of the wzx and wzy genes; DebRoy C et al.; PCR-based assays for detecting enterohemorrhagic Escherichia coli serogroups O26 and O113 were developed by targeting the wzx (O-antigen flippase) and the wzy (O-antigen polymerase) genes found in the O-antigen gene cluster of each organism . The PCR assays were specific for the respective serogroups, as there was no amplification of DNA from non-O26 and non-O113 E . coli serogroups or from other bacterial genera tested . Using the PCR assays, we were able to detect the organisms in seeded apple juice inoculated at concentration levels as low as < or =10 CFU/ml . The O26- and O113-specific PCR assays can potentially be used for typing E . coli O26 and O113 serogroups; these assays will offer an advantage to food and environmental microbiology laboratories in terms of identifying these non-O157 serogroups by replacing antigen-based serotyping. Appl Environ Microbiol, 2004 Mar, 70(3), 1307 - 14 Physiology and nutrition of Treponema primitia, an H2/CO2-acetogenic spirochete from termite hindguts; Graber JR et al.; Treponema primitia strains ZAS-1 and ZAS-2, the first spirochetes to be isolated from termite hindguts (J . R . Leadbetter, T . M . Schmidt, J . R . Graber, and J . A . Breznak, Science 283:686-689, 1999), were examined for nutritional, physiological, and biochemical properties relevant to growth and survival in their natural habitat . In addition to using H(2) plus CO(2) as substrates, these strains were capable of homoacetogenic growth on mono- and disaccharides and (in the case of ZAS-2) methoxylated benzenoids . Cells were also capable of mixotrophic growth (i.e., simultaneous utilization of H(2) and organic substrates) . Cell extracts of T . primitia possessed enzyme activities of the Wood/Ljungdahl (acetyl coenzyme A) pathway of acetogenesis, including tetrahydrofolate-dependent enzymes of the methyl group-forming branch . However, a folate compound was required in the medium for growth . ZAS-1 and ZAS-2 growing on H(2) plus CO(2) displayed H(2) thresholds of 650 and 490 ppmv, respectively . Anoxic cultures of ZAS-1 and ZAS-2 maintained growth after the addition of as much as 0.5% (vol/vol) O(2) to the headspace atmosphere . Cell extracts exhibited NADH and NADPH peroxidase and NADH oxidase activities but neither catalase nor superoxide dismutase activity . Results indicate that (i) T . primitia is able to exploit a variety of substrates derived from the food of its termite hosts and in so doing contributes to termite nutrition via acetogenesis, (ii) in situ growth of T . primitia is likely dependent on secretion of a folate compound(s) by other members of the gut microbiota, and (iii) cells possess enzymatic adaptations to oxidative stress, which is likely to be encountered in peripheral regions of the termite hindgut. Mech Dev, 2004 Mar, 121(3), 213 - 24 The Caenorhabditis elegans eukaryotic initiation factor 5A homologue, IFF-1, is required for germ cell proliferation, gametogenesis and localization of the P-granule component PGL-1; Hanazawa M et al.; Eukaryotic initiation factor 5A (eIF-5A) was originally isolated as a translation initiation factor . However, this function has since been reconsidered, with recent studies pointing to roles for eIF-5A in mRNA metabolism and trafficking {Microbiol . Mol . Biol . Rev . 66 (2002) 460; Eur . Mol . Biol . Org . J . 17 (1998) 2914} . The Caenorhabditis elegans genome contains two eIF-5A homologues, iff-1 and iff-2, whose functions in vivo were examined in this study . The iff-2 mutation causes somatic defects that include slow larval growth and disorganized somatic gonadal structures in hermaphrodites . iff-2 males show disorganized tail sensory rays and spicules . On the other hand, iff-1 mRNA is expressed in the gonad, and the lack of iff-1 activity causes sterility with an underproliferated germline resulting from impaired mitotic proliferation in both hermaphrodites and males . In spite of underproliferation, meiotic nuclei are observed, as revealed by presence of immunoreactivity to the anti-HIM-3 antibody; however, no gametogenesis occurs in the iff-1 gonads . These phenotypes are in part similar to the mutants affected in the components of P granules, which are the C . elegans counterparts of germ granules {Curr . Top Dev . Biol . 50 (2000) 155} . We found that localization of the P-granule component PGL-1 to P granules is disrupted in the iff-1 mutant . In summary, the two C . elegans homologues of eIF-5A act in different tissues: IFF-2 is required in the soma, and IFF-1 is required in the germline for germ cell proliferation, for gametogenesis after entry into meiosis, and for proper PGL-1 localization on P granules. Curr Opin Plant Biol, 2004 Apr, 7(2), 137 - 47 What can bacterial genome research teach us about bacteria-plant interactions? Puhler A, Arlat M, Becker A, Gottfert M, Morrissey JP, O'Gara F. Biological research is changing dramatically . Genomic and post-genomic research is responsible for the accumulation of enormous datasets, which allow the formation of holistic views of the organisms under investigation . In the field of microbiology, bacteria represent ideal candidates for this new development . It is relatively easy to sequence the genomes of bacteria, to analyse their transcriptomes and to collect information at the proteomic level . Genome research on symbiotic, pathogenic and associative bacteria is providing important information on bacteria-plant interactions, especially on type-III secretion systems (TTSS) and their role in the interaction of bacteria with plants. Mycoses, 2004 Feb, 47(1-2), 82 - 4 Chronic bilateral otomycosis caused by Aspergillus niger; Mishra GS et al.; Aspergillus niger, an opportunistic filamentous fungus, was identified as the cause of chronic bilateral otomycosis in a 46-year-old female patient who was unresponsive to different drugs . The patient showed signs of erythema, otalgia, itching, otorrhoea and presence of greyish black coloured mass in both the ear canals . The direct microscopical examination of the ear debris in potassium hydroxide preparations, Giemsa, phase contrast and Gram revealed many thin, branched septate hyphae, condia and conidiophores morphologically indistinguishable from Aspergillus spp . The histopathological section of the ear wax mass by haematoxylin and eosin and periodic acid-Schiff techniques also showed similar fungal elements . The patient responded to 1% solution of mercurochrome . The use of mercurochrome in developing countries like India may be recommended to treat the fungal otitis in patients . We also emphasize that 'Narayan' stain should be routinely employed by microbiology and public health laboratories to study the morphology of pathogenic fungi. Clin Cornerstone, 2003, Suppl 3, S12 - 20 Antibiotics and collateral damage; Hoban DJ; The precarious stability of the normal indigenous flora of the human gastrointestinal tract may be disturbed by a number of factors, but frequently and crucially by antibiotic therapy . This article explores recent insights on the collateral and ecologic effects of antibiotics on the microbiologic flora of the human body and the possible consequences of those effects, which are just beginning to be better understood . New data on this subject will not only help in designing better clinical trials but also begin to answer key questions about collateral damage. Sci Total Environ, 2004 Mar 5, 320(1), 25 - 35 In vivo studies on lead content of deciduous teeth superficial enamel of preschool children; Gomes VE et al.; The amount of lead in the superficial enamel of deciduous teeth from 4- and 5-year-old children was determined by means of an enamel biopsy followed by lead analysis by graphite furnace atomic absorption spectrometry . Children examined in this study (n=329) attended public preschools in the city of Piracicaba, SP, Brazil . The selection of preschools was based on geographical location: some were located in an industrial area of the city, whereas others were located in an area devoid of industries . The objectives of this study were (1) to test whether enamel biopsies were an appropriate method for lead content population screening in young children; (2) to verify the lead content of superficial enamel of preschool children's deciduous teeth and to compare the results obtained from children exposed to different environments; (3) to analyze how biopsy depth correlated with lead content and affected comparisons between groups . An adhesive tape with a central hole (diameter=1.6 mm) was placed on one of the upper central incisors . Five microliters of 1.6 M l(-1) HCl in 70% glycerol (v/v) were applied to this area for 20 s, followed by a rinse with 5-microl ultrapurified water for 10 s . It was shown that the technique was feasible for population studies on young children . The lead concentration in enamel biopsies from the industrial area children (median: 169 ppm Pb) was significantly higher than that from the children living far from industries (118.1 ppm Pb) (P<0.0001) . Analysis of the data showed that biopsies deeper than 3.9 microm gave more reliable results for comparisons between groups . To our knowledge, this study is the first to use an enamel microbiopsy to characterize a large sample of young children in vivo for lead monitoring in teeth. Enferm Infecc Microbiol Clin, 2004 Mar, 22(3), 183 - 91; quiz 192 {Multiplex PCR in clinical microbiology}; Mendez-Alvarez S et al.; The introduction of molecular biology methods in clinical microbiology laboratories brings important insights to obtain sensitive and specific diagnoses as fast as possible . These methods are not intended for replacement but for complement of the already applied microbiologic methods . The integrated analyses of all of them is bringing to the most feasible and efficient results . Within the molecular techniques applied, polymerase chain reaction (PCR) has acquired a great diagnostic value, permitting the identification of etiologic agents and the fast and sensitive detection of their virulence and resistance genotypes . Since some years ago, the development of the so called multiplex PCRs has gained deep interest . Those are reactions that get the simultaneous single tube amplification of different target sequences, allowing the simultaneous detection and identification of various genes of interest . In the present article, the most relevant applications of multiplex PCR for clinical microbiology are summarized. Endoscopy, 2004 Mar, 36(3), 217 - 9 A teaching model for endorectal ultrasound-guided biopsy and drainage of pararectal tumors; Bussen D et al.; For determination of histology of small solid and microbiology of liquid pararectal tumors there is no other method than endorectal ultrasound-guided biopsy . Also for therapy of abscesses or seromas drainage application is possible . With this technique an operation may be prevented . In awareness of possible complications such as major injury of rectal wall, bleeding, persistent fistula and sepsis, teaching and training in this method is consequently necessary . For a training program a lifelike model of the rectum was developed . It has been used by 780 participants in 20 training courses organized by the Surgical Department of University Hospital Wurzburg. J Antimicrob Chemother, 2004 Apr, 53(4), 567 - 76 Epub 2004 Feb 25. Antibiotic resistance in general dental practice--a cause for concern? Sweeney LC, Dave J, Chambers PA, Heritage J. This review examines the contribution dental prescribing makes to the selection of antibiotic resistance in bacteria of the oral flora . The antibiotics commonly used in dental prescribing in the UK are discussed, together with the problems of resistance in members of the oral flora . The antibiotic prescribing habits of general dental practitioners are then reviewed with respect to therapeutic prescriptions and those drugs that are prescribed prophylactically . Not all antibiotic prescriptions for dental problems are written by dentists; prescribing outside the dental profession is also considered . The review then considers the support available to dentists from clinical diagnostic microbiology laboratories . It concludes that better use of diagnostic services, surveillance and improvements in dental education are required now to lessen the impact of antibiotic resistance in the future. Int J Food Microbiol, 2004 Mar 15, 91(3), 261 - 77 Estimating the bacterial lag time: which model, which precision? Baty F, Delignette-Muller ML. The objective of this work was to explore the large number of bacterial growth models recently proposed in the field of predictive microbiology, concerning their capacity to give reliable estimates of the lag phase duration (lambda) . We compared these models on the basis of their underlying biological explanations of the lag phenomenon, their mathematical formulation and their statistical fitting properties . Results show that a variety of biological interpretations of the lag phase exists, although different biological hypotheses sometimes converge to give identical mathematical equations . The fit of the different models provides relatively close lambda estimates, especially if we consider that the imprecision of the lambda estimates is generally larger than the differences between the models . In addition, the consistency of the lambda estimates closely depends on the quality of the dataset on which models were fitted. Biosci Biotechnol Biochem, 2004 Feb, 68(2), 413 - 20 Purification, characterization, and subsite affinities of Thermoactinomyces vulgaris R-47 maltooligosaccharide-metabolizing enzyme homologous to glucoamylases; Ichikawa K et al.; A maltooligosaccharide-metabolizing enzyme from Thermoactinomyces vulgaris R-47 (TGA) homologous to glucoamylases does not degrade starch efficiently unlike most glucoamylases such as fungal glucoamylases (Uotsu-Tomita et al., Appl . Microbiol . Biotechnol., 56, 465-473 (2001)) . In this study, we purified and characterized TGA, and determined the subsite affinities of the enzyme . The optimal pH and temperature of the enzyme are 6.8 and 60 degrees C, respectively . Activity assays with 0.4% substrate showed that TGA was most active against maltotriose, but did not prefer soluble starch . Kinetic analysis using maltooligosaccharides ranging from maltose to maltoheptaose revealed that TGA has high catalytic efficiency for maltotriose and maltose . Based on the kinetics, subsite affinities were determined . The A1+A2 value of this enzyme was highly positive whereas A4-A6 values were negative and little affinity was detected at subsites 3 and 7 . Thus, the subsite structure of TGA is different from that of any other GA . The results indicate that TGA is a metabolizing enzyme specific for small maltooligosaccharides. Front Biosci, 2004 May 01, 9, 1743 - 58 Proteasomes: perspectives from the Archaea; Maupin-Furlow JA et al.; The development of whole systems approaches to microbiology (e.g . genomics and proteomics) has facilitated a global view of archaeal physiology . Surprisingly, as archaea respond to environmental signals, the majority of protein concentration changes that occur are not reflected at the mRNA level . This incongruity highlights the importance of post-transcription control mechanisms in these organisms . One of the central players in proteolysis is the proteasome, a multicatalytic energy-dependent protease . Proteasomes serve both proteolytic and non-proteolytic roles in protein quality control and in the regulation of cell function . The proteolytic active sites of these enzymes are housed within a central chamber of an elaborate nanocompartment termed the 20S proteasome or core particle . Axial gates, positioned at each end of this particle, restrict the type of substrate that can access the proteolytic active sites . Assortments of regulatory AAA complexes are predicted to recognize/bind and unfold substrate proteins, open the axial gates, and translocate substrate into the 20S core particle. Infez Med, 1996, 4(3), 149 - 52 {Bacterial endocarditis in an infectious diseases department: our experience}; Urbani C et al.; The infective endocarditis remains a serious disease . The microbiologic, immunologic and technologic researches have contributed to the diagnosis and cure of the disease . The Authors report the results on ten cases of infective endocarditis that came to their observation in the last five years . They emphasize the more important epidemiologic aspects, the role of the laboratory and instrumental results to define an early diagnosis and the best results about of the therapies used Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2004 Feb, 97(2), 204 - 7 Efficacy of chlorhexidine gluconate rinse for treatment and prevention of oral candidiasis in HIV-infected children: a pilot study; Barasch A et al.; PURPOSE: We evaluated the effect of chlorhexidine (CHX) 0.12% rinses on the clinical and microbiologic manifestations of oral candidiasis in HIV-infected children . STUDY DESIGN: This was a cross-sectional, clinical intervention study of 38 HIV-positive children . Inclusion in the study was based on oral examination and positive oral culture for Candida . At baseline, subjects with no clinical lesions but who were culture-positive for Candida (N = 9) were placed on preventive therapy of CHX q.d . for 90 days . Subjects with clinical oral candidiasis (N = 9) were placed on therapeutic CHX b.i.d . All 38 subjects received oral exams at monthly intervals . At 90 days oral mucosal samples were again taken for Candida . Colony-forming units (CFU) were determined before and after CHX treatment . RESULTS: Of 18 culture-positive subjects, 12 were included in the CFU analyses . After 3 months of CHX oral rinse therapy, Candida was undetectable in 3 children; another 8 showed an average 2-fold reduction in CFU . In 1 child the number of CFU increased modestly . Overall, the average pre- and posttreatment mean CFU was 6.18 +/- 2.19 and 2.73 +/- 3.15, respectively (P = .009) . Five patients with clinical oral candidiasis at baseline, including all 3 who had pseudomembranous candidiasis, were free of signs of disease at the end of the study . CONCLUSIONS: This study suggests that the topical disinfectant CHX may be a promising agent for treating and preventing oral candidiasis in HIV-infected children. New Microbiol, 2004 Jan, 27(1), 49 - 53 Non tuberculous mycobacteria (NTM) in patients with underlying diseases: results obtained by using polymerase chain reaction-restriction enzyme analysis between 1997-2002; Ergin A et al.; In this study, we aimed to evaluate the frequency of non-tuberculous mycobacteria (NTM) isolated from clinical specimens using Polymerase Chain Reaction-Restriction Enzyme Analysis (PCR-REA) and to investigate the patients who had clinically significant NTM infections in our hospital through the five year period from May 1997 to June 2002 . A total of 364 mycobacterial strains isolated from clinical specimens which gave positive growth index in the BACTEC 460 radiometric system in Hacettepe University Hospital Clinical Microbiology Laboratory were evaluated by PCR-REA and clinical data were obtained from the patient records . Three hundred and one of the strains (82.7%) were identified as Mycobacterium tuberculosis and 63 (17.3%) were identified as nontuberculous mycobacteria . Seven (11.1%) of 63 NTM patients were regarded as having clinical mycobacteriosis . Chronic obstructive pulmonary disease and other pre-existing lung diseases were seen in 39 (61.9%) of the patients, 11 (17.5%) of'the patients had chronic renal failure . Four (6.3%) and 9 (14.3%) of them had AIDS and carcinomas, respectively . PCR-REA was found to be a reliable method for typing of our mycobacterial isolates to the species level . These data may shed light on the epidemiology of the mycobacterial species and help to select a proper treatment regimen. Am J Ophthalmol, 2004 Feb, 137(2), 370 - 2 Ocular pythiosis: is it under-diagnosed? Krajaejun T, Pracharktam R, Wongwaisayawan S, Rochanawutinon M, Kunakorn M, Kunavisarut S. PURPOSE: To increase awareness of ocular pythiosis by presenting a typical case and summarizing clinical data of 11 ocular pythiosis cases in Ramathibodi Hospital . DESIGN: Interventional case report . METHODS: A 48-year-old healthy woman with a history of 3-week painful corneal ulcer of left eye was treated with enucleation . RESULTS: The histopathology of enucleated eye revealed endophthalmitis and ulcerative keratitis with numerous hyphae in full-thickness of corneal stroma . The culture identification of the causative organism was Pythium insidiosum . The final diagnosis was ocular pythiosis . CONCLUSIONS: Pythium insidiosum is a causative agent of pythiosis and is distributed worldwide . Ocular pythiosis may not be uncommon, as it may be underdiagnosed due to unfamiliarity among clinicians and microbiologists . Diagnosis of pythiosis is difficult . The disease has high morbidity, as evidenced by nearly evisceration or enucleation among all patients at Ramathibodi Hospital . Early detection and effective treatment are needed for possible cure. Chest, 2004 Feb, 125(2), 712 - 22 Role of flexible bronchoscopy in immunocompromised patients with lung infiltrates; Jain P et al.; STUDY OBJECTIVE: To study the diagnostic role of flexible bronchoscopy (FB) in immunocompromised patients with pulmonary infiltrates . DESIGN: Prospective, observational study . SETTING: Tertiary care hospital . PATIENTS: A total of 104 consecutive non-HIV-infected immunocompromised patients with lung infiltrates in whom FB was performed . METHODS: The primary outcome measure was the diagnostic yield of FB, which was derived as the number of the diagnoses made using FB results divided by all final diagnoses . Final diagnoses were established using data from FB, surgical lung biopsy (SLB), and microbiology and serology testing, and by the clinical response to empiric therapy . We also studied the diagnostic yields of individual sampling procedures such as BAL, transbronchial biopsy (TBB), and protected-specimen brush (PSB) sampling . RESULTS: Overall, 128 diagnoses were made in 104 patients . The overall diagnostic yield of FB was 56.2% (95% confidence interval {CI}, 47 to 64%) . FB provided at least one diagnosis in 53 of 104 patients (51%; 95% CI, 40 to 62%) . FB was more likely to establish the diagnosis when the lung infiltrate was due to an infectious agent (81%; 95% CI, 67 to 90%) than to a noninfectious process (56%; 95% CI, 43 to 67%; p = 0.011) . The diagnostic yields of BAL (38%; 95% CI, 30 to 47%) and TBB (38%; 95% CI, 27 to 51%) were similar (p = 0.94) . The diagnostic yield of PSB sampling was lower (13%; 95% CI 6 to 24%; p = 0.001) than that of BAL . The combined diagnostic yield of BAL and TBB (70%; 95% CI, 57 to 80%) was higher than that of BAL alone (p < 0.001) . Finally, the diagnostic yield of FB with PSB sampling, BAL, and TBB was similar to that of FB with BAL and TBB . The complication rate from FB was 21% (95% CI, 15 to 31%) . Minor bleeding (13%) and pneumothorax (4%) were the most common complications . CONCLUSIONS: FB has a high diagnostic yield in immunocompromised patients with pulmonary infiltrates . Based on our results, we recommend performing TBB in these patients, whenever possible. Med Lav, 2003 Nov-Dec, 94(6), 506 - 20 {Effectiveness of interventions for the control and prevention of tuberculosis in health care facilities: a review of ATS,CDC, OSHA recommendations}; Franchi A et al.; BACKGROUND: Health care workers (HCWs) are occupationally exposed to a multitude of biological hazards, and among these to the risk of tuberculosis (TB) infection, especially involving individuals working in specific workplace (TB and Chest divisions, Infectious Diseases wards, Microbiology laboratories) and performing thoracic endoscopy and "cough-inducing" procedures . According to national legislation (title VIII D.lgs . 626/94, 1998 Health Minister guide lines document) concerning the control and prevention of TB transmission among HCWs, health care facilities are required to (i) perform an accurate risk assessment and (ii) implement an exposure control plan and worker health surveillance program, thus involving the occupational health professionals . OBJECTIVES: The aim of this paper is to provide a general view of the epidemiological and scientific evidence related to the effectiveness of health interventions in the prevention of occupational TB infection . METHODS: Comparative evaluation and critical review of U.S . CDC (1994) guidelines, OSHA (1997) rules, and the most recent ATS and CDC (2000) "statement" documents . RESULTS: In low risk groups TCT shows decreased positive predictive value, high variability, and can be confounded by other factors (age, BCG, MNT), thus reducing its diagnostic value for latent TB infection . CONCLUSIONS: Recent recommendations on the control of TB infection in health care settings underline the need of implementing accurate risk evaluation in all hospital units, compared to the epidemiological profile in the community, and "targeted tuberculin testing" programs among high risk HCWs. Proc Natl Acad Sci U S A, 2004 Feb 17, 101(7), 1981 - 6 Epub 2004 Feb 06. Aberrant expansion of segmented filamentous bacteria in IgA-deficient gut; Suzuki K et al.; The mechanism to maintain homeostasis of the gut microbiota remains largely unknown despite its critical role in the body defense . In the intestines of mice with deficiency of activation-induced cytidine deaminase (AID), the absence of hypermutated IgA is partially compensated for by the presence of large amounts of unmutated IgM and normal expression levels of defensins and angiogenins . We show here a predominant and persistent expansion of segmented filamentous bacteria throughout the small intestine of AID(-/-) mice . Reconstitution of lamina propria IgA production in AID(-/-) mice recovered the normal composition of gut flora and abolished the local and systemic activation of the immune system . The results indicate that secretions of IgAs rather than innate defense peptides are critical to regulation of commensal bacterial flora and that the segmented filamentous bacteria antigens are strong stimuli of the mucosal immune system. J Clin Microbiol, 2004 Feb, 42(2), 881 - 3 Development, implementation, and impact of acceptability criteria for serologic tests for infectious diseases; Crump JA et al.; Serologic testing is essential for the diagnosis of some infectious diseases and yet is fraught with potential pitfalls . All parts of the diagnostic process must be optimized to ensure that serologic tests perform adequately . Recognizing that a lack of clinical data and correctly timed, paired sera frequently led to uninterpretable serology results at our laboratory, we developed and implemented simple acceptability criteria for serologic tests . We assessed the impact of these criteria by comparing submissions and results for the year before and the year after implementation of the criteria . The number of serologic tests performed declined by 25% after implementation of the acceptability criteria, despite an increase in requests for serologic tests . Inappropriate testing of acute-phase sera alone fell from 49 to 0% (P < 0.001) for the tests monitored . Appropriate submission of paired sera rose from 9 to 19% (P = 0.006) . The proportion of results classified as interpretable rose from 52 to 100% (P < 0.001) . We recommend that acceptability criteria be developed and applied to samples submitted to clinical microbiology laboratories for serologic testing. Adv Perit Dial, 2003, 19, 191 - 4 Severe peritoneal mononucleosis associated with icodextrin use in continuous ambulatory peritoneal dialysis; Martin J et al.; Icodextrin is a glucose polymer obtained from starch hydrolysis . It is used as an osmotic agent at 7.5% for peritoneal dialysis (PD) . Its use in PD has been associated with several side effects separate from the one reported here, the most frequent being sterile peritonitis . Recently, three mechanisms have been proposed to explain the occurrence of sterile peritonitis: allergy to dextrin, production of anti-dextran antibodies, and impurities introduced during manufacture . Here, we report a peritoneal mononucleosis outbreak that is highly suggestive of being a consequence of the last-mentioned mechanism . During the period December 2001 to May 2002, a group of 8 Spanish hospitals whose individual PD programs regularly share information and activity reported 29 cases of sterile peritonitis associated with icodextrin use in continuous ambulatory peritoneal dialysis (CAPD) patients {mean age: 60.7 +/- 14.47 years; 8 women (27.59%), 21 men (72.41%); mean time on PD: 25.21 +/- 35.31 months; mean time on icodextrin: 15.17 +/- 11.03 months} . Of the 29 patients, 51.8% showed no symptoms . The remainder presented with mild abdominal discomfort and anorexia . Only 2 patients showed general malaise, severe nausea, fever, and abdominal pain . The initial white cell count in peritoneal effluent was 512 +/- 386 cells/mL (45.0% +/- 28% neutrophils, 44.92% +/- 32.6% mono-nuclear cells, 7.75% +/- 12% eosinophils) . In 5 of the patients, we performed an immunophenotype (CD14) study, demonstrating the monocyte nature of 60%-80% (mean: 70.6%) of the cells . Microbiology cultures were always negative . A rechallenge with the same batches of PD fluid was tried . In 100% of the patients, the clinical and cellular patterns relapsed . No short-term changes in peritoneal function have been observed . The manufacturer informed us that the icodextrin was contaminated with a peptidoglycan . In this sterile peritonitis outbreak with a simultaneous, similar clinical presentation in a group of patients treated with icodextrin solution (presumably contaminated with peptidoglycan), clinical outcome was, for the most part, mild-to-moderate . Symptoms disappeared immediately after icodextrin withdrawal and relapsed after rechallenge with the relevant fluid batches . Monocyte cell counts predominated during the episode . Although we cannot rule out an allergic cause, the massive peritoneal mononuclear cell recruitment suggests a particular mechanism . This is a new mechanism for peritoneal cell recruitment in PD. Pesqui Odontol Bras, 2003 Jul-Sep, 17(3), 217 - 22 Epub 2003 Dec 16. Yeasts from the oral cavity of children with AIDS: exoenzyme production and antifungal resistance; Bosco VL et al.; The oral fungal microbiota of 30 children with AIDS, of both genders, aged from two to six years, receiving outpatient treatment, was evaluated and compared with that of a control group composed of 30 healthy subjects with matching ages and genders . Virulence factors, such as exoenzyme production, and susceptibility to five antifungal agents using an E-Test kit were evaluated . C . albicans predominated over other species in the AIDS group, showing a higher production of proteinase and phospholipase when compared with that observed in the control group . In this study few clinical manifestations of and low selectivity for C . albicans (23.3%) were observed in the AIDS group . The enzymatic studies showed that 53.8% of the AIDS strains were strongly positive whereas only 33.3% of the non-AIDS strains were positive . Amphotericin B was the most effective drug among the antifungal agents tested against C . albicans . The frequency, selectivity and level of exoenzyme production by C . albicans suggest a higher pathogenicity in the AIDS children than in the control children. J Clin Periodontol, 2003 Nov, 30(11), 996 - 1002 Effect of smoking on gingival crevicular fluid cytokine profile during experimental gingivitis; Giannopoulou C et al.; BACKGROUND: Cigarette smoking is a significant risk factor in the pathogenesis of periodontal disease, able to influence both the subgingival microbiota and host responses . AIM: The aim of the present study was to determine the influence of smoking on the amount of IL-1beta, IL-4 and IL-8 in gingival crevicular fluid (GCF) during experimental gingivitis in humans . MATERIAL AND METHODS: Twenty-two healthy subjects, 10 smokers and 12 non-smokers, participated in the study . After professional cleaning, they performed optimal hygiene to reach perfect clinical gingival health . Oral hygiene measures were ceased for a period of 10 days . Clinical indices, including plaque index (PI), gingival index (GI), probing pocket depth (PPD) and bleeding on probing (BOP), were assessed 2 days before (day -2), at the beginning (day 0) and at the end of the experimental gingivitis period (day 10) . At the same time, GCF was collected from 12 sites in each patient, by means of durapore filter membranes . Total amounts of IL-1beta, IL-4 and IL-8 were determined by enzyme-linked immunoadsorbent assay . RESULTS: Clinical data revealed that both smokers and non-smokers showed an increase in PI, GI and BOP scores during the experiment . Although no differences were noted with regard to PI at day 10, the GI and BOP were significantly less pronounced in smokers than non-smokers (p < 0.005) . Non-smokers showed higher total amounts of IL-4 but lower amounts of IL-8 than smokers, throughout the experiment . Total amounts of IL-1beta and IL-8 increased significantly during plaque accumulation in both groups . IL-4 remained stable for the smoker group and decreased for the non-smoker group . CONCLUSIONS: The present results indicate that smoking interferes with cytokine production . When performing studies regarding the pathogenesis of periodontitis, the smoking status of the participants needs to be taken into consideration. Saudi Med J, 2004 Jan, 25(1), 34 - 7 Intestinal parasites among presumably healthy individuals in Lebanon; Saab BR et al.; OBJECTIVE: To determine the prevalence period of intestinal parasites among presumably healthy subjects in Lebanon . METHODS: One stool specimen from 2634 presumably healthy Lebanese subjects, mean age 32.1 years with a range of 14-71 years, resident of different areas in Lebanon was examined for the presence of parasites . The analysis took place in the Clinical Microbiology Laboratory, Department of Pathology and Laboratory Medicine, American University of Beirut Medical Center, Beirut, Lebanon, over 25-months between 1995-1997 . RESULTS: The prevalence of intestinal parasites was 12.4% . The most common parasites identified were Entamoeba coli (38%), Giardia lamblia (3.1%) and Entamoeba histolytica (2.3%) {corrected} . CONCLUSION: The data presented shows the need to improve hygienic conditions to contain the problem of intestinal infections with parasites in Lebanon. Pathology, 2004 Feb, 36(1), 45 - 50 Validation of assembled nucleic acid-based tests in diagnostic microbiology laboratories; Dimech W et al.; Medical microbiology and virology laboratories use nucleic acid tests (NAT) to detect genomic material of infectious organisms in clinical samples . Laboratories choose to perform assembled (or in-house) NAT if commercial assays are not available or if assembled NAT are more economical or accurate . One reason commercial assays are more expensive is because extensive validation is necessary before the kit is marketed, as manufacturers must accept liability for the performance of their assays, assuming their instructions are followed . On the other hand, it is a particular laboratory's responsibility to validate an assembled NAT prior to using it for testing and reporting results on human samples . There are few published guidelines for the validation of assembled NAT . One procedure that laboratories can use to establish a validation process for an assay is detailed in this document . Before validating a method, laboratories must optimise it and then document the protocol . All instruments must be calibrated and maintained throughout the testing process . The validation process involves a series of steps including: (i) testing of dilution series of positive samples to determine the limits of detection of the assay and their linearity over concentrations to be measured in quantitative NAT; (ii) establishing the day-to-day variation of the assay's performance; (iii) evaluating the sensitivity and specificity of the assay as far as practicable, along with the extent of cross-reactivity with other genomic material; and (iv) assuring the quality of assembled assays using quality control procedures that monitor the performance of reagent batches before introducing new lots of reagent for testing. J Neurosci Methods, 2004 Feb 15, 133(1-2), 49 - 55 Large-scale microarray gene expression analysis in discrete electrophysiologically identified neuronal clusters; Bragin A et al.; The normal processes of learning and memory as well as the pathological progress of various neurological diseases may result in changes in gene expression in small, local populations of neurons in any given brain area, leading to the occurrence of specific patterns of electrical activity without easily detectable changes in the morphology of this brain area . One way of identifying these changes might be the comparison of gene expression of areas which generate and areas which do not generate specific patterns of electrical activity . A method for microbiopsy of limited (0.5-1.0 mm3) tissue samples from electrophysiologically identified areas of neurons generating epileptiform activity in the rat brain is described . Here we demonstrate that total RNA isolated from individual microbiopsy samples might be successfully used for microarray based gene expression analysis of any discretely localized neuronal group which can be identified electrophysiologically, including neurons in cortical columns, cell assemblies or other functional units. Rev Med Chir Soc Med Nat Iasi, 2003 Oct-Dec, 107(4), 772 - 80 {Fever of unknown origin in Romania . II . Diagnostic Procedures . Prospective multicenter study of 164 patients}; Baicus C et al.; BACKGROUND: The Diagnostic workup of patients with fever of unknown origin is a challenge, due to the great number of possible etiologies . After we studied the etiologic spectrum of fever of unknown origin in Romania, we tried to evaluate the diagnostic procedures used and their efficiency . METHODS: A multicenter cohort study of two years, with another two years of follow-up was carried out on 164 consecutive patients who met the classic, modified criteria of fever of unknown origin . We used a standardised diagnostic protocol . MAIN OUTCOME MEASURED: The role of every diagnostic procedure in establishing the final diagnosis . RESULTS: The diagnosis was made by microbiology and serology in 41 cases (25%), by histopathology in 22 cases (18%), with the help of imaging techniques in 30 cases (1.3%), based on the clinical evolution and response to treatment in 54 cases (33%) and by other methods in 12 cases (7.3%) . The abdomino-pelvic ultrasonography had a sensitivity of 60%, a specificity of 70%, a positive likelihood ratio of 2.02 and a negative likelihood ratio of 0.57, while the scanner had a sensitivity of 81%, a specificity of 64%, a positive likelihood ratio of 2.23 and a negative likelihood ratio of 0.29 . CONCLUSIONS: Of all the diagnostic procedures used, none had a good sensitivity/specificity . The clinical evolution and the treatment response had an important role in the diagnostic workup. J Comp Physiol {B}, 2004 May, 174(4), 293 - 7 Epub 2004 Feb 04. Energetics and torpor of a South American "living fossil", the microbiotheriid Dromiciops gliroides; Bozinovic F et al.; We examined the energetics of the living fossil microbiotheriid Dromiciops gliroides, a nocturnal and rare small marsupial, endemic to the northern portion of the temperate forest of southern South America . We investigated the effects of changes at ambient temperature and food restriction on the energetics and patterns of torpor . We determined whether they exhibit shallow daily torpor or deep prolonged torpor like some Australian marsupials . Thermal conductance was 92.5% of the expected value for a similarly sized eutherian and basal metabolic rate was 82.9 and 58.6% of the predicted value for standard metatherians and eutherians, respectively . Euthermic D . gliroides showed daily fluctuations in body temperature, being significantly higher during the night . Dromiciops gliroides entered torpor and aroused spontaneously . The duration of torpor bouts increased in response to decreasing ambient temperature; torpor bout duration ranged from 10 h at 20 degrees C to 120 h at 12.5 degrees C . This study is the first record of deep torpor or hibernation for a South American mammal . Torpor in this species as well as in marsupials in general appears to be an opportunistic response to unpredictable biotic and abiotic conditions. J Pediatr, 2004 Feb, 144(2), 206 - 12 Chronic rhinosinusitis in young children differs from adults: a histopathology study; Chan KH et al.; OBJECTIVE: A histopathologic study of children with chronic rhinosinusitis (CRS) was undertaken to compare the sinus mucosa in pediatric and adult CRS . STUDY DESIGN: CRS has been defined as persistent or recurrent sinusitis symptoms for >or=12 weeks despite conventional medical therapy, with abnormal computed tomography of the maxillary sinuses . Maxillary mucosal biopsies were obtained from pediatric CRS subjects for inflammatory cell and morphologic studies . Archival sinus mucosal tissues from adults with CRS were used as histologic controls . Sinus lavages were performed on children with and without CRS for microbiologic studies . RESULTS: Sinus mucosal biopsies were obtained from 19 children with CRS (median age, 3.0 years; range 1.4-8.2 years) . Pediatric CRS biopsies, as compared with adult CRS controls, had a higher density of submucosal lymphocytes (median 469 versus 294 cells/mm(2) per 5 high-power fields {HPF}; P=.02), lower density of submucosal eosinophils (medians 13 versus 82 cells/mm(2) per 5 HPF; P=.01), thinner and more intact epithelium (P=.01 and.07, respectively), thinner basement membranes (P=.002), and fewer submucosal mucous glands (P=.004) . CONCLUSION: The sinus mucosa of young children with CRS has less eosinophilic inflammation, basement membrane thickening, and mucus gland hyperplasia characteristic of adult CRS. Am J Reprod Immunol, 2004 Feb, 51(2), 130 - 7 Extracellular Matrix Protein-dependent Apoptosis of T Cells in Women with a History of Recurrent Spontaneous Abortion; Jerzak M et al.; PROBLEM: The purpose of the study was to determine the role of T-cell apoptosis in extracellular matrix (ECM) environment in pregnancy maintenance in women with a history of recurrent spontaneous abortion (RSA) . METHOD OF STUDY: Thirty-nine non-pregnant women with the history of RSA (anatomic, genetic, endocrine and microbiologic causes were excluded) and 22 healthy women with the previous successful pregnancy outcome were studied . In addition, 21 women with the history of RSA were also studied at the beginning of their next pregnancy . We studied apoptosis of peripheral blood T cells after culture with monoclonal antibody (mAb) OKT-3 alone or with mAb OKT-3 following ECM proteins: collagen IV (C-IV) or fibronectin (Fn) . We used Cell Death Detection ELISA for studying cell death in cell population . In addition, apoptotic peripheral blood T cells were identified by annexin V-PE staining protocol using flow cytometry . CD29+ and CD95+ T-cell surface receptors were also analyzed by flow cytometry . RESULTS: The significantly higher values of enrichment factor: mU of the sample (dying/dead cells) per mU of the corresponding control (viable cells) were observed after peripheral blood T-cell culture with C-IV (P = 0.0002) or Fn (P = 0.004) in samples of non-pregnant women with the history of RSA when compared with control women . The significantly higher values of enrichment factor were observed after peripheral blood T-cell culture with C-IV in samples of pregnant women with the history of RSA with successful pregnancy outcome when compared with pregnant women with the history of RSA with failed pregnancy outcome (P = 0.01) . However, the percentage of apoptotic T cells stained by annexin V was significantly lower in non-pregnant RSA women compared with control (P = 0.0001) . CD95 expression was significantly lower in non-pregnant RSA women compared with control (P = 0.01) . CONCLUSIONS: Apoptosis of T cells might be an interesting possible explanation of successful pregnancy outcome in women with the history of RSA. Biochem J, 2004 May 1, 379(Pt 3), 833 - 40 A combination of both arginine- and lysine-specific gingipain activity of Porphyromonas gingivalis is necessary for the generation of the micro-oxo bishaem-containing pigment from haemoglobin; Smalley JW et al.; The black pigment of Porphyromonas gingivalis is composed of the mu-oxo bishaem complex of Fe(III) protoporphyrin IX (mu-oxo oligomer, dimeric haem), namely {Fe(III)PPIX}2O . P . gingivalis W50 and Rgp (Arg-gingipain)- and Kgp (Lys-gingipain)-deficient mutants K1A, D7, E8 and W501 {Aduse-Opoku, Davies, Gallagher, Hashim, Evans, Rangarajan, Slaney and Curtis (2000) Microbiology 146, 1933-1940} were grown on horse blood/agar for 14 days and examined for the production of mu-oxo bishaem . Mu-oxo Bishaem was detected by UV-visible, Mossbauer and Raman spectroscopies in wild-type W50 and in the black-pigmented RgpA- and RgpB-deficient mutants (W501 and D7 respectively), whereas no haem species were detected in the straw-coloured colonies of Kgp-deficient strain K1A . The dark brown pigment of the double RgpA/RgpB knockout mutant (E8) was not composed of mu-oxo bishaem, but of a high-spin monomeric Fe(III) protoporphyrin IX species (possibly a haem-albumin complex) . In vitro incubation of oxyhaemoglobin with cells of the W50 strain and the RgpA- and RgpB-deficient mutants (W501 and D7) resulted in the formation of mu-oxo bishaem via methaemoglobin as an intermediate . Although the Kgp-deficient strain K1A converted oxyhaemoglobin into methaemoglobin, this was not further degraded into mu-oxo bishaem . The double RgpA/RgpB knockout was also not capable of producing mu-oxo bishaem from oxyhaemoglobin, but instead generated a haemoglobin haemichrome . Inhibition of Arg-X protease activity of W50, W501, D7 and K1A with leupeptin, under conditions where Lys-X protease activity was unaffected, prevented the production of mu-oxo bishaem from oxyhaemoglobin, but resulted in the formation of a haemoglobin haemichrome . These results show that one or both of RgpA and RgpB gingipains, in addition to the lysine-specific gingipain, is necessary for the production of mu-oxo bishaem from haemoglobin by whole cells of P . gingivalis. Cesk Slov Oftalmol, 2003 Nov, 59(6), 373 - 80 {Importance of diagnostic and therapeutic pars plana vitrectomy in endogenous uveitis}; Rihova E et al.; OBJECTIVE: To evaluate the importance of diagnostic-therapeutic pars plana vitrectomy in patients with clinical manifestations of endogenous uveitis . PATIENTS AND METHODS: The diagnostic-therapeutic pars plana vitrectomy (DT PPV) has been performed at the Eye Clinic of General Faculty Hospital and 1st Medical Faculty on Prague on 62 eyes in 49 patients at the mean age of 45 years . The DT PPV was indicated in patients with uveitis of unusual clinical picture or poorly reacting to treatment, suspected infection or tumor etiology . In the beginning of PPV, 0.5 to 1.5 ml of undiluted vitreous body was samples and subsequently examined by microbiology, immunology and molecular genetic methods . PPV was performed on both eyes in 10 patients, on one eye twice in each of three patients . The authors evaluated 62 samples of vitreous body on the whole . The patients were observed for up to 96 months . RESULTS: Laboratory examination of the samples of vitreous body from 22 eyes (35.4%) supported our notion on infection etiology of intraocular inflammation, manifestations of intraocular tumor were in 12 eyes (19.3%), vascular or vitreoretinal pathology was present in 6 eyes (9.6%) . Nonspecific inflammatory changes were detected in 22 eyes (35.4%) . Drugs were administered into the vitreous body in 16 eyes . In the observation period the authors detected marked late complications in connection with DD PPV in 18 eyes (29%) . Visual acuity improved by two or more lines of the Snellen optotypes in 43 eyes (69.3%), remained unchanged in 11 (17.7%) eyes . The vision deteriorated in 8 eyes (12.9%) . CONCLUSION: Based on this experience the authors are of the opinion that DT PPV is indicated in patients with uveitis for giving more precision to the diagnosis of intraocular inflammation or a masking syndrome . DT PPV also represents a therapeutic procedure to cure complications connected with a severe form of inflammation and makes it possible to save central visual acuity. Virus Genes, 2004 Jan, 28(1), 135 - 43 Complexity of dsRNA mycovirus isolated from Fusarium graminearum; Chu YM et al.; Fusarium graminearum is the causal agent of a serious scab disease of small grains in Korea . We screened 827 isolates of F . graminearum from diseased barley and maize and tested for the presence of double-stranded RNA (dsRNA) mycovirus . Of them, 19 isolates contained various sizes of dsRNAs . A dsRNA associated with pronounced morphological changes including reduction in mycelial growth, increase in dark orange to red pigmentation, reduced sporulation and virulence was previously observed in nine dsRNA-containing Fusarium isolates (Chu et al., Appl Env Microbiol 68, 2529-2534, 2002) . Ten additional isolates were found infected with dsRNA mycoviruses . These mycoviruses contain 2-4 different segments of dsRNAs with the size-range of approximately 1.7-10 kbp in length . The presence of dsRNAs did not affect colony morphology and were transmissible through conidia and ascospore with incidence of 30-100% . Interestingly, dsRNA mycovirus found in F . graminearum isolates, JB33 and JNKY19, that show the pattern of mixed infection of two different viruses were transmitted to all progeny conidia and ascospores . These results indicate that there is genomic diversity of dsRNA mycoviruses that infect F . graminearum isolates and that impact of virus infection on host's morphology and virulence is determined by the interaction between dsRNAs and the fungal host, not by the mere presence of the dsRNAs. J Med Screen, 2003, 10(4), 172 - 5 Testing for toxoplasmosis in pregnancy: a study of the routines in primary antenatal care; Eskild A et al.; OBJECTIVES: The question of benefit of screening for antibodies against Toxoplasma gondii in pregnancy to prevent impairment in the infant has not been settled . Despite this, it is believed that such testing is common . The aim of this study was to describe the test routines and to identify factors associated with testing in primary antenatal care . SETTING: Ulleval University Hospital, Oslo, Norway . METHODS: All women who gave birth at Ulleval University Hospital, Oslo, Norway, in May 2000 were included . The number of antibody tests during pregnancy was obtained from the patient registry at the department of microbiology at the same hospital . RESULTS: 81% (361/446) had been tested for antibodies against Toxoplasma gondii during pregnancy . Women attending primary antenatal care at a specialist in obstetrics and gynaecology were most likely to be tested . Among women with no signs of prior toxoplasmosis, 35% (110/317) were tested only once . DISCUSSION: The majority of pregnant women are being tested for toxoplasmosis even though such testing has no documented preventive effect and has not been recommended. J Laryngol Otol, 2003 Dec, 117(12), 946 - 50 Infection, allergy and the hygiene hypothesis: historical perspective; Kim DS et al.; The 'hygiene hypothesis' was popularized in the late 1980s to explain the high prevalence of atopic disorders in the developed countries . It links atopic disorders and the lack of early life infections . An association between the two is not novel and dates back to the beginnings of allergy, immunology and microbiology . Allergy and infection have always been closely related and the study of one has often provided new insights into the pathobiology of the other . Early research into bacterial infections led to the discovery of the human immune system and the concept of allergy . An important relationship exists between parasite infections and the development of atopic disorders . This review traces the long and intimate historical relationship between infection and allergy. N Engl J Med, 2004 Jan 22, 350(4), 342 - 50 The detection of monkeypox in humans in the Western Hemisphere; Reed KD et al.; BACKGROUND: During May and June 2003, an outbreak of febrile illness with vesiculopustular eruptions occurred among persons in the midwestern United States who had had contact with ill pet prairie dogs obtained through a common distributor . Zoonotic transmission of a bacterial or viral pathogen was suspected . METHODS: We reviewed medical records, conducted interviews and examinations, and collected blood and tissue samples for analysis from 11 patients and one prairie dog . Histopathological and electron-microscopical examinations, microbiologic cultures, and molecular assays were performed to identify the etiologic agent . RESULTS: The initial Wisconsin cases evaluated in this outbreak occurred in five males and six females ranging in age from 3 to 43 years . All patients reported having direct contact with ill prairie dogs before experiencing a febrile illness with skin eruptions . We found immunohistochemical or ultrastructural evidence of poxvirus infection in skin-lesion tissue from four patients . Monkeypox virus was recovered in cell cultures of seven samples from patients and from the prairie dog . The virus was identified by detection of monkeypox-specific DNA sequences in tissues or isolates from six patients and the prairie dog . Epidemiologic investigation suggested that the prairie dogs had been exposed to at least one species of rodent recently imported into the United States from West Africa . CONCLUSIONS: Our investigation documents the isolation and identification of monkeypox virus from humans in the Western Hemisphere . Infection of humans was associated with direct contact with ill prairie dogs that were being kept or sold as pets . Infection, 2003 Dec, 31(6), 417 - 20 Fever of unknown origin in Turkey; Tabak F et al.; BACKGROUND: The etiology of fever of unknown origin (FUO) includes primarily infectious, collagen-vascular and neoplastic diseases . The distribution of the disorders causing FUO may differ according to the geographic area and the socioeconomical status of the country . Moreover, the developments in radiographic and microbiologic methods have changed the spectrum of diseases causing FUO . MATERIALS AND METHODS: We reviewed 117 cases that fulfilled the criteria of FUO followed in our department during the period 1984 to 2001 . RESULTS: The etiology of FUO was infectious diseases in 34% of the patients, collagen-vascular diseases in 23%, neoplasms in 19% and miscellaneous diseases in 10% . In 14% of the cases the etiology could not be found . The three leading diseases were tuberculosis (24%), lymphomas (19%) and adult-onset Still's disease (11%) . Tuberculosis was found to be a more common cause of FUO than reported in studies in developed countries . Invasive procedures helped to establish the diagnosis in 50 out of 92 patients (43%) . As a final diagnostic procedure, laparotomy aided the establishment of a diagnosis in 15 out of 20 patients (75%) . CONCLUSION: Although the relative rate of infectious disease as etiologic category is less commonly encountered, infectious disease, especially tuberculosis, remains a common cause of FUO . Although several diseases may lead to FUO, lymphomas, adult-onset Still's disease and particularly tuberculosis should be considered in the differential diagnosis of a patient admitted with FUO. Pneumologie, 2004 Jan, 58(1), 17 - 22 {Spread of a drug-resistant strain of mycobacterium tuberculosis among homeless people in a German city}; Thielen H et al.; BACKGROUND: Cases of active pulmonary tuberculosis are much more common among homeless people because of their personal risk factors than in the average population; it is assumed that there are about 500 000 homeless people in the Federal Republic of Germany . Several sputum samples were sent from the tuberculosis advice center of the public health office in Hannover to the governmental institute of public health services of Lower Saxony in fall 1996 in order to carry out laboratory tests for mycobacteria . The isolates found revealed an unusual pattern of drug resistance to streptomycin and rifampin . METHODS: The sputum samples were tested according to the usual standard test procedures as described in DIN 58 943-3 (DIN standard of the German Institute for Standardization) and in MiQ 5/1998 (Quality standards in microbiology-infectiologic diagnostics issued by the German Society for Hygiene and Microbiology {DGHM} . DNA-fingerprinting was carried out by the IS 6110-RFLP-technique . PATIENTS: In Hannover a total of 12 patients with an active pulmonary tuberculosis, whose M . tuberculosis-strains showed resistance to streptomycin and rifampin, were investigated . 9 of the 12 patients were homeless and lodged in a homeless shelter of the city; in one case a healthcare worker acquired a tuberculosis infection while caring for these patients . RESULTS: The identity of the 12 isolated mycobacteria strains could be proved by IS 6110-fingerprinting . The result clearly indicates the existence of a tuberculosis cluster among the homeless people in a German city . The treatment of the homeless persons proved to be difficult because of the lack of compliance; in 4 cases compulsory isolation with therapy in a closed clinic was unavoidable and beforehand 2 of the patients even had to be tracked down by the police . CONCLUSIONS: The German law on the prevention of infectious diseases offers some new possibilities to the public health offices concerning the control and prevention of tuberculosis among homeless persons . For example the public health offices are now allowed to run a "preventive visiting services" prevention and to practice ambulant therapies; the payment of the costs for poor patients is prescribed, too . These possibilities should be used in an extended way, as the efforts to control the spread to tuberculosis among homeless people need to be intensified. Comp Med, 2003 Dec, 53(6), 663 - 70 Clinical and microbiologic characterization of hemorrhagic pneumonia due to extraintestinal pathogenic Escherichia coli in four young dogs; Handt LK et al.; Over a 21-month period, three Beagle dogs and one mixed-breed dog at our facility developed fatal pneumonia . The four dogs, all purpose bred, came from three vendors and had received the standard canine vaccines prior to shipment . In each instance, the affected dog had been shipped to our facility within the past 10 days . Three cases presented as a peracute clinical syndrome, and all had gross and microscopic findings consistent with hemorrhagic pneumonia . Escherichia coli was isolated from the lungs of all four dogs . Results of testing of lung tissue for canine parainfluenza virus and canine adenovirus were negative . Escherichia coli was also isolated from blood of three of the four dogs . Serotyping of the E . coli isolates indicated that two were serotype 06 and two were 04 . Isolates from all four dogs were positive for the virulence factors alpha hemolysin and cytotoxic necrotizing factor 1 and for the adhesin factor class-III papG allele . These traits place the isolates in the class of extraintestinal pathogenic E . coli, which is being increasingly implicated as a cause of extraintestinal infections in animals and humans and may represent a zoonotic risk to humans working with research dogs. Biochimie, 2003 Nov, 85(11), 1043 - 52 Four decades of DNA repair: from early insights to current perspectives; Hanawalt PC; A brief history of the evolution of the DNA repair field over the past four decades is presented, as documented through the Proceedings from a selected series of five scientific meetings, beginning with the 1965 Radiation Microbiology Conference, held at the University of Chicago with only 40 participants, and extending through the 1988 UCLA Symposium on "Mechanisms and Consequences of DNA Damage Processing", convened in Taos, New Mexico, with over 400 participants . The published proceedings and recorded discussions from these early conferences contain notable insights, of which many have turned out to be remarkably clairvoyant while others must be reevaluated in light of recent discoveries and developments in the field. Med Mycol, 2003 Dec, 41(6), 457 - 67 Status of medical mycology education; Steinbach WJ et al.; The number of immunocompromised patients and subsequent invasive fungal infections continues to rise . However, the education of future medical mycologists to engage this growing problem is diminishing . While there are an increasing number of publications and grants awarded in mycology, the time and detail devoted to teaching medical mycology in United States medical schools are inadequate . Here we review the history in medical mycology education and the current educational opportunities . To accurately gauge contemporary teaching we also conducted a prospective survey of microbiology and immunology departmental chairpersons in United States medical schools to determine the amount and content of contemporary education in medical mycology. Mikrobiol Z, 2003 Sep-Oct, 65(5), 46 - 65 {Actual problems of bacteria phylogenetic classification}; Romanovskaia VA et al.; Historical aspects of development of phylogenetic bacteria taxonomy and essence of species in systematics of bacteria are considered . Comparative analysis of phenotypic and phylogenic classification of bacteria is conducted . Basic results of 16S rRNA gene sequence for bacteria are adduced . Comparative estimation of molecular biological methods in microbiologic researches and also estimation of potentialities and shortcomings of 16S rRNA gene sequence-analysis in systematics of bacteria are conducted. J Clin Microbiol, 2004 Jan, 42(1), 119 - 25 Molecular analysis of Mycobacterium kansasii isolates from the United States; Zhang Y et al.; We studied the population genetics of Mycobacterium kansasii isolates from the United States by PCR restriction enzyme analysis (PRA) of the 441-bp Telenti fragment of the hsp-65 gene and pulsed-field gel electrophoresis (PFGE) of genomic DNA with the restriction endonucleases AseI, DraI, and XbaI, and we compared the patterns to those previously reported from France and Japan . By PRA, 78 of 81 clinical isolates (96%) from the United States belonged to subspecies I . With PFGE, 28 AseI patterns, 32 DraI patterns, and 35 XbaI patterns were produced . PFGE showed marked clonality of the U.S . isolates, with differences between genotypes involving only one or two bands . Isolates within Texas showed lower pattern diversity than those from different states . With DraI, 31 of 71 isolates (44%) had the same common PFGE pattern, which matched the predominant pattern in France (pattern Ia), determined by Picardeau et al . (M . Picardeau, G . Prod'hom, L . Raskine, M . P . LePennec, and V . Vincent, J . Clin . Microbiol . 35:25-32, 1997), and in Japan (type M), determined by Iinuma et al . (Y . Iinuma, S . Ichiyama, Y . Hasegawa, K . Shimokata, S . Kawahara, and T . Matsushima, J . Clin . Microbiol . 35:596-599, 1997) . With AseI, 42% of isolates produced a common pattern indistinguishable from the common pattern seen in French isolates (Ia) and with only one band difference from the common pattern (type M) in Japan . This study demonstrates that subspecies I is the predominant subspecies of M . kansasii among clinical isolates in the United States, as it is in Europe and Japan, and that genotype I is highly clonal worldwide, with the same major genotype responsible for human infection . The fact that a single clone of M . kansasii is responsible for most cases of human disease suggests that specific virulence factors may be associated with this specific genotype. J Clin Microbiol, 2004 Jan, 42(1), 106 - 14 Expression and immunogenicity of proteins encoded by sequences specific to Mycobacterium avium subsp . paratuberculosis; Bannantine JP et al.; The development of immunoassays specific for the diagnosis of Johne's disease in cattle requires antigens specific to Mycobacterium avium subsp . paratuberculosis . However, because of genetic similarity to other mycobacteria comprising the M . avium complex, no such antigens have been found . Through a comparative genomics approach, 21 potential coding sequences of M . avium subsp . paratuberculosis that are not represented in any other mycobacterial species tested (n = 9) were previously identified (J . P . Bannantine, E . Baechler, Q . Zhang, L . Li, and V . Kapur, J . Clin . Microbiol . 40:1303-1310, 2002) . Here we describe the cloning, heterologous expression, and antigenic analysis of these M . avium subsp . paratuberculosis-specific sequences in Escherichia coli . Nucleotide sequences representing each unique predicted coding region were amplified and cloned into two different E . coli expression vectors encoding polyhistidine or maltose binding protein (MBP) affinity purification tags . All 21 of the MBP fusion proteins were successfully purified under denaturing conditions and were evaluated in immunoblotting studies with sera from rabbits and mice immunized with M . avium subsp . paratuberculosis . These studies showed that 5 of the 21 gene products are produced by M . avium subsp . paratuberculosis and are antigenic . Immunoblot analysis with a panel of sera from 9 healthy cattle and 10 cattle with clinical disease shows that the same five M . avium subsp . paratuberculosis proteins are also detected within the context of infection . Collectively, these studies have used a genomic approach to identify novel M . avium subsp . paratuberculosis antigens that are not present in any other mycobacteria . These findings may have a major impact on improved diagnostics for Johne's disease. Adv Genet, 2003, 50, 13 - 21; discussion 507-10 The evolution of gene patents viewed from the United States Patent Office; Mossinghoff GJ; In my remarks here, I have necessarily limited myself to general principles of patent law that are applicable to a consideration of gene-related inventions . Were I still Commissioner of Patents and Trademarks, I would not let electrical engineers anywhere near the examination of applications for patents for gene-related inventions, just as I would not let the USPTO's many Ph.D.s in microbiology or genetics near the examination of complex applications on computer architecture or programming . The same should hold true for attorneys who prosecute patent applications . That is why the U.S . patent system works so well . There can be no discrimination in the system by the field of technology--a principle enshrined in Article 27 of the World Trade Organization's Trade-Related Aspects of Intellectual Property . But we quite properly depend upon specialists to apply across-the-board general principles to very specialized technological fields of endeavor . This works very well in a system in which in the U.S., "everything under the sun made by humans" is patentable. Presse Med, 2003 Dec 13, 32(39), 1841 - 8 {Management of acute community-acquired pneumonia in a health centre . Assessment of 101 cases using the retrospective clinical audit method.}; Labarere J et al.; OBJECTIVE: To assess the conformity of practitioners' practices in the management of community acquired pneumonia with the French Agence Nationale d'Accreditation et d'Evaluation en Sante (Anaes) guidelines . METHODS: We retrospectively reviewed a random sample of 210 medical records which included a principal or associated diagnosis of pneumonia in a French university hospital . RESULTS: A hundred and one medical records were assessable . Sixty-two patients were high risk (Pneumonia Severity Index class IV or V of the prediction rule of Fine et al.), and 10 patients were admitted into an intensive care unit . The overall in-hospital mortality was 14 patients {8-22} . The level of care was appropriate according to the guidelines in 40 cases ({30-50)} . Seven patients did not require hospitalisation, 31 patients required admission into a medical department, 56 patients into an intensive care unit and 7 patients were managed in non specified conditions . Eighteen patients ({11-27}) had appropriate microbiologic investigations . Forty-three patients (({33-53}) received antibiotics within 8 hours of arrival . Empirical antibiotic treatment (dosage and molecule) was appropriate in 38 patients ({28-48}) . There was no significant relationship between compliance with the guidelines and in-hospital mortality . CONCLUSION: The rate of conformity of practitioners' practices with the Anaes guidelines for management of community-acquired pneumonia is low in our hospital . It could be improved by active implementation of these guidelines. Novartis Found Symp, 2003, 254, 234 - 47; discussion 247-52 Viral bioinformatics: computational views of host and pathogen; Kellam P et al.; Wherever cellular life occurs, viruses are also found . As a result, complex organism and cellular antiviral responses co-evolve with virally encoded countermeasures . Since viruses co-opt or interfere with specific cellular pathways during their replication, knowledge of viral genome sequences has helped fundamental understanding of host biology . During viral infection, shifts in the balance between host and viral biological processes result in acute or chronic viral disease pathology accompanied with either active viral replication, viral containment/persistence or viral clearance . Studying host-virus interactions at the level of single gene effects, however, fails to produce a global systems-level understanding . This should now be achievable in the context of complete host and pathogen genome sequences . New experimental methods and computational approaches are rapidly developing, allowing global views of dynamic viral and cellular molecular mechanisms . Systems level virology using DNA microarrays and specific viral data resources will reveal the detailed cellular context in which viruses replicate, highlighting common and distinct antiviral mechanisms, the effect of different host cell gene expression programs, and the response of cells to similar or diverse virus types . Ultimately, microbiology and immunology will tend towards a systems-level view of how host and pathogen interact. Expert Rev Vaccines, 2003 Aug, 2(4), 561 - 9 Prospects for vaccine development against Buruli disease; Huygen K; Buruli disease, caused by Mycobacterium ulcerans, is an emerging infectious disease in tropical areas, particularly West Africa, which can cause deep necrotizing skin lesions, called Buruli ulcer . Buruli disease affects all age groups but about 50% of the cases are diagnosed in children . There is no evidence that Buruli disease is transmitted by direct person-to-person contact and it is very likely that contaminated water of rivers, swamps and lakes serves as the wildlife reservoir of M . ulcerans . This review briefly discusses the epidemiology, microbiology, pathology and treatment of the disease . It describes in detail the current knowledge of the immune response and focuses on the studies that have dealt with vaccination . Finally, experimental approaches for future immunoprophylaxis are discussed. J Chemother, 2003 Nov, 15 Suppl 2, 16 - 27 Scedosporium species infections and treatments; Steinbach WJ et al.; Scedosporium species are now increasingly isolated from immunocompromised and immunocompetent patients . Unfortunately, Scedosporium species infections are generally resistant to amphotericin B, and S . prolificans strains are particularly resistant to presently-available antifungal agents . Here we review the microbiology, expanding epidemiology, numerous clinical presentations, and diagnostic tools available for Scedosporium species infections . Finally, we detail the available in vitro, animal model, and clinical data on the treatment of Scedosporium species infections with special emphasis on the role of newer antifungal therapies for these recalcitrant infections. Rev Esp Quimioter, 2003 Sep, 16(3), 308 - 12 {Resistance of HIV-1 to antiretroviral drugs in Valencia (Spain): mutations and susceptibility}; Molina JM et al.; Resistance of HIV to antiretroviral drugs was studied in 210 samples taken in the last two years from patients at the Molecular Biology Unit of the Microbiology Department of the Hospital La Fe in Valencia, Spain . Once the viral load in plasma was determined, resistance was detected using complete gene sequencing for protease until position 3464 of the HIV-1 inverse transcriptase gene . The results were analyzed using the programs Omiga 1.2 (Oxford Molecular Group) and HR-ASAP 1.0 (Stanford University) . The protease inhibitors the least affected by the presence of mutations leading to resistance were amprenavir (68.96% activity), and lopinavir (70.69% activity), and of the inverse transcriptase inhibitors, tenofovir (94.02% activity), D4T (74.62% activity) and 3TC (76.12% activity) . The treatment combination with the greatest activity, based on the different mutations, was D4T + 3TC + NNRTI . To justify the persistence of viremia with relatively low genotypic resistance to antiretroviral drugs other variables must be considered, such as treatment compliance and the pharmacokinetics of the drugs. Jpn J Infect Dis, 2003 Oct-Dec, 56(5-6), 193 - 9 Encephalitis in Taiwan: a prospective hospital-based study; Lee TC et al.; To investigate encephalitis in Taiwan, a multicenter study was conducted with patients who had acute severe neurological dysfunction and suspected encephalitis from May 2000 to December 2001 . Demographic data such as age, sex, and seasons were analyzed . Polymerase chain reaction analyses were performed to determine the microbiologic diagnosis . The patients included 73 males and 54 females, with a peak age of 10-40 years old . Microbiologic diagnoses in 86 (69%) of 124 cases involved herpes simplex virus (HSV, 45 cases), varicella zoster (16 cases), Mycobacterium tuberculosis (10 cases), cytomegalovirus (8 cases), adenovirus (5 cases), influenza (1 case), and enterovirus (1 case) . Pathogens were found in 69% of the cases . Encephalitis was most likely to occur in June and July . Based on the results, HSV is still the major viral cause of encephalitis in Taiwan. Pharmacotherapy, 2003 Dec, 23(12), 1627 - 33 Optimizing antibiotic use in hospitals: the role of population-based antibiotic surveillance in limiting antibiotic resistance . Insights from the society of infectious diseases pharmacists; Madaras-Kelly K; To minimize antibiotic resistance, pharmacists increasingly are becoming involved in antibiotic surveillance, formulation of antibiotic use policies, and day-to-day control of problematic antibiotic use . Population-based antibiotic surveillance has become common with the proliferation of electronic databases . The most widely applied measure of antibiotic consumption is the defined daily dose/1000 patient days . Most studies correlating antibiotic consumption with resistance have focused on antibiogram-related end points; antibiogram data generally reflect institutional nosocomial infection patterns . Most study designs have been derived from traditional epidemiology such as case-control with regression modeling or simple linear regression; however, these methods have limitations . Several experimental designs show promise . Many historical-control studies, including a multicentered study, suggest that population-based antibiotic surveillance and policy intervention can decrease antibiotic resistance in hospitals . Further research on the relationships among antibiotic surveillance, structured antibiotic policy interventions, and other microbiologic, patient-oriented, and economic end points is needed. Compend Contin Educ Dent, 2003 Sep, 24(9 Suppl), 10 - 3; quiz 42 In vitro efficacy of Colgate Total advanced fresh; Pilch S et al.; Generally, clinical studies using organoleptic judges, gas chromatography, or a sulfide monitor have been employed in the assessment of treatments for the control of oral malodor . However, these studies can be expensive and time consuming . Also, for agents whose safety has not been proven, these methods are not appropriate . Therefore, in vitro assessment is a critical step toward developing a new technology or implementing changes to an existing formula . The in vitro methods employed in this study combined basic microbiology methods, such as growth inhibition and zone of inhibition experiments . In addition, a newly developed, in vitro, volatile sulfur compounds (VSC) experiment that combined the essential elements of zone of inhibition, head space analysis, and dynamic flow cell techniques was also employed . The in vitro VSC method has been validated using technologies clinically proven to control oral malodor . All in vitro experiments have demonstrated that Colgate Total Advanced Fresh toothpaste has efficacy similar to that of the original Colgate Total toothpaste and that both variants showed superior efficacy to the control toothpaste . These results have been corroborated by clinical study results. Int Rev Immunol, 2004 Jan-Apr, 23(1-2), 61 - 74 Poxvirus infection and apoptosis; Pogo BG et al.; The following excellent reviews have been published on poxviruses and apoptosis during the last few years: P.C . Turner and R.W . Moyer, Semin . Virology, 8: 453-469, 1998; J.L . Shisler and B . Moss, Semin . Immunol., 13: 67-72, 2001; and H . Everett and G . McFadden, Curr . Opin . Microbiol., 5: 395-402, 2002 . These articles dealt with the viral products and the mechanisms by which they interfere with apoptosis . In this review, we summarize new and old information and also introduce a new approach to explore interactions between the host cell and the replicating virus. Int J Infect Dis, 2004 Jan, 8(1), 39 - 45 Well diffusion for antifungal susceptibility testing; Magaldi S et al.; INTRODUCTION: The increasing clinical and microbiologic resistance of Candida spp . isolates to several antifungal agents is becoming a serious problem . It is now reasonable to propose the use of antifungal susceptibility testing in Candida spp . isolates from patients who have failed conventional therapy, before the selection of an empirical therapy . METHODS: One hundred and fifty eight isolates of Candida spp . were evaluated simultaneously by broth microdilution (NCCLS standard) and well diffusion testing (WD), a diffusion method similar to disc diffusion . RESULTS: According to the Wilcoxon Signed Ranks test performed, there was no significant difference (p>0.05) between both methodologies for all antifungal agents tested (fluconazole, itraconazole, posaconazole, caspofungin and amphotericin B, with C . tropicalis, C . krusei, C . dubliniensis, C . guillermondii, C . parapsilosis, C . albicans and C . glabrata) . A significant difference was observed when comparing well diffusion with NCCLS for fluconazole WD 80% (p=0.008) in C . glabrata, as well as WD 80% (p=0.002) and WD 50% (p=0.002) in C . albicans . CONCLUSIONS: The well diffusion test is simple, easy to reproduce, inexpensive, easy both to read and interpret, and has a good correlation to the reference NCCLS microdilution test and may represent an alternative method for antifungal drug susceptibility testing of Candida spp., mainly in laboratories with few resources. J Pediatr (Rio J), 1998 Jul, 74(Suppl 1), S76 - 94 {Cystic fibrosis}; Reis FJ et al.; OBJECTIVE: Due to the great advances recently achieved in the treatment of Cystic Fibrosis as well as to the fact that pediatricians need to have a better understanding of this disease, the authors propose an extensive review of the subject . METHODS: We selected the most outstanding publications on Cystic Fibrosis in the international literature of the recent years, with the purpose of being up-to-date and at the same time offering a practical synthesis for the readers . RESULTS: We elaborated an extensive review about Cystic Fibrosis covering the following topics: historical remarks, genetics, physiopathogenesis, microbiology of pulmonary infections, clinical manifestations, clinical and laboratorial criteria for diagnosis, differential diagnosis, treatment and prognosis. AJR Am J Roentgenol, 2004 Jan, 182(1), 119 - 22 MRI findings of septic arthritis and associated osteomyelitis in adults; Karchevsky M et al.; OBJECTIVE: The purpose of this study was to describe the soft-tissue, synovial, and osseous MRI findings of septic arthritis . MATERIALS AND METHODS: At 1.5 T (T1-weighted, T2-weighted or STIR, and contrast-enhanced images), 50 consecutive cases of septic arthritis were evaluated by two observers for synovial enhancement, perisynovial edema, joint effusion, fluid outpouching, fluid enhancement, and synovial thickening . The marrow was assessed for abnormal signal on T1- and T2-weighted images or after contrast enhancement . We noted whether the marrow signal was diffuse or abnormal in bare areas . MRI findings were compared with microbiologic, clinical, and surgical data and diagnoses . RESULTS: The frequency of MRI findings in septic joints was as follows: synovial enhancement (98%), perisynovial edema (84%), joint effusions (70%), fluid outpouching (53%), fluid enhancement (30%), and synovial thickening (22%) . The marrow showed bare area changes (86%), abnormal T2 signal (84%), abnormal gadolinium enhancement (81%), and abnormal T1 signal (66%) . Associated osteomyelitis more often showed T1 s