Bacteriological

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 signal abnormalities and was diffuse . CONCLUSION: Synovial enhancement, perisynovial edema, and joint effusion had the highest correlation with the clinical diagnosis of a septic joint . However, almost a third of patients with septic arthritis lacked an effusion . Abnormal marrow signal-particularly if it was diffuse and seen on T1-weighted images-had the highest association with concomitant osteomyelitis.

Am J Surg, 2003 Nov 28, 186(5A), 44S - 54S; discussion 61S-64S
An evidence-based approach to diabetic foot infections; Frykberg RG; Foot infections are a major complication of diabetes mellitus and contribute to the development of gangrene and lower extremity amputation . Recent evidence indicates that persons with diabetes are at greater risk for infection because of underlying neuropathy, peripheral vascular disease, and impaired responses to infecting organisms . This article reviews the underlying pathophysiology, causes, microbiology, and current management concepts for this potentially limb-threatening complication . Multidisciplinary management consisting of teams of specialists with a focus on limb preservation can make significant improvements in outcomes, including a reduction in rates of lower extremity amputation.

Mycopathologia, 2003, 156(4), 309 - 12
Phaeohyphomycosis caused by Chaetomium globosum in an allogeneic bone marrow transplant recipient; Teixeira AB et al.; Bone marrow transplant recipients are highly susceptible to opportunistic fungal infections . This is the report, of the first case of a Chaetomium systemic infection described in Brazil . A 34 year-old patient with chronic myeloid leukemia underwent an allogeneic sibling matched bone marrow transplant . Seven months later, he developed systemic infection with enlargement of the axillary and cervical lymph nodes . Culture of the aspirates from both lymph nodes yielded Chaetomium globosum . The infection was successfully treated with amphotericin B . The increasing population of immunosupressed patients requires a careful microbiologic investigation for uncommon fungal infections.

J Clin Epidemiol, 2003 Dec, 56(12), 1218 - 23
A prospective, community-based study on virologic assessment among elderly people with and without symptoms of acute respiratory infection; Graat JM et al.; BACKGROUND/OBJECTIVES: Community-based elderly studies concerning microbiology of acute respiratory infections are scarce . Data on subclinical infections are even totally absent, although asymptomatic persons might act as a source of respiratory infections . METHODS: In a 1-year community-based study, we prospectively investigated the possible virologic cause of acute respiratory infections in 107 symptomatic case episodes and 91 symptom-free control periods . Participants, persons >/=60 years, reported daily the presence of respiratory symptoms in a diary . Virologic assessment was performed by polymerase chain reaction (PCR) and serology . RESULTS: In 58% of the case episodes a pathogen was demonstrated, the most common being rhinoviruses (32%), coronaviruses (17%), and influenzaviruses (7%) . The odds ratio for demonstrating a virus in cases with symptoms vs . controls without symptoms was 30.0 (95% confidence interval 10.2-87.6) . In 4% of the symptom-free control periods a virus was detected . CONCLUSIONS: This study supports the importance of rhinovirus infections in community-dwelling elderly persons, whereas asymptomatic elderly persons can also harbor pathogens as detected by PCR, and thus might be a source of infection for their environment.

Ann Biol Clin (Paris), 2003 Sep-Oct, 61(5), 571 - 5
{Contribution of the laboratory to the evaluation of brain-dead patients for obtaining organs from dead individuals}; Bouhsain S et al.; The removal organ in order to transplantation is realized often for patient deceased . It was regulation activity that passes in urgency context where laboratory play an important role . The biochemistry, immunology, haematology, toxicology and microbiology analysis allows the validation of the donors, the supervision of the functional quality of organ and the definition of the best couple donor-recipient.

Clin Chem, 2004 Feb, 50(2), 423 - 32 Epub 2003 Dec 11.
Determination of folate vitamers in human serum by stable-isotope-dilution tandem mass spectrometry and comparison with radioassay and microbiologic assay; Pfeiffer CM et al.; BACKGROUND: Current clinical methods for folate give different results and cannot measure the various forms of folate . We developed an isotope-dilution tandem mass spectrometric method coupled to liquid chromatography (LC/MS/MS) as a candidate reference method for 5-methyltetrahydrofolic acid (5MeTHF), 5-formyltetrahydrofolic acid (5FoTHF), and folic acid (FA) in human serum . METHODS: We quantitatively isolated folates from 275 microL of serum with a phenyl solid-phase extraction cartridge, then detected and quantified them in stabilized serum extracts by positive-ion electrospray ionization LC/MS/MS . We used an isocratic mobile phase of acetic acid in organic solvent on a C(8) analytical column . (13)C-labeled folates were used as internal standards . RESULTS: Limits of detection in serum were 0.13 (5MeTHF), 0.05 (5FoTHF), and 0.07 (FA) nmol/L . Within- and between-run imprecision (CV) was <7% for 5MeTHF and <10% for 5FoTHF at concentrations >0.5 nmol/L, and <10% for FA at concentrations >2.0 nmol/L . Total folate (TFOL) concentrations determined by competitive protein binding radioassay were approximately 9% lower than results obtained with LC/MS/MS . The microbiologic assay gave approximately 15% higher TFOL results with FA calibrator and no difference with 5MeTHF calibrator . The mean (SD) {range} TFOL in 42 sera was 35.5 (17.8) {6.5-75.6} nmol/L . Thirty-two samples with TFOL <50 nmol/L had, on average, 93.3% 5MeTHF, 2.3% FA, and 4.4% 5FoTHF . Ten samples with TFOL >50 nmol/L had, on average, 81.7% 5MeTHF, 15.7% FA, and 2.5% 5FoTHF . CONCLUSIONS: This stable-isotope-dilution LC/MS/MS method can quantify 5MeTHF, 5FoTHF, and FA in serum . Currently used clinical assays agree with this candidate reference method.

Gac Sanit, 2003 Nov-Dec, 17(6), 453 - 7
{Incidence of invasive pneumococcal disease in Cantabria, Spain, {1995-2001} and implications for the childhood inmunization schedule}; Gonzalez A et al.; OBJECTIVE: To describe the incidence of invasive pneumococcal disease in Cantabria (Spain) between 1995 and 2001 . METHOD: We reviewed the records of the Minimum Data Set (MDS) of public hospitals in Cantabria, discharges from private hospitals and the registry of diseases of mandatory reporting, as well as the microbiologic diagnoses and medical records of children discharged from the Pediatric Service of the Cantabria Hospital (the tertiary care hospital in our autonomous community) . RESULTS: We obtained a meningitis incidence of 5.55, 5.03 and 0.76/100,000 in children < 2 years, > or = 2 and < 5 years, and > or = 5 years respectively, and an incidence of invasive disease of 11.11, 11.32 and 1.49/100,000 in the same age groups . CONCLUSIONS: The incidence of meningitis and invasive pneumococcal disease in Cantabria is low . We discuss factors that should be taken into account when introducing the pneumococcal conjugate vaccine in the childhood immunization schedule of Cantabria.

Microbiol Mol Biol Rev, 2003 Dec, 67(4), 550 - 73
On the evolution of structure in aminoacyl-tRNA synthetases; O'Donoghue P et al.; The aminoacyl-tRNA synthetases are one of the major protein components in the translation machinery . These essential proteins are found in all forms of life and are responsible for charging their cognate tRNAs with the correct amino acid . The evolution of the tRNA synthetases is of fundamental importance with respect to the nature of the biological cell and the transition from an RNA world to the modern world dominated by protein-enzymes . We present a structure-based phylogeny of the aminoacyl-tRNA synthetases . By using structural alignments of all of the aminoacyl-tRNA synthetases of known structure in combination with a new measure of structural homology, we have reconstructed the evolutionary history of these proteins . In order to derive unbiased statistics from the structural alignments, we introduce a multidimensional QR factorization which produces a nonredundant set of structures . Since protein structure is more highly conserved than protein sequence, this study has allowed us to glimpse the evolution of protein structure that predates the root of the universal phylogenetic tree . The extensive sequence-based phylogenetic analysis of the tRNA synthetases (Woese et al., Microbiol . Mol . Biol . Rev . 64:202-236, 2000) has further enabled us to reconstruct the complete evolutionary profile of these proteins and to make connections between major evolutionary events and the resulting changes in protein shape . We also discuss the effect of functional specificity on protein shape over the complex evolutionary course of the tRNA synthetases.

Chest, 2003 Dec, 124(6), 2105 - 11
Polymerase chain reaction of pleural biopsy is a rapid and sensitive method for the diagnosis of tuberculous pleural effusion; Hasaneen NA et al.; BACKGROUND: Tuberculous pleural effusion occurs in 30% of patients with tuberculosis (TB) . Rapid diagnosis of a tuberculous pleural effusion would greatly facilitate the management of many patients . Polymerase chain reaction (PCR) has been used to detect Mycobacterium tuberculosis in pleural fluid with highly variable sensitivity . OBJECTIVE: To improve our laboratory diagnosis of tuberculous pleural effusion . METHODS: We applied PCR to detect DNA specific for M tuberculosis in 33 of the studied pleural biopsy specimens using an IS986-based primer that was specific for mycobacterium complex, and compared it to the results of pleural fluid and biopsy cultures performed on either Lowenstein-Jensen (LJ) medium or BACTEC 12B liquid medium (Becton Dickinson Microbiology Systems; Cockeysville, MD), Ziehl-Neelsen (ZN) staining, and histopathology in 45 patients with pleural effusion . RESULTS: Of the 45 patients with pleural effusion who were studied, 26 patients received diagnoses of tuberculous pleural effusion that had been confirmed by either culture and or histopathology, 10 patients received diagnoses of exudative effusion due to causes other than TB, and 9 patients received diagnoses of transudative effusion . Histopathology of the pleural biopsy specimen had a sensitivity of 53.8% . The sensitivity of the ZN staining of pleural fluid and biopsy specimens was 0.0% and 3.8%, respectively . The sensitivity of the culture on both BACTEC 12B liquid medium and LJ medium was higher in pleural biopsy specimens (92.3%) than in pleural fluid specimens (15.4%; p > 0.001) . The improvements of the BACTEC culture system improved and shortened the detection time of M tuberculosis in pleural biopsy specimens . PCR of pleural biopsy specimens had 90% sensitivity and 100% specificity . The positive predictive value and the negative predictive value for pleural biopsy specimen cultures were 100% and 90.5% vs 100% and 86.7% for pleural biopsy specimen PCRs . CONCLUSION: The overall accuracy of PCR of pleural biopsy was similar to the results of pleural biopsy culture, however, PCR of the pleural biopsy was much faster in reaching diagnosis . PCR of pleural biopsy is a useful method when used in combination with the BACTEC culture system and histopathologic examination of pleural biopsy to reach a rapid diagnosis of tuberculous pleural effusion.

J Agric Food Chem, 2003 Dec 17, 51(26), 7607 - 10
Antifungal clerodane diterpenes from Macaranga monandra (L) Muell . et Arg . (Euphorbiaceae); Salah MA et al.; Hexane and ethyl acetate phases of the methanol extract of Macaranga monandra showed fungal growth inhibition of Colletotrichum acutatum, C . fragariae and C . gloeosporioides, Fusarium oxysporum, Botrytis cinerea, Phomopsis obscurans, and P . viticola . Bioassay-guided fractionation led to the isolation of two active clerodane-type diterpenes that were elucidated by spectroscopic methods (1D-, 2D-NMR, and MS) as kolavenic acid and 2-oxo-kolavenic acid . A 96-well microbioassay revealed that kolavenic acid and 2-oxo-kolavenic acid produced moderate growth inhibition in Phomopsis viticola and Botrytis cinerea.

J Clin Microbiol, 2003 Dec, 41(12), 5747 - 9
DNA fingerprinting of Ralstonia paucula by infrequent-restriction-site PCR and randomly amplified polymorphic DNA analysis; Moissenet D et al.; Ralstonia paucula (formerly CDC group IV c-2) is an environmental organism that can cause serious human infections, occasionally clusters of nosocomial infections . In the present work, 26 strains of R . paucula (4 from the American Centers for Disease Control and Prevention collection, 10 from the Belgian Laboratorium voor Microbiologie {LMG} collection, and 12 French clinical isolates) were analyzed with infrequent-restriction-site PCR and randomly amplified polymorphic DNA analysis . Both techniques accurately distinguished between collection strains . Two close patterns obtained for all the French isolates suggested a clonal strain . Two LMG collection strains originating from human sources in the United States also showed patterns close to those of French isolates.

J Clin Microbiol, 2003 Dec, 41(12), 5699 - 708
rpoB-based identification of nonpigmented and late-pigmenting rapidly growing mycobacteria; Adekambi T et al.; Nonpigmented and late-pigmenting rapidly growing mycobacteria (RGM) are increasingly isolated in clinical microbiology laboratories . Their accurate identification remains problematic because classification is labor intensive work and because new taxa are not often incorporated into classification databases . Also, 16S rRNA gene sequence analysis underestimates RGM diversity and does not distinguish between all taxa . We determined the complete nucleotide sequence of the rpoB gene, which encodes the bacterial beta subunit of the RNA polymerase, for 20 RGM type strains . After using in-house software which analyzes and graphically represents variability stretches of 60 bp along the nucleotide sequence, our analysis focused on a 723-bp variable region exhibiting 83.9 to 97% interspecies similarity and 0 to 1.7% intraspecific divergence . Primer pair Myco-F-Myco-R was designed as a tool for both PCR amplification and sequencing of this region for molecular identification of RGM . This tool was used for identification of 63 RGM clinical isolates previously identified at the species level on the basis of phenotypic characteristics and by 16S rRNA gene sequence analysis . Of 63 clinical isolates, 59 (94%) exhibited <2% partial rpoB gene sequence divergence from 1 of 20 species under study and were regarded as correctly identified at the species level . Mycobacterium abscessus and Mycobacterium mucogenicum isolates were clearly distinguished from Mycobacterium chelonae; Mycobacterium mageritense isolates were clearly distinguished from "Mycobacterium houstonense." Four isolates were not identified at the species level because they exhibited >3% partial rpoB gene sequence divergence from the corresponding type strain; they belonged to three taxa related to M . mucogenicum, Mycobacterium smegmatis, and Mycobacterium porcinum . For M . abscessus and M . mucogenicum, this partial sequence yielded a high genetic heterogeneity within the clinical isolates . We conclude that molecular identification by analysis of the 723-bp rpoB sequence is a rapid and accurate tool for identification of RGM.

J Clin Microbiol, 2003 Dec, 41(12), 5478 - 87
Genetic Heterogeneity in the rRNA Gene Locus of Trichophyton tonsurans; Gaedigk A et al.; Trichophyton tonsurans is the major pediatric pathogen in tinea capitis, causing disparate disease presentations . Little is known about genetic variation, which may ultimately be linked to divergent disease status . This investigation was aimed at identifying genetic variants of T . tonsurans by methods that can facilitate strain discrimination in population-based studies . Ninety-two isolates were acquired from six U.S . microbiology laboratories, and genomic DNA was isolated from mature colonies . The nontranscribed spacer (NTS) was amplified by PCR, and products from isolates with various amplicon sizes were fully sequenced . Nested amplification, targeting a variable internal repeat (VIR) region, allowed assignment of variant type by fragment size . Subvariant type was assigned by a combination of PCR-restriction fragment length polymorphism-based assays . Five variants differing in size (348 to 700 bp) and sequence were identified within the VIR region comprised of several large repeats (104, 140, and 194 bp) arranged in tandem . Seven single-nucleotide polymorphisms (SNPs) were detected across the NTS, with five occurring in the constant regions flanking the VIR region and two occurring in the VIR region . Additionally, a 10-bp insertion and a 14-bp deletion were identified upstream of the VIR region . The combination of SNPs revealed seven haplotype patterns which were stable upon serial passage over 1 year . No sequence variations were identified within the internal transcribed spacer regions . Unique NTS sequences were utilized to develop a duplex PCR assay that discriminated T . tonsurans from other dermatophytes . Of the 92 isolates evaluated, this genotyping scheme distinguished 12 distinct strains, providing evidence of genetic heterogeneity in T . tonsurans.

J Clin Microbiol, 2003 Dec, 41(12), 5372 - 6
Surge capacity for response to bioterrorism in hospital clinical microbiology laboratories; Shapiro DS; Surge capacity is the ability to rapidly mobilize to meet an increased demand . While large amounts of federal funding have been allocated to public health laboratories, little federal funding has been allocated to hospital microbiology laboratories . There are concerns that hospital laboratories may have inadequate surge capacities to deal with a significant bioterrorism incident . A workflow analysis of a clinical microbiology laboratory that serves an urban medical center was performed to identify barriers to surge capacity in the setting of a bioterrorism event and to identify solutions to these problems . Barriers include a national shortage of trained medical technologists, the inability of clinical laboratories to deal with a dramatic increase in the number of blood cultures, a delay while manufacturers increase production of critical products and then transport and deliver these products to clinical laboratories, and a shortage of class II biological safety cabinets . Federal funding could remedy staffing shortages by making the salaries of medical technologists comparable to those of similarly educated health care professionals and by providing financial incentives for students to enroll in clinical laboratory science programs . Blood culture bottles, and possibly continuous-monitoring blood culture instruments, should be added to the national antibiotic stockpile . Federal support must ensure that companies that manufacture essential laboratory supplies are capable of rapidly scaling up production . Hospitals must provide increased numbers of biological safety cabinets and amounts of space dedicated to clinical microbiology laboratories . Laboratories should undertake limited cross-training of technologists, ensure that adequate packaging supplies are available, and be able to move to a 4-day blood culture protocol.

Nat Rev Immunol, 2004 Jan, 4(1), 1 - 23
Immunity to fungal infections; Romani L; The topic of immunity to fungal infections is of interest to a wide range of disciplines, from microbiology to immunology . It is of particular interest in terms of therapy of HIV-infected individuals, and patients with cancer or individuals who have received transplants . Understanding the nature and function of the immune response to fungi is an exciting challenge that might set the stage for new approaches to the treatment of fungal diseases, from immunotherapy to vaccines . The past decade has witnessed the development of a wide range of new approaches to elucidate events that occur at the host-fungus interface.

Appl Environ Microbiol, 2003 Dec, 69(12), 7242 - 7
Transduction of porcine enteropathogenic Escherichia coli with a derivative of a shiga toxin 2-encoding bacteriophage in a porcine ligated ileal loop system; Toth I et al.; In this study, we have investigated the ability of detoxified Shiga toxin (Stx)-converting bacteriophages Phi3538 (Deltastx(2)::cat) (H . Schmidt et al., Appl . Environ . Microbiol . 65:3855-3861, 1999) and H-19B::Tn10d-bla (D . W . Acheson et al., Infect . Immun . 66:4496-4498, 1998) to lysogenize enteropathogenic Escherichia coli (EPEC) strains in vivo . We were able to transduce the porcine EPEC strain 1390 (O45) with Phi3538 (Deltastx(2)::cat) in porcine ligated ileal loops but not the human EPEC prototype strain E2348/69 (O127) . Neither strain 1390 nor strain E2348/69 was lysogenized under these in vivo conditions when E . coli K-12 containing H-19B::Tn10d-bla was used as the stx1 phage donor . The repeated success in the in vivo transduction of an Stx2-encoding phage to a porcine EPEC strain in pig loops was in contrast to failures in the in vitro trials with these and other EPEC strains . These results indicate that in vivo conditions are more effective for transduction of Stx2-encoding phages than in vitro conditions.

Trends Microbiol, 2003 Dec, 11(12), 570 - 7
Hard labour: bacterial infection of the skeleton; Henderson B et al.; The skeleton is the largest mammalian organ system, containing a myriad of blood vessels, tissue surfaces and bone cells for bacterial colonization . Although rock-like, the skeleton is a dynamic structure that is undergoing constant remodelling . This is the result of the opposing actions of two key cells: the osteoblast, which produces bone, and the osteoclast, a multinucleate cell that 'eats' bone . It is not generally realized that the most prevalent chronic bacterial diseases of Homo sapiens afflict the skeleton . Several pathogens, and members of the normal microbiota, have evolved specific cellular and molecular mechanisms for invading bone, including its cellular constituents . The host cellular pathways that are activated and lead to destruction or loss of the bone matrix will be described.

Med Mycol, 2003 Oct, 41(5), 369 - 81
Rapid identification of fungi by sequencing the ITS1 and ITS2 regions using an automated capillary electrophoresis system; Pryce TM et al.; We developed a standardized DNA sequence-based approach for the accurate and timely identification of medically important fungi by sequencing polymerase chain reaction (PCR) products with a rapid automated capillary electrophoresis system . A simple DNA extraction method and PCR amplification using universal fungal primers was used to amplify ribosomal DNA from a range of clinical isolates and reference strains . The entire internal transcribed spacer (ITS) 1-5.8S-ITS2 ribosomal DNA region was sequenced using automated dye termination sequencing for 89 clinical isolates . These had previously been identified by traditional methods and included 12 ascomycetous yeast species, three basidiomycetous yeast species, eight dermatophyte species and two thermally dimorphic fungi, Scedosporium prolificans and S . apiospermum . Furthermore, 21 reference strains representing 19 different Candida species, Geotrichum candidum and Malassezia furfur were also sequenced as part of this study and were used either as standards for sequence-based comparisons, or as assay controls . Sequence-based identification was compared to traditional identification in a blinded manner . Of the clinical isolates tested, 88/89 had DNA sequences that were highly homologous to those of reference strains accessioned in GenBank, and 87/89 gave a sequence-based identification result that correlated with the traditional identification . In contrast to relatively slow conventional methods of identification, a sequence-based identification from a pure culture can be obtained within 24 h of a DNA extraction carried out after a minimal period of culture growth . We conclude that this approach is rapid, and may be a more accurate cost-effective alternative than most phenotypic methods for identification of many medically important fungi frequently encountered in a routine diagnostic microbiology laboratory.

Biochimie, 2003 Sep, 85(9), 823 - 40
NMR and microbiology: from physiology to metabolomics; Grivet JP et al.; Recent advances in the application of NMR to microbiology are reviewed . Instrumental and methodological developments are described . The advantages and the constraints of NMR are illustrated with examples from the literature . Recent work from the authors' laboratories, taken as representative of current research in the field, is described in more detail.

Mol Microbiol, 2004 Jan, 51(1), 3 - 5
Restricting restriction; Bickle TA; Systems biology is a new, fashionable and well-funded discipline, which to quote from a recent review aims to 'examine the structure and dynamics of cellular and organismal function, rather than the characteristics of isolated parts of a cell or organism em leader ' (Kitano, H . (2002) Science 295:1662-1664) . Systems biology will do this by profiting from the vast amounts of biological information that are available in the genomics era and make extensive use of computer modelling . But: 'many breakthroughs in experimental devices, advanced software and analytical methods are required before the achievements of system biology can live up to their much-touted potential' . This edition of Molecular Microbiology contains a paper that is the product of traditional experimental biology but which could serve as a test case for systems biology . The paper shows how bacteria integrate such disparate subsystems as DNA restriction, homologous recombination and regulated proteolysis to protect their chromosomes from degradation . When systems biology can predict this level of choreography, it will be a mature discipline.

Med Sci (Paris), 2003 Nov, 19(11), 1128 - 36
{Bacterial cyclostatin, or how do bacteria manipulate the eukaryotic cell cycle}; De Rycke J et al.; Several bacterial proteins have been recently described that share the ability to inhibit the proliferation of cells in culture without causing early signs of cytotoxicity . Such observations suggest the existence of bacterial mechanisms of control of the eukaryotic cell cycle contributing to pathogenicity or adaptation to the host . This emerging concept of cellular microbiology is critically analyzed considering as a model the cytolethal distending toxins (CDT), a family of toxins whose mode of action on the cell cycle has been thoroughly studied over the last few years . CDTs activate a physiological G2 checkpoint in exposed cells, probably from an initial DNA alteration whose precise molecular nature has not yet been determined . Experimental data are lacking to extrapolate in vivo the antiproliferative effect of these bacterial proteins that we tentatively propose to call cyclostatins.

J Bacteriol, 2003 Dec, 185(24), 7273 - 8
The Streptomyces coelicolor polynucleotide phosphorylase homologue, and not the putative poly(A) polymerase, can polyadenylate RNA; Sohlberg B et al.; A protein containing a nucleotidyltransferase motif characteristic of poly(A) polymerases has been proposed to polyadenylate RNA in Streptomyces coelicolor (P . Bralley and G . H . Jones, Mol . Microbiol . 40:1155-1164, 2001) . We show that this protein lacks poly(A) polymerase activity and is instead a tRNA nucleotidyltransferase that repairs CCA ends of tRNAs . In contrast, a Streptomyces coelicolor polynucleotide phosphorylase homologue that exhibits polyadenylation activity may account for the poly(A) tails found in this organism.

Infect Control Hosp Epidemiol, 2003 Nov, 24(11), 801 - 6
A survey of latent tuberculosis infection among laboratory healthcare workers in New York City; Garber E et al.; OBJECTIVE: To determine the prevalence of positive tuberculin skin tests (TSTs), incidence of TST conversion, risk factors for positive TSTs, and history of active TB among HCWs in microbiology laboratories in New York City . DESIGN: Two-year survey from May 1999 to June 2001 . SETTING: Nineteen microbiology laboratories . RESULTS: During the first year, interviews were conducted with 345 laboratory HCWs (mean, 18 HCWs per site; range, 2 to 51) to assess the prevalence of positive TSTs, but 3 (1%) could not recall their result and were excluded from further analyses . The mean age of the remaining 342 HCWs was 48 years; 68% (n = 233) were female, 54% (n = 183) received bacille Calmette-Guerin (BCG) vaccination, and 71% (n = 244) were foreign born . The prevalence of a positive TST was 57% (n = 196), but only 20% (n = 39) of the HCWs received isoniazid . The incidence of TST conversion in the second year of the study was 1% (1 of 108) . Multivariate analysis identified age (odds ratio {OR} per year, 1.05; 95% confidence interval {CI95}, 1.02-1.08), foreign birth (OR, 3.80; CI95, 1.98-7.28), BCG immunization (OR, 4.89; CI95, 2.72-8.80), and employment in a mycobacteriology laboratory (OR, 2.14; CI95, 1.25-3.68) as risk factors for a positive TST . Only one HCW had been treated for active TB . CONCLUSIONS: The prevalence of positive TSTs was high among laboratory HCWs, but the TST conversion rate was low . Higher rates of treatment for latent TB infection are desirable.

J Biotechnol, 2003 Dec 5, 106(1), 23 - 32
The role of the Aspergillus niger furin-type protease gene in processing of fungal proproteins and fusion proteins . Evidence for alternative processing of recombinant (fusion-) proteins; Punt PJ et al.; We have characterized growth and protein processing characteristics of Aspergillus niger strains carrying a disrupted allele of the previously cloned and characterized kexB gene {Appl . Environ . Microbiol . 66 (2000) 363} encoding a furin-type endoprotease . Deletion of the single-copy gene confirms it to be non-essential but disruptant strains exhibit a morphologically distinct phenotype characterized by hyperbranching . Processing of homologous pro-proteins and fusion proteins comprised of a heterologous protein fused down-stream of glucoamylase and separated at the fusion junction by an endoproteolytic cleavage site was compared in wildtype and mutant strains of A . niger . We show that maturation of the native glucoamylase requires KexB, whereas maturation of aspergillopepsin does not . The processing of fusion proteins carrying Lys-Arg requires KexB, although alternative endoproteases are capable of cleaving protein fusions at sites adjacent to Lys-Arg.

J Med Virol, 2004 Jan, 72(1), 156 - 61
Genetic variability of human rotavirus strains isolated from Eastern and Northern India; Das S et al.; An epidemiological study was conducted in Eastern and Northern India to determine the genomic diversity of rotaviruses in these parts of the country . In 2001, a total of 126 Group A rotavirus positive samples were detected from children below 4 years of age with diarrhoea from Kolkata, Dibrugarh and Bhubaneswar in Eastern India, and Chandigarh, a city in Northern India . All the samples were genotyped for VP7 (G-type) and VP4 (P-type) gene by reverse transcription (RT) and multiplex PCR using different type specific primers . The strains with G1P{8} (32.5%) was predominant as reported earlier {Das et al . (2002) J Clin Microbiol 40:146-149} followed by G2P{4}(4.7%) and only one sample was of G4P{8} specificity . Along with these common types some rare strains like G1P{6}, G2P{8}, G2P{6}, G4P{4}, and G4P{6} were also detected in 14.3% of cases . Thirty percent of samples in this study were mixed infections and 21 (16.7%) specimens remained untypeable either for the VP7 or for the VP4 gene . After sequencing of the VP7 gene, two G9 strains (RMC321 and ISO-3) were identified with P{8} and P{19} specificities . Sequence analysis revealed that they have much lower homology to the G9 strains (116E, INL1, and G16) isolated earlier from Indian subcontinent, but have much higher homology to isolates from Argentina, Brazil, Malawi, Taiwan, and USA suggesting a separate progenitor for these strains .

J Appl Microbiol, 2003, 95(5), 1034 - 8
Influence of inoculum preparation on the growth of Penicillium chrysogenum; Sautour M et al.; AIMS: The influence of the spore preparation on subsequent fungal growth of Penicillium chrysogenum was assessed . METHODS AND RESULTS: The influence of four factors {the nature of the diluting solution (physiological water and physiological water added with Tween-80), the age of the sporulating culture (4, 8 and 12 days), the strain (737, 738 and 740) and the inoculum size (102, 103, 104 and 105 spores ml(-1)} on two responses (i.e . the radial growth rate, mu, and the lag time, lambda) was studied using an experimental screening methodology . CONCLUSIONS: The main conclusion was the strong effect of the inoculum size on lambda . In contrast, the diluting solution had no effect on both the experimental responses . In order to obtain the highest growth rates, it is recommended to use 4-day-old sporulating cultures with an inoculum size of 102 spores ml(-1) . SIGNIFICANCE AND IMPACT OF THE STUDY: There is a need for standardizing spore preparation in predictive mycology . The screening methodology is a powerful tool to determine the influence of qualitative and quantitative factors on various biological responses and can be applied widely in microbiology.

J AOAC Int, 2003 Sep-Oct, 86(5), 1038 - 44
ISO 17025: practical benefits of implementing a quality system; Honsa JD et al.; As a laboratory certified to ISO 9001:2000 and accredited to ISO 17025, rtech laboratories has incorporated an overall system for technical and quality management, which results in benefits observed in daily laboratory practices . Technical requirements were updated to include the addition of formal personnel training plans and detailed records, method development and validation procedures, measurement of method uncertainty, and a defined equipment calibration and maintenance program . In addition, a stronger definition of the sample preparation process was documented to maintain consistency in sampling, and a more rigorous quality control monitoring program was implemented for chemistry and microbiology . Management quality improvements focused on document control to maintain consistent analytical processes, improved monitoring of supplier performance, a contract review process for documenting customer requirements, and a system for handling customer comments and complaints, with continuous improvement through corrective and preventive action procedures and audits . Quarterly management review of corrective actions, nonconforming testing, and proficiency testing aid in determining long-term trending . The practical benefits of these technical and management quality improvements are seen on a daily basis in the laboratory . Faster identification and resolution of issues regarding methods, personnel or equipment, improved customer satisfaction, meeting quality requirements of specialized customers, and overall increased laboratory business are all the result of implementing an effective quality system.

Hist Sci Med, 2003 Jul-Sep, 37(3), 349 - 55
{What future for neo Hippocratism?}; Salomon-Bayet C; To vindicate the title of his article the author gives three noticeable dates: 1802/1902 are the dates of starting for clinical and physiological medicine and the recognition of microbiology and effective legal policy for public health . In 2002 the molecular biology with its ability to intervene on living matter puts new kinds of questions but who will be able to give answers? The scientists? The medical doctors? The politicians? Will be the Hippocratic Oath still relevant to the modern scientific spirit?

J Virol Methods, 2003 Dec, 114(2), 151 - 8
Validation of a modified commercial assay for the detection of rubella-specific IgG in oral fluid for use in population studies; Ben Salah A et al.; Matching serum and oral fluid (saliva) samples were collected from 369 subjects in Tunisia in 2002, from a city in the north and a rural district in the south . Rubella-specific IgG was detected in sera by commercial ELISA (Dade Behring) and in matching oral fluids by two methods, a previously described IgG-capture ELISA (GACELISA) {J . Clin . Microbiol . 37 (1999) 391} and the Dade Behring ELISA with the assay protocol modified for use with oral fluids . Total IgG concentration of oral fluids was also measured . Rubella-specific IgG was detected in 289 (78.3%) sera overall . Differences in the age distribution of the study population in the north and south led to a higher prevalence being found in the north (86.2%) than in the south (71.8%) . This difference was reflected in the oral fluid rubella-specific IgG results . With GACELISA, rubella-specific IgG was detected in 79.4% of oral fluids from the north and 69.7% from the south and with the modified Dade Behring assay, in 81.4% of oral fluids from the north and in 64.9% from the south . The sensitivity and specificity of GACELISA in comparison to results from the matching sera were 92.4 and 93.2%, respectively . The sensitivity and specificity of the modified Dade Behring ELISA were 89.8 and 92.0%, respectively . Total IgG concentration in oral fluid showed a weak correlation (r=0.19) with the modified Dade Behring results and with samples where total IgG was >7.5 mg/l, the sensitivity and specificity were 94.4 and 90.0%, respectively . Twenty-nine oral fluids, which gave false negative rubella-specific IgG results with the modified Dade Behring ELISA, had a lower mean total IgG concentration than 256 oral fluids which gave concordant positive results (7.0mg/l versus 15.8 mg/l, P<0.001) . The study validated the modified Dade Behring ELISA, providing an alternative to the GACELISA for assessing levels of rubella immunity for population studies using oral fluid samples.

Mol Microbiol, 2003 Nov, 50(4), 1229 - 39
Drug-induced alterations in gene expression of the asexual blood forms of Plasmodium falciparum; Gunasekera AM et al.; The complete genome sequence of the malarial parasite, Plasmodium falciparum {Gardner, M.J., Hall, N., Fung, E., White, O., Berriman, M., and Hyman, R.W . (2002) Nature 419: 498-510; Hyman, R.W., Fung, E., Conway, A., Kurdi, O., Mao, J., Miranda, M . et al . (2002) Nature 419: 534-537}, has provided researchers with the informational base for establishing genomic {Volkman, S.K., Hartl, D.L., Wirth, D.F., Nielsen, K.M., Choi, M., Batalov, S., et al . (2002) Science 298: 216-218}, proteomic {Florens, L., Washburn, M.P.J.D.R., Anthony, R.M., Grainger, M., Haynes, J.D., et al . (2002) Nature 419: 520-526; Lasonder, E., Ishihama, Y., Anderson, J.S., Vermunt, A.M.W., Pain, A., Sauerwein, R.W., et al . (2002) Nature 419: 537-542} and genome-wide RNA expression {Ben Mamoun, C., Gluzman, I.Y., Hott, C., MacMillan, S.K., Amarakone, A.S., Anderson, D.L., et al . (2001) Mol Microbiol 39: 26-36; Hayward, R.E., Derisi, J.L., Alfadhli, S., Kaslow, D.C., Brown, P.O., and Rathod, P.K . (2000) Mol Microbiol 35: 6-14} analyses in this system . In fact, we have previously utilized SAGE (serial analysis of gene expression) to identify abundant loci that probably constitute part of the active metabolome {Patankar, S., Munasinghe, A., Shoaibi, A., Cummings, L.M., and Wirth, D.F . (2001) Mol Biol Cell 12: 3114-3125}, as well as to characterize antisense transcription on a global scale in Plasmodium . In the present study, the comprehensive annotation of SAGE libraries derived from an asexual stage population exposed to drug and its matched control was used to assess the modulation of gene expression by chloroquine . Here, we observed a constellation of changes, with the differential regulation of over 100 transcripts, and have confirmed the data by alternate methods . A few responsive loci, including PfMDR1, have previously been implicated in the mechanism of chloroquine action/resistance . Several others, however, were derived from unexpected categories, including a large number of unknown open reading frames (ORFs), whose induction after drug exposure may provide first hints to their possible function.

Water Sci Technol, 2003, 48(5), 59 - 66
Seasonal variation in phosphorus removal processes within reed beds--mass balance investigations; Headley TR et al.; The phosphorus (P) removal processes in two pairs of High and Low Loaded reed beds were investigated during five periods within a 27-month study . The uptake/release of P was measured in seven mass balance compartments . With the exception of the first year of operation, the reed beds consistently removed over 96% of the influent P load, with total phosphorus (TP) concentrations being reduced from 0.5 mg/L to generally less than 0.005 mg/L across the range of loading rates and seasons studied . During the first year, uptake by Phragmites australis accounted for greater than 75% of P removed, and was equally distributed between above and below-ground biomass . During the second and third years, three seasonal stages were identified in the uptake and cycling of P by P . australis . A period of rapid above-ground growth and uptake occurred during spring fuelled partly by P reserves accumulated in rhizomes during the previous year . During summer, uptake by above-ground biomass was governed by the influent P loading rate, while the amount of P held in below-ground biomass remained relatively stable . During autumn and winter, P appeared to be translocated from senescent shoots to reserves in the rhizomes . Approximately 85% of the below-ground biomass P occurred in the top 20 cm of the substrate . Gravel fixation increased in importance from 12% in the first year to approximately 30% of P removed in the second year, with a highly significant correlation between the influent P loading rate and P fixed by the gravel . The weakly-bound P fraction from a sequential extraction was the dominant form of P fixed by the gravel . HCI extracts were inappropriate for the examination of sorption processes as they dissolved large amounts of mineral P from within the basaltic gravel . The bottom 30 cm of the substrate became the most important site for gravel fixation during the second year . Incorporation of P into the detritus/microbiota/other compartment increased after the first year to become one of the most important P removal processes, probably consisting mainly of leaf litter and slowly accreted organic sediments.

Eur Respir J Suppl, 2003 Nov, 47, 57s - 64s
Current status of lung transplantation; Lau CL et al.; Two decades have passed since the first successful clinical lung transplant was performed in 1983, and, in the interim, lung transplantation has become the preferred treatment option for a variety of end-stage pulmonary diseases . Remarkable progress has been made in the field through refinement of technique and improved understanding of transplant immunology and microbiology . Unfortunately, donor shortages continue to limit the more widespread application of lung transplantation . In order to address this issue, marginal donors, living lobar and split lung donor techniques, and nonheartbeating donors have been used clinically to increase the number of donor lungs available . Chronic rejection of the lung allograft is currently the major hurdle limiting longterm survival . To date, prevention of known risk factors and treatment strategies have not lessened the devastating toll this process has on lung transplant survival . Better understanding of the cause of chronic rejection is needed in order to develop novel strategies for its treatment . Promotion of immune tolerance is a promising area that could potentially eliminate chronic rejection . The present article discusses recent advances in lung transplantation . It also details the major issues facing the field today . Only through continued clinical and experimental investigation will lung transplantation eventually reach its full potential.

Kaohsiung J Med Sci, 2003 Oct, 19(10), 510 - 5
Splenic abscesses: review of 29 cases; Chiang IS et al.; Splenic abscess is an unusual and potentially life-threatening disease . Due to the nonspecific clinical picture, it remains a diagnostic challenge . Splenic abscess should be suspected in febrile patients with left upper quadrant tenderness and leukocytosis, and diagnosis confirmed based mostly on imaging studies, microbiologic and / or pathologic evidence, or by response to antibiotic or antifungal treatment . We present 29 cases of splenic abscess treated in our hospital from 1990 to 2001 . There were 18 male patients (62%) and 11 female patients (38%) . Ages ranged from 4 to 85 years, with a median of 44 years . There were five pediatric patients (17%) and 24 adults (83%) . The most common associated condition was leukemia . Most patients were immunocompromised (72%) . The more common signs and symptoms were fever (90%), chills (41%), abdominal pain (31%), and leukocytosis (38%) . Ultrasonography of the abdominal cavity was positive in 27 cases (93%); computerized tomography or magnetic resonance imaging was used in 26 patients (90%) and was positive in all patients . The abscess was solitary in 21 cases (72%) and multiple in eight cases (28%) . Positive blood cultures were found in only seven patients (24%) . According to the literature, the treatment of choice is still splenectomy, but in our study, the success rate of 75% with antibiotics alone indicates that antibiotic therapy should be considered an important alternative treatment modality in patients not suitable for percutaneous drainage and splenectomy.

Ophthalmologe, 2003 Oct, 100(10), 802 - 7
{Practical advice for recovery and successful processing of anterior chamber puncture and vitrectomy samples}; Thurau S; For the diagnosis of unclear intraocular inflammation which threatens visual acuity and does not respond to therapy, the collection of intraocular fluid from the anterior chamber or the vitreous body is sometimes required . After surgery the correct and speedy processing of the rapidly degrading fluid determines the quality and reliability of the laboratory results . This article describes the different steps which are important after surgery to ensure optimal results for the different diagnostic laboratory procedures . If specimens are to be sent to cytology or microbiology laboratories, processing directly after surgery is necessary, while material which has to undergo PCR analysis, should remain untreated.

Int J Immunopathol Pharmacol, 2003 Sep-Dec, 16(3), 261 - 8
Enzyme-linked immunosorbent assay, immunofluorescent assay, and recombinant immunoblotting in the serodiagnosis of early Lyme borreliosis; Christova I; Serum samples from Bulgarian patients with physician-diagnosed erythema migrans (EM) (n=105) were examined using Borrelia burgdorferi ELISA (Boehring, Germany) after previous absorption with Treponema phagedenis . For IgM antibody detection sera were additionally pretreated with anti-IgG serum (RF absorbent) . Serum samples of 93% of persons from healthy control group were IgM negative and all were IgG negative . Out of 105 patients with EM, 49% were IgM positive and 14 % were borderline . IgG ELISA showed positive results for 17% and borderline for 6% of the patients . Positive and borderline serum samples were examined further by immunofluorescent assay (IFA) and immunoblot test with recombinant B . burgdorferi proteins from strain PKo (B . afzelii) - p100, flagellin, OspA and OspC, and internal flagellin fragments from strains PKo and PBi (B . garinii) {B.Wilske, V.Fingerle, P.Herzer et al . 1993 . Med . Microbiol . Immunol . 182:255} . IFA detected IgM antibodies against B . burgdorferi in 47 % of the positive and in none of the borderline by IgM ELISA serum samples as well as IgG antibodies in 83% of the positive and in 50% of the borderline by IgG ELISA samples . Presence of specific antibodies was confirmed by immunoblot in 71 % of the IgM ELISA postive and in 67 % of the IgG ELISA positive sera . In addition, anti-B . burgdorferi antibodies were detected in 60 % of the borderline by IgM ELISA serum samples . IgM serum reactivity was directed mainly against OspC antigen and flagellin and IgG antibodies were directed mainly against flagellin and p100 . These findings clearly showed advantages of the ELISA test based on previous pretreatment of sera and capable to detect specific antibodies in more than half of patients with early Lyme borreliosis despite the well-known delayed immune response . IFA was less sensitive than ELISA in detection of anti-B . burgdorferi antibodies . An additional examination of ELISA borderline sera by immunoblot revealed more positive results . Serum reactivity to a single OspC antigen seems to be a sufficient criterion for positive IgM immunoblot.

J Immunol, 2003 Nov 15, 171(10), 5380 - 8
Alveolar macrophage apoptosis contributes to pneumococcal clearance in a resolving model of pulmonary infection; Dockrell DH et al.; The role of alveolar macrophages (AM) in host defense against pulmonary infection has been difficult to establish using in vivo models . This may reflect a reliance on models of fulminant infection . To establish a unique model of resolving infection, with which to address the function of AM, C57BL/6 mice received low-dose intratracheal administration of pneumococci . Administration of low doses of pneumococci produced a resolving model of pulmonary infection characterized by clearance of bacteria without features of pneumonia . AM depletion in this model significantly increased bacterial outgrowth in the lung . Interestingly, a significant increase in the number of apoptotic AM was noted with the low-dose infection as compared with mock infection . Caspase inhibition in this model decreased AM apoptosis and increased the number of bacteremic mice, indicating a novel role for caspase activation in pulmonary innate defense against pneumococci . These results suggest that AM play a key role in clearance of bacteria from the lung during subclinical infection and that induction of AM apoptosis contributes to the microbiologic host defense against pneumococci.

J Biol Chem, 2004 Jan 30, 279(5), 3318 - 26 Epub 2003 Nov 07.
Determination of transmembrane topology of the Escherichia coli natural resistance-associated macrophage protein (Nramp) ortholog; Courville P et al.; The natural resistance-associated macrophage protein (Nramp) defines a conserved family of secondary metal transporters . Molecular evolutionary analysis of the Nramp family revealed the early duplication of an ancestral eukaryotic Nramp gene, which was likely derived from a bacterial ortholog and characterized as a proton-dependent manganese transporter MntH (Makui, H., Roig, E., Cole, S . T., Helmann, J . D., Gros, P., and Cellier, M . F . (2000) Mol . Microbiol . 35, 1065-1078) . Escherichia coli MntH represents a model of choice to study structure function relationship in the Nramp protein family . Here, we report E . coli MntH transmembrane topology using a combination of in silico predictions, genetic fusion with cytoplasmic and periplasmic reporters, and MntH functional assays . Constructs of the secreted form of beta-lactamase (Blam) revealed extra loops between transmembrane domains 1/2, 5/6, 7/8, and 9/10, and placed the C terminus periplasmically; chloramphenicol acetyltransferase constructs indicated cytoplasmic loops 2/3, 6/7, 8/9, and 10/11 . Two intra loops for which no data were produced (N terminus, intra loop 4/5) both display composition bias supporting their deduced localization . The extra loops 5/6 and 6/7 and periplasmic exposure of the C terminus were confirmed by targeted reporter insertion . Three of them preserved MntH function as measured by a disk assay of divalent metal uptake and a fluorescence assay of divalent metal-dependent proton transport, whereas a truncated form lacking transmembrane domain 11 was inactive . These results demonstrate that EcoliA is a type III integral membrane protein with 11 transmembrane domains transporting both divalent metal ions and protons.

Rev Esp Anestesiol Reanim, 2002 Apr, 49(4), 209 - 12
{Candida parapsilosis endocarditis after prolonged antibiotic therapy}; Castillo Caparros A et al.; Infective endocarditis is still a very serious disease whose prognosis has improved in recent years, even though advanced technologies have brought a greater number of cases attributable to increased growth of fungi in more virulent forms . We describe a case of endocarditis caused by Candida parapsilosis on a previously healthy aortic valve in a man who had received prolonged intravenous antibiotic treatment a few months earlier . The initial presentation included acute arterial ischaemia of the lower limbs . The diagnosis of endocarditis was then confirmed by echocardiography and microbiology . Specific antifungal therapy was started, and embolectomy and valve replacement were performed; however, the infection could not be controlled and the patient died a few days later.

Euro Surveill, 2003 Oct, 8(10), 189 - 95
Seventh International Meeting of the European Laboratory Working Group on Diphtheria - Vienna, June 2002; Kelly C et al.; The Seventh International Meeting of the European Laboratory Working Group on Diphtheria (ELWGD) was held in Vienna, Austria in June 2002 and brought together the microbiologists and epidemiologists responsible for diphtheria in many countries throughout the world . The ELWGD was formed in 1993 in response to the epidemics of diphtheria in Russia and the Newly Independent States (NIS) of the former Soviet Union (1,2) . These epidemics, and others elsewhere, highlighted the importance of strengthening and maintaining both epidemiological surveillance and reliable laboratory screening . Since December 2001, these areas have been progressed and strengthened by European Commission (EC) funding to undertake a feasibility study for diphtheria surveillance amongst European Union (EU) member states and accession countries, and to establish a definitive and official European diphtheria surveillance network called DIPNET (EC DG SANCO Agreement No . S12.324473 (2001 CVG4-012)) . This was the first international meeting of the ELWGD to be organised and funded under the auspice of the DIPNET (European Diphtheria Surveillance Network).

J Clin Microbiol, 2003 Nov, 41(11), 5085 - 93
Evaluation and field validation of PCR tests for detection of Actinobacillus pleuropneumoniae in subclinically infected pigs; Fittipaldi N et al.; Eight PCR tests were evaluated for their abilities to detect Actinobacillus pleuropneumoniae in swine tonsils . At first they were compared regarding their specificities by using A . pleuropneumoniae and related bacterial species and their analytical sensitivities by using tonsils experimentally infected in vitro . PCRs were carried out both directly with tonsil homogenates (direct PCR) and after culture of the sample (after-culture PCR) . Most tests demonstrated good specificities; however, some tests gave false-positive results with some non-A . pleuropneumoniae species . High degrees of variation in the analytical sensitivities among the tests were observed for the direct PCRs (10(9) to 10(2) CFU/g of tonsil), whereas those of most of the after-culture PCRs were similar (10(2) CFU/g of tonsil) . In a second phase, the effects of sample storage time and storage conditions were evaluated by using tonsils from experimentally infected animals . Storage at -20 degrees C allowed the detection of the organism for at least 4 months . Finally, the omlA PCR test described by Savoye et al . (C . Savoye et al., Vet . Microbiol . 73:337-347, 2000) and the commercially available Adiavet App PCR test were further validated with field samples . Their effectiveness was compared to those of standard and immunomagnetic separation-based methods of bacterial isolation . In addition, a comparison of tonsil biopsy specimens (from living animals) and whole tonsils (collected at the slaughterhouse) was also conducted . A . pleuropneumoniae was neither isolated nor detected by PCR from a herd serologically negative for A . pleuropneumoniae . PCR was more sensitive than the standard isolation method with whole tonsils from three infected herds . After-culture PCR offered the highest degree of sensitivity (93 and 83% for the omlA and Adiavet App PCRs, respectively) . The PCR detection rate was higher with whole tonsils than with tonsil biopsy specimens . Good agreement (kappa = 0.65) was found between the presence of A . pleuropneumoniae in tonsils and the individual serological status of the animals.

Ann Thorac Surg, 2003 Nov, 76(5), 1733 - 5
Tuberculosis presenting as Pancoast tumor; Beshay M et al.; A 48-year-old man presented with pain in his left shoulder radiating to the left scapula and a tingling sensation of the left arm with involvement of the fourth and fifth finger . Based on the clinical and radiologic findings, the diagnosis of Pancoast tumor of the left lung was made . Computed tomographic guided fine needle biopsy was not conclusive . A video-assisted thoracoscopic surgery was performed to obtain a biopsy . The histologic and microbiologic examinations established the diagnosis of tuberculosis (TB).

Indian J Ophthalmol, 2003 Sep, 51(3), 269 - 72
Ophthalmic photography using a digital camera; Fogla R et al.; PURPOSE: To report the application of a digital camera for ophthalmic photography in routine clinical use . METHODS: A digital camera (Nikon Coolpix 995) was used both for external macrophotography of the eye and ocular adnexa, and slitlamp photography of the anterior segment of the eye . RESULTS: We were able to take external macrophotographs under high magnification of the eye and ocular adnexa . Slitlamp photography could be performed under diffuse, slit beam, and retroillumination . The structures of the angle, the optic disc and surrounding retina could be photographed using appropriate lenses . The attachment to the operating microscope allowed intraoperative photography . It could also be attached to the laboratory microscope to capture images of various histopathology and microbiology slides . CONCLUSIONS: A digital camera is a versatile instrument for ophthalmic photography . It is easy to use in routine clinical practice and provides good quality photographs.

Gene, 2003 Nov 13, 319, 99 - 106
The pyruvate dehydrogenase complex of Mycoplasma hyopneumoniae contains a novel lipoyl domain arrangement; Matic JN et al.; The genes encoding the pyruvate dehydrogenase (PDH) complex (pdhA, pdhB, pdhC and pdhD) from Mycoplasma hyopneumoniae have been cloned and sequenced . The genes are arranged into two operons, designated pdhAB and pdhCD, which are not found together in the chromosome . The pdhA, pdhB, pdhC and pdhD genes encode proteins of predicted molecular masses of 44.2 kDa (pyruvate dehydrogenase major subunit; E1alpha), 36.6 kDa (pyruvate dehydrogenase minor subunit; E1beta), 33.1 kDa (dihydrolipoyl acetyltransferase; E2) and 66.3 kDa (dihydrolipoyl dehydrogenase; E3), respectively . Sequence analysis of the pdhCD operon revealed the presence of a lipoyl-binding domain in pdhD but not in pdhC . The lipoyl domain is believed to act as a "swinging arm" that spans the gaps between the catalytic domains of each of the subunits . Portions of the N-terminal regions of pdhA and pdhD were expressed as 6xHis-tag fusion proteins in Escherichia coli and purified by nickel affinity chromatography . The purified proteins were used to raise antibodies in rabbits, and Western blot analysis was performed with the polyclonal rabbit antiserum . Both the pdhA and pdhD genes were expressed among various strains of M . hyopneumoniae as well as the porcine mycoplasmas, Mycoplasma hyorhinis and Mycoplasma flocculare . Southern hybridisation analysis using probes from pdhA and pdhD detected one copy of each gene in the chromosome of M . hyopneumoniae . Since previous studies have shown pyruvate dehydrogenase activity in M . hyopneumoniae {J . Gen . Microbiol . 134 (1988) 791}, it appears likely that a functional lipoyl-binding domain in the N terminus of PdhC is not an absolute prerequisite for pyruvate dehydrogenase enzyme activity . We hypothesise that the lipoyl-binding domain of PdhD is performing the enzymatic function normally attributed to the PdhC lipoyl-binding domain in other organisms . Searches of pyruvate dehydrogenase gene sequences derived from other Mycoplasma species showed that a putative lipoyl domain was absent in the pdhC gene from Mycoplasma pulmonis . However, like other bacterial species, pdhC gene sequences from Mycoplasma capricolum, Mycoplasma genitalium and Mycoplasma pneumoniae contain a putative lipoyl domain.

Pediatrics . 2003 Nov;112(5):e380.
Increased diagnosis of Lemierre syndrome and other Fusobacterium necrophorum infections at a Children's Hospital; Ramirez S et al.; OBJECTIVE: To assess the apparent increase in the diagnosis of Lemierre syndrome (LS) and other Fusobacterium necrophorum infections at a large children's hospital . Infections with F necrophorum ranged from peritonsillar abscess to potentially fatal LS . LS is an oropharyngeal infection characterized by septic thrombophlebitis of head and neck veins, complicated by dissemination of septic emboli to pulmonary and systemic sites . METHODS: Review of the medical and laboratory records was conducted of all patients who were seen at or admitted to the Children's Hospital of Wisconsin with the diagnosis of LS and/or isolation of F necrophorum from a clinical specimen between January 1995 and January 2002 . RESULTS: During the 7-year period of the study, there was an increase in the isolation of F necrophorum from patients who were seen at Children's Hospital of Wisconsin, as well as the number of cases of LS . There was 1 isolation of F necrophorum from clinical specimens per year from 1996 to 1999, which increased to 10 isolates of the organism from January 2000 to January 2002 . During the most recent period, January 2001-January 2002, 5 cases of LS were diagnosed, a distinctive entity not recognized previously at the institution . CONCLUSIONS: The cause for the recent increase in the number of serious infections caused by F necrophorum infection diagnosed at our institution is unclear but does not seem to be related to changes in microbiologic techniques or patient demography . We speculate that it could be attributable, in part, to alterations in antibiotic usage patterns in our region . Clinicians need to be aware of the increasing clinical importance of F necrophorum infections and the life-threatening nature of LS.

Fed Regist, 2003 Oct 31, 68(211), 62007 - 8
Medical devices; immunology and microbiology devices; classification of the endotoxin assay . Final rule; Food and Drug Administration et al.; The Food and Drug Administration (FDA) is classifying the endotoxin assay into class II (special controls) . The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990 (SMDA), the Food and Drug Administration Modernization Act of 1997 (FDAMA), and the Medical Device User Fee and Modernization Act of 2002 (MDUFMA) . The agency is classifying this device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device . Elsewhere in this issue of the Federal Register, FDA is announcing the availability of a guidance document that will serve as the special control for the device.

J Infect Dis, 2003 Nov 1, 188(9), 1394 - 403 Epub 2003 Oct 15.
Association of mannose-binding lectin polymorphisms with sepsis and fatal outcome, in patients with systemic inflammatory response syndrome; Garred P et al.; Genetic factors may predispose critically ill patients to increased risk of developing sepsis . Mannose-binding lectin (MBL) is an important factor in innate immune defense . We investigated whether MBL gene polymorphisms causing low levels of MBL are associated with the development and progression of sepsis in adult patients in intensive care units . In 272 prospectively monitored patients with systemic inflammatory response syndrome, different MBL genotypes were compared, with respect to microbiology, sepsis development, and survival . The presence of MBL variant alleles was associated with the development of sepsis, severe sepsis, and septic shock . An increased risk of fatal outcome was observed in patients carrying variant alleles . These data show that MBL insufficiency plays an important role in the susceptibility of critically ill patients to the development and progression of sepsis and confers a substantial risk of fatal outcome.

Fed Regist, 2003 Oct 30, 68(210), 61743 - 5
Medical devices; immunology and microbiology devices; classification of the West Nile Virus IgM capture Elisa assay . Final rule; Food and Drug Administration et al.; The Food and Drug Administration (FDA) is classifying the West Nile Virus IgM Capture Elisa assay into class II (special controls) . The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997 (FDAMA) . The agency is classifying this device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device . Elsewhere in this issue of the Federal Register, FDA is announcing the availability of a guidance document that will serve as the special control for the device.

Eur Respir J, 2003 Oct, 22(4), 589 - 91
Diagnostic tools in tuberculous pleurisy: a direct comparative study; Diacon AH et al.; Thoracoscopy is the most accurate yet most expensive tool for establishing the diagnosis of tuberculous (TB) pleurisy . However, most high TB-incidence regions have limited financial resources, lack the infrastructure needed for routine thoracoscopy and require an alternative, cost-effective diagnostic approach for pleural effusions . Altogether, 51 patients with undiagnosed exudative pleural effusions were recruited for a prospective, direct comparison between bronchial wash, pleural fluid microbiology and biochemistry (adenosine deaminase (ADA) and cell count), closed needle biopsy, and medical thoracoscopy . The final diagnosis was TB in 42 patients (82%), malignancy in five (10%) and idiopathic in four patients (8%) . Sensitivity of histology, culture and combined histology/culture was 66, 48 and 79%, respectively for closed needle biopsy and 100, 76 and 100%, respectively for thoracoscopy . Both were 100% specific . Pleural fluid ADA of > or = 50 U x L(-1) was 95% sensitive and 89% specific . Combined ADA, lymphocyte/neutrophil ratio > or = 0.75 plus closed needle biopsy reached 93% sensitivity and 100% specificity . A combination of pleural fluid adenosine deaminase, differential cell count and closed needle biopsy has a high diagnostic accuracy in undiagnosed exudative pleural effusions in areas with high incidences of tuberculosis and might substitute medical thoracoscopy at considerably lower expense in resource-poor countries.

Allergol Immunopathol (Madr), 2003 Sep-Oct, 31(5), 294 - 6
Familial presentation of occupational hypersensitivity pneumonitis caused by aspergillus-contaminated esparto dust; Moreno-Ancillo A et al.; Esparto grass (Stipa tenacissima), which is commonly found in the Mediterranean area, has a wide variety of uses . Five plaster workers from the same family developed cough, dyspnea, malaise, and fever after exposure to the esparto fiber used in their work for the previous few years . They showed a significant decrease in symptoms when away from work . Precipitating antibodies against an esparto extract were found in the sera of all patients . Specific IgG antibodies against Aspergillus fumigatus were detected . A . fumigatus was identified after microbiologic evaluation of esparto fiber samples . The dust derived from fungi-contaminated esparto fibers can cause hypersensitivity pneumonitis in exposed subjects . The causative antigen is A . fumigatus . When esparto fibers were strongly contaminated by fungi, all the workers developed a clinical picture compatible with hypersensitivity pneumonitis . The coincidental finding of an occupational and a familiar condition is unusual.

Enferm Infecc Microbiol Clin, 2003 Nov, 21(9), 498 - 508
{Guidelines for the treatment of invasive fungal infection . Invasive fungal infection by Candida spp . Invasive Fungal Infection Study Group (MICOMED) and Infection in Transplantation Study Group (GESITRA) of the Spanish Society for Infectious Diseases and Clinical Microbiology (SEIMC)}; Gavalda J et al.; These guidelines resume different issues related to the clinical management of invasive candidosis in the immunossuppressed patients . They are based on the recommendations of the different Drug's Agencies.

Orv Hetil, 2003 Jul 27, 144(30), 1481 - 7
{Accreditation of medical laboratories}; Horvath AR et al.; In Hungary, the National Accreditation Body was established by government in 1995 as an independent, non-profit organization, and has exclusive rights to accredit, amongst others, medical laboratories . The National Accreditation Body has two Specialist Advisory Committees in the health care sector . One is the Health Care Specialist Advisory Committee that accredits certifying bodies, which deal with certification of hospitals . The other Specialist Advisory Committee for Medical Laboratories is directly involved in accrediting medical laboratory services of health care institutions . The Specialist Advisory Committee for Medical Laboratories is a multidisciplinary peer review group of experts from all disciplines of in vitro diagnostics, i.e . laboratory medicine, microbiology, histopathology and blood banking . At present, the only published International Standard applicable to laboratories is ISO/IEC 17025:1999 . Work has been in progress on the official approval of the new ISO 15189 standard, specific to medical laboratories . Until the official approval of the International Standard ISO 15189, as accreditation standard, the Hungarian National Accreditation Body has decided to progress with accreditation by formulating explanatory notes to the ISO/IEC 17025:1999 document, using ISO/FDIS 15189:2000, the European EC4 criteria and CPA (UK) Ltd accreditation standards as guidelines . This harmonized guideline provides 'explanations' that facilitate the application of ISO/IEC 17025:1999 to medical laboratories, and can be used as a checklist for the verification of compliance during the onsite assessment of the laboratory . The harmonized guideline adapted the process model of ISO 9001:2000 to rearrange the main clauses of ISO/IEC 17025:1999 . This rearrangement does not only make the guideline compliant with ISO 9001:2000 but also improves understanding for those working in medical laboratories, and facilitates the training and education of laboratory staff . With the official acceptance of ISO 15189 the clauses of this harmonized guideline fulfill the requirements of the new international standard as well . Accreditation of medical laboratories in Hungary may not only facilitate quality improvement of laboratory services, but also the development of a quality-based purchasing and reimbursement policy of the health insurance fund.

J Med Libr Assoc, 2003 Oct, 91(4), 434 - 41
Bibliometric study of grey literature in core veterinary medical journals; Pelzer NL et al.; OBJECTIVES: Grey literature has been perceived by many as belonging to the primary sources of information and has become an accepted method of nonconventional communication in the sciences and medicine . Since little is known about the use and nature of grey literature in veterinary medicine, a systematic study was done to analyze and characterize the bibliographic citations appearing in twelve core veterinary journals . METHODS: Citations from 2,159 articles published in twelve core veterinary journals in 2000 were analyzed to determine the portion of citations from grey literature . Those citations were further analyzed and categorized according to the type of publication . RESULTS: Citation analysis yielded 55,823 citations, of which 3,564 (6.38%) were considered to be grey literature . Four veterinary specialties, internal medicine, pathology, theriogenology, and microbiology, accounted for 70% of the total number of articles . Three small-animal clinical practice journals cited about 2.5-3% grey literature, less than half that of journals with basic research orientations, where results ranged from almost 6% to approximately 10% grey literature . Nearly 90% of the grey literature appeared as conferences, government publications, and corporate organization literature . CONCLUSIONS: The results corroborate other reported research that the incidence of grey literature is lower in medicine and biology than in some other fields, such as aeronautics and agriculture . As in other fields, use of the Internet and the Web has greatly expanded the communication process among veterinary professionals . The appearance of closed community email forums and specialized discussion groups within the veterinary profession is an example of what could become a new kind of grey literature.

Uisahak, 2003 Jun, 12(1), 1 - 12
{A study about Il Chun Yu}; Lee GS; Il Chun Yu was the first Korean microbiologist and the Korean student who studied in Germany . He got the two doctor degree: one in Freiburg University of Germany in 1924, and the other Keio University of Japan in 1926 . He became a professor of Kyungsung Medical Junior College in 1926, and as the first grade scholar in the Korean microbiology field . He issued many essays in several mass communication media to cultivate people and played an important role in the field of health hygiene by contributing to the improvement of health hygiene.

Am J Clin Pathol, 2003 Oct, 120(4), 560 - 6
A high proportion of novel mycobacteria species identified by 16S rDNA analysis among slowly growing AccuProbe-negative strains in a clinical setting; Pauls RJ et al.; Sequencing of the 16S ribosomal DNA (rDNA) for identification of nontuberculous mycobacteria (NTM) has contributed to the establishment of more than 35 new species during the last decade . Increasingly, NTM are accepted as potential or proven pathogens . We identified, by 16S rDNA sequence analysis, slowly growing NTM isolates negative by AccuProbe (GenProbe, San Diego, CA) that previously were identified by using conventional biochemical techniques, to determine the accuracy of reporting AccuProbe-negative NTM prior to sequence-based identification . Of 82 strains, 30 were deemed novel . An attempt was made to determine the clinical importance of previously misidentified novel species . Clinical cases are described for a number of strains previously identified as Mycobacterium terrae complex, Mycobacterium scrofulaceum, and Mycobacterium avium complex . As sequence-based identification methods become more commonplace in clinical microbiology laboratories, there is a need to understand the significance of previously undescribed species, which often mimic and subsequently are identified as well-established species.

Dtsch Tierarztl Wochenschr, 2003 Sep, 110(9), 388 - 94
{Intestinal microecology in human and veterinary medicine--a short excursion through the human and animal gut}; Ruffer A et al.; In honour to a great gut microbiologist the authors descend into the abdomen of humans and animals . By the way through the gut and excrements they illustrate the medical, culture-historical and literary importance of the gut and the gut flora.

Clin Infect Dis, 2003 Nov 1, 37(9), 1234 - 43 Epub 2003 Oct 03.
A prospective, randomized trial examining the efficacy and safety of clarithromycin in combination with ethambutol, rifabutin, or both for the treatment of disseminated Mycobacterium avium complex disease in persons with acquired immunodeficiency syndrome; Benson CA et al.; This multicenter, randomized, open-label phase 3 clinical trial compared the safety and efficacy of 3 clarithromycin-containing combination regimens for the treatment of disseminated Mycobacterium avium complex (MAC) disease in persons with acquired immunodeficiency syndrome . A total of 160 eligible patients with bacteremic MAC disease were randomized to receive clarithromycin with either ethambutol (C+E), rifabutin (C+R), or both (C+E+R) for 48 weeks . After 12 weeks of treatment, the proportion of subjects with a complete microbiologic response was not statistically significantly different among treatment arms: the proportion was 40% in the C+E group, 42% in the C+R group, and 51% in the C+E+R group (P=.454) . The proportion of patients with complete or partial responses who experienced a relapse while receiving C+R (24%) was significantly higher than that of patients receiving C+E+R (6%; P=.027) and marginally higher than that of patients receiving C+E (7%; P=.057) . Subjects in the C+E+R group had improved survival, compared with the C+E group (hazard ratio {HR}, 0.44; 95% confidence interval {CI}, 0.23-0.83) and the C+R group (HR, 0.49; 95% CI, 0.26-0.92).

Clin Microbiol Rev, 2003 Oct, 16(4), 713 - 29
Laboratory diagnosis of amebiasis; Tanyuksel M et al.; The detection of Entamoeba histolytica, the causative agent of amebiasis, is an important goal of the clinical microbiology laboratory . To assess the scope of E . histolytica infection, it is necessary to utilize accurate diagnostic tools . As more is discovered about the molecular and cell biology of E . histolytica, there is great potential for further understanding the pathogenesis of amebiasis . Molecular biology-based diagnosis may become the technique of choice in the future because establishment of these protozoa in culture is still not a routine clinical laboratory process . In all cases, combination of serologic tests with detection of the parasite (by antigen detection or PCR) offers the best approach to diagnosis, while PCR techniques remain impractical in many developing country settings . The detection of amebic markers in serum in patients with amebic colitis and liver abscess appears promising but is still only a research tool . On the other hand, stool antigen detection tests offer a practical, sensitive, and specific way for the clinical laboratory to detect intestinal E . histolytica . All the current tests suffer from the fact that the antigens detected are denatured by fixation of the stool specimen, limiting testing to fresh or frozen samples.

Infection, 2003 Oct, 31(5), 336 - 40
Spectrum of opportunistic infections and profile of CD4+ counts among AIDS patients in North India; Vajpayee M et al.; BACKGROUND: As the number of AIDS cases increases in India, information available among clinicians about the prevalence of opportunistic infections (OIs) is scarce . The aim of the present study was to document the characteristic OIs of HIV-infected North Indian patients along with their CD4+ counts . PATIENTS AND METHODS: The study group consisted of subjects with confirmed serodiagnosis of HIV, attending the medical clinics at a tertiary health care center in North India . The CD4+ counts were estimated by FACS Calibur (BD) flow cytometer . Simultaneously, routine microbiology smears, cultures and serology were performed to confirm OI . RESULTS: In this retrospective study of 421 subjects, the predominant OI was tuberculosis (47%, 189 cells/ micro l), followed by parasitic diarrhea (43.5%, 227 cells/ micro l) and oral candidiasis (25.2%, 189 cells/ micro l) . CONCLUSION: Tuberculosis was the most frequent OI in the HIV-infected patients studied, and the major mode of transmission of HIV was by sexual route . The median CD4+ counts observed were lower when compared to other studies.

FEMS Microbiol Lett, 2003 Sep 26, 226(2), 373 - 8
Concurrence of losing a chromosome and the ability to produce destruxins in a mutant of Metarhizium anisopliae; Wang C et al.; In a previous study, a spontaneous subtilisin pr1A and pr1B gene-deficient mutant of the entomopathogenic fungus Metarhizium anisopliae strain V275 has been identified {Wang, C.-S . et al . (2002) FEMS Microbiol . Lett . 213, 251-255} . The insecticidal metabolites of this mutant were studied further . High-performance liquid chromatography (HPLC) analysis indicated that the mutant isolate lost the ability to produce cyclic peptide toxins, destruxins, both in vitro and in vivo . Pulsed-field gel electrophoresis revealed that the mutant concurrently lost a 1.05 Mb (approximately) chromosome, demonstrating for the first time that a conditionally dispensable (CD) chromosome exists in the insect pathogenic fungus, M . anisopliae . Concurrence of losing the ability to produce destruxins and a CD chromosome in the mutant suggests that the toxin synthetase genes of M . anisopliae are located on this CD chromosome, as similarly described for plant pathogenic fungi . Semi-quantitative api ZYM analysis showed more biochemical disparities between the mutant and the wild-type strain.

FEMS Microbiol Lett, 2003 Sep 26, 226(2), 209 - 14
Localizing cell division in spherical Escherichia coli by nucleoid occlusion; Zaritsky A et al.; The spatial relationship between FtsZ localization and nucleoid segregation was followed in Escherichia coli thyA cells, made spheroidal by brief exposure to mecillinam and after manipulating chromosome replication time using changes ('steps') in thymine concentration {Zaritsky et al., Microbiology 145 (1999) 1015-1022} . In such cells, fluorescent FtsZ-GFP arcs did not overlap the DAPI-stained nucleoids . It is concluded that FtsZ rings are deposited between segregating nucleoids, consistent with the nucleoid occlusion model {Woldringh et al., J . Bacteriol . 176 (1994) 6030-6038}.

FEMS Microbiol Rev, 2003 Oct, 27(4), 525 - 45
Bacterial hemoglobins and flavohemoglobins: versatile proteins and their impact on microbiology and biotechnology; Frey AD et al.; In response to oxygen limitation or oxidative and nitrosative stress, bacteria express three kinds of hemoglobin proteins: truncated hemoglobins (tr Hbs), hemoglobins (Hbs) and flavohemoglobins (flavo Hbs) . The two latter groups share a high sequence homology and structural similarity in their globin domain . Flavohemoglobin proteins contain an additional reductase domain at their C-terminus and their expression is induced in the presence of reactive nitrogen and oxygen species . Flavohemoglobins detoxify NO in an aerobic process, termed nitric oxide dioxygenase reaction, which protects the host from various noxious nitrogen compounds . Only a small number of bacteria express hemoglobin proteins and the best studied of these is from Vitreoscilla sp . Vitreoscilla hemoglobin (VHb) has been expressed in various heterologous hosts under oxygen-limited conditions and has been shown to improve growth and productivity, rendering the protein interesting for biotechnology industry . The close interaction of VHb with the terminal oxidases has been shown and this interplay has been proposed to enhance respiratory activity and energy production by delivering oxygen, the ultimate result being an improvement in growth properties.

J Vet Med B Infect Dis Vet Public Health, 2003 Sep, 50(7), 332 - 8
Diagnosis of porcine proliferative enteropathy: detection of Lawsonia intracellularis by pathological examinations, polymerase chain reaction and cell culture inoculation; Dittmar M et al.; A total of 21 pigs aged 7-17 weeks with clinical symptoms suggestive for Porcine Proliferative Enteropathy were examined for Lawsonia intracellularis by analysing the following parameters: (i) intestinal gross and histological lesions, (ii) presence of comma-shaped bacteria in enterocytes by Warthin-Starry and a modified Ziehl-Neelsen stain, (iii) PCR amplification of L . intracellularis DNA from intestinal mucosa by using two oligonucleotide primer pairs targeting a 255-bp DNA fragment of the 16S rDNA-gene and a 319-bp DNA fragment of the L . intracellularis chromosome . Specificity of PCR reactions was confirmed by using DNA extracted from the L . intracellularis reference strain N343 (ATCC 55672) as well as by DNA sequence comparisons of PCR amplification products with data bank entries . Intestinal gross lesion indicative for PPE were observed in 20 pigs (95.2%) . For all 21 pigs, the L . intracellularis aetiology was confirmed by histological as well as bacterioscopical examinations . Specific PCR amplification products were obtained from 20 pigs (95.2%) . Taking PCR positivity as the definite criterion, L . intracellularis was diagnosed in 20 pigs from 11 herds in seven Swiss cantons (Argovia, Berne, Fribourg, Grisons, Lucerne, Schwyz, Thurgovia) . To grow L . intracellularisin vitro, the cell culture method of Lawson et al . (J . Clin . Microbiol . 1993: 31, 1136-1142) was adopted . Inocula prepared from heavily infected fresh and frozen ileal mucosa of 15 pigs were cultured in rat enterocytic IEC-18 cells (ATCC CRL 1589) . Six cell culture passages of 10 days each were completed . The reference strain N343 was examined for cultivability, accordingly . Except for occasional specific PCR amplifications from cell cultures up to the second passage, any indications for growth of L . intracellularis in IEC-18 cells were not found.

J Clin Microbiol, 2003 Oct, 41(10), 4671 - 5
Comparison of a shiga toxin enzyme-linked immunosorbent assay and two types of PCR for detection of shiga toxin-producing Escherichia coli in human stool specimens; Pulz M et al.; Shiga toxin (Stx)-producing Escherichia coli (STEC) is a major cause of sporadic cases of disease as well as serious outbreaks worldwide . The spectrum of illnesses includes mild nonbloody diarrhea, hemorrhagic colitis, and hemolytic-uremic syndrome . STEC produces one or more Stxs, which are subdivided into two major classes, Stx1 and Stx2 . The ingestion of contaminated food or water, person-to-person spread, and contact with animals are the major transmission modes . The infective dose of STEC may be less than 100 organisms . Effective prevention of infection is dependent on rapid detection of the causative bacterial pathogen . In the present study, we examined 295 stool specimens for the presence of Stx-producing E . coli by three different methods: an Stx enzyme-linked immunosorbent assay, a conventional PCR assay, and a LightCycler PCR (LC-PCR) assay protocol recently developed by our laboratory at the Institute of Medical Microbiology at Hannover Medical School . Our intent was to compare these three methods and to examine the utility of the STEC LC-PCR protocol in a clinical laboratory . The addition of a control DNA to each sample to clearly discriminate inhibited specimens from negative ones enhanced the accuracy of the LC-PCR protocol . From our results, it can be concluded that LC-PCR is a very useful tool for the rapid and safe detection of STEC in clinical samples.

J Microbiol Methods, 2003 Nov, 55(2), 383 - 92
Comparison of different PCR tests for detecting Shiga toxin-producing Escherichia coli O157 and development of an ELISA-PCR assay for specific identification of the bacteria; Fach P et al.; In an attempt to develop a standard for ELISA-PCR detection of Shiga toxin producing Escherichia coli (STEC) O157, six published PCR tests were tested in a comparative study on a panel of 277 bacterial strains isolated from foods, animals and humans . These tests were based on the detection of the genes rfbE {J . Clin . Microbiol . 36 (1998) 1801} and rfbB {Appl . Environ . Microbiol . 65 (1999) 2954}, the 3' end of the eae gene {Epidemiol . Infect . 112 (1994) 449}, the region immediately flanking the 5' end of the eae gene {Int . J . Food . Microbiol . 32 (1996) 103}, the flicH7 gene {J . Clin . Microbiol . 35 (1997) 656}, or a part of the recently described 2634-bp Small Inserted Locus (SILO(157) locus) of STEC O157 {J . Appl . Microbiol . 93 (2002) 250} . Unlike the other PCR assays, those amplifying the rfb sequences were unable to distinguish toxigenic from nontoxigenic O157 . These assays were relatively specific to STEC O157, giving essentially a cross reaction with clonally related E . coli O55 and to a lesser extent with E . coli O145, O125, O126 . They also detected the Shiga toxin (stx)-negative derivatives of STEC O157 . Based on these results, an ELISA-PCR assay consisting of the solution hybridization of amplicons with two probes that ensured the specificity of the amplification was developed . The ELISA-PCR assay, which used an internal control (IC) of inhibition, was able to detect 1 to 10 copies of STEC O157 in the PCR tube . Adaptation of PCR into ELISA-PCR assay format facilitates specific and sensitive detection of PCR amplification products and constitutes a method of choice for screening STEC O157.

J Am Med Inform Assoc, 2004 Jan-Feb, 11(1), 71 - 7 Epub 2003 Oct 05.
Comparative impact of guidelines, clinical data, and decision support on prescribing decisions: an interactive web experiment with simulated cases; Sintchenko V et al.; OBJECTIVE: The aim of this study was to compare the clinical impact of computerized decision support with and without electronic access to clinical guidelines and laboratory data on antibiotic prescribing decisions . DESIGN: A crossover trial was conducted of four levels of computerized decision support-no support, antibiotic guidelines, laboratory reports, and laboratory reports plus a decision support system (DSS), randomly allocated to eight simulated clinical cases accessed by the Web . MEASUREMENTS: Rate of intervention adoption was measured by frequency of accessing information support, cost of use was measured by time taken to complete each case, and effectiveness of decision was measured by correctness of and self-reported confidence in individual prescribing decisions . Clinical impact score was measured by adoption rate and decision effectiveness . RESULTS: Thirty-one intensive care and infectious disease specialist physicians (ICPs and IDPs) participated in the study . Ventilator-associated pneumonia treatment guidelines were used in 24 (39%) of the 62 case scenarios for which they were available, microbiology reports in 36 (58%), and the DSS in 37 (60%) . The use of all forms of information support did not affect clinicians' confidence in their decisions . Their use of the DSS plus microbiology report improved the agreement of decisions with those of an expert panel from 65% to 97% (p=0.0002), or to 67% (p=0.002) when antibiotic guidelines only were accessed . Significantly fewer IDPs than ICPs accessed information support in making treatment decisions . On average, it took 245 seconds to make a decision using the DSS compared with 113 seconds for unaided prescribing (p<0.001) . The DSS plus microbiology reports had the highest clinical impact score (0.58), greater than that of electronic guidelines (0.26) and electronic laboratory reports (0.45) . CONCLUSION: When used, computer-based decision support significantly improved decision quality . In measuring the impact of decision support systems, both their effectiveness in improving decisions and their likely rate of adoption in the clinical environment need to be considered . Clinicians chose to use antibiotic guidelines for one third and microbiology reports or the DSS for about two thirds of cases when they were available to assist their prescribing decisions.

BMC Public Health . 2003 Oct 06;3(1):31.
An evaluation of completeness of tuberculosis notification in the United Kingdom; Pillaye J et al.; BACKGROUND: There has been a resurgence of tuberculosis worldwide, mainly in developing countries but also affecting the United Kingdom (UK), and other Western countries . The control of tuberculosis is dependent on early identification of cases and timely notification to public health departments to ensure appropriate treatment of cases and screening of contacts . Tuberculosis is compulsorily notifiable in the UK, and the doctor making or suspecting the diagnosis is legally responsible for notification . There is evidence of under-reporting of tuberculosis . This has implications for the control of tuberculosis as a disproportionate number of people who become infected are the most vulnerable in society, and are less likely to be identified and notified to the public health system . These include the poor, the homeless, refugees and ethnic minorities . METHOD: This study was a critical literature review on completeness of tuberculosis notification within the UK National Health Service (NHS) context . The review also identified data sources associated with reporting completeness and assessed whether studies corrected for undercount using capture-recapture (CR) methodology . Studies were included if they assessed completeness of tuberculosis notification quantitatively . The outcome measure used was notification completeness expressed between 0% and 100% of a defined denominator, or in numbers not notified where the denominator was unknown . RESULTS: Seven studies that met the inclusion and exclusion criteria were identified through electronic and manual search of published and unpublished literature . One study used CR methodology . Analysis of the seven studies showed that undernotification varied from 7% to 27% in studies that had a denominator; and 38%-49% extra cases were identified in studies which examined specific data sources like pathology reports or prescriptions for anti-tuberculosis drugs . Cases notified were more likely to have positive microbiology than cases not notified which were more likely to have positive histopathology or be surgical in-patients . Collation of prescription data of two or more anti-tuberculosis drugs increases case ascertainment of tuberculosis . CONCLUSION: The reporting of tuberculosis is incomplete in the UK, although notification is a statutory requirement . Undernotification leads to an underestimation of the disease burden and hinders implementation of appropriate prevention and control strategies . The notification system needs to be strengthened to include education and training of all sub-specialities involved in diagnosis and treatment of tuberculosis.

J Bacteriol, 2003 Oct, 185(20), 6005 - 15
Characterization of the two Mycobacterium tuberculosis recA promoters; Gopaul KK et al.; The recA gene of Mycobacterium tuberculosis is unusual in that it is expressed from two promoters, one of which, P1, is DNA damage inducible independently of LexA and RecA, while the other, P2, is regulated by LexA in the classical way (E . O . Davis, B . Springer, K . K . Gopaul, K . G . Papavinasasundaram, P . Sander, and E . C . Bottger, Mol . Microbiol . 46:791-800, 2002) . In this study we characterized these two promoters in more detail . Firstly, we localized the promoter elements for each of the promoters, and in so doing we identified a mutation in each promoter which eliminates promoter activity . Interestingly, a motif with similarity to Escherichia coli sigma(70) -35 elements but located much closer to the -10 element is important for optimal expression of P1, whereas the sequence at the -35 location is not . Secondly, we found that the sequences flanking the promoters can have a profound effect on the expression level directed by each of the promoters . Finally, we examined the contribution of each of the promoters to recA expression and compared their kinetics of induction following DNA damage.

J Biol Chem, 2003 Dec 19, 278(51), 51068 - 74 Epub 2003 Oct 02.
Glycation and post-translational processing of human interferon-gamma expressed in Escherichia coli; Mironova R et al.; Until recently, nonenzymatic glycosylation (glycation) was thought to affect the proteins of long living eukaryotes only . However, in a recent study (Mironova, R., Niwa, T., Hayashi, H., Dimitrova, R., and Ivanov, I . (2001) Mol . Microbiol . 39, 1061-1068), we have shown that glycation takes place in Escherichia coli as well . In the present study, we demonstrate that the post-translational processing (proteolysis and covalent dimerization) observed with cysteineless recombinant human interferon-gamma (rhIFN-gamma) is tightly associated with its in vivo glycation . Our results show that, at the time of isolation, rhIFN-gamma contained early (but not advanced) glycation products . Using reverse phase high performance liquid chromatography in conjunction with fluorescence measurements, enzyme-linked immunosorbent assay, and mass spectrometry, we found that advanced glycation end products arose in rhIFN-gamma during storage . The latter were identified mainly in the Arg/Lys-rich C terminus of the protein, which was also the main target of proteolysis . Mass spectral analysis and N-terminal sequencing revealed four major (Arg140/Arg141, Phe137/Arg138, Met135/Leu136, and Lys131/Arg132) and two minor (Lys109/Ala110 and Arg90/Asp91) cleavage sites in this region . Tryptic peptide mapping indicated that the covalent dimers of rhIFN-gamma originating during storage were formed mainly by lateral cross-linking of the monomer subunits . Antiviral assay showed that proteolysis lowered the antiviral activity of rhIFN-gamma, whereas covalent dimerization completely abolished it.

J Biol Regul Homeost Agents, 2003 Jul-Sep, 17(3), 213 - 22
Past and present concepts in flow cytometry: a European perspective; Valet G; The development of flow cytometric instrumentation, methods and research concepts in Europe has been a continuous driving force for the general scientific advancement in this area over the years . This review addresses early European concepts of continuing interest with regard to instrumentation, data analysis, clinical and eperimental DNA analysis, cell function and microbiology at their worldwide first appearence while flow cytometric immunology and immunophenotyping will be covered separately . Flow cytometry represents an efficient approach to the enormous complexity of molecular cell architecture and cell function by the analysis of apparent molecular cell phenotypes in heterogeneous cell samples . The present merger of flow and image cytometry into the method independent cytomics discipline increases the potential of cell analysis very significantly . It opens the way for predictive medicine as well as for predictive cytopathology and predictive cytology in everyday clinical and medical practice . Current progress is driven by joint advances in molecular fluorescence technologies and instrument development . This complements the analysis of genome sequence information in an efficient way.

Pediatrics . 2003 Oct;112(4):e261.
Children hospitalized with severe acute respiratory syndrome-related illness in Toronto; Bitnun A et al.; OBJECTIVE: An outbreak of severe acute respiratory syndrome (SARS) occurred in the greater Toronto area between February and June 2003 . We describe the clinical, laboratory, and epidemiologic features of children who were admitted to the Hospital for Sick Children, Toronto, with a presumptive diagnosis of suspect or probable SARS . METHODS: A prospective investigational study protocol was established for the management of children with a presumptive diagnosis of suspect or probable SARS . All were ultimately classified as having probable SARS, suspect SARS, or another cause on the basis of their epidemiologic exposure, clinical and radiologic features, and results of microbiologic investigations . RESULTS: Twenty-five children were included; 10 were classified as probable SARS and 5 were classified as suspect SARS, and in 10 another cause was identified . The exposure consisted of direct contact with at least 1 adult probable SARS case in 11 children, travel from a World Health Organization-designated affected area in Asia in 9 children, and presence in a Toronto area hospital in which secondary SARS spread had occurred in 5 children . The predominant clinical manifestations of probable cases were fever, cough, and rhinorrhea . With the exception of 1 teenager, none of the children developed respiratory distress or an oxygen requirement, and all made full recoveries . Mild focal alveolar infiltrates were the predominant chest radiograph abnormality . Lymphopenia; neutropenia; thrombocytopenia; and elevated alanine aminotransferase, aspartate aminotransferase, and creatine kinase were present in some cases . Nasopharyngeal swab specimens were negative for the SARS-associated coronavirus by an in-house reverse transcriptase-polymerase chain reaction in all 25 children . CONCLUSIONS: Our results indicate that SARS is a relatively mild and nonspecific respiratory illness in previously healthy young children . The presence of fever in conjunction with a SARS exposure history should prompt one to consider SARS as a possible diagnosis in children irrespective of the presence or absence of respiratory symptoms . Reverse-transcriptase polymerase chain reaction analysis of nasopharyngeal specimens seems to be of little utility for the diagnosis of SARS during the early symptomatic phase of this illness in young children.

Pediatrics . 2003 Oct;112(4):e257.
A young infant with severe acute respiratory syndrome; Sit SC et al.; Severe acute respiratory syndrome (SARS), a new contagious respiratory disease associated with a novel coronavirus, has spread worldwide and become a global health concern after its first outbreak in Guangdong Province of the People's Republic of China in November 2002 . The clinical presentation and the radiologic, hematologic, biochemical, and microbiologic findings of a 56-day-old male infant with SARS are described . Some clinical and laboratory features are similar to those reported in adult and pediatric patients . However, this infant had a more severe clinical course as compared with the older children . This is the youngest patient with symptomatic SARS reported to date.

Diagn Microbiol Infect Dis, 2003 Oct, 47(2), 431 - 3
Pericarditis as primary manifestation of Mycobacterium bovis SSP . caprae infection; Blaas SH et al.; A 76-year-old white male presented with progressive malaise, weight loss and dyspnea at rest . Echocardiography revealed a circular pericardial effusion and global hypokinesia . Pericardiocentesis showed a purulent exudate and microbiologic examination revealed Mycobacterium bovis fully sensitive to isoniazid, streptomycin, ethambutol, rifampin, and pyrazinamide . By spoligotyping the isolate could be further differentiated to M . bovis ssp . caprae . Antimycobacterial therapy was initiated but 3 weeks later the patient's circulation and renal function deteriorated and he died with clinical signs of sepsis despite intensive care treatment . Pericarditis is a rare manifestation of tuberculosis and can be fatal even when diagnosed and treated appropriately . In low incidence countries diagnosis is often delayed and even overlooked.

Hosp Med, 2003 Sep, 64(9), 539 - 42
Helicobacter pylori: a microbiologist's view; Williams C; Helicobacter pylori infection is known to cause a number of gastrointestinal diseases . From the standpoint of a microbiologist there are several important facets to this organism, which include the nature of the organism, it's diagnosis and the effectiveness of eradication therapy.

J Antimicrob Chemother, 2003 Nov, 52(5), 849 - 52 Epub 2003 Sep 30.
Variability of treatment duration for bacteraemia in the critically ill: a multinational survey; Corona A et al.; OBJECTIVES: No definitive evidence is available to inform 'best' antibiotic practice for treating bacteraemia in the critically ill patient, either in terms of duration of therapy, or the use of mono- versus combination therapy . We therefore undertook a large-scale international survey to assess the variability of current practice . METHODS: A questionnaire was sent to membership lists of national and international intensive care societies . RESULTS: Responses from 254 intensive care units in 34 countries revealed a wide variation in antibiotic strategy for all types of bacteraemia, ranging from short course (<or=5 days) therapy with restricted-spectrum antibiotics, to long course (>or=10 days) use of broad-spectrum combinations . Two factors were significantly associated with antibiotic prescribing practice, namely the country of origin (in those with >or=10 responders) and the level of microbiologist and/or infectious diseases specialist input . The greater the specialist input, the shorter the duration of therapy (P < 0.0001) . CONCLUSIONS: The wide variability in antibiotic prescribing patterns suggests an urgent need to produce high-quality evidence to identify optimal antibiotic prescribing policies for bacteraemia in the critically ill patient.

Emerg Infect Dis, 2003 Sep, 9(9), 1163 - 7
Microbiologic characteristics, serologic responses, and clinical manifestations in severe acute respiratory syndrome, Taiwan; Hsueh PR et al.; The genome of one Taiwanese severe acute respiratory syndrome-associated coronavirus (SARS-CoV) strain (TW1) was 29,729 nt in length . Viral RNA may persist for some time in patients who seroconvert, and some patients may lack an antibody response (immunoglobulin G) to SARS-CoV <21 days after illness onset . An upsurge of antibody response was associated with the aggravation of respiratory failure.

Expert Opin Investig Drugs, 2003 Oct, 12(10), 1615 - 20
The Lyme disease controversies continue; Pavia CS; Within both the biomedical and lay community, Lyme disease continues to evoke considerable disagreements, especially regarding its diagnosis, patient management and treatment . A recent article of mine, published in this journal, dealing primarily with therapy for Lyme disease, subsequently led to a highly critical commentary on some of its content . Too often, however, the authors of these criticisms came to premature conclusions based mostly on little or no published data . Instead, they relied almost exclusively on highly speculative theories and on citing the uncorroborated published or unpublished findings and anecdotal reports of others in an attempt to substantiate their weak claims . In this reply account of mine, I respond by providing compelling evidence, derived primarily from the extensive peer-reviewed published literature, that runs counter to most of their key critical concerns . Another goal here is to further clarify certain inconsistencies, related to the epidemiology and microbiology of Lyme disease and its causative agent, including the dubious 'cyst forms', that have an important bearing on therapy.

J Microsc, 2003 Oct, 212(Pt 1), 3 - 12
A rapid microbiopsy system to improve the preservation of biological samples prior to high-pressure freezing; Vanhecke D et al.; A microbiopsy system for fast excision and transfer of biological specimens from donor to high-pressure freezer was developed . With a modified, commercially available, Promag 1.2 biopsy gun, tissue samples can be excised with a size small enough (0.6 mm x 1.2 mm x 0.3 mm) to be easily transferred into a newly designed specimen platelet . A self-made transfer unit allows fast transfer of the specimen from the needle into the specimen platelet . The platelet is then fixed in a commercially available specimen holder of a high-pressure freezing machine (EM PACT, Leica Microsystems, Vienna, Austria) and frozen therein . The time required by a well-instructed (but not experienced) person to execute all steps is in the range of half a minute . This period is considered short enough to maintain the excised tissue pieces close to their native state . We show that a range of animal tissues (liver, brain, kidney and muscle) are well preserved . To prove the quality of freezing achieved with the system, we show vitrified ivy leaves high-pressure frozen in the new specimen platelet.

Croat Med J, 2003 Oct, 44(5), 626 - 9
Serologically detected "new" tick-borne zoonoses in eastern Croatia; Topolovec J et al.; AIM: To establish serologically a contact with causative agents of human monocytic and granulocytic ehrlichiosis, human babesiosis, recently detected rickettsioses, and Lyme disease in individuals with a history of tick bite from three counties in eastern Croatia . METHODS: Seroepidemiologic testing was performed in 102 subjects with a history of tick bite, who either requested examination for a tick bite or were suspected of having tick-borne disease . The study was carried out during the 1998-1999 period in the area of the Vukovar-Srijem, Osijek-Baranya, and Brod-Posavina counties . Serum analysis was performed by indirect immunofluorescence assay for the detection of antibodies to causative agents of human monocytic and granulocytic ehrlichiosis (Ehrlichia chaffeensis and human granulocytic ehrlichiosis agent), human babesiosis (Babesia divergens), and rickettsiosis . Enzyme-linked immunosorbent assay was used for the detection of antibodies to the spirochete Borrelia burgdorferi, the cause of Lyme disease . The assays were performed at the Department of Microbiology, Osijek Public Health Institute in Osijek, and their results were confirmed at the Department of Microbiology and Immunology, School of Medicine in Ljubljana, Slovenia . RESULTS: Ehrlichia chaffensis antibodies were detected in 5 sera, and antibodies to the causative agent of human granulocytic ehrlichiosis in 7 sera . A low titer of antibodies to the etiologic agent of babesiosis (Babesia microti) was detected only in a single serum . Eight sera that were positive for rickettsial antibodies contained rather high titers of antibodies against Rickettsia conorii, the agent of Mediterranean fever, and Rickettsia rickettsii, the agent of Rocky Mountain spotted fever . In six out of these 8 sera, antibodies to Rickettsia typhi, the cause of murine typhus, were detected possibly as a cross-reaction with some "newly detected" rickettsia circulating in this part of Europe, most likely Rickettsia slovaca . Positive titer of antibodies to Borrelia burgdorferi was detected in 15 sera . CONCLUSION: The agents of human monocytic and granulocytic ehrlichiosis and of possibly newly detected rickettsiae were indirectly demonstrated to circulate in eastern parts of Croatia . The results obtained by IFA failed to provide definite evidence for the circulation of the human babesiosis agent, because the IFA used in our study detected Babesia microti, which prevails in the USA, but not Babesia divergens, which is the predominant cause of the disease in Europe . Serologic evidence for Borrelia burgdorferi infection was demonstrated in 80% of the subjects suspected of having the skin manifestation of Lyme disease.

Br J Dermatol, 2003 Sep, 149(3), 620 - 3
Surveillance of nosocomial infections in dermatology patients in a German university hospital; Dettenkofer M et al.; BACKGROUND: Nosocomial infections (NIs) are a growing problem in healthcare today . Thus, surveillance of NIs is an important aspect of modern infection control, which aims to improve the quality of care . OBJECTIVES: To identify overall and site-specific NI rates in dermatology patients in a German university hospital . METHODS: In a prospective study, 1450 patients were surveyed for NIs according to criteria laid down by the Centers for Disease Control and Prevention . Case records were reviewed twice a week, microbiology reports were assessed and the ward staff was consulted . RESULTS: Altogether, 37 NIs were identified in 35 patients, of whom two had two NIs . The overall incidence was 2.5 NIs per 100 patients, and the incidence density was 1.9 NIs per 1000 patient days . Twenty-one patients developed superficial surgical site infections (SSIs) . Thirteen of the 21 SSIs occurred after surgical removal of basal cell carcinoma (BCC; 172 in total) . This represents an infection rate of 7.6% after surgery for BCC . CONCLUSIONS: Our data suggest that routine surveillance in dermatological wards is not accorded a high priority . However, surveillance of SSIs, especially following surgery for BCC, may be indicated.

Expert Rev Mol Diagn, 2003 Sep, 3(5), 649 - 55
PNA FISH: an intelligent stain for rapid diagnosis of infectious diseases; Stender H; Fluorescence in situ hybridization using peptide nucleic acid probes (PNA FISH) is a novel diagnostic technique combining the simplicity of traditional staining procedures with the unique performance of PNA probes to provide rapid and accurate diagnosis of infectious diseases; a feature that makes PNA FISH well suited for routine application and enables clinical microbiology laboratories to report important information for patient therapy within a time frame not possible using classic biochemical methods . Having transitioned from an academic curiosity into an advanced diagnostic tool, PNA probes are now debuting on the infectious disease stage, representing the new generation of therapy-directing diagnostics.

Afr J Med Med Sci, 2001 Dec, 30(4), 333 - 5
Hepatitis B surface antigen (HbsAg) in the sera of medical, nursing and microbiology students in Ibadan, Nigeria; Odemuyiwa SO et al.; A total of 331 serum samples collected from medical students, student nurses, microbiology students, and patients presenting with Pyrexia of Unknown Origin (PUO) were tested for the presence of Hepatitis B surface Antigen (HbsAg) . While only seven (14.0%) of 50 microbiology students (mean age 24.0 years) tested positive for HbsAg, six (6.7%) of 89 student nurses (mean age 21.6 years) and 13 (13.5%) of 95 medical students (mean age 24.3 years) in the clinical phase of their study were found to have HbsAg in their sera . Also, 10 (10.3%) of 97 patients with PUO (mean age 25.4 years), a group of patients from whom medical personnel are most likely to often collect blood for laboratory studies, were found to have HbsAg in their sera . No significant difference was found in the prevalence of HbsAg among the different groups examined in this study (P>0.05) . The result of the study thus shows that medical and nursing students, unlike what is known for practising nurses, physicians and surgeons are not at a higher risk of HBV transmission than students of botany and microbiology . Likewise, patients with PUO do not constitute a group that is more likely to transmit HBV to medical personnel than other groups of patients . Vaccination against hepatitis B virus during the early period of medical and nursing training may therefore go a long way to reduce the high prevalence of hepatitis B virus infection previously reported among practising health personnel in Nigeria.

Curationis, 2003 May, 26(1), 57 - 61
Integration of study material in the problem-based learning method; Becker S et al.; Throughout the world educators are starting to realise that integrated curricula reflect the "real world" . Students learn how to integrate theory appropriately with practice and it stimulates higher order thinking skills . Problem-based learning (PBL) was used as a learning strategy in an integrated undergraduate programme . A quantitative non-experimental design, a survey, was used to determine how problem-based learning influences integration of learning by the students . Using a purposive sampling method all the first-year nursing students (43) at the University of the Free State participating in the programme were included in the study . Data was collected by means of a student perception questionnaire (SPQ) . PBL does contribute to an integration of learning . All the students agreed that community problems were brought to the classroom and that theory and practice were complementary . Students also indicated that PBL enhanced gathering of information from various sources and a holistic view of a situation . Problem-based learning also increased the ability to consider problems from various viewpoints and taught students clinical reasoning . Certain aspects of integration still seem to be a problem as 42% of the students indicated that subjects were not related, 21% that subjects were not integrated and 37% that scenarios were not realistic . This might be attributed to the fact that only the nursing subjects were taught by means of PBL . It is recommended that scenarios be revised to make them more realistic and that traditional subjects such as Anatomy, Microbiology and Chemical Science be integrated with nursing subjects by means of problem-solving themes, to form a meaningful core curriculum.

J Prosthodont, 2003 Sep, 12(3), 187 - 91
The effect of a denture adhesive on the colonization of Candida species in vivo; Kim E et al.; PURPOSE: The purpose of this study was to evaluate the effect of a single denture adhesive on oral quantities of Candida species in vivo by determination of absolute and proportional counts of Candida species on dentures and in saliva of individuals who used this denture adhesive for a period of 14 days . MATERIALS AND METHODS: Samples were collected from saliva and maxillary dentures of 12 patients who wore existing dentures without adhesives for 2 weeks, then wore dentures with adhesive (Poly Grip Free; Glaxo Smith Kline, U.K.) for 2 weeks . Periodically, maxillary dentures were sampled by adding saline to the intaglio surface, dispersing by sonication and removing aliquots for culturing . These aliquots and saliva were diluted and plated in duplicate on bismuth sulfite, glycerine, and glucose yeast agar (Biggy) for recovery of Candida and on trypticase soy agar for total viable counts (TVC) of the microbiota . After 72 hours of incubation at 37 degrees C in air with 10% CO(2), colony-forming units were enumerated . In each individual, absolute counts of Candida and TVC, and proportional counts of Candida relative to TVC were compared and statistically evaluated during the periods of no adhesive use (control) and adhesive use (test) . RESULTS: There were no statistically significant differences between the test and control periods for recovery of total Candida or TVC in saliva or on dentures, or the percent of Candida relative to TVC in saliva or on dentures . CONCLUSION: Within the limitations of this study, the data suggested that the denture adhesive tested did not significantly alter the denture microbiota during the 14-day trial period.

Mol Microbiol, 2003 Oct, 50(1), 153 - 66
Negative feedback regulation of dnaK, clpB and lon expression by the DnaK chaperone machine in Streptomyces coelicolor, identified by transcriptome and in vivo DnaK-depletion analysis; Bucca G et al.; The dnaK operon of Streptomyces coelicolor encodes the DnaK chaperone machine and the negative autoregulator HspR, which confers repression of the operon by binding to several inverted repeat sequences in the promoter region, dnaKp . Previous in vitro studies demonstrated that DnaK forms a specific complex with HspR bound to its operator sequences in dnaKp, and a model was proposed in which DnaK functions as a corepressor of the dnaK operon (Bucca, G., Brassington, A., Schonfeld, H.J., and Smith, C.P . (2000) Mol Microbiol 38: 1093-1103) . Here we report in vivo DnaK depletion experiments which demonstrate that DnaK is a negative regulator of the dnaK operon . Cellular depletion of the DnaK chaperone leads to high-level transcription from dnaKp at the normal growth temperature . DNA microarray-based analysis of gene expression in wild-type and hspR-disruption mutant strains has identified a core cluster of genes regulated by HspR: the dnaK and clpB-SCO3660 operons and lon . These three transcription units are considered to be the direct targets of HspR . Significantly, analysis of the entire genome sequence revealed that the promoter regions of dnaK, clpB and lon are the only sequences that contain the HspR consensus binding sequence 5'-TTGAGY-N7-ACTCAA . S1 nuclease mapping confirmed that transcription of both clpB and lon is substantially enhanced at ambient temperature in strains depleted of DnaK, providing further evidence that these genes are members of the DnaK-HspR regulon . From transcriptome analysis, 17 genes were shown to be upregulated more than twofold in an hspR disruption mutant . This included the seven genes encoded by the dnaK, clpB and lon transcription units . Significantly, the other 10 genes are not heat-shock inducible in the wild type and their upregulation in the hspR mutant is considered to be an indirect consequence of enhanced synthesis of one or more components of the HspR regulon (the DnaK chaperone machine, ClpB and Lon protease).

Ann R Australas Coll Dent Surg, 2002 Oct, 16, 141 - 4
Oral malodour and its clinical management; Yeung S; Oral malodour or halitosis is one of the most common complaints by dental patients . It often creates serious personal and social embarrassment for the afflicted individual . Dental health care workers can play a pivotal role in helping these patients by eliminating dental diseases and improving oral hygiene . This paper reviews the current understanding in the microbiology and aetiology of oral malodour and presents a clinical management strategy, which is both rational and effective.

Pathology, 2003 Jun, 35(3), 240 - 3
Reappraisal of C-reactive protein as a screening tool for neonatal sepsis; Garland SM et al.; AIMS: To assess the usefulness of C-reactive protein (CRP) {either alone or in combination with a full blood examination (FBE) and microbiology of gastric aspirate} in predicting the diagnosis of neonatal sepsis compared with routinely available markers of infection . METHODS: A prospective study of quantitative CRP, FBE, gastric aspirate and surface swab microscopy and culture in predicting neonatal sepsis in a level III perinatal referral hospital was performed . RESULTS: Of 301 episodes evaluated over a 5-month period, there was a 5% rate of major (culture proven) sepsis (3% early onset, <72 hours) and 10% rate of modified sepsis . No single test alone was sufficiently reliable as an indicator of infection to be a satisfactory screening tool for early onset sepsis {CRP sensitivity 67%, negative predictive value (NPV) 86%, FBE sensitivity 63%, NPV 80%, gastric aspirate sensitivity 57%, NPV 83%} . CONCLUSION: The three-test combination had a sensitivity of 97%, NPV 98% and likelihood ratio of 49, thus providing a useful diagnostic tool.

Environ Toxicol, 2003 Oct, 18(5), 347 - 52
Cyst-based toxicity tests XV--application of ostracod solid-phase microbiotest for toxicity monitoring of contaminated soils; Chial B et al.; A new "culture/maintenance-free" microbiotest has recently been developed for "direct-contact" toxicity determination of contaminated sediments . The 6-day Ostracodtoxkit F makes use of the neonates of the ostracod crustacean Heterocypris incongruens hatched from dormant eggs (cysts) . The new low-cost assay has already been applied in three studies on river sediment from Flanders (Belgium) and from Canada and was found to perform comparably to the 10-day Hyalella azteca and Chironomus riparius tests in detecting and quantifying sediment toxicity . Taking into account that sediments are in fact underwater soils, the ostracod microbiotest was tentatively applied on 15 contaminated soils from Flanders (Belgium), and its sensitivity (based on mortality as the test criterion) was evaluated in comparison to the 28-day reproduction inhibition assay with the springtail Folsomia candida . The results revealed that the ostracod test species was as sensitive as or, in several samples, even more sensitive than the springtails . Leachate experiments on the same soil samples also showed that in most cases ostracod mortality was a result of the (nonsoluble) toxicants bound to the solid-phase particles, rather than of those that had dissolved in the water phase . Providing confirmation of these first findings through additional studies, the new culture/maintenance-free ostracod microbiotest seems to have good potential as a low-cost and user-friendly tool for routine toxicity monitoring of contaminated soils .

Curr Opin Otolaryngol Head Neck Surg, 2003 Oct, 11(5), 316 - 23
Skull base osteomyelitis secondary to malignant otitis externa; Sreepada GS et al.; PURPOSE OF REVIEW: Skull base osteomyelitis secondary to malignant otitis externa was first described in 1959 . Since then, advances have been made in the diagnosis, treatment, and clinical outcomes of this condition . RECENT FINDINGS: This review discusses the pathophysiology and microbiology of malignant otitis externa . The review highlights the sometimes subtle presenting symptoms and recent advances in imaging and their practical application to diagnosing and monitoring the disease . Therapy for malignant otitis externa has changed since this entity was first described; this article reviews the medical, surgical, and adjuvant therapies and the relevant controversies . SUMMARY: The review discusses the history, pathogenesis, diagnosis, and treatment of skull base osteomyelitis in the context of malignant otitis externa with particular emphasis on HIV, children, and other immunodeficient states.

J Microbiol Methods, 2003 Oct, 55(1), 257 - 72
A new and sensitive Co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water; Caruso P et al.; Three primers from 16S rRNA were successfully assayed simultaneously in one reaction for sensitive detection of Ralstonia solanacearum in watercourses . The protocol is a modification of the Co-operational polymerase chain reaction (Co-PCR), which allows the simultaneous and co-operational action of the primers . It specifically amplified R . solanacearum strains belonging to biovars 1, 2 and 4 . No products were obtained from any of the 162 unidentified isolates from river water . The sensitivity of the assay was <1 cfu/ml as determined by analysis of heat-treated water samples spiked with R . solanacearum, also containing indigenous microbiota up to 10(5) cfu/ml . The developed Co-PCR assay was more sensitive than other standard PCR assays in the analysis of 51 Spanish environmental water samples . Namely 31.3% of the samples were positive using the newly developed assay, whereas 13.7% or less positive samples were found with the other protocols . The Co-PCR improves the detection sensitivity of R . solanacearum and provides an important tool for its routine detection from environmental water samples and for epidemiological studies.

Clin Infect Dis, 2003 Oct 1, 37 Suppl 3, S225 - 64
Allergic bronchopulmonary aspergillosis in cystic fibrosis--state of the art: Cystic Fibrosis Foundation Consensus Conference; Stevens DA et al.; Because of the difficulties of recognizing allergic bronchopulmonary aspergillosis (ABPA) in the context of cystic fibrosis (because of overlapping clinical, radiographic, microbiologic, and immunologic features), advances in our understanding of the pathogenesis of allergic aspergillosis, new possibilities in therapy, and the need for agreed-upon definitions, an international consensus conference was convened . Areas addressed included fungal biology, immunopathogenesis, insights from animal models, diagnostic criteria, epidemiology, the use of new immunologic and genetic techniques in diagnosis, imaging modalities, pharmacology, and treatment approaches . Evidence from the existing literature was graded, and the consensus views were synthesized into this document and recirculated for affirmation . Virulence factors in Aspergillus that could aggravate these diseases, and particularly immunogenetic factors that could predispose persons to ABPA, were identified . New information has come from transgenic animals and recombinant fungal and host molecules . Diagnostic criteria that could provide a framework for monitoring were adopted, and helpful imaging features were identified . New possibilities in therapy produced plans for managing diverse clinical presentations.

Drug News Perspect, 2003 Jul-Aug, 16(6), 399 - 401
Highlights from the 13th European Congress of Clinical Microbiology and Infectious Diseases in Glasgow, Scotland, May 10-13, 2003 . The complex world of infectious diseases; Jack D; At the 13th European Congress of Clinical Microbiology and Infectious Diseases, held in Glasgow, Scotland, May 10-13, 2003, the latest developments in clinical microbiology and the treatment of infectious diseases were presented alongside recent progress on molecular aspects of diagnosis and emerging patterns of infection . Around 5,000 delegates from more than 80 countries attended the congress, which saw the presentation of more than 400 oral communications and 1,700 posters . In addition to a historical session looking at Scotland's own contribution to the control of infectious diseases, the meeting involved up to six parallel sessions a day, looking at all the major aspects of infectious diseases, treatment, surveillance, epidemiology and drug pharmacodynamics and pharmacokinetics . The organizers also organized a Late Breaker symposium on severe acute respiratory syndrome . The topics likely to be of most interest to Drug News and Perspectives readers are described here . (c) 2003 Prous Science . All rights reserved.

Int J Tuberc Lung Dis, 2003 Sep, 7(9), 879 - 85
Investigation of biological risk in mycobacteriology laboratories: a multicentre study; Vaquero M et al.; SETTING: Microbiology laboratories constitute a special working environment that may pose a risk of infectious disease to persons in or near them . OBJECTIVE: To assess whether clinical mycobacteriology laboratories in Spanish hospitals comply with occupational health and safety guidelines, and if laboratory workers are protected against the risk of exposure to biological agents in the workplace . DESIGN: A transversal epidemiological study was carried out in 26 hospitals from October to December 2000 by means of a standardised survey questionnaire to evaluate the workers, workload, training and information and safety practices in mycobacteria laboratories . RESULTS: Less than half of employees receive periodic information on the health hazards involved in their work . More than a third of the workers in mycobacteriology laboratories stated that the laboratories lack effective air filtering systems and more than half stated that negative pressurisation was not maintained in the work area . Biosafety masks are used by little more than half of the workers . The most frequently used Biological Safety Cabinets are Class IIB . CONCLUSION: Our findings demonstrate that clinical microbiology laboratories in Spanish hospitals comply poorly with the prevention measures outlined in the applicable directives and regulations.

Chest, 2003 Sep, 124(3), 909 - 14
Combined use of the polymerase chain reaction and detection of adenosine deaminase activity on pleural fluid improves the rate of diagnosis of pleural tuberculosis; Lima DM et al.; STUDY OBJECTIVES: Evaluation of the combined use of polymerase chain reaction (PCR) and adenosine deaminase (ADA) activity on the diagnosis of pleural tuberculosis (pTB) in a region of high prevalence of tuberculosis . PATIENTS: PCR and determination of ADA activity were performed on the pleural fluid of every patient presenting with pleural effusion suspected to be associated with tuberculosis . The case definition of pTB involved parameters including the combination of clinical and radiologic findings; biochemical, microbiologic, and cytologic examination of the pleural fluid; and the histopathologic findings of pleural fragments obtained by biopsy . The diagnosis of pTB was confirmed in any patient presenting with positive culture findings of Mycobacterium tuberculosis, either on the pleural fluid or other biological material, or the presence of histopathologic findings suggestive of pTB on pleural biopsy, and also, in the absence of negative laboratory results, those patients with clinical improvement after empirical treatment . RESULTS: We studied 45 patients with pleural effusion . Of these, 16 patients met the diagnosis of pTB by our broad case definition . PCR findings were positive in six patients . The reaction was also positive in a patient whose diagnosis of tuberculosis could not be confirmed . ADA activity was considered positive in 11 patients with pTB . The combined use of PCR and ADA activity confirmed pTB in 14 patients; however, when analyzed in combination with the conventional methods, diagnosis of pTB was achieved in all 16 patients . CONCLUSION: Our results show that, even in a highly endemic area, neither PCR nor ADA activity should be relied on as a single test that substitutes for the diagnostic methods already available, but rather they should be used as an extra tool in the diagnosis of pTB . The combined analysis of PCR and ADA activity, however, is a very useful diagnostic approach to achieve a more rapid and precise diagnosis in the cases of pTB.

Proc R Soc Lond B Biol Sci, 2003 Aug 22, 270(1525), 1703 - 12
Host-based divergence in populations of the pea aphid: insights from nuclear markers and the prevalence of facultative symbionts; Simon JC et al.; In North America, the pea aphid Acyrthosiphon pisum encompasses ecologically and genetically distinct host races that offer an ideal biological system for studies on sympatric speciation . In addition to its obligate symbiont Buchnera, pea aphids harbour several facultative and phylogenetically distant symbionts . We explored the relationships between host races of A . pisum and their symbiotic microbiota to gain insights into the historical process of ecological specialization and symbiotic acquisition in this aphid . We used allozyme and microsatellite markers to analyse the extent of genetic differentiation between populations of A . pisum on pea, alfalfa and clover in France . In parallel, we examined: (i) the distribution of four facultative symbionts; and (ii) the genetic variation in the Buchnera genome across host-associated populations of A . pisum . Our study clearly demonstrates that populations of A . pisum on pea, clover and alfalfa in France are genetically divergent, which indicates that they constitute distinct host races . We also found a very strong association between host races of A . pisum and their symbiotic microbiota . We stress the need for phylogeographic studies to shed light on the process of host-race formation and acquisition of facultative symbionts in A . pisum . We also question the effects of these symbionts on aphid host fitness, including their role in adaptation to a host plant.

Analyst, 2003 Aug, 128(8), 995 - 7
Targeting cancer treatment: the challenge of anatomical pathology to the analytical chemist; Demetrick DJ; Anatomical pathology involves the scrutiny of specimens of human tissue for the diagnosis of disease . Included in this field is cytopathology which addresses the evaluation of small groups of cells . Doug Demetrick of the Department of Pathology of the University of Calgary argues that analytical chemistry has had minimal impact in such medical diagnostic endeavours, unlike the situation for hematology, microbiology and molecular genetics . Furthermore, it is stressed that new strategies are required for sample handling, increasing the cost effectiveness of instrumentation, and decreasing the subjectiveness of data processing . In tandem with drug development, it is more important to predict the response of disease to treatment than to concentrate on new methods for diagnosis.

Ann Clin Lab Sci, 2003 Summer, 33(3), 329 - 33
Case report: Nocardia asteroides mycetoma; Lum CA et al.; Primary cutaneous infections with Nocardia asteroides are rare and have been reported in immunocompromised patients . Herein, we report a case of primary cutaneous Nocardia asteroides mycetoma of the skin in an immunocompetent individual . The infection was treated successfully with trimethoprim-sulfamethoxazole . Because a prolonged incubation time is required for the cultures and since additional biochemical tests are necessary for identification of this species, the clinician should alert the microbiology laboratory when such an infection is suspected clinically.

Gastroenterology, 2003 Sep, 125(3), 755 - 64
Infection and the progression of hepatic encephalopathy in acute liver failure; Vaquero J et al.; BACKGROUND & AIMS: Progression of hepatic encephalopathy (HE) is a major determinant of outcome in acute liver failure (ALF) . Our aim was to identify predictive factors of worsening HE, including the relation of encephalopathy with the systemic inflammatory response (SIRS) and infection . METHODS: We included 227 consecutive patients with stage I-II HE prospectively enrolled in the U.S . Acute Liver Failure Study . Univariate and multivariate analysis of 27 variables at admission were performed separately for acetaminophen (n = 96) and nonacetaminophen (n = 131) etiologies . RESULTS: On multivariate analysis, acquisition of infection during stage I-II HE (P < 0.01), increased leukocyte levels at admission (P < 0.01), and decreased platelet count (P < 0.05) were predictive factors of worsening HE in the acetaminophen group . By contrast, only increased pulse rate (P < 0.05) and AST levels (P < 0.05) at admission were predictors in nonacetaminophen patients . In patients who progressed to deep HE, the first confirmed infection preceded progression in 15 of 19 acetaminophen patients compared with 12 of 23 nonacetaminophen patients . In patients who did not demonstrate positive microbiologic cultures, a higher number of components of SIRS at admission was associated with more frequent worsening of HE (25% vs . 35% vs . 50% for 0, 1, and >or=2 components of SIRS, P < 0.05) . CONCLUSIONA: This prospective evaluation points to infection and/or the resulting systemic inflammatory response as important factors contributing to worsening HE in ALF, mainly in patients with acetaminophen- induced ALF . The use of prophylactic antibiotics in these patients and the mechanisms by which infection triggers hepatic encephalopathy require further investigation.

J Clin Neurosci, 2003 Sep, 10(5), 596 - 601
Effects of immunonutrition on intestinal mucosal apoptosis, mucosal atrophy, and bacterial translocation in head injured rats; Ulusoy H et al.; BACKGROUND: Immunonutrition has been demonstrated to modulate gut function, reduce infectious complications, hospital stay, and ventilator days in the critical patients.AIM OF THE STUDY: We assessed the effect of immunonutrition for the prevention of intestinal mucosal atrophy, apoptosis, and bacterial translocation in head injured rats.METHODS: Thirty five rats were randomised into 5 groups . Following moderate closed head injury, in Group 1; Standard Enteral Nutrition, Group 2; Immunonutrition, Group 3; TPN, Group 4; pe . saline were applied . Group 5 was control group (chow-fed) . The rats were sacrificed and segments of the ileum were removed for histologic examination, and samples of tissues taken for microbiologic evaluation.RESULTS: Both intestinal apoptosis and mucosal atrophy were significantly lower in Group 2 and Group 5 (p<0.008) . Bacterial translocation was significantly lower in Group 2 than Group 1 (p<0.008).CONCLUSION: The enteral immunonutrition prevents intestinal barrier function in brain injured rats.

Rev Biol Trop, 2002 Sep-Dec, 50(3-4), 941 - 9
{Biomedical research in Revista de Biologia Tropical}; Gutierrez JM; The contributions published in Revista de Biologia Tropical in the area of Biomedical Sciences are reviewed in terms of number of contributions and scope of research subjects . Biomedical Sciences, particularly Parasitology and Microbiology, constituted the predominant subject in the Revista during the first decade, reflecting the intense research environment at the School of Microbiology of the University of Costa Rica and at Hospital San Juan de Dios . The relative weight of Biomedicine in the following decades diminished, due to the outstanding increment in publications in Biological Sciences; however, the absolute number of contributions in Biomedical Sciences remained constant throughout the last decades, with around 80 contributions per decade . In spite of the predominance of Parasitology as the main biomedical subject, the last decades have witnessed the emergence of new areas of interest in the Revista, such as Pharmacology of natural products, Toxinology, especially related to snake venoms, and Human Genetics . This retrospective analysis evidences that Biomedical Sciences, particularly those related to Tropical Medicine, were a fundamental component during the first years of Revista de Biologia Tropical, and have maintained a significant presence in the scientific output of this journal, the most relevant scientific publication in biological sciences in Central America.

Arch Pathol Lab Med, 2003 Sep, 127(9), 1089 - 97
Genomics, transcriptomics, proteomics, and numbers; Kiechle FL et al.; OBJECTIVE: To review the advances in clinically useful molecular biologic techniques and to identify their applications in clinical practice, as presented at the 11th Annual William Beaumont Hospital DNA Symposium . DATA SOURCES: The 8 manuscripts submitted were reviewed, and their major findings were compared with literature on the same or related topics . STUDY SELECTION: Manuscripts address the use of molecular techniques in microbiology to evaluate infectious disease and epidemiology; molecular microbiology methods, including rapid-cycle real-time polymerase chain reaction; peroxisome proliferator-activated receptor gamma as a potential therapeutic target in inflammatory bowel disease or colon cancer; the effect of nonapoptotic doses of the bisbenizamide dye Hoechst 33342 on luciferase expression in plasmid-transfected BC3H-1 myocytes; the routine use of cystic fibrosis screening and its challenges; and the use of flow cytometry and/or chromosomal translocation in the diagnostic evaluation of hematopoietic malignancies . DATA SYNTHESIS: Three current issues related to the use of molecular tests in clinical laboratories are (1) the restriction on introducing new tests secondary to existing patents or licenses; (2) the preanalytic variables for the different specimen types currently in use, including whole blood, plasma, serum, fresh or frozen tissues, and free-circulating DNA; and (3) the interpretation of studies evaluating the association of complex diseases with a single mutation or single-nucleotide polymorphism . Molecular methods have had a major impact on infectious disease through the rapid identification of organisms, the evaluation of outbreaks, and the characterization of drug resistance when compared with standard culture techniques . The activation of peroxisome proliferator-activated receptor gamma stimulated by thiazolidinedione is useful in the treatment of type II diabetes mellitus and may have value in preventing inflammatory bowel disease or colon cancer . Hoechst 33342 binding to adenine-thymine-rich regions in the minor groove of DNA is a fluorescent stain for DNA and initiates apoptosis at >10 microg/mL . Lower doses of Hoechst 33342 promote luciferase expression by a mechanism that may involve binding to cryptic promoters facilitated by dye-associated misalignment of the tertiary structure of DNA . The routine use of cystic fibrosis screening is complicated by the more than 1000 mutations associated with the disease . The use of 4-color flow cytometry and the detection of chromosomal translocation are both invaluable aids in establishing the diagnosis of lymphoid or myeloid hematopoietic malignancies . CONCLUSIONS: The current postgenomic era will continue to emphasize the use of microarrays and database software for genomic, transcriptomic, and proteomic screening in the search for useful clinical assays . The number of molecular pathologic techniques will expand as additional disease-associated mutations are defined.

Arch Pathol Lab Med, 2003 Sep, 127(9), 1098 - 105
The role of molecular biology and nucleic Acid technology in the study of human infection and epidemiology; Zaidi N et al.; OBJECTIVE: In this review, we will focus on recent molecular typing methods that can be applied to different pathogens and assess their values and limitations . BACKGROUND: Resistant subgroups within a species of pathogenic organisms often emerge as dominant strains under the selective pressures of antibiotics, vaccinations, and new health care practices . The emergence of resistant organisms has added to the burden and cost of health care-related infections . Pathogens derived from a common ancestry are often difficult to distinguish by conventional methods, and the practice of clinical microbiology and infectious disease epidemiology must adapt to this problem . DISCUSSION: Conventional strain typing methods provide a limited means of distinguishing epidemic from endemic or sporadic isolates of pathogens . Nucleic acid-based methods complement conventional and serologic methods of organism isolation and typing . Often, these genomic methods offer more discrimination and details than the phenotype-based conventional methods . RESULTS AND CONCLUSIONS: Highly sensitive molecular techniques are capable of detecting single base pair substitutions and resolving the mechanism of underlying complex variation.

Arch Pathol Lab Med, 2003 Sep, 127(9), 1106 - 11
Introducing a molecular test into the clinical microbiology laboratory: development, evaluation, and validation; Forbes BA; CONTEXT: In the mid-1980s, the polymerase chain reaction methodology for the amplification of minute amounts of target DNA was successfully developed and then introduced into clinical use; such technology has led to a revolution in diagnostic testing . Despite enormous advances in the detection of infectious agents by amplification methods, there are also limitations that must be addressed . OBJECTIVE: To highlight the pertinent steps and issues associated with the introduction of an amplification assay into a clinical microbiology laboratory as well as the subsequent ongoing activities following its introduction into routine laboratory use . DATA SOURCES: Data were obtained from literature searches from 1990 through September 2002 using the subject headings "polymerase chain reaction," "molecular assays," and "amplification" as well as publications of the National Committee for Clinical Laboratory Standards . DATA EXTRACTION AND SYNTHESIS: Using the findings obtained from these studies and publications, the process of introducing a molecular assay into the clinical microbiology laboratory was broken down into 4 major components: (1) initial phase of assay development, (2) polymerase chain reaction assay verification in which analytic sensitivity and specificity is determined, (3) assay validation to determine clinical sensitivity and specificity, and (4) interpretation of results and ongoing, required activities . The approach, as well as the advantages and limitations involved in each step of the process, was highlighted and discussed within the context of the published literature . CONCLUSIONS: The application of molecular testing methods in the clinical laboratory has dramatically improved our ability to diagnose infectious diseases . However, the clinical usefulness of molecular testing will only be maximized to its fullest benefit by appropriate and careful studies correlating clinical findings with assay results.

Ann Chir, 2003 Jul, 128(6), 368 - 72
{Role of microbiopsy in diagnostic and therapeutic approach of mammary suspect microcalcification}; Benoit L et al.; PURPOSE: To determine the sensibility, the specificity, the positive and negative predictive values of microcalcifications detection by core needle biopsy and intra-operative pathologic examination . MATERIALS AND METHODS: A hundred and one patients (between 1998 and 1999) were investigated in this retrospective study . The initial presentation was breast microcalcifications without palpable tumour . The mean age of patients was 55 (34-79) years . Mammography was performed in 3 standard projections . All suspect microcalcifications were recommended for surgical excision . In the others cases, 5 core biopsies were taken of the lesion . Needle guidance was accomplished by means of either dedicated stereotaxic device or ultrasound equipment . All biopsies were performed with a biopsy device fitted with 14 G needles . The mean follow-up period was 3 years . RESULTS: Clinical or surgical follow-up was available in 101 lesions . Only 4 benign lesions did not have surgery . The 97 remaining were subsequently excised . Pathologic study showed cancer in 38 (39%) lesions, carcinoma in-situ in 14 lesions, and benign disease in 45 lesions . There was 1 false-negative biopsy . The specificity and sensitivity of percutaneous biopsy diagnosis were 73,6 and 93,7% respectively . Intra-operative pathologic diagnoses were concordant in 77% of 30 cases . Discordance occurred in 2 cases of atypical hyperplasia with a single false-negative result for a carcinoma that led to an additional surgical procedure . Positive and negative predictive values of intra-operative pathologic diagnosis were 82 and 100% respectively . CONCLUSION: Needle biopsy findings are accurate and allow definitive therapeutic surgery, including mastectomy.

South Med J, 2003 Jun, 96(6), 621 - 3
Pulmonic valve endocarditis; Tariq M et al.; Pulmonic valve endocarditis is an extremely rare infection that shares epidemiologic, clinical, radiologic, microbiologic, and prognostic features with tricuspid valve endocarditis . We report a case of pulmonic valve infection on a structurally normal heart and review the English-language literature on this subject.

Braz J Med Biol Res, 2003 Sep, 36(9), 1227 - 32 Epub 2003 Aug 19.
Ethanol-induced colitis prevents oral tolerance induction in mice; Andrade MC et al.; The gut mucosa is a major site of contact with antigens from food and microbiota . Usually, these daily contacts with natural antigens do not result in inflammatory reactions; instead they result in a state of systemic hyporesponsiveness named oral tolerance . Inflammatory bowel diseases (IBD) are associated with the breakdown of the immunoregulatory mechanisms that maintain oral tolerance . Several animal models of IBD/colitis are available . In mice, these include targeted disruptions of the genes encoding cytokines, T cell subsets or signaling proteins . Colitis can also be induced by intrarectal administration of chemical substances such as 2,4,6-trinitrobenzene sulfonic acid in 50% ethanol . We report here a novel model of colitis induced by intrarectal administration of 50% ethanol alone . Ethanol-treated mice develop an inflammatory reaction in the colon characterized by an intense inflammatory infiltrate in the mucosa and submucosa of the large intestine . They also present up-regulation of both interferon gamma (IFN-gamma) and interleukin-4 (IL-4) production by cecal lymph node and splenic cells . These results suggest a mixed type of inflammation as the substrate of the colitis . Interestingly, cells from mesenteric lymph nodes of ethanol-treated mice present an increase in IFN-gamma production and a decrease in IL-4 production indicating that the cytokine balance is altered throughout the gut mucosa . Moreover, induction of oral tolerance to ovalbumin is abolished in these animals, strongly suggesting that ethanol-induced colitis interferes with immunoregulatory mechanisms in the intestinal mucosa . This novel model of colitis resembles human IBD . It is easy to reproduce and may help us to understand the mechanisms involved in IBD pathogenesis.

Med Tr Prom Ekol, 2003, (6), 20 - 5
{Laboratory studies at an occupational disease clinic}; Ivanova LA et al.; The authors provide results of hematologic, cytochemical, cytogenetical, microbiologic, cytologic and immunologic studies and describe their role in studying pathogenesis for main occupational diseases of internal organs, nervous system, skin, ENT and other systems . Prospects of clinical laboratory examinations development are using new cellular technologies and novel methods evaluating influence of chemical and physical factors on vital cytoplasmic, nuclear and membrane structures.

Clin Microbiol Infect, 2003 Jul, 9(7), 724 - 6
Informal consultations in infectious diseases and clinical microbiology practice; Leblebicioglu H et al.; Informed consultation between physicians is an important part of medical practice . The aim of the study was to evaluate the nature and frequency of such consultations in infectious diseases and clinical microbiology practice . This study was done in five university hospitals . Twenty-one infectious diseases and clinical microbiology specialists participated in informal ('curbside') consultations . In a written questionnaire, physicians were asked to report the number and nature of the informal consultations (ICs) they were asked to provide . A total three hundred and sixty-two such consultations were carried out during a three-month period . The ICs occurred most frequently in the hospital (82.3%) . Most of the ICs from outside the hospital were by telephone . Most of the ICs (54.4%) were requested by fellows of specialists . 78.7% of the ICs were requested during working hours . 58.8% of consultations took less than 5 min, 18.8% took 6-10 min, 15.2% took 11-20 min, and 7.2% took over 20 min . The four most common reasons for obtaining ICs were to: help to select an appropriate treatment plan (41.4%), help to select an appropriate prophylaxis (19.3%), interpret laboratory data (10.2%), and provide information about antibiotics (10.2%) . 30.1% of ICs resulted in subsequent formal consultation and only four patients (1.1%) were transferred to the consultants' clinics . Informal consultations are a frequent occurrence in the practice of infectious diseases and clinical microbiology (ID&CM) . Physicians use this sort of consultation to select an appropriate treatment plan and obtain medical information . This study confirms the importance of the ID&CM specialists as a resource for medical personnel.

Ann Rheum Dis, 2003 Sep, 62(9), 807 - 11
Normal intestinal microbiota in the aetiopathogenesis of rheumatoid arthritis; Toivanen P; A series of observations have led to the hypothesis that normal intestinal microbiota in patients with rheumatoid arthritis may harbour, for genetic reasons, bacteria with cell walls capable of inducing arthritis . Differences occur between bacterial species, and even between strains of a single species, because some cell walls induce experimental chronic arthritis, whereas some others induce only a transient acute arthritis or no arthritis at all . In susceptible subjects, with continuous seeding of bacterial products from the gut, the synovial inflammation is followed by erosion, exposition of cartilage antigens, and self perpetuating chronic arthritis.

Anal Bioanal Chem, 2003 Oct, 377(3), 434 - 45 Epub 2003 Aug 12.
Biosensors for environmental pollutants and food contaminants; Baeumner AJ; This review article provides an overview of the most recent literature on biosensors for environmental pollutants and food contaminants . Due to the large number of publications, only papers published between 2000 and January 2003 were considered . Also, while not all of the published literature could be reviewed here, over 200 references are cited to provide a good overview of research undertaken in the last two years . Older publications are covered by a number of earlier review articles . This article provides an introduction into the field including specific consideration of the application areas, describes the typical biosensor assay format used, and is subsequently structured according to the biorecognition elements used (i.e., nucleic acids, enzymes, whole cells, tissue and whole organisms, antibodies and receptors, and biomimetic materials) . In addition, a section on microbiosensing systems is provided . Since only very few microbiosensors with applications in environmental and food systems have been published, enabling technology is also covered in this article.

Ther Apher Dial, 2003 Apr, 7(2), 225 - 31
Long-term catheters for apheresis and dialysis with surface treatment with infection resistance and low thrombogenicity; Bambauer R et al.; Infection, thrombosis, and stenosis are among the most frequent complications associated with blood-contacting catheters . These problems are usually related to surface properties of the base catheter material . Surface treatment processes, such as ion implantation and ion beam assisted deposition (IBAD) and microdomain structured surfaces, can be used to mitigate such complications . This study evaluated silver coated and implanted large bore catheters used for extracorporeal detoxification . In a 186 patient prospective study, 225 large bore catheters were inserted into the internal jugular or subclavian veins . Eighty-five surface-treated catheters (Spi-Argent, Spire Corporation, Bedford, MA, USA) and 28 catheters with surface treatment (Spi-Silicone, Spire Corporation) were inserted in 90 patients . One hundred and twelve untreated catheters placed in 96 patients served as controls, After removal, the catheters were cultured for bacterial colonization using standard microbiologic assays . They also were examined using a scanning electron microscope (SEM) . Bacterial colonization was observed in 8% of the treated catheter compared with 46.4% of untreated catheters . The SEM investigations showed all treated catheters to possess low thrombogenicity . Catheters with microdomain structured surfaces showed same results in preliminary observation . The surface treatments of the large bore catheters can be used to improve thrombus and infection resistance of blood contacting catheters.

Ophthal Plast Reconstr Surg, 2003 May, 19(3), 244 - 6
Ringworm causing childhood preseptal cellulitis; Rajalekshmi PS et al.; A 4-year-old girl had redness and swelling of the left upper eyelid of 1 week's duration . She had been treated with topical fusidic add and oral flucloxacillin without resolution . Ophthalmic examination showed a diffuse, erythematous, tender lesion affecting the entire upper eyelid . There was a central area of skin ulceration with marked bleeding and discharge . Also found were small vesicles under the left eyebrow . The child's mother had noticed a scaly rash on her own forearm . Examination under anesthesia was carried out to assess the globe and to obtain samples for microbiology investigation . Microbiology scrapings showed fungal hyphae of the Dermatophyte group . Treatment with Griseofulvin was commenced, and complete resolution of the lesion occurred . To our knowledge there is only one similar case reported in the literature describing ring-worm as a cause of preseptal cellulitis . This case highlights the importance of considering alternative diagnoses when standard antibiotic treatment has failed.

Arch Esp Urol, 2003 Jun, 56(5), 536 - 8
{Pneumopyocystitis in a patient with adult polycystic kidney disease}; Lanzas Prieto JM et al.; OBJECTIVES: We present a case of adult polycystic kidney disease, also known as autosomal dominant polycystic kidney, complicated by infection of the cysts and the formation of gas within them . METHODS/RESULTS: A 59 year old patient diagnosed of adult polycystic kidney disease with chronic renal failure on treatment with haemodialysis, who presented sepsis secondary to infection of the renal cysts . The CT scan demonstrated the presence of gas within the cysts and the microbiology revealed E . coli in one of them . Urgent nephrectomy was performed . A histological specimen of the excised organ is also presented . CONCLUSIONS: Infection of one or more cysts in adult polycystic kidney disease is a rare and serious complication which may require immediate nephrectomy, particularly if gas appears within the cysts.

Ophthalmology, 2003 Aug, 110(8), 1593 - 600
Persistently culture positive acanthamoeba keratitis: in vivo resistance and in vitro sensitivity; Perez-Santonja JJ et al.; PURPOSE: To characterize the risk factors, clinical course, treatment outcome and the association between in vivo resistance and in vitro sensitivity for subjects with persistently culture-positive Acanthamoeba keratitis . DESIGN: Retrospective noncomparative case series . PARTICIPANTS: Eleven subjects with repeatedly positive cultures for Acanthamoeba treated between January 1990 and December 2000, were reviewed . Only subjects with 2 or more positive cultures, availability of the clinical data, and availability of the last Acanthamoeba isolate were included in this study . METHODS: The medical records were analyzed, and the last isolate from each case was tested in vitro for the antiamoebic drugs used clinically: polyhexamethylene biguanide (PHMB), chlorhexidine, propamidine and hexamidine . MAIN OUTCOME MEASURES: Risk factors, the clinical outcome and in vitro cysticidal drug sensitivity assay . RESULTS: Eleven subjects (11/180, 6.1%) had 2 or more positive cultures of whom 8 eyes of 8 subjects (8/180, 4.45%) were included in this study . Seven of eight (87%) subjects were diagnosed over 1 month from onset (late diagnosis) . The most common presenting findings were diffuse stromal infiltrate (5/8, 62.5%), ring infiltrate (5/8, 62.5%), and corneal ulceration (3/8, 37.5%) . The clinical course of the disease in all subjects consisted of recurrent episodes of corneal and scleral inflammation, with a mean duration of 13.4 +/- 9 months . All subjects received PHMB, and 5/8 (62.5%) chlorhexidine too; hexamidine was used in combination in 6/8 (75%), and propamidine in 1/8 (12.5%) . All subjects had topical steroids, and 5/8 (62.5%) systemic immunosuppression . The disease resolved with corneal scarring in 3/8 (37.5%) subjects, corneal (or impending) perforation treated with therapeutic keratoplasty in 4/8 (50%), and enucleation in 1/8 (12.5%) . Final visual acuity was 0.43 +/- 0.37 . In vitro most isolates were resistant to propamidine, hexamidine was cysticidal in high concentrations, and PHMB and chlorhexidine had excellent sensitivity profiles . CONCLUSIONS: In our large series of Acanthamoeba keratitis with a positive microbiologic diagnosis at presentation, nearly 5% developed recurrent episodes of corneal and scleral inflammation with viable Acanthamoeba in the cornea despite prolonged treatment with biguanides and/or diamidines . There was no correlation between in vitro drug sensitivities and the in vivo response for biguanides.

J Appl Microbiol, 2003, 95(3), 612 - 20
Analysis of environmental Escherichia coli isolates for virulence genes using the TaqMan PCR system; Davis KC et al.; AIMS: To assess the presence of virulence genes in environmental and foodborne Escherichia coli isolates using the TaqMan PCR system . METHODS AND RESULTS: Three TaqMan pathogen detection kits called O157:H7, StxI and StxII were used to investigate the presence of virulence genes in Escherichia coli isolates . All 54 foodborne E . coli O157:H7 isolates showed expected results using these kits . Ninety (15%) of 604 environmental isolates gave positive amplification with an O157:H7-specific kit . TaqMan PCR amplification products from these 90 isolates were analysed by agarose gel electrophoresis, and 90% (81 of 90) of the environmental samples contained the expected PCR product . Sixty-six of these 90 were chosen for serotyping tests and only 35% (23 of 66) showed agglutination with both anti-O157 and anti-H7 antibodies . Further ribotyping of 16 sero-positive isolates in an automated Riboprinter did not identify these to be O157:H7 . Multiplex PCR with primers for eaeA, stxI and stxII genes was used to confirm the TaqMan results in 10 selected environmental isolates . CONCLUSIONS: All three TaqMan pathogen detection kits were useful for virulence gene analysis of prescreened foodborne O157:H7 isolates, while the O157:H7-specific kit may not be suitable for virulence gene analysis of environmental E . coli isolates, because of high false positive identification . SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to rapidly identify the presence of pathogenic E . coli in food or environmental samples is essential to avert outbreaks . These results are of importance to microbiologists seeking to use TaqMan PCR to rapidly identify pathogenic E . coli in environmental samples . Furthermore, serotyping may not be a reliable method for identification of O157:H7 strains.

Przegl Epidemiol, 2003, 57(2), 381 - 9
{Molecular methods in the diagnosis of infectious diseases}; Bartkowiak J; In the long term, many of the conventional diagnostic approaches to detection and characterisation of infectious diseases is complemented or even replaced by recognition of DNA/RNA specific sequences . The molecular procedures are based on two distinct types of methods--nucleic acid hybridization techniques with specific molecular probes, and DNA amplification by the polymerase chain reaction PCR . In the first method DNA fragments generated by restriction endonucleases are separated by electrophoresis, transferred to proper membrane and annealed with specific oligonucleotides which are labelled with 32P or with a non-radioactive marker . PCR reaction utilizes a DNA extension enzyme (polymerase) which can add nucleotide bases to primers once a template is provided . Oligonucleotides primers have to recognize target molecule . The single cycle is then repeated and each time DNA segments are doubled . Multiplex polymerase chain reactions have been designed to identify an organism as well as its virulence-specific sequences or antibiotic resistance plasmids simultaneously . The major drawback of PCR procedure which result from its exquisite sensitivity have to be consider . The problem is contamination and many strategies have been developed to avoid it . To these research tools were added probes for in situ hybridization . Advantages provided by this technique include the ability to detect latent (non-replicating) viruses and to localize their genomes to nuclear or cytoplasmic regions within cells . Nucleic acid probes or the hybridization conditions can be manipulated so that a broad spectrum of genotypes could be detectable . This is particularly valuable in those emerging infections where the individual serotypes are unknown . Recombinant DNA approaches have now been described for detection of a wide range of infectious agents . Some remain research activities, others are more appropriate to the routine diagnostic laboratory . An understanding of a pathogen's life cycle and the host's responses to the infectious agent is enhanced by characterisation of the former's genome using molecular technology . The spread of epidemics or hospital-acquired (nonsocomial) infections is followed and characterised with more accuracy by the identification of unique DNA fingerprints for individual pathogens . The prospects for molecular medicine in microbiology are vast and will have profound effects in laboratory and clinical practice.

AACN Clin Issues, 2003 Aug, 14(3), 295 - 301
Clinical decision support in critical care nursing; Lyons A et al.; A clinical decision support system (CDSS) is a computerized application that helps clinicians detect and prevent untoward clinical events such as drug interactions, errors of omission, and trends in symptomatology . A CDSS in healthcare usually is built around an alerting system based on rules of logic . The alerting system of a CDSS can notify clinicians immediately on clinical data entry, or it can generate alerts over time after relating data from multiple sources . A CDSS for nurses and patients offers immediate benefits for nurses and patients by detecting potential drug-laboratory and drug-drug combinations and impending pharmacologic complications, monitoring microbiology results, and helping nurses relate symptoms to pharmacology and medication side effects . Other benefits include savings in time and money and reductions in morbidity and mortality . A CDSS presents an opportunity for nursing informatics and critical care nursing to collaborate for the benefit of the patient and the profession.

Hist Cienc Saude Manguinhos, 2002, 9 Suppl, 67 - 87
{Economics, politics, and public health in Porfirian Mexico (1876-1910)}; Carrillo AM; The article examines the scientific, political, and economic elements that permitted the birth of modern public health in Mexico under the Porfirio Diaz administration (1876-1910) . Firstly, a portion of Mexican physicians were open to the discoveries of microbiology, immunology, and epidemiology . Secondly, the State's growing concentration of power in public health matters ran parallel to its concentration of disciplinary political power and enabled this new knowledge to be placed at the service of collective health problem prevention . Lastly, both imperialism and the Porfirian elite needed to protect their business interests . The article evaluates public health achievements and limitations during the Porfirian period, abruptly interrupted by the revolution begun in 1910.

Chest, 2003 Aug, 124(2), 554 - 9
Low hospital mortality in patients with acute interstitial pneumonia; Quefatieh A et al.; STUDY OBJECTIVES: To compare the presenting features and outcome of patients with acute interstitial pneumonia (AIP) with other patients with diffuse alveolar damage (DAD) and with historical control subjects . DESIGN: Retrospective chart review . SETTING: A large, urban, teaching hospital . INTERVENTIONS: Patients were classified into idiopathic (AIP group) and secondary causes of DAD (ARDS group) according to available clinical and microbiology data . AIP and ARDS cases were compared, and ARDS cases were analyzed for long-term outcome . Measurements and results: Twenty patients with pathologic diagnosis of DAD were identified . Four cases were excluded; eight cases of ARDS due to known etiologies were identified . These etiologies included pneumonia and sepsis (n = 6), cocaine use (n = 1), and carmustine chemotherapy (n = 1) . Eight cases of AIP were found . When compared with the ARDS group, patients in the AIP group had a longer time from the onset of symptoms until hospital admission (16.8 +/- 15.7 days vs 2.2 +/- 1.0 days, p = 0.0015) and a shorter time from hospital admission to open-lung biopsy (8.3 +/- 3.0 days vs 15.5 +/- 9.5 days, p = 0.02) {mean +/- SD} . Seven of eight patients with AIP and four of eight patients with ARDS survived to hospital discharge (p = not significant) . The 12.5% mortality rate for patients with AIP reported in this series was significantly lower than the previously reported cumulative rate of 69.5% (p = 0.0025) . Follow-up in five AIP survivors for a mean of 7.6 +/- 3.5 years (range, 3 to 11 years) showed all to be without shortness of breath or relapse despite mild residual fibrosis on chest radiograph and mild-to-moderate restriction on pulmonary function tests (mean total lung capacity, 68.5 +/- 6.2% predicted) . CONCLUSIONS: Our data support a favorable hospital and long-term outcome for patients with AIP, with no evidence of recurrence or progression to chronic interstitial lung disease.

J Clin Microbiol, 2003 Aug, 41(8), 3864 - 7
Improved molecular detection of dengue virus serotype 1 variants; Reynes JM et al.; The dengue virus molecular typing method described by Lanciotti and coworkers (R . S . Lanciotti, C . H . Calisher, D . J . Gubler, G . J . Chang, and A . Vance-Vorndam, J . Clin . Microbiol . 30:545-551, 1992) is used worldwide for diagnosis and surveillance . However, it failed to detect DENV-1 variants in Cambodia due to a point mutation . We describe an improvement of the method that allows the detection of additional DENV-1 strains, including potential variants.

J Clin Microbiol, 2003 Aug, 41(8), 3641 - 8
Penicillin susceptibility and epidemiological typing of invasive pneumococcal isolates in the Republic of Ireland; Bennett D et al.; A national study was undertaken to investigate the incidence of invasive pneumococcal disease in the Republic of Ireland and to examine the associated isolates . In 1999, 144 S . pneumoniae isolates, all recovered from cases of invasive disease, were received from 12 microbiology laboratories . The incidence of invasive pneumococcal disease was estimated to be 6.6/100000 population . All isolates were analyzed for serotype, penicillin susceptibility, chromosomal relatedness (by using pulsed-field gel electrophoresis {PFGE}), and penicillin-binding protein (pbp) fingerprinting . Several findings of note were observed regarding the pneumococcal population in Ireland . First, isolates of 25 different serotypes were represented, with serotypes 14, 9V, 8, 5, 4, and 3 being the most common . This finding, together with the pbp fingerprinting and PFGE typing results, indicated the clonal spread of strains of these serotypes in Ireland . Second, 27 (18.7%) isolates had reduced susceptibility to penicillin, and 74% of these were serotype 9V . Of these, 80% appeared to belong to the same clone . This could suggest the spread of the international Spanish/French 9V penicillin-resistant clone into Ireland . Third, nine different pbp genotypes were identified, four of which were new . Two pbp genotypes accounted for the majority of isolates dividing them according to their penicillin susceptibility status but irrespective of serotype and PFGE type . This is strong evidence for the occurrence of horizontal transfer of pbp genes between strains, observed with both penicillin-susceptible and penicillin-nonsusceptible isolates . Fourth, there was evidence of serotype transformation since isolates, indistinguishable by pbp fingerprinting and PFGE typing, expressed different capsular types.

Appl Environ Microbiol, 2003 Aug, 69(8), 4662 - 9
PCR-restriction fragment length polymorphism method for detection of Cyclospora cayetanensis in environmental waters without microscopic confirmation; Shields JM et al.; We developed an alternative nested-PCR-restriction fragment length polymorphism (RFLP) protocol for the detection of Cyclospora cayetanensis in environmental samples that obviates the need for microscopic examination . The RFLP method, with the restriction enzyme AluI, differentiates the amplified target sequence from C . cayetanensis from those that may cross-react . This new protocol was used to reexamine a subset (121 of 180) of surface water samples . Samples previously positive when the CYCF3E and CYCR4B primers (33) and RFLP with MnlI (20) were used were also PCR positive with the new primers; however, they were RFLP negative . We verified, by sequencing these amplicons, that while two were most likely other Cyclospora species, they were not C . cayetanensis . We can detect as few as one oocyst seeded into an autoclaved pellet flocculated from 10 liters of surface water . This new protocol should be of great use for environmental microbiologists and public health laboratories.

Curr Hematol Rep, 2003 Jul, 2(4), 302 - 9
Prevention and treatment of fungal infections in bone marrow transplantation; Mossad SB; There has not been as much success in the prevention and treatment of invasive fungal infections, particularly aspergillosis, compared to the prevention and treatment of cytomegalovirus infection and graft-versus-host disease in bone marrow transplant (BMT) recipients . Allogeneic BMT recipients who develop graft-versus-host disease and remain immunosuppressed for long periods are at major risk for development of these infections . Prevention of environmental exposure, antifungal chemoprophylaxis, and attempts at early diagnosis are essential for the reduction of mortality from invasive fungal infections . Chest computerized axial tomography is extremely useful in diagnosing pulmonary aspergillosis . However, microbiologic or histologic identification of infection remains essential . Unfortunately, the response to therapy in BMT recipients remains suboptimal . With the development of the lipid formulations of amphotericin B, the newer azoles, and the echinocandins, safer and more efficacious options have become available . The optimal use of antifungal agents or their combinations remains to be determined.

J Pediatr Surg, 2003 Aug, 38(8), 1244 - 5
Methylene blue: dangerous dye for neonates; Albert M et al.; Methylene blue is a basic dye commonly used in histologic microbiologic, and tissue staining . This report describes an instance of methylene blue toxicity in a premature neonate.

Commun Dis Public Health, 2003 Jun, 6(2), 87 - 96
The Communicable Disease Surveillance Centre 1977-2002: an overview; Bartlett CL; The Communicable Disease Surveillance Centre (CDSC) is now firmly established within the new Health Protection Agency (HPA), which was inaugurated on 1st April 2003 . In 2002, CDSC celebrated its 25th anniversary . Its achievements over those 25 years, a period when it evolved from a small unit with three staff to an international centre of excellence with over 250 staff, are recalled in this paper . The development of the Centre is reviewed, as are the ways in which it identified and responded to changing patterns of communicable disease over the quarter century . The considerable benefits of placing CDSC within the Public Health Laboratory Service are demonstrated through several examples . The fact that the expansion of CDSC took place during an era when public expenditure was constrained indicates that government valued the service it provided . The elements required and put in place for effective control and prevention of communicable diseases are highlighted and several themes emerge including the close working relationship needed between the various agencies and disciplines, particularly those between epidemiologists, microbiologists and clinicians . Another theme is the commitment to training, (which is necessary to ensure that work is practiCed to the highest standards), as well as to innovation through research and development . A rigorous approach to field investigation is essential, not just to enable the most appropriate control measures to be applied but to increase the knowledge base on infections and the means of prevention . Good surveillance and reference microbiology stand out as the vital components needed to produce the timely and precise information required to influence practice and policy.

Am J Infect Control, 2003 Aug, 31(5), 302 - 8
A prospective study of surgical site infections in a pediatric hospital in Mexico City; Porras-Hernandez JD et al.; BACKGROUND: Pediatric surgical site infection (SSI) rates in the United States range from 2.5% to 4.4% . There is little data regarding their risk factors among children . We quantified SSI rates and identified risk factors of SSI in a tertiary care pediatric teaching hospital in Mexico City . METHODS: All neurosurgical, cardiovascular, and general surgical patients who underwent operation between Aug 1, 1998, and Jan 31, 1999, were followed-up daily during hospitalization . On postoperative day 30, a full review of microbiology reports and medical records was performed . Univariate and multivariate analyses were done to identify risk factors . RESULTS: Four hundred twenty-eight of 530 children completed follow-up . The overall SSI rate was 18.7% . Forty percent of SSI were superficial incisional, 21% were deep incisional, and 39% were organ/space infections . For clean, clean-contaminated, contaminated, and dirty procedures, SSI infection rates were 12.4%, 24.4%, 14.3%, and 32.4%, respectively . Open drains (OR = 2.3; 95% CI = 1.3-4.2; P <.005) and surgery that lasted 90 or more minutes (OR = 2.9; 95% CI = 1.6-5.1; P <.001) were associated with infection . CONCLUSIONS: Our rates are greater than comparable reported data among children . Duration of surgery and use of open drains were associated with SSI.

Med Hypotheses, 2003 Aug, 61(2), 314 - 7
SARS: just another viral acronym?
Broxmeyer L.
Recent observations and experimental evidence have purported that a virus causes SARS, but such viruses have been isolated in only less than half of SARS patients in some studies and virologist Vincent Plummer of Winnipeg's National Microbiology Laboratory found that indeed 1 in 5 perfectly healthy Canadians with a history of recent travel to Asia had the virus . Therefore SARS microbiologic origins remain unclear . Outbreaks of multi-drug resistant (MDR) tuberculosis and the atypical mycobacteria simulate SARS on clinical, radiologic, epidemiologic, and diagnostic laboratory grounds and it is only logical then to include them in the differential to find a definitive cause and cure for SARS.

Mol Cell, 2003 Jul, 12(1), 125 - 34
Control of rRNA expression by small molecules is dynamic and nonredundant; Murray HD et al.; The control of ribosomal RNA transcription is one of the most enduring issues in molecular microbiology, having been subjected to intense scrutiny for over 50 years . Rapid changes in rRNA expression occur during transitions in the bacterial growth cycle and following nutritional shifts during exponential growth . Genetic approaches and measurements of initiating nucleoside triphosphate (iNTP) and guanosine 5'-diphosphate, 3'-diphosphate (ppGpp) concentrations and of rRNA promoter activities showed that rapid changes in the concentrations of iNTPs and ppGpp account for the rapid changes in rRNA expression . The two regulatory signals are nonredundant: changes in iNTP concentration dominate regulation during outgrowth from stationary phase, whereas changes in ppGpp concentration are responsible for regulation following upshifts and downshifts during exponential phase . The results suggest a molecular logic for the use of two homeostatic regulatory mechanisms to monitor different aspects of ribosome activity and provide general insights into the nature of overlapping regulatory circuits.

C R Biol, 2003 May, 326(5), 459 - 66
Modelling autocatalytic networks with artificial microbiology; Demarty M et al.; Cells can usefully be equated to autocatalytic networks that increase in mass and then divide . To begin to model relationships between autocatalytic networks and cell division, we have written a program of artificial chemistry that simulates a cell fed by monomers . These monomers are symbols that can be assembled into linear (non-branched) polymers to give different lengths . A reaction is catalysed by a particular polymer or 'enzyme' that may itself be a reactant of that reaction (autocatalysis) . These reactions are only studied within the confines of the 'cell' or 'reaction chamber' . There is a flux of material through the cell and eventually the mass of polymers reaches a threshold at which we analyse the cell . Our results indicate a similarity between the connectivity of the reaction network and that of real metabolic networks . Developing the model will entail attributing increased probabilities of reactions to polymers that are colocalised to evaluate the consequences of the dynamics of large assemblies of diverse molecules (hyperstructures) and of cell division.

Zh Mikrobiol Epidemiol Immunobiol, 2003 May-Jun, (3), 39 - 43
{The possibility of the detection of one more tick-borne infection--babesiosis--on the territory of Russia}; Alekseev AN; A total of 739 taiga ticks of ixodes persulcatus species, obtained in the recreational zone of St . Petersburg, were studied for the presence of Babesia sp . with polymerase chain reaction . All these ticks underwent the preliminary examination for the presence of Borrelia (3 species), Ehrlichia (2 species) and tick-borne encephalitis (TBE) virus . In 7 cases Babesia were detected among 413 ticks containing other pathogens . Among 326 ticks no Babesia were detected, as well as no other pathogens . All ticks having Babesia were also found to contain Borrelia species: B . afzelii, B . garinii, or both (1 case) . In one female tick, in addition to Babesia, also B . garinii and TBE virus were determined . The data thus obtained should draw special attention not only of parasitologists, epidemiologists and microbiologists studying ticks obtained from natural sources, but also of clinicists who should consider the possibility of mixed infection, when one infection may mask the presence of some other infection, in particular babesiosis . Due to rare occurrence of Babesia in ticks and the presence of mixed infection difficulties may arise in the detection of Babesia.

An Sist Sanit Navar, 1999 Sep-Dec, 22(3), 393 - 9
{AIDS/HIV epidemiological surveillance in Navarra}; Registro de VIH/SIDA de Navarra; The incidence of AIDS in the European Union has been declining since 1996 as a result, at least in part, of the use of more efficient treatments for HIV infection . The same tendency can be observed in Navarra . In 1998 the incidence of AIDS in Navarra, 75 cases per million inhabitants, was situated at an intermediate level in the ranking of Spanish autonomous communities . In the same year, the highest rates of incidence in Europe were recorded in Spain (93.3 cases per million inhabitants) and Portugal (81.8), and the lowest in Ireland and Finland, with less than 5 cases per million inhabitants . With respect to the incidence of HIV infection, 2,240 cases were diagnosed in Navarra between 1985 and December 1998 . The available data indicate that at least 1,864 of these cases were resident in Navarra . The annual number of new diagnoses of HIV infection show two high points in the years 1987 and 1991, with over 200 annual cases . From 1993 onwards, a progressive decline can be observed in the annual number, with 171, 145, 19, 86 and 63 cases in the final year, 1998 . The figure of over 2,000 cases of HIV infection contrast with the 694 cases of AIDS registered in this autonomous community in the same period and help to resituate the epidemic in more accurate terms . The development of AIDS in 40% of the persons with HIV infection and the death of 26% have been confirmed, although lethality has been declining in recent years . The use of intravenous drugs in 73.2% of the AIDS cases is the most frequent category of transmission . 13.4% are due to transmission by heterosexual contact and 6.1% to homosexual practices between males . Although the cases of AIDS have been declining in Navarra since 1996, the data from the microbiology laboratories and from the hospital services that treat persons with HIV infection indicate that a considerable number of new infections will continue to occur, which justifies the need for maintaining the prevention programs in order to avoid new cases of a disease whose prognosis is of such seriousness

An Sist Sanit Navar, 2000 Sep-Dec, 23(3), 467 - 72
{Epidemiology of HIV and AIDS infection in Navarra}; Registro de VIH/SIDA de Navarra; The incidence of AIDS has been declining in the European Union since 1996 as a result, at least in part, of the use of more efficient treatments for HIV infection . The same tendency can be observed in Navarra, where the average has dropped from 70 annual cases in the first five years of the 1990s to 30 cases in the year 1999 . In Navarra, the most frequent category of transmission was the consumption of intravenous drugs, a practice referred to by 72% of those diagnosed with AIDS between 1985 and 1999 . 21% were attributed to sexual transmission (7% to homosexual practices and 14% to heterosexual), 2.5% to other mechanisms (mother-child, hemoderivates, etc.) and in the remaining 4.6% information was unavailable on the probable mechanism of transmission . With respect to the incidence of HIV infection, between 1985 and December 1999, 2,379 cases were diagnosed in Navarra . The annual number of new diagnoses of HIV infection showed two maximum peaks in the years 1987 and 1991, with over 200 annual cases . A progressive decline can be observed from 1993 onwards, falling to 40 cases in 1999 . The figure of over 2,000 cases of HIV infection contrasts with the 672 cases of AIDS registered in this autonomous community up until the year 2000, and resituates the epidemic in more certain terms . Although the cases of AIDS and HIV have fallen in Navarra in recent years, the data from the microbiology laboratories and the hospital services that treat persons with HIV infection indicate that a considerable number of new cases of infection will continue to occur, which justifies the need for maintaining the prevention programs.

J Med Libr Assoc, 2003 Jul, 91(3), 303 - 10
Mapping the literature of clinical laboratory science; Delwiche FA; This paper describes a citation analysis of the literature of clinical laboratory science (medical technology), conducted as part of a project of the Nursing and Allied Health Resources Section of the Medical Library Association . Three source journals widely read by those in the field were identified, from which cited references were collected for a three-year period . Analysis of the references showed that journals were the predominant format of literature cited and the majority of the references were from the last eleven years . Applying Bradford's Law of Scattering to the list of journals cited, three zones were created, each producing approximately one third of the cited references . Thirteen journals were in the first zone, eighty-one in the second, and 849 in the third . A similar list of journals cited was created for four specialty areas in the field: chemistry, hematology, immunohematology, and microbiology . In comparing the indexing coverage of the Zone 1 and 2 journals by four major databases, MEDLINE provided the most comprehensive coverage, while the Cumulative Index to Nursing and Allied Health Literature was the only database that provided complete coverage of the three source journals . However, to obtain complete coverage of the field, it is essential to search multiple databases.

Ostomy Wound Manage, 2003 Jul, 49(7A Suppl), 1 - 7
The wound infection continuum and its application to clinical practice; Kingsley A; Four basic conditions exist in open wounds resulting from the level of bioburden present (bacterial contamination--normal but short-lived state, colonization--normal state, critical colonization--abnormal state, and infection--abnormal state) . The two abnormal states have the potential to disrupt the orderly healing sequence, which results in the development of a chronic wound . In addition, the impact of the wound s anatomical position, duration, shape, or presentation; the patient s level of health and control of underlying pathologies; the presence of infection-potentiating factors such as foreign bodies, hematoma, and necrotic tissue; the sources and frequency of exogenous contamination; the considered virulence of the individual micro-flora species; and the potential synergism between different species all require consideration . This article discusses the states of bioburden and other aspects of microbiology relating to wound infection and their application to clinical practice.

Mol Phylogenet Evol, 2003 Aug, 28(2), 186 - 96
Nuclear gene sequences provide evidence for the monophyly of australidelphian marsupials; Amrine-Madsen H et al.; Relationships among the seven extant orders of marsupials remain poorly understood . Most classifications recognize a fundamental split between Ameridelphia, which contains the American orders Didelphimorphia and Paucituberculata, and Australidelphia, which contains four Australasian orders (Dasyuromorphia, Diprotodontia, Notoryctemorphia, and Peramelina) and the South American order Microbiotheria, represented by Dromiciops gliroides . Ameridelphia and Australidelphia are each supported by key morphological characters with dichotomous character states . To date, molecular studies indexing all marsupial orders have reported inconclusive results . However, several studies have suggested that Dromiciops is nested within Australidelphia . This result has important implications for understanding the biogeographic history of living marsupials . To address questions in higher-level marsupial systematics, we sequenced portions of five nuclear genes (Apolipoprotein B gene; Breast and Ovarian cancer susceptibility gene 1; Recombination activating gene 1; Interphotoreceptor retinoid binding protein gene; and von Willebrand factor gene) for representatives of all orders of marsupials, as well as placental outgroups . The resulting 6.4kb concatenation was analyzed using maximum parsimony, distance methods, maximum likelihood, and Bayesian methods . tests were used to examine a priori hypotheses . All analyses provided robust support for the monophyly of Australidelphia (bootstrap support=99-100%; posterior probability=1.00) . Ameridelphia received much lower support, although this clade was not rejected in statistical tests . Within Diprotodontia, both Vombatiformes and Phalangeriformes were supported at the 100% bootstrap level and with posterior probabilities of 1.00.

J Feline Med Surg, 2003 Aug, 5(4), 217 - 26
An epizootic of highly virulent feline calicivirus disease in a hospital setting in New England; Schorr-Evans EM et al.; This article reports an outbreak of 24 cases of an unusually virulent feline calicivirus (FCV) infection in a small animal hospital . The circumstances and disease signs were very similar to those recently described in an outbreak of FCV hemorrhagic disease in Northern California (Vet . Microbiol . 73 (2000) 281) . The virus entered the facility through shelter cats showing upper respiratory signs . Affected cats manifested high fever, anorexia, labored respirations, oral ulceration, facial and limb edema, icterus, and pancreatitis . The infection spread rapidly among the patients by contaminated animal caretakers and hospital equipment . One case of fomite transmission from an employee to a housecat was documented . Prior vaccination, even with multiple doses of FCV-F9-based live calicivirus vaccine, was not protective . Affected cats often required extensive supportive care for 7-10 days, and the overall mortality from death and euthanasia was 32% . The strain of FCV responsible for this outbreak was genetically and serologically distinct from the FCV strain responsible for a similar epizootic and the FCV-F9 strain contained in most vaccines . Outbreaks of this type are being reported with increasing frequency, and are often associated with the practice of treating sick shelter cats in private practices . Similar to the present epizootic, outbreaks of FCV hemorrhagic disease have been self-limiting, but require prompt application of strict quarantine, isolation, personnel sanitation, and disinfection procedures.

Ann Allergy Asthma Immunol, 2003 Jul, 91(1), 3 - 12; quiz 12-5, 91
Primary dietary prevention of food allergy; Fiocchi A et al.; OBJECTIVE: To present research and clinical evidence on the use of primary dietary prevention in food allergy management . DATA SOURCES: We conducted MEDLINE searches for pertinent articles published between January 1986 and October 2001 with use of the following keywords or phrases: prevention and diet and allergy, food allergy and prevention, and dietary prevention and food allergy or allergens . Also included are information and commentary reflecting the authors' cumulative clinical experience in an allergy unit of a city hospital . RESULTS: We define as "proactive" those strategies centered on "success factors," such as the early postnatal environment, prolonged breast-feeding, and use of formula and probiotic supplementation, in contrast to earlier "prohibitionist" approaches to prevention of food allergy . These two approaches are not antagonistic and may even be synergistic . We introduce this distinction in light of epidemiologic evidence and out of concern about compliance and the quality of life for patients . CONCLUSIONS: Inasmuch as nutritional and immune maturation are implicated, the prohibitionist measures that are most effective in primary prevention of food allergy are exclusive breast-feeding for at least 6 months (for lifelong immunity and other benefits), delayed (after the sixth month) introduction of solid foods, and on-indication use of "hypoallergenic" formulas . Whether proactive strategies can be contemplated remains a debatable issue . Evidence for and against the scientific use of probiotics as well as microbiologic, epidemiologic, and clinical data are discussed . Review of published epidemiologic studies and randomized clinical trials is essential before planning dietary intervention or prevention.

An Sist Sanit Navar, 2001 May, 24(2), 213 - 8
{Epidemiological vigilance for HIV/AIDS in Navarra}; Registro de VIH/SIDA de Navarr; Between 1985 and December 2000, 2,395 new cases of HIV infection were diagnosed in Navarra . The years showing the highest incidence were 1987 with 250 cases, and 1991 with 232 cases . Starting in 1993 a progressive decline can be observed, reaching 35 cases in the year 2000 . In the same period, 1985-2000, 690 cases of AIDS were diagnosed amongst residents in Navarra . The cases of AIDS have undergone a similar decline in recent years, dropping from an average of about 60 cases per year in the first half of the 1990s to the 19 cases diagnosed in the year 2000 . This tendency, which became evident from 1996 onwards in most countries of our area, can be attributed, at least in part, to the employment of more efficient treatments for HIV infection . In Navarra, the most frequent category of transmission between patients with AIDS was the use of drugs by intravenous means, a practice referred to by 71.2% of the persons diagnosed between 1985 and 2000 . Twenty point seven per cent of the cases were attributed to sexual transmission (7% to homosexual practices and 14% to heterosexual practices) and 2.5% to other mechanisms (mother-child, hemoderivatives and transfusions) . Although the cases of AIDS and HIV have declined in Navarra in recent years, the data from microbiology laboratories and the hospital services that treat persons with HIV infection indicate that a considerable number of new cases will continue to occur, which justifies the need to maintain the prevention programmes.

Cardiol Clin, 2003 May, 21(2), 253 - 71, vii
Infections of intracardiac devices; Karchmer AW et al.; Intracardiac devices are increasingly used to correct hemodynamically dysfunctional valves and electrophysiologic abnormalities . These devices become infected at relatively low rates . Nevertheless, when these low rates are applied to widely used devices, significant numbers of infections result . Additionally, these infections have been associated with high degrees of morbidity and high mortality rates . This article reviews the epidemiology, microbiology, clinical presentation, and medical as well as surgical therapy of intracardiac device infections.

Cardiol Clin, 2003 May, 21(2), 207 - 17
Diagnostic methods . Current best practices and guidelines for identification of difficult-to-culture pathogens in infective endocarditis; Houpikian P et al.; IE is a serious, life-threatening disease . Because treatment must often be adapted to the pathogen involved, rapid identification of the etiologic agent is critical to successful management of each patient . When difficult-to-culture pathogens are involved, routine microbiologic tests, including blood culture, may remain negative . Because such cases may account for up to 31% of all IE cases, alternative diagnostic approaches are necessary . Among the etiologic agents of culture-negative endocarditis, C burnetii and Bartonella spp play a major role; each is responsible for up to 3% of episodes of IE . The authors therefore recommend the systematic use of specific serologies in all cases of clinically suspected endocarditis . The cross-reactivity between C burnetii, Bartonella spp, and Chlamydia spp is of diagnostic importance because all are potential etiologic agents of endocarditis . However, given that the levels of specific antibodies observed in Bartonella endocarditis are extremely high, low-level cross-reactions with other antigens should not lead to misdiagnosis, provided serology for all suspected agents is performed . When serologic test results are negative for both Bartonella spp and C burnetii, special staining by the Gram, Giemsa, Gimenez, PAS, Warthin-Starry, and Grocott methods may guide the use of new diagnostic tools such as PCR and tissue culture for isolation and identification of the causative agent . Such novel approaches may lead to more comprehensive patient evaluations and the discovery of new etiologic agents of IE.

Clin Otolaryngol, 2003 Aug, 28(4), 371 - 3
The natural history of recurrent acute tonsillitis and a clinical trial of azithromycin for antibiotic prophylaxis; Lildholdt T et al.; The pharmacokinetics of azithromycin (Zitromax), Pfizer Inc., USA) in tonsil tissue warranted the present trial . In 110 patients eligible for tonsillectomy because of recurrent acute tonsillitis, surgery was replaced by randomized medication with azithromycin 500 mg or placebo once per week for 6 months . Subsequently, their clinical condition and microbiology was monitored for 12 months . Acute tonsillitis developed in 40% of the patients who received azithromycin and in 49% of the patients in the placebo group (P > 0.05) . Accordingly, 45% of all patients developed acute tonsillitis . Resistance to azithromycin was not detected . In this trial long-term medication with azithromycin was not efficacious in recurrent acute tonsillitis . As all patients were eligible for tonsillectomy according to current criteria, it is surprising that only 45% developed acute tonsillitis during the trial period . Therefore, the criteria for tonsillectomy in recurrent acute tonsillitis must be revised.

J Med Microbiol, 2003 Aug, 52(Pt 8), 685 - 91
Development of broad-range 16S rDNA PCR for use in the routine diagnostic clinical microbiology service; Harris KA et al.; The aim of the present study was to develop a broad-range PCR based on bacterial 16S rDNA for use in the routine diagnostic clinical microbiology service . The optimization and validation of the assay for use on clinical specimens from normally sterile sites is described, and preliminary results are reported on the use of the assay in the clinical diagnosis of bacterial infection in 382 paediatric specimens over a 2-year period . These results are compared to those obtained by standard culture techniques and show increased diagnosis of bacterial infection when both culture and PCR are used together; 16S rDNA PCR provided the sole evidence of pathogenic infection in 71 cases . Key stages in the assay development and potential pitfalls of the technique are highlighted and the improvement the assay offers in the diagnosis of infection in the paediatric setting is discussed.

J Med Microbiol, 2003 Aug, 52(Pt 8), 609 - 13
Severe acute respiratory syndrome (SARS): breath-taking progress; Hawkey PM et al.; Reports of a new severe respiratory disease, now defined as severe acute respiratory syndrome (SARS), began to emerge from Guangdong, in southern China, in late 2002 . The condition came to international attention through an explosive outbreak in Hong Kong in March 2003 . Cases appeared throughout South-East Asia and in Toronto, the spread of SARS being accelerated by international air travel . A global emergency was declared by the World Health Organization, bringing together an international team of epidemiologists, public health physicians and microbiologists to study and contain the disease . This response has enabled the nature of the infectious agent to be identified, its mode of transmission to be established and diagnostic tests to be created rapidly.

Med Clin (Barc), 2003 Jun 28, 121(4), 139 - 41
{Imported hookworm infection in African immigrants in Spain: study of 285 patients}; Roca C et al.; BACKGROUND AND OBJECTIVE: Hookworm infection is a worldwide intestinal parasitic disease affecting more than one billion people . It represents an important public health problem in rural areas of developing countries . In our environment, it is generally considered an imported disease due to the immigration process . PATIENTS AND METHOD: Retrospective observational study of African immigrants diagnosed with hookworm infection at the Immigration and Tropical Medicine Unit of the Hospital of Mataro over the period 1984-1999 . RESULTS: We identified 285 patients, mainly young males, from Gambia or Senegal, with a precarious job who had arrived in Spain 3 years earlier or less . Abdominal pain was the commonest reason for consultation (28.8%) cases . Non digestive symptoms were 35.6% and 4.6% remained assymptomatic . 60% had a concomitant infectious disease . Laboratory tests showed iron-deficiency anemia in 28.4% and eosinophilia in 52.3% . 70% of patients did not come to visit after treatment . CONCLUSIONS: Microbiologic stool examination is recommended as part of the health assessment of immigrants from countries where hookworm infection is highly prevalent, even in the absence of abdominal symptoms or abnormalities of the red and white blood series.

J Vet Med B Infect Dis Vet Public Health, 2003 Jun, 50(5), 214 - 20
Virulence of recent and former classical swine fever virus isolates evaluated by their clinical and pathological signs; Floegel-Niesmann G et al.; The clinical diagnosis of classical swine fever (CSF) still caused problems to the veterinarians during the last decade . The primary CSF outbreak was often detected too late and, meanwhile, the virus had spread . Consequently, the recent classical swine fever virus isolates (CSFV) were suspected to be of low virulence . The purpose of the study was to quantify the virulence of four recent CSFV by evaluating the clinical and pathological signs caused by different CSFV . Pigs of the same breed and age group were inoculated intranasally with CSFV from recent epidemics in European Union (EU) member states . The CSFV used are registered in the data base of the EU Reference Laboratory for CSF and belong to different genotypes: 2.1, 2.2 and 2.3 respectively . Clinical signs of CSF were evaluated by using a score system suggested previously (Mittelholzer et al., 2000: Vet . Microbiol . 74, 293) . For the evaluation of pathological lesions, a new pathological score was introduced . The four CSFV tested here were classified as moderately virulent in general, although one CSFV may cause different clinical courses, ranging from highly virulent to avirulent . This indicates the importance of additional factors in the host animal for virulence . Differences in the clinical and pathological signs between these four recent CSFV were rather minor, emphasizing that the genetic typing of CSFV is absolutely essential . Differences towards former CSFV (e.g . reference virus strain Alfort 187) were more pronounced, especially regarding the onset and duration of the disease, the occurrence of skin haemorrhages and pathological lesions of kidney, subcutis and serosae . It is concluded that clinical diagnosis of CSF is rather difficult in pigs up to 14 days post-CSFV infection using these four CSFV, emphasizing the need for careful differential diagnosis and the laboratory investigation for CSF at an early stage.

J Infect, 2003 Aug, 47(2), 170 - 3
Tropheryma whippelii as a cause of afebrile culture-negative endocarditis: the evolving spectrum of Whipple's disease; Richardson DC et al.; With the advent of molecular diagnostics culture-negative endocarditis caused by the organism Tropheryma whippelii is an increasingly described entity . We describe two patients with afebrile, culture-negative endocarditis caused by T . whippelii who had neither the gastrointestinal nor arthritic manifestations of Whipple's disease . Whipple's disease is a systemic illness caused by the organism Tropheryma whippelii and is typically characterized by diarrhea, weight loss, and arthropathy {Clin . Microbiol . Rev . 2001;14:561-583; Medicine (Baltimore) 1997;76:170-184} . Whipple's endocarditis is relatively common in autopsy studies {Can . J . Cardiol . 1996;12:831-834} but has rarely been diagnosed before death . With the advent of molecular diagnostic tools such as the polymerase chain reaction (PCR), Tropheryma whippelii as a cause of culture-negative endocarditis has become increasingly recognized {Clin . Infect . Dis . 2001;33:1309-1316; Ann . Intern . Med . 1999;131:112-116; Infection 2001;29:44-47; Ann . Intern . Med . 2000;132:595} . With this increased recognition has come the realization that Whipple's endocarditis can occur without other common manifestations of Whipple's disease {Ann . Intern . Med . 1999;131:112-116; Infection 2001;29:44-47; Ann . Intern . Med . 2000;132:595} . We report here two cases of Whipple's endocarditis without discrete febrile illness, gastrointestinal manifestations, or arthritic manifestations, diagnosed by PCR of resected valvular material.

J Infect, 2003 Aug, 47(2), 99 - 103
How should laboratories communicate with primary care? Obtaining general practitioners' views; McNulty CA et al.; AIMS: Recognising the importance of communication with our primary care colleagues, focus groups were held with GPs to determine how they perceived the current lines of communication with their local microbiology laboratory and the PHLS, and how they could be improved . METHODS: Focus groups were held in Plymouth, Gloucester, Bristol and Hereford . Between four and 10 GPs and/or PCG Board members attended each workshop . The modes of communication i.e . websites, face-to-face contact, laboratory reporting, telephone advice, newsletters, guidance and surveillance were discussed . RESULTS: Microbiology websites should be user friendly, with clear labelling as to whom the page is directed . They should contain locally relevant data, antibiotic guidance and information leaflets . Despite great variation in laboratory reporting protocols GPs were mostly happy with reports received . Results, especially serology, should contain a clear conclusion and could refer to a website for further information . Electronic reporting was enthusiastically awaited . All GPs felt they had excellent access to telephone advice . GPs would value data and guidance on their use of diagnostic tests . CONCLUSION: These workshops highlight the variation in laboratory reporting protocols that should be addressed . Website development for GPs should include locally relevant data . GPs would value details of their laboratory use and costs.

Clin Diagn Lab Immunol, 2003 Jul, 10(4), 647 - 51
Epitope mapping of the Brucella melitensis BP26 immunogenic protein: usefulness for diagnosis of sheep brucellosis; Seco-Mediavilla P et al.; Sequencing of bp26, the gene encoding the Brucella sp . immunogenic BP26 periplasmic protein, was performed in the reference strains of Brucella abortus, B . suis, and B . ovis . The three bp26 sequences were almost identical to that published for B . melitensis 16M bp26, and only minor nucleotide substitutions, without modifying the amino acid sequence, were observed between species . The bp26 genes of the seven B . abortus biovar reference strains and B . abortus S19 and RB51 vaccine strains were also sequenced . Again, only minor differences were found . Surprisingly, the bp26 nucleotide sequence for B . abortus S19 was almost identical to that found for B . melitensis 16M and differed from the sequence described previously by others (O . L . Rossetti, A . I . Arese, M . L . Boschiroli, and S . L . Cravero, J . Clin . Microbiol . 34:165-169, 1996) for the same B . abortus strain . The epitope mapping of BP26, performed by using a panel of monoclonal antibodies and recombinant DNA techniques, allowed the identification of an immunodominant region of the protein interesting for the diagnosis of B . melitensis and B . ovis infection in sheep . A recombinant fusion protein containing this region of BP26 reacted indeed, in Western blotting, as the entire recombinant BP26 against sera from B . melitensis- or B . ovis-infected sheep while it avoided false-positive reactions observed with sera from Brucella-free sheep when using the entire recombinant BP26 . Thus, use of this recombinant fusion protein instead the entire recombinant BP26 could improve the specific serological diagnosis of B . melitensis or B . ovis infection in sheep.

G Ital Nefrol, 2003 May-Jun, 20 Suppl 22, S43 - 8
{Disinfection is prevention}; Conti F et al.; Prevention of outbreaks is very important in haemodialysis units because of the high risk of infectious complications caused by contaminated water or equipment . Our aim was to evaluate the efficiency of a chloride based disinfectant for haemodialysis machines . Five different machines were tested for infection by culturing water drawn from both inlet (point A) and outlet (point B) of the dialyser and from the waste site (point C) . Six series of tests were analysed during a 3-week period . In the first week water was drawn from point A before and after disinfection on two different days; during the second and third weeks the same procedure was performed at points B and C, respectively . Three non-contaminated samples of 2 mL each were sent to the microbiology department for cultures . Samples from point B grew a number of bacteria even after the disinfectant for the water system was changed . We conclude that machine contamination is due to patients and, therefore, a very accurate cleaning and disinfection of system components is essential . This procedure can avoid isolation of infected patients . Prevention of infection is a complex procedure and should engage water systems, machines, as well as environment and nurses.

Am J Respir Crit Care Med, 2003 Jul 15, 168(2), 165 - 72
Current epidemiology of septic shock: the CUB-Réa Network; Annane D et al.; To update the epidemiology of septic shock we analyzed clinical, microbiologic, and outcome variables from 100,554 intensive care unit admissions on the College des Utilisateurs de Bases de donnees en Reanimation (CUB-Rea) database, collected from 22 hospitals over a 8-year period, 1993 to 2000 . The overall frequency of septic shock was 8.2 per 100 admissions (i.e., 8,251 stays) . It increased from 7.0 (in 1993) to 9.7 per 100 admissions (in 2000) . The distribution analysis of the sites of infection and of the types of pathogens showed an increase in the rate of pulmonary infection (p = 0.001) and of multiresistant bacteria-related septic shock (p = 0.001) . The crude mortality was 60.1% and declined from 62.1% (in 1993) to 55.9 (in 2000) (p = 0.001) . As compared with matched intensive care unit admissions without sepsis, the excess risk of death due to septic shock was 25.7 (95% confidence interval, 24.0-27.3) and the matched odds ratio of death was 3.9 (95% confidence interval, 3.5-4.3) . The frequency of septic shock is increasing with more multiresistant strains . Its crude mortality rate is decreasing, but patients with septic shock still have a high excess risk of death than critically ill patients who are nonseptic.

Obstet Gynecol, 2003 Jul, 102(1), 162 - 6
Tumor necrosis factor-alpha promoter gene polymorphism -308 and chorioamnionitis; Simhan HN et al.; OBJECTIVE: A single-base polymorphism within the promoter of the gene for tumor necrosis factor alpha at position -308 relative to the transcriptional start site of the gene is correlated with differences in production of tumor necrosis factor alpha . There are two allelic forms: TNFA1 and TNFA2 . The TNFA2 allele is associated with increased production of tumor necrosis factor alpha . The purpose of this study is to assess the relationship between carriage of the TNFA2 allele and clinical chorioamnionitis . METHODS: In this retrospective cohort study, previously banked deoxyribonucleic acid from 149 women who had spontaneous labor from 37 to 42 weeks' gestation was used . Polymerase chain reaction was used for polymorphism assay . Demographic and clinical information was obtained from the medical record . Clinical chorioamnionitis was defined as at least one temperature elevation above 38C combined with at least two of the following signs: maternal or fetal tachycardia, uterine tenderness greater than expected, foul-smelling vaginal discharge, and white blood cell count more than 18,000 . RESULTS: Chorioamnionitis was present in 18 women (12.1%) . Among women who did not carry TNFA2, the chorioamnionitis rate was 7.4% . Among women who carried TNFA2, the chorioamnionitis rate was 24.4% . The relative risk for chorioamnionitis with carriage of TNFA2 was 3.3 (95% confidence interval 1.3, 7.1) . This increased risk was not altered after adjustment for race, type of rupture of membranes, intrauterine pressure catheter use, smoking, and prolonged rupture of membranes . CONCLUSION: Carriage of the TNFA2 allele is associated with a more than three-fold increased risk of clinical chorioamnionitis, even when accounting for important clinical and microbiologic risk factors.

J Clin Microbiol, 2003 Jul, 41(7), 3192 - 7
Simplifying collection of corneal specimens in cases of suspected bacterial keratitis; Kaye SB et al.; Identification of the causative organisms in suspected bacterial keratitis traditionally involves collecting multiple corneal scrapes, which are plated directly onto different solid agar culture media . Difficulties have been reported with this practice, so the development of a simpler diagnostic method in suspected bacterial keratitis would be useful . It is unclear whether a single corneal scrape sent to the microbiology laboratory in a liquid transport culture medium (indirect method) is as reliable for the diagnosis of bacterial keratitis as inoculation of multiple scrapes directly onto agar plates (direct method) . To investigate this, bacterial recovery was assessed following transfer and transport of different concentrations and types of bacteria from an artificially contaminated surgical blade into brain heart infusion (BHI) . Bacterial recovery rates between the proposed (indirect) and standard (direct) method were then compared after the in vitro inoculation of pig corneas and following specimen collection in patients with presumed bacterial ulcerative keratitis . Recovery of bacteria from contaminated surgical blades was found to be the same from both solid and liquid culture media . There was no significant difference in the numbers of positive cultures from solid (direct) and liquid (indirect) culture media, both in the experimental pig cornea inoculation study (P = 0.34) and in experiments with patients with clinical infections (P = 0.4), with an 85.2% agreement between methods (kappa = 0.61, P < 0.0001) . In conclusion, therefore, the collection of two corneal scrapes, one used for Gram staining and the other transported in BHI followed by plating and subculturing in an enrichment medium, provides a simple method for the investigation of presumed bacterial keratitis.

J Indian Med Assoc, 2003 Feb, 101(2), 93 - 5
Head and neck manifestations of HIV infection: a preliminary study; Deb T et al.; The study deals with evaluation of the head and neck manifestations of human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) . It is a preliminary, prospective study, conducted in the department of ENT and microbiology, Regional Institute of Medical Sciences, Imphal, Manipur . The clinical presentations for HIV infection and AIDS with head and neck involvement are shown in this paper . Forty patients with HIV infection and various head and neck manifestations are included in this study The median age of diagnosis was 33 years with male to female ratio of 3.4: 1 . The predominant mode of transmission of HIV infection among the patients of this series was found to be intravenous drug use (IDU) in 65% of cases . Rhinosinusitis was found to be the most common presenting feature constituting 27.5% of the cases followed by oral candidiasis in 22.5% of the cases . After consideration and observation of all the facts and findings, this study concludes and proposes that it will be well for all clinicians, including otolaryngologists especially, to bear a high level of suspicion for HIV infections in their day to day practice.

Appl Environ Microbiol, 2003 Jul, 69(7), 3826 - 32
Two-step freezing procedure for cryopreservation of rumen ciliates, an effective tool for creation of a frozen rumen protozoa bank; Nsabimana E et al.; The present study aimed at the long-term storage of rumen protozoa as living cells in liquid nitrogen . The two-step or interrupted slow freezing procedure was used to cryopreserve six of the dominant species of rumen ciliates isolated from monofaunated animals, Dasytricha ruminantium, Entodinium caudatum, Epidinium ecaudatum caudatum, Eudiplodinium maggii, Isotricha prostoma, and Polyplastron multivesiculatum . We optimized the first step in the interrupted slow freezing procedure, from the extracellular ice nucleation temperature to the holding temperature, and studied the effects of the cooling rates on survival . In addition to the nature of the cryoprotectant (dimethyl sulfoxide), the equilibration temperature and equilibration time (25 degrees C and 5 min, respectively), and the holding time at subzero temperature (45 min) recommended previously (S . Kisidayova, J . Microbiol . Methods 22:185-192, 1995), we found that a holding temperature of -30 degrees C, a cooling rate from extracellular ice nucleation temperature to holding temperature of between 1.2 degrees C/min and 2.5 degrees C/min, depending on the ciliate, and rumen juice as the freezing and thawing medium markedly improved the survival rate . Survival rates determined after 2 weeks in liquid nitrogen were 100% for Isotricha, 98% for Dasytricha, 85% for Epidinium, 79% for Polyplastron, 63% for Eudiplodinium, and 60% for Entodinium . They were not significantly modified after a period of 1 year in liquid nitrogen . Four of the five ciliate species cryopreserved for 8 months in liquid nitrogen successfully colonized the rumen when inoculated into defaunated animals . These results have made it possible to set up a bank of cryopreserved rumen protozoa.

Int J Infect Dis, 2003 Jun, 7(2), 152 - 5
Treating obsessive compulsive disorder: a new role for infectious diseases physicians?
Hambridge J, Loewenthal M.
BACKGROUND: Patients with psychiatric disorders are often seen by infectious disease physicians . Sometimes the psychiatric condition is the primary disorder, and the physician's main task is the early identification of the disorder and referral to specialist psychiatric services . On other occasions, the psychiatric condition will need to be addressed in addition to the infectious disease, and the physician aims to treat in conjunction with a psychiatrist . It is rare for referrals to be made from psychiatry to infectious diseases physicians . METHOD: A single case study is used to describe a modification of Danger Ideation Reduction Therapy (DIRT), a novel intervention for obsessive compulsive disorder (OCD) . In our modification the infectious diseases physician plays a key, collaborative role in the psychological treatment of the patient . RESULTS: Although an uncontrolled trial, results from the modified DIRT protocol are encouraging and warrant replication in a randomised controlled trial . CONCLUSIONS: A collaborative approach by the infectious diseases physician, the microbiology laboratory and the psychologist can provide a valuable means of retaining patients with OCD in treatment and in the management of this common, disabling condition.

Infect Dis Obstet Gynecol, 2003, 11(1), 53 - 7
Pregnancy-associated pyogenic sacroiliitis: case report and review; Almoujahed MO et al.; BACKGROUND: Pyogenic sacroiliitis occurs infrequently during the peripartum period . CASE: A case at our institution and a review of the literature were analyzed . A total of 15 cases were discovered . The onset of illness was during pregnancy (40% of cases), within 3 weeks postpartum (40%) or post-abortion (20%), and the presentation was usually acute (< 7 days in 67% of cases) . Frequent manifestations included localized pain in the hips or buttock, sacroiliac joint tenderness and fever . Computed tomography or magnetic resonance imaging revealed joint involvement in all cases tested . Microbiology was confirmed by blood (40%) or joint aspirate (75%), and most patients were treated with antibiotics . Surgical intervention took place in five cases . Preterm labor was reported in only one case . All patients responded well to therapy without locomotive disability, and persistent pain was uncommon . CONCLUSION: Septic sacroiliitis should be considered in peripartum patients who present with fever and severe localized pain . Medical management is usually curative, and without an adverse effect on pregnancy.

Urology, 2003 Jul, 62(1), 30 - 4
Counting leukocytes in expressed prostatic secretions from patients with chronic prostatitis/chronic pelvic pain syndrome; Krieger JN et al.; OBJECTIVES: The evaluation of WBCs in EPS is recommended for classifying patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) but no agreement has been reached on the optimal method . We sought to determine the relationship between the expressed prostatic secretions (EPS) leukocyte (WBC) count per high-power field (evaluated by a more quantitative wet mount method and the traditional gram-stained smear method used in clinical microbiology laboratories) and the EPS WBC concentration to determine whether quantitative methods are necessary for accurate patient classification . METHODS: EPS collected from 94 patients with CP/CPPS were evaluated by gram-stained smear, a standardized wet mount, and a hemocytometer method . RESULTS: The gram-stained smear detected EPS WBCs in 21 (22%) of 94 subjects compared with 78 (83%) by the standardized wet mount and 57 (60%) by the hemocytometer method . The gram-stained EPS WBC count correlated poorly with the WBC concentration by hemocytometer (R(2) = 0.051, P = 0.03) . Although the standardized EPS WBC count correlated better with the concentration by hemocytometer, the correlation coefficient remained low (R(2) = 0.244, P <0.0001) . CONCLUSIONS: The standardized wet mount proved superior to the gram-stained smear, but both methods lacked precision . Quantitative determination of the EPS WBC concentration by a counting chamber method proved to be the superior evaluation for research studies of CP/CPPS.

Anal Bioanal Chem, 2003 Aug, 376(7), 1062 - 8 Epub 2003 Jun 27.
A microfluidic biosensor based on nucleic acid sequence recognition; Kwakye S et al.; The development of a generic semi-disposable microfluidic biosensor for the highly sensitive detection of pathogens via their nucleic acid sequences is presented in this paper . Disposable microchannels with defined areas for capture and detection of target pathogen RNA sequence were created in polydimethylsiloxane (PDMS) and mounted onto a reusable polymethylmethacrylate (PMMA) stand . Two different DNA probes complementary to unique sequences on the target pathogen RNA serve as the biorecognition elements . For signal generation and amplification, one probe is coupled to dye encapsulated liposomes while the second probe is coupled to superparamagnetic beads for target immobilization . The probes hybridize to target RNA and the liposome-target-bead complex is subsequently captured on a magnet . The amount of liposomes captured correlates directly to the concentration of target sequence and is quantified using a fluorescence microscope . Dengue fever virus serotype 3 sequences and probes were used as a model analyte system to test the sensor . Probe binding and target capture conditions were optimized for sensitivity resulting in a detection limit of as little as 10 amol microL(-1) (10 pmol L(-1)) . Future biosensors will be designed to incorporate a mixer and substitute the fluorescence detection with an electrochemical detection technique to provide a truly portable microbiosensor system.

Curr Pain Headache Rep, 2003 Aug, 7(4), 297 - 302
Recent advances in temporal arteritis; Redillas C et al.; Temporal arteritis was first described in the late nineteenth century . Despite considerable progress in understanding the disease, its rarity in the young and in those who are not of Scandinavian ethnicity remains unexplained . Microbiologic agents and immunologic mechanisms have been implicated as causative factors . Although steroids remain the drug of choice, the use of other immunologic therapies has been proposed . This paper reviews the disease's history, probable etiologies, clinical manifestations, and its diagnostic and treatment challenges.

Pediatr Int, 2003 Jun, 45(3), 311 - 3
Incidence of giardiasis among siblings in Turkey; Balcioglu IC et al.; BACKGROUND: Giardiasis, a common infection among children, is caused by a flagellated protozoan called Giardia lamblia . It is well known to be contagious in common living places . This is an epidemiologic study investigating the incidence of giardiasis among the siblings of patients with giardiasis living in the same household . METHODS: Stool samples of 50 patients with a G . lamblia positive sibling in the same household, and 50 patients with a G . lamblia negative sibling in the same household, were examined for giardiasis by saline-Lugol, formalin-ethyl acetate concentration and trichrome staining methods . Other causes of diarrhea were excluded by microbiologic laboratory tests . RESULTS: Thirty-eight per cent of siblings of G . lamblia positive patients and 8% of siblings of G . lamblia negative children were found to be positive for G . lamblia cysts and/or trophozoites . The differences between the groups were statistically significant (P < 0.001) . CONCLUSION: The results suggested that the examination of G . lamblia among the siblings of patients with giardiasis both in the same age group and living in the same household should be considered.

Ann Emerg Med, 2003 Jul, 42(1), 9 - 13
Stability and microbiology of inhalant N-acetylcysteine used as an intravenous solution for the treatment of acetaminophen poisoning; Dribben WH et al.; STUDY OBJECTIVE: Intravenous N-acetylcysteine has been used as an antidote for acetaminophen poisoning for more than 25 years in Europe and Canada . In the United States, only the oral administration of N-acetylcysteine is approved by the US Food and Drug Administration . Many physicians routinely use the inhalant preparation as an intravenous formulation; however, no stability, microbiology, or pyrogen studies have been performed . In this study, we evaluate the stability and microbiology of inhalational N-acetylcysteine compounded as an intravenous formulation . METHODS: A total of 8 N-acetylcysteine solutions (solution A through H) were prepared by injecting 150 mL of 20% solution through a 22-microm filter to 1 L of 5% dextrose (D(5)W; 2.6% solution) . Solutions A through C were prepared at ambient conditions (25 degrees C {77 degrees F}, 65% relative humidity), and solution D was prepared at accelerated conditions (40 degrees C {104 degrees F}, 75% relative humidity) for stability testing . The assays were performed by means of high-performance liquid chromatography at 0, 4, 8, 12, 24, 36, 48, 60, and 72 hours according to US Pharmacopeia XXIV methodology . Solutions E through G were assessed for bacterial growth, and solution H underwent pyrogen testing by using a Limulus amebocyte lysate method . RESULTS: Solutions A through C remained stable for at least 60 hours (<10% decomposition), but at 72 hours, there was a 10.3%, 14.9%, and 13.4% degradation, respectively . Under accelerated conditions (solution D), stability lasted for more than 72 hours . Solutions E through G remained free from bacterial growth at 72 hours, and solution H tested negative for endotoxins-pyrogens . CONCLUSION: Inhalational N-acetylcysteine prepared as an intravenous solution meets US Pharmacopeia standards for stability up to 60 hours and is free from bacteria and their byproducts, offering a viable alternative to the traditional use of oral N-acetylcysteine.

Anadolu Kardiyol Derg, 2003 Jun, 3(2), 137 - 43
Surgical treatment of isolated cardiac echinococciasis: report of five cases; Djoshibaev S et al.; OBJECTIVE: Cardiac echinococcosis is a rare pathology and in about half of cases it is not accompanied by other organs involvement . We report on our experience of surgical treatment of isolated cardiac echinococcosis (ICE) . METHODS: Among 5 patients with ICE operated in our clinic 1 had pericardial and epicardial involvement, 1 patient had the single cyst in the apicolateral wall of the left ventricle and 3 patients had singular cysts of interventricular septum . All patients underwent chest X-Ray, transthoracic echocardiography, and ultrasound examinations of the internal organs, hemagglutinin tests and microbiologic evaluation of operative specimens . RESULTS: Two operations were performed on a beating heart and three - using cardiopulmonary bypass . In cases of intramyocardial localizations the cyst enucleation and closure of residual cavities by plication were performed and in a case of peri- and epicardial involvement the extirpation of cysts and partial pericardiectomy were carried out . All patients postoperatively received therapy with albendazole . No intra- and postoperative complications were observed . Control echocardiography did not reveal any disturbances of myocardial performance . Two years follow-up showed absence of cyst recurrence . CONCLUSION: The prevailed localization of echinococci cysts in ICE is the myocardium of the LV and IVST, places with the highest myocardial mass and good perfusion . Ideal echinococcectomy is an effective surgical treatment of isolated cardiac echinococciasis.

Rev Iberoam Micol, 2003 Mar, 20(1), 24 - 8
{Acute pulmonary histoplasmosis in a Spanish traveller to Nicaragua: an imported disease case}; Flor A et al.; Pulmonary histoplasmosis is a rare disease in Spain . Moreover, it is difficult to diagnose due to unspecific clinical and radiological symptoms . The isolation of the fungus is essential for a proper diagnosis . Nevertheless, it is very difficult to identify the fungus itself in respiratory stains and we usually need invasive techniques . For all these reasons and taking into account the increase in journeys and immigration, we believe that it is probably underdiagnosed in our country . We report a case of acute pulmonary histoplasmosis in a Spanish traveller and emphasize the importance of the anamnesis and the value of the microbiologist's experience to obtain the diagnosis.

Eur J Clin Nutr, 2003 Jul, 57(7), 816 - 8
n-3 fatty acids in human fat tissue aspirates are stable for up to 6 y; Katan MB et al.; OBJECTIVES: The content of n-3 (omega-3) polyunsaturated fatty acids in fat tissue is a valid indicator of their long-term consumption . We studied the stability of n-3 fatty acids in human subcutaneous fat microbiopsies after 6 and 11 y of storage . DESIGN: Microbiopsies were taken from a lump of human adipose tissue and stored at +20 and -80 degrees C . SETTING: Laboratory study . RESULTS: After 5.6 y at -80 degrees C the proportion of six out of seven highly polyunsaturated fatty acids varied between 91 and 102% (mean 97%) of their baseline values . Storage at +20 degrees C yielded recoveries between 82 and 105% . After 11 y at -80 degrees C the proportions in the original lump of tissue ranged from 88 to 101% (mean 94%) . CONCLUSION: n-3 fatty acids in stored fat tissue aspirates are stable for 6-11 y, and are suitable markers of baseline diet in long-term epidemiological studies . SPONSORSHIP: Wageningen Centre for Food Sciences.

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