Microbial

Science, 2005 Jan 14, 307(5707), 217 - 8
Cell signaling . Stat acetylation--a key facet of cytokine signaling?
O'Shea JJ, Kanno Y, Chen X, Levy DE.
National Institute of Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

Clin Chem Lab Med, 2005 Jan, 43(1), 54 - 63
External failures as descriptor of quality in clinical microbiology laboratory services; Saarela U et al.; This study employed customer feedback surveys to assess the quality of the laboratory services provided by an accredited clinical microbiology laboratory . The purpose of the study was to classify external failures and determine the quantity of such errors, and to establish how much time was spent processing and correcting these errors, together with the costs arising . The study indicates that the quality of laboratory test results can be regarded as good based on the accuracy of the result . Customer needs were also well taken into account in practice because of the good quality of the service . However, improvements are required in providing customers with test results without delay . The cost of processing and correcting such errors was negligible . The use of a customer feedback system allows systematic monitoring of external failures . In the accredited laboratory under study, it is unnecessary to monitor continually the costs and working time involved in processing and correcting external failures . The most essential point is to monitor the effects of the quality measures taken to reduce the number of failures.

Clin Diagn Lab Immunol, 2005 Jan, 12(1), 122 - 4
Identification of another B-cell epitope in the type-specific region of equine herpesvirus 4 glycoprotein g; Maeda K et al.; Recently, a novel 12-mer B-cell epitope, MKNNPIYSEGSL, in the type-specific region of equine herpesvirus 1 (EHV-1) glycoprotein G (gG) was identified and used as an antigen for enzyme-linked immunosorbent assay (Maeda et al., J . Clin . Microbiol . 42:1095-1098, 2004) . Although our prototype strain, TH20p, possesses two repeat sequences containing the B-cell epitope, the EHV-4 NS80567 strain has two repeat sequences that are not identical . One repeat sequence stretch contained the B-cell epitope, while the other contained the 11-mer, MKNNPVYSESL (underlining indicates a different amino acid) . In this study, heterogeneity of the type-specific region was compared among Japanese EHV-4 isolates . The 11-mer peptide, MKNNPVYSESL, specifically reacted with sera from horses naturally infected with EHV-4 but not with sera from horses experimentally infected with EHV-4 TH20p . The 11-mer peptide may be another B-cell epitope in the type-specific region.

J Clin Periodontol, 2005 Jan, 32(1), 68 - 74
Evaluation of the relationship between smoking during pregnancy and subgingival microbiota; Buduneli N et al.; Buduneli N, Baylas H, Buduneli E, Turkoglu O, Dahlen G: Evaluation of the relationship between smoking during pregnancy and subgingival microbiota . J Clin Periodontol 2004; doi: 10.1111/j.1600-051X.2004.00633.x . (c) Blackwell Munksgaard, 2004 . Abstract Background: Numerous studies have shown that smoking negatively affects periodontal health . Hormonal changes, which occur during pregnancy have also been reported to have adverse effects on the periodontal tissues or indirectly through alterations in the subgingival bacterial flora . At present, no knowledge exists concerning possible effects of smoking on the composition of subgingival plaque in pregnancy . The purpose of the present study was to evaluate the effects of smoking during pregnancy on the subgingival plaque bacteria most commonly associated with periodontal disease . Methods: A total number of 181 women were examined within 72 h post-partum . Smoking status was recorded by means of a self-reported questionnaire and the study population was divided into three groups; non-smokers, light smokers, and heavy smokers . In each woman, two subgingival plaque samples were obtained from mesio- or disto-buccal aspect of randomly selected one molar and one incisor tooth by sterile paperpoints . Clinical periodontal recordings comprising presence of dental plaque, bleeding on probing (BOP), and probing pocket depth (PPD) were performed at six sites per each tooth at all teeth . Plaque samples were analysed by checkerboard DNA-DNA hybridization with respect to 12 bacterial species . In all analyses, the individual subject was the computational unit . Thus, mean values for all clinical parameters were calculated and bacterial scores from each individual sample were averaged . Statistical methods included chi(2) test, Kruskal-Wallis test and Mann-Whitney U-test . Results: Mean ages were similar in the study groups . Plaque, BOP and PPD recordings were lower in the heavy-smoker group, but the differences were not statistically significant (p>0.05) . The detection rates and bacterial loads of the specific subgingival bacteria exhibited no significant differences between the groups . No correlation could be found between smoking status and detection rates and bacterial loads of various bacterial species . Conclusion: The present findings suggest that smoking during pregnancy does not have a significant effect on the composition of subgingival plaque bacteria.

Infect Control Hosp Epidemiol, 2004 Dec, 25(12), 1050 - 5
Mycobacterium simae outbreak associated with a hospital water supply; Conger NG et al.; OBJECTIVE: Mycobacterium simiae is found primarily in the southwestern United States, Israel, and Cuba, with tap water as its suspected reservoir . Our institution saw an increase in M . simiae isolates in 2001 . An investigation into possible contaminated water sources was undertaken . DESIGN: Environmental cultures were performed from water taps in the microbiology laboratory, patient rooms, points in the flow of water to the hospital, and patients' homes . Patient and environmental M . simiae were compared by PFGE . SETTING: Military treatment facility in San Antonio, Texas . PATIENTS: All patients with cultures positive for M . simiae between January 2001 and April 2002 . Medical records were reviewed . RESULTS: M . simiae was recovered from water samples from the hospital, patients' home showers, and a well supplying the hospital . A single PFGE clone was predominant among water isolates (9 of 10) and available patient isolates (14 of 19) . There was an association between exposure to hospital water and pulmonary samples positive for the clonal M . simiae strain (P = .0018) . Only 3 of 22 culture-positive patients met criteria for M . simiae pulmonary disease . Of them, two had indistinguishable M . simiae strains from tap water to which they were routinely exposed . CONCLUSIONS: This represents an outbreak of M . simiae colonization with one nosocomial infection . It is only the second time that M . simiae has been recovered from hospital tap water and its first presentation in municipal water . This study raises issues about the need and feasibility of eliminating or avoiding exposure to M . simiae from tap water.

J Clin Microbiol, 2005 Jan, 43(1), 382 - 6
Aspergillus hyphae in infected tissue: evidence of physiologic adaptation and effect on culture recovery; Tarrand JJ et al.; Microbiologic cultures of fungi are routinely incubated at ambient temperatures in room air, and the rate of recovery of Aspergillus species from clinical specimens is poor . Failure of current culture methods to mimic the physiologic temperature and low-oxygen environment found in hypha-laden infected tissue may underlie this poor recovery . Experiments were performed to compare the recovery of Aspergillus spp . incubated at 35 degrees C in 6% O(2)-10% CO(2) with that at 25 degrees C in room air . The samples tested included Aspergillus-infected tissue specimens from a dog model and human autopsies, experimental anaerobically stressed Aspergillus inocula, and 10,062 consecutive clinical specimens . Culture at 35 degrees C in 6% O(2)-10% CO(2) significantly enhanced the recovery of Aspergillus spp . from the infected autopsy tissue samples . Incubation at 35 degrees C alone resulted in approximately 10-fold-improved culture recovery from the experimentally stressed hyphae, and the 6% O(2)-10% CO(2) atmosphere independently favored growth under temperature-matched conditions . Finally, incubation at 35 degrees C (in room air) improved the overall recovery of Aspergillus spp . from clinical specimens by 31% . Culture at 35 degrees C in a microaerobic atmosphere significantly enhances the recovery of Aspergillus spp . from various sources . Aspergillus hyphae growing in infected tissue appear to be adapted to the physiologic temperature and hypoxic milieu.

Epidemiol Mikrobiol Imunol, 2004 Nov, 53(4), 169 - 73
{External quality control system in medical microbiology and parasitology in the Czech Republic}; Slosarek M et al.; The External Quality Control System (EQAS) of laboratory activities in medical microbiology and parasitology was implemented in the Czech Republic in 1993 with coded sera samples for diagnosis of viral hepatitis and bacterial strains for identification distributed to first participating laboratories . The number of sample types reached 31 in 2003 and the number of participating laboratories rised from 79 in 1993 to 421 in 2003 . As many as 15.130 samples were distributed to the participating laboratories in 2003 . Currently, almost all microbiology and parasitology laboratories in the Czech Republic involved in examination of clinical material participate in the EQAS . Based on the 11-year experience gained with the EQAS in the Czech Republic, the following benefits were observed: higher accuracy of results in different tests, standardisation of methods and the use of most suitable test kits.

Clin Lab Sci, 2001 Winter, 14(1), 21 - 6
Predictive value of a senior comprehensive examination as to performance on a national certification examination; Wiggers TB et al.; OBJECTIVE: To determine what value a senior departmental comprehensive examination holds in predicting the future success of a student on the MT(ASCP) certification examination . DESIGN: Part 1: To evaluate the efficacy of the comprehensive examination, scores were obtained in the examination categories of hematology, clinical chemistry, immunohematology, and microbiology for all dinical laboratory science students who have graduated from the University of Mississippi Medical Center since 1993 . The data were analyzed to determine if a correlation exists between student performance on the senior comprehensive, and their future performance on the MT(ASCP) national certification examination . Part 2: To determine the extent to which a senior comprehensive examination was required for graduation at other university-based clinical laboratory science programs, a simple survey was e-mailed to members of the clinical laboratory science educators forum . SETTING: 2+2 university-based dinical laboratory science program PARTICIPANTS: Part 1: Previous graduates of the Clinical Laboratory Science Program at the University of Mississippi Medical Center since 1993 . Part 2: Program directors who are members of the Clinical Laboratory Sciences Educator's Forum . RESULTS: Part 1: Results indicated a distinct division between participants who scored higher than 74.36% (Group A) on the senior comprehensive examination, and those scoring below 74.36% (Group B) . In Group A, 100% of participants passed the MT(ASCP) national certification examination on the first attempt . Results were mixed for Group B . Part 2: The survey indicated that of the 40 respondents, most were similar to the University of Mississippi Medical Center Clinical Laboratory Science Program in that they require a comprehensive to be taken, that the grade received is part of another course grade, and that the examination is prepared using questions submitted by the faculty . CONCLUSIONS: Part 1: The senior departmental comprehensive examination is of value in predicting the future success of a student on the MT(ASCP) national certification examination . Part 2: Unlike the University of Mississippi Medical Center, 16 of the 40 respondents stated that passage of the comprehensive examination was a requirement for graduation . In those programs, the comprehensive was a major part of a course grade.

Vector Borne Zoonotic Dis, 2004 Fall, 4(3), 249 - 60
Update on spotted fever group Rickettsiae in South Africa; Pretorius AM et al.; Until very recently, Mediterranean spotted fever caused by Rickettsia conorii was the only spotted fever group (SFG) rickettsioses recognized in southern Africa . However, increasing medical awareness of tick-borne infections, together with the introduction of improved isolation methods and the availability of molecular techniques, have led to the identification of several new SFG rickettsioses in the region . African tick bite fever, caused by Rickettsia africae, is currently the most important of these new rickettsioses, affecting large numbers of international travellers each year, but infections due to Rickettsia aeschlimannii and Rickettsia mongolotimonae have also been recently encountered . In this review, we describe the current status of the epidemiology, microbiology, clinical presentation, diagnosis, treatment, and prevention of SFG rickettsioses in southern Africa.

Klin Khir, 2004 Oct, (10), 48 - 51
{Microbiocenosis of alimentary canal under the influence of the enteral tube feeding in patients with choledocholithiasis}; Impact of ventilator-associated pneumonia in patients with severe head injury; From the Critical Care and Emergency Department (M.D.R-F., J.M.F-C., S.R.L-N., F.M-C., M.A . M-S., J.I.S-O.) and the Supportive Research Unit (A.C.), University Hospital Virgen del Rocio, Seville, SpainBACKGROUND:: The impact of ventilator-associated pneumonia (VAP) on outcome seems to vary depending on the critically ill patients we analyze . Our objective, therefore, has been to evaluate the influence of VAP on the mortality and morbidity in patients with severe head injury (Glasgow Coma Scale score </= 8) . METHODS:: A prospective, matched, case-control study was conducted in our intensive care unit (ICU) for a 3-year period (1998-2000) . Seventy-two patients with severe head injury (HI) who developed VAP were matched with 72 patients with severe HI without VAP . The matching criteria were as follows: age (+/- 5 years); category of HI based on computed tomographic scanning; Acute Physiology and Chronic Health Evaluation II (+/- 4 points) score; Injury Severity Score (+/- 4 points); and duration of mechanical ventilation . VAP was diagnosed on the basis of quantitative microbiologic criteria . RESULTS:: Mortality did not differ significantly between cases and matched control subjects (15 {20.8%} vs . 11 {15.3%}, p = 0.54) . However, patients with VAP had a significantly longer duration of mechanical ventilation (median, 14 vs . 10 days; p = 0.015) and ICU stay (median, 21 vs . 15.5 days; p = 0.008) . The occurrence of multiple organ failure was also significantly more frequent among the case group (33.3% vs . 12.5%, p = 0.004) during the overall ICU stay . CONCLUSION:: VAP does not seem to be associated with a significantly increased risk of death in patients with severe HI, but it may be associated with greater morbidity during the ICU stay.

Ying Yong Sheng Tai Xue Bao, 2004 Oct, 15(10), 1953 - 7
{Degradation mechanism of Chinese fir plantation}; Chen L et al.; Chinese fir (Cunninghanzia lanceolata) is a fast-growing species native to China, and has a very important status in production of commercial timber . In response to the growing need for timber, pure Chinese fir plantations have been extensively replanted on the same site in successive rotations, which has resulted in serious problems of soil fertility degradation and productivity decline, and these problems have caused considerable attention from foresters, microbiologists, soil scientists and ecologists . They have also done a lot of work on these problems and studied the mechanism of productivity decline of replanted Chinese fir plantation . In this article, the author described the characteristics of productivity decline and soil degradation, and summed up the factors caused lower productivity, and analyzed the degradation mechanism of replanted Chinese fir plantation . In sum, productivity decline and soil physical and chemical characteristics deterioration were the main properties of replanted Chinese fir plantation . The factors summarized primarily as two kinds of reasons . One was the unreasonable management that was the edogenic factor, and another was the biotic self-characteristics of Chinese fir that was the intrinsic factor of poor establishment of replanted Chinese fir plantation . The combination of the edogenic and intrinsic factors reduced the productivity decline and soil degradation of the replanted Chinese fir plantation.

Rev Med Liege, 2004 Oct, 59(10), 570 - 6
{The return of pertussis}; Demaret P et al.; Pertussis (whooping cough) is a contagious respiratory tract bacterial infection due to Bordetella pertussis who makes global comeback despite routine vaccination . Bordetella pertussis produces a series of virulence factors that are involved in one of several steps of the pathogenesis of whooping cough (adhesins, toxins) . Unusual presentations are now most common . Clinical diagnosis of whooping cough is often difficult, and microbiologic evaluation is helpful . Treatment is based on antibiotherapy and sometimes intensive care management . Complications are most common in children younger than 6 months (death, malignant pertussis, respiratory and neurological complications) . Pertussis can be controlled by treatment of exposed people and by vaccination.

Obstet Gynecol Surv, 2005 Jan, 60(1), 26 - 28
Vulvovaginal Symptoms in Women With Bacterial Vaginosis; Klebanoff MA et al.; To investigate the symptoms among women with bacterial vaginosis, the authors conducted a longitudinal study comparing symptoms experienced by women with and without a diagnosis of bacterial vaginosis confirmed by Gram stain and Amsel clinical criteria . Subjects were recruited from women making routine healthcare visits to clinics in Birmingham, Alabama.Participating patients were interviewed intensively for information about lower genital tract symptoms, including vaginal wetness, vaginal discharge, vaginal odor, persistent vaginal itch, pain with urination, and abdominal or pelvic pain . All subjects underwent an initial clinical assessment with pelvic examination and lower genital tract microbiologic evaluation, as well as detailed questioning, which included demographic factors, obstetric and gynecologic history, dental symptoms and practices, feminine hygienic and health behaviors, sexual history and practices, history of genital tract infections or sexually transmitted diseases, alcohol and drug use, and psychosocial status.There were 2888 eligible patients who had Gram stain results available . These women were predominantly black, young, parous, unmarried, and of low income . Thirty-seven percent (1063 patients) were positive for bacterial vaginosis by Gram stain . Positivity was more common in black women and women who smoked and increased with decreasing general health status . Women who were married, of higher income, and less than 20 years of age were less likely to have bacterial vaginosis.Few of the total study population reported pelvic or abdominal pain . More women who were positive for bacterial vaginosis reported vaginal wetness and vaginal odor, and more women who were free of bacterial vaginosis reported dysuria and vaginal irritation . The differences were significant, but the numbers of women in either group who had symptoms were small . The most common symptom was vaginal odor, but only 25% of the patients with and 18% of those without bacterial vaginosis noticed vaginal odor . In all, 58% and 57% of the positive and negative patients, respectively, reported experiencing wetness, discharge, and/or vaginal odor.Among sexually active participants, those who were positive for bacterial vaginosis were more likely to have any odor or fishy odor with sexual intercourse, but less than 20% of either group reported these symptoms . When analyses were performed using Amsel criteria to define bacterial vaginosis, the pattern of reported symptoms was largely unchanged.Participants were asked to describe their usual vaginal odor to clarify their reports of symptoms . No one description was more common than another, but all descriptive terms were more frequent among patients with bacterial vaginosis . The characteristic "fishy" odor was reported by only 6% and 4% of the positive and negative patients, respectively.Thirty-seven percent of the women with no reported symptoms, 26.3% of those with vaginal irritation alone, and 19.4% of those with both irritation and discharge or wetness were positive for bacterial vaginosis . Women who reported vaginal irritation were less likely to have bacterial vaginosis no matter what other symptoms or odor they reported . Vaginal discharge did not indicate the presence of bacterial vaginosis when experienced alone or with other symptoms . Vaginal odor was the only symptom with even a moderate association with bacterial vaginosis.At clinical examination, 35% of patients who reported no discharge, and 49% of those who said they had severe discharge, were described by the examining nurse as having a thin, homogeneous discharge . Similarly, 36% of those reporting no vaginal odor, and 48% of those who felt they had severe vaginal odor, were positive for the "whiff" test.

BMC Ophthalmol . 2004 Dec 24;4(1):16 {Epub ahead of print}
Keratocyte loss in corneal infection through apoptosis: a histologic study of 59 cases; Vemuganti GK et al.; BACKGROUND: Keratocyte loss by apoptosis following epithelial debridement is a well-recognized entity . In a study of corneal buttons obtained from patients of corneal ulcer undergoing therapeutic keratoplasty, we observed loss of keratocytes in the normal appearing corneal stroma, surrounding the zone of inflammation . Based on these observations, we hypothesized that the cell loss in the inflammatory free zone of corneal stroma is by apoptosis that could possibly be a non-specific host response, independent of the nature of infectious agent . METHODS: To test our hypothesis, in this study, we performed Terminal deoxyribonucleotidyl transferase- mediated d-Uridine 5" triphosphate Nick End Labelling (TUNEL) staining on 59 corneal buttons from patients diagnosed as bacterial, fungal, viral and Acanthamoeba keratitis . The corneal sections were reviewed for morphologic changes in the epithelium, stroma; type, degree and depth of inflammation, loss of keratocytes in the surrounding stroma (posterior or peripheral) . TUNEL positivity was evaluated in the corneal sections, both in the zone of inflammation as well as the surrounding stroma . A correlation was attempted between the keratocyte loss, histologic, microbiologic and clinical features . RESULTS: The corneal tissues were from 59 patients aged between 16 years and 85 years (mean 46 years) and included fungal (22), viral (15), bacterial (14) and Acanthamoeba (8) keratitis . The morphological changes in corneal tissues noted were: epithelial ulceration (52, 88.1%), destruction of Bowman's layer (58, 99%), mild to moderate (28;47.5%) to severe inflammation (31;52.5%) . Morphologic evidence of disappearance or reduced number of keratocytic nuclei in the corneal stroma was noted in 49 (83%) cases; while the TUNEL positive brown cells were identified in all cases 53/54 (98%), including cases of fungal (19), bacterial (14), viral (13), and Acanthamoeba keratitis . TUNEL staining was located mostly in the deeper stroma and in few cases the peripheral stroma . TUNEL positivity was also noted with the polymorphonuclear infiltrates and in few epithelial cells (10 of 59, 17%) cases, more with viral infections (6/10; 60%) . CONCLUSIONS: We report apoptotic cell death of keratocytes in the corneal stroma in infectious keratitis, a phenomenon independent of type of infectious agent . The inflammatory cells in the zone of inflammation also show evidence of apoptotic cell death . It could be speculated that the infective process possibly triggers keratocyte loss of the surrounding stroma by apoptosis, which could possibly be a protective phenomenon . It also suggests that necrotic cell death and apoptotic cell deaths could occur simultaneously in infective conditions of the cornea.

Drug News Perspect, 1998 Jun, 11(5), 310 - 9
Medical mycology and current trends in antifungal chemotherapy; Fromtling RA; The Medical Mycological Society of the Americas (MMSA) and the American Society for Microbiology (ASM) are both premier scientific organizations which held their annual conferences in Atlanta, Georgia, May 17-21, 1998 . The MMSA scientific session focused on the role of animal models in drug development and on the immunology and molecular biology of the medically important fungi . The use of current molecular tools in identifying new fungal targets for the development of antifungal agents and for the study of fungal epidemiology was reviewed . At the ASM, several sessions were held which focused on the pathogenesis of fungal disease, the in vitro susceptibility testing of new (voriconazole, SCH-56592, LY-303366, metalloporphyrins) and established antifungal agents, the experimental in vivo activity of new antifungal drugs in various animal models of fungal disease and pediatric mycology; the clinical activity of fluconazole, itraconazole, liposomal nystatin and liposomal amphotericin B formulations was also reviewed . (c) 1998 Prous Science . All rights reserved.

Clin Infect Dis, 2005 Jan 1, 40(1), 123 - 6 Epub 2004 Dec 06.
Global Infectious Diseases and Epidemiology Network (GIDEON): a world wide Web-based program for diagnosis and informatics in infectious diseases; Edberg SC; The Global Infectious Diseases and Epidemiology Network (GIDEON) consists of 4 modules . The first is designed to generate a ranked differential diagnosis list for any infectious diseases scenario in any of 220 countries . The second follows the country-specific epidemiology of 337 individual diseases . The third presents a comprehensive encyclopedia of 308 generic anti-infective drugs and vaccines, including a listing of >9500 trade names . The fourth generates a ranked identification list based on the phenotype of bacteria, mycobacteria, and yeasts . The program performs well and serves as a useful paradigm for World Wide Web-based informatics . GIDEON is an eclectic program that can serve the needs of clinicians, epidemiologists, and microbiologists working in the fields of infectious diseases and geographic medicine.

Clin Infect Dis, 2005 Jan 1, 40(1), 114 - 22 Epub 2004 Dec 06.
A new paradigm for clinical investigation of infectious syndromes in older adults: assessment of functional status as a risk factor and outcome measure; High KP et al.; Adults aged >or=65 years comprise the fastest-growing segment of the United States population, and older adults experience greater morbidity and mortality due to infection than do young adults . Although age is well established as a risk factor for infection, most clinical investigations of infectious diseases in older adults focus on microbiology and on crude end points of clinical success, such as cure rates or death; however, they often fail to assess functional status, which is a critical variable in geriatric care . Functional status can be evaluated either as a risk factor for infectious disease or as an outcome of interest after specific interventions using well-validated instruments . This article outlines the currently available data that suggest an association between infection, immunity, and impaired functional status in elderly individuals, summarizes the instruments commonly used to determine specific aspects of functional status, and provides recommendations for a new paradigm in which clinical trials that involve older adults include assessment of functional status.

Oral Microbiol Immunol, 2005 Feb, 20(1), 56 - 9
Down-regulation of interleukin-1alpha-induced matrix metalloproteinase-13 expression via EP receptors by prostaglandin E in human periodontal ligament cells; Noguchi K et al.; Noguchi K, Ruwanpura SMPM, Yan M, Yoshida N, Ishikawa I . Down-regulation of interleukin-1alpha-induced matrix metalloproteinase-13 expression via EP(1) receptors by prostaglandin E(2) in human periodontal ligament cells . Oral Microbiol Immunol 2005: 20: 56-59 . (c) Blackwell Munksgaard, 2005 . In the present study, we investigated the effect of prostaglandin (PG) E(2) on matrix metalloproteinase (MMP)-13 production in human periodontal ligament cells stimulated with interleukin (IL)-1alpha . IL-1alpha enhanced both MMP-13 and PGE(2) production . Indomethacin, a nonselective cyclooxygenase inhibitor, and NS-398, a specific cyclooxygenase-2 (COX-2) inhibitor, significantly enhanced IL-1alpha-induced MMP-13 production in periodontal ligament cells, although both the agents completely inhibited IL-1alpha-induced PGE(2) production . Exogenous PGE(2) reduced IL-1alpha-induced MMP-13 mRNA and protein production in a dose-dependent manner . 17-phenyl-omega-trinor PGE(2), a selective EP(1) receptor agonist, mimicked the inhibitory effect of PGE(2) on IL-1alpha-induced MMP-13 mRNA and protein production . On the basis of these data, we suggest that COX-2-dependent PGE(2) down-regulates IL-1alpha-elicited MMP-13 production via EP(1) receptors in human periodontal ligament cells . PGE(2) may be involved in the regulation of destruction of extracellular matrix components in periodontal lesions.

Oral Microbiol Immunol, 2005 Feb, 20(1), 51 - 5
Recognition and phagocytosis of multiple periodontopathogenic bacteria by anti-Porphyromonas gingivalis heat-shock protein 60 antisera; Choi JI et al.; Choi J-I, Choi K-S, Yi N-N, Kim U-S, Choi J-S, Kim S-J . Recognition and phagocytosis of multiple periodontopathogenic bacteria by anti-Porphyromonas gingivalis heat-shock protein 60 antisera . Oral Microbiol Immunol 2005: 20: 51-55.(c) Blackwell Munksgaard, 2005 . The present study has been performed to evaluate Porphyromonas gingivalis heat shock protein (HSP) 60 as a candidate vaccine to protect against multiple putative periodontopathic bacteria . Mouse anti-P . gingivalis HSP antisera demonstrated the elevated IgG antibody titers against the multiple bacteria tested and cross-reacted with heat-induced bacterial proteins of the target bacteria . The antisera also demonstrated a significantly higher opsonophagocytosis function against all the target bacteria than the control sera (P < 0.01) . We concluded that P . gingivalis HSP 60 could potentially be developed as a vaccine against multiple periodontopathic bacteria.

Oral Microbiol Immunol, 2005 Feb, 20(1), 47 - 50
Retinoic acid-inducible gene-I is induced by interleukin-1beta in cultured human gingival fibroblasts; Sakaki H et al.; Sakaki H, Imaizumi T, Matsumiya T, Kusumi A, Nakagawa H, Kubota K, Nishi N, Nakamura T, Hirashima M, Satoh K, Kimura H . Retinoic acid-inducible gene-I is induced by interleukin-1beta in cultured human gingival fibroblasts . Oral Microbiol Immunol 2005: 20: 47-50.(c) Blackwell Munksgaard, 2005 . Retinoic acid-inducible gene-I (RIG-I) is a member of the DExH box family protein, and details of its biological function are not known . We have studied the mechanism of the interleukin-1beta (IL-1beta)-induced RIG-I expression in human gingival fibroblasts in culture . We also addressed the possibility of enhanced expression of COX-2, RANTES and galectin-9 in fibroblasts overexpressed RIG-I . We stimulated cultured human gingival fibroblasts with IL-1beta and examined the expression of RIG-I mRNA and protein by reverse transcriptase-mediated polymerase chain reaction and Western blot analysis . The effect of cycloheximide, a protein synthesis inhibitor, on the IL-1beta-induced expression of RIG-I was examined . The expression of COX-2, RANTES, galectin-9 and monocyte chemoattractant protein-1 in gingival fibroblasts transfected with RIG-I cDNA was also examined . IL-1beta stimulated the expressions of mRNA and protein for RIG-I, in cultured fibroblasts, in a time- and concentration-dependent manner . Cycloheximide did not suppress the IL-1beta-induced RIG-I expression . Introduction of RIG-I cDNA into fibroblasts resulted in enhanced expression of COX-2 mRNA, and slightly enhanced the expression of mRNA for RANTES and galectin-9 . In contrast, RIG-I overexpression did not alter the level of mRNA for monocyte chemoattractant protein-1 . We conclude that IL-1beta stimulates RIG-I expression in human gingival fibroblasts.

Oral Microbiol Immunol, 2005 Feb, 20(1), 25 - 30
Immunohistological analysis of Tannerella forsythia-induced lesions in a murine model; Gosling PT et al.; Gosling PT, Gemmell E, Carter CL, Bird PS, Seymour GJ . Immunohistological analysis of Tannerella forsythia-induced lesions in a murine model . Oral Microbiol Immunol 2005: 20: 25-30.(c) Blackwell Munksgaard, 2005 . Tannerella forsythia has been implicated as a defined periodontal pathogen . In the present study a mouse model was used to determine the phenotype of leukocytes in the lesions induced by subcutaneous injections of either live (group A) or nonviable (group B) T . forsythia . Control mice (group C) received the vehicle only . Lesions were excised at days 1, 2, 4, and 7 . An avidin-biotin immunoperoxidase method was used to stain infiltrating CD4(+) and CD8(+) T cells, CD14(+) macrophages, CD19(+) B cells, and neutrophils . Hematoxylin and eosin sections demonstrated lesions with central necrotic cores surrounded by neutrophils, macrophages and lymphocytes in both group A and group B mice . Lesions from control mice exhibited no or only occasional solitary leukocytes . In both groups A and B, neutrophils were the dominant leukocyte in the lesion 1 day after injection, the numbers decreasing over the 7-day experimental period . There was a relatively low mean percent of CD4(+) and CD8(+) T cells in the lesions and, whereas the percent of CD8(+) T cells remained constant, there was a significant increase in the percent of CD4(+) T cells at day 7 . This increase was more evident in group A mice . The mean percent of CD14(+) macrophages and CD19(+) B cells remained low over the experimental period, although there was a significantly higher mean percent of CD19(+) B cells at day 1 . In conclusion, the results showed that immunization of mice with live T . forsythia induced a stronger immune response than nonviable organisms . The inflammatory response presented as a nonspecific immune response with evidence of an adaptive (T-cell) response by day 7 . Unlike Porphyromonas gingivalis, there was no inhibition of neutrophil migration.

Arch Med Res, 2004 Sep-Oct, 35(5), 435 - 41
Extrapulmonary tuberculosis: clinical and epidemiologic spectrum of 636 cases; Ilgazli A et al.; BACKGROUND: The aim of this study was to evaluate the percentage and characteristics of extrapulmonary tuberculosis (EPTB) cases in a Turkish industrial city, Kocaeli (population: 1,203,335) and to illustrate its extent as a serious health problem for this city and country . METHODS: We investigated the results of microbiologic, radiologic, and histopathologic findings of patients with extrapulmonary tuberculosis, retrospectively, who were admitted to four Tuberculous Dispensaries between 1996 and 2000 . RESULTS: Six hundred thirty six cases were diagnosed with EPTB . Three hundred forty five were males (54.2%) and 291, females (45.8%) . Mean age of patient cases was 22.5+/-17.1 years (range, 1-86 years); 41.4% of cases were <15 years of age and 30.9%, between 20 and 39 years of age . Contact history with tuberculosis cases was determined in 242 cases (38%); of these, 194 were <15 years of age (80.2%) . The most common form of EPTB was observed to be lymph node tuberculosis (56.3%); of these cases, 256 (71.5%) had involvement in intrathoracic, 92 (25.7%) in cervical, and 10 (2.8%) in axillary lymph nodes . The second most frequent extrapulmonary form was pleural tuberculosis (31.1%) . EPTB was diagnosed by histopathologic methods in 229 patients (36.0%) and by microbiologic methods in 27 (4.2%); tuberculin skin test was significantly positive (>10 mm) in 95.5% of patients . When cases were classified according to severity, 87 cases (13.7%) were found severe and 549 (86.3%) as less severe EPTB . Five hundred sixty one cases (88.2%) lived in urban areas and 75 (11.8%), in rural areas . CONCLUSIONS: EPTB is very common in early adulthood in the Kocaeli region, with lymph nodes the most common localization.

Mol Biochem Parasitol, 2005 Jan, 139(1), 99 - 106
Multiple replication origins within the inverted repeat region of the Plasmodium falciparum apicoplast genome are differentially activated; Singh D et al.; The 35kb circular genome (plDNA) of the Plasmodium falciparum apicoplast replicates by the bidirectional ori/D-loop mechanism . PlDNA replication was previously shown to initiate within the inverted repeat (IR) region of the apicoplast genome {Williamson DH, Preiser PR, Moore PW, McCready S, Strath M, Wilson RJM (Iain) . The plastid DNA of the malaria parasite Plasmodium falciparum is replicated by two mechanisms . Mol Microbiol 2002;45:533-42; Singh D, Chaubey S, Habib S . Replication of the Plasmodium falciparum apicoplast DNA initiates within the inverted repeat region . Mol Biochem Parasitol 2003;126:9-14.} and the presence of at least two ori within each segment of the IR was postulated . Using 5' end-labelled nascent DNA as probe, we now demonstrate the utilization of several putative ori located within the IR for plDNA replication . Quantitation of signals obtained for different segments of the IR as well as determination of the number of molecules emanating from two ori regions by competitive PCR analysis indicated differential strengths of ori during plDNA replication prior to schizogony.

Springer Semin Immunopathol . 2004 Dec 18; {Epub ahead of print}
Surveillance B lymphocytes and mucosal immunoregulation; Velazquez P et al.; Mucosal lymphocyte homeostasis involves the dynamic interaction of enteric microbiota, the intestinal host epithelium, and the mucosal immune system . Dysregulation of mucosal lymphocyte homeostasis results in a variety of intestinal disorders, notably inflammatory bowel diseases like ulcerative colitis and Crohn's disease . One key cellular component regulating homeostasis are B lymphocytes that reside in gut-associated lymphoid tissue . This compartment includes Peyer's patches, isolated lymphoid follicles, lamina propria, and mesenteric lymph nodes . Recent data have pointed to two new and exciting aspects of B cells in the gut . First, there has been progress on identification and functional analysis of abundant isolated lymphoid follicle B cells that are key mediators of IgA genesis . Second, several groups have now clarified the functional identification and characterization of immunoregulatory B cells in the gut . This review examines the novel aspects of these B cells, and examines how each plays a role in mediating mucosal homeostasis in this bacteria-laden compartment.

Nucleic Acids Res, 2005 Jan 1, 33 Database Issue, D321 - 4
Metagrowth: a new resource for the building of metabolic hypotheses in microbiology; Ogata H et al.; Metagrowth is a new type of knowledge base developed to guide the experimental studies of culture conditions of obligate parasitic bacteria . We have gathered biological evidences giving possible clues to the development of the axenic (i.e . 'cell-free') growth of obligate parasites from various sources including published literature, genomic sequence information, metabolic databases and transporter databases . The database entries are composed of those evidences and specific hypotheses derived from them . Currently, 200 entries are available for Rickettsia prowazekii, Rickettsia conorii, Tropheryma whipplei, Treponema pallidum, Mycobacterium tuberculosis and Coxiella burnetii . The web interface of Metagrowth helps users to design new axenic culture media eventually suitable for those bacteria . Metagrowth is accessible at http://igs-server.cnrs-mrs.fr/axenic/.

Clin Microbiol Infect, 2004 Dec, 10(12), 1108 - 32
Guidelines for the diagnosis of tick-borne bacterial diseases in Europe; Brouqui P et al.; Ticks are obligate haematophagous acarines that parasitise every class of vertebrate (including man) and have a worldwide distribution . An increasing awareness of tick-borne diseases among clinicians and scientific researchers has led to the recent description of a number of emerging tick-borne bacterial diseases . Since the identification of Borrelia burgdorferi as the agent of Lyme disease in 1982, 11 tick-borne human bacterial pathogens have been described in Europe . Aetiological diagnosis of tick-transmitted diseases is often difficult and relies on specialised laboratories using very specific tools . Interpretation of laboratory data is very important in order to establish the diagnosis . These guidelines aim to help clinicians and microbiologists in diagnosing infection transmitted by tick bites and to provide the scientific and medical community with a better understanding of these infectious diseases.

Mil Med, 2004 Nov, 169(11), 863 - 5
Bullis fever: a vanished infection of unknown etiology; Murray CK et al.; Camp Bullis, Texas, is an active training facility for the U.S . Army and Air Force with a storied history dating back to the late 19th century . In the early 1940s, an epidemic of a seasonal tick-borne rickettsial-like illness occurred at Camp Bullis; the last case was reported in 1947 . To date, the etiology of this disease has remained elusive . In an attempt to retrospectively identify the causal pathogen, we surveyed Camp Bullis for the presumed tick vector with intent to screen molecularly for Rickettsia and Ehrlichia . However, no ticks were recovered from primary dragging attempts in the spring or from harvested deer in the fall . Moreover, pathologic and microbiologic materials obtained during the epidemic are no longer extant, making them unavailable for analysis . In this study, we review potential circumstances that impact emerging and, in this case, vanishing infections . The etiology of Bullis fever will probably remain undetermined, and this once-emerging infection may have vanished into history . However, given Camp Bullis' status as an active medical training site, awareness of and surveillance for the disease should continue.

Ann Dermatol Venereol, 2004 Nov, 131(11), 979 - 83
{Fahr syndrome associated to hypoparathyroidy, revealed by dermatologic manifestation}; Khadir K et al.; BACKGROUND: Fahr syndrome is defined by the presence at the brain base of strio-pallido-notched non-arteriosclerotic bilateral and symmetric calcifications . We report an observation of Fahr syndrome revealed by a growing pyodermitis in direct relation with hypocalcemia . OBSERVATION: A 17 year-old girl was admitted to the Dermatology Department for a growing pyodermitis of the large folds . The patient was known to be epileptic and had undergone surgery for a left eye cataract a year before . Microbiologic and mycologic examinations of the pus and of the cutaneous fragments were negative . During hospitalization, the patient had an attack of tetany, a laryngeal spasm and a generalized erythema thrust studded with pustules . The phosphocalcic assessment showed severe hypocalcemia and hyperphosphoremia . Parathormone serum rate was decreased . The brain tomodensitometry conveyed bilateral calcifications of the central gray nucleus . The diagnosis of a growing amicrobial pustulosis secondary to an idiopathic hypoparathyroidism and responsible for the Fahr syndrome was maintained . Treatment was composed of calcium (2 g/day) and vitamin D3 . The evolution was favorable with a relapse at 2 years . DISCUSSION: We report an idiopathic hypoparathyroidism associated with Fahr syndrome conveyed by growing amicrobial pustulosis . We discuss the dermatologic manifestations of Fahr syndrome associated with hypoparathyroidism and the physiopathologic mechanisms of the dermatologic lesions during hypocalcemia.

BMC Infect Dis . 2004 Dec 15;4(1):59 {Epub ahead of print}
Third generation cephalosporin use in a tertiary hospital in Port of Spain, Trinidad: need for an antibiotic policy; Pinto Pereira LM et al.; BACKGROUND: Tertiary care hospitals are a potential source for development and spread of bacterial resistance being in the loop to receive outpatients and referrals from community nursing homes and hospitals . The liberal use of third-generation cephalosporins (3GCs) in these hospitals has been associated with the emergence of extended-spectrum beta- lactamases (ESBLs) presenting concerns for bacterial resistance in therapeutics . We studied the 3GC utilization in a tertiary care teaching hospital, in warded patients (medical, surgical, gynaecology, orthopedic) prescribed these drugs . METHODS: Clinical data of patients (at or above 13 years) admitted to the General Hospital, Port of Spain (POSGH) from January to June 2000, and who had received 3GCs based on the Pharmacy records were studied . The Sanford Antibiotic Guide 2000, was used to determine appropriateness of therapy . The agency which procures drugs for the Ministry of Health supplied the cost of drugs . RESULTS: The prevalence rate of use of 3GCs was 9.5 per 1000 admissions and was higher in surgical and gynecological admissions (21/1000) compared with medical and orthopedic (8 /1000) services (p<0.05) . Ceftriaxone was the most frequently used 3GC . Sixty-nine (36%) patients without clinical evidence of infection received 3Gcs and prescribing was based on therapeutic recommendations in 4% of patients . At least 62% of all prescriptions were inappropriate with significant associations for patients from gynaecology (p<0.003), empirical prescribing (p<0.48), patients with undetermined infection sites (p <0.007), and for single drug use compared with multiple antibiotics (p <0.001) . Treatment was twice as costly when prescribing was inappropriate CONCLUSIONS: There is extensive inappropriate 3GC utilization in tertiary care in Trinidad . We recommend hospital laboratories undertake continuous surveillance of antibiotic resistance patterns so that appropriate changes in prescribing guidelines can be developed and implemented . Guidelines for rational antibiotic use though developed have not been re-visited or encouraged, suggesting urgent antibiotic review of the hospital formulary and instituting an infection control team . Monitoring antibiotic use with microbiology laboratory support can promote rational drug utilization, cut costs, halt inappropriate 3GC prescribing, and delay the emergence of resistant organisms . An ongoing antibiotic peer audit is suggested.

Mikrobiologiia, 2004 Sep-Oct, 73(5), 674 - 86
{Mycology at the Institute of Microbiology, Russian Academy of Sciences: history and prospects for the future}
{History of the study of biodiversity of photosynthesizing bacteria}
{No authors listed}

The tendencies in the study of anoxygenic photosynthesizing bacteria (APB) are considered in the review in the historical aspect, since APB discovery till present day . The contribution made by the researchers of the Institute of Microbiology, Russian Academy of Sciences, to the study of the phylogeny, ecology, and morphophysiological diversity of APB is noted . At present, the molecular-biological approaches play a decisive role in ecology and taxonomy . The most important task at the modem stage of the development of microbiology is to maintain the continuity of the historically formed classical approaches in the study of APB diversity.

Acta Odontol Latinoam, 2004, 17(1-2), 23 - 7
Long-term sterility of orthodontic-surgical appliances; Brusca MI et al.; The aim of this study was to evaluate the long-term sterility of new dental appliances according to the non rigid wrapping employed and assess the effectiveness of sterilization in a steam autoclave at 134 degrees for 20 minutes using physical, chemical, and biological indicators . All the experimental (E) samples and the control samples (C) were assigned to one of three groups according to the type of packaging: paper bag (E1), paper/plastic pouch (E2), nylon tubing bag (E3) . Each bag contained standardized orthodontic wires and brackets and sterility indicators . The samples were evaluated at the following experimental times: immediately, and 6, 12, 24 and 30 months post-sterilization . The samples were analyzed under aerobic and anaerobic conditions in keeping with the protocol currently in use at the Department of Microbiology, School of Dentistry, University of Buenos Aires . The group of control, non-sterilized samples (C1, C2, C3) were analyzed prior to the onset of the study, and were found to be contaminated . None of the sterilized samples in any of the three experimental groups evidenced contamination at any of the experimental times . The results showed that, under the present conditions, the packages and orthodontic appliances remained sterile for 30 months . These results show the importance of controlling sterility and the storage conditions over time for all the orthodontic/surgical appliances used in invasive treatments.

Bone Marrow Transplant . 2004 Dec 06; {Epub ahead of print}
What is the clinical significance of positive blood cultures with Aspergillus sp in hematopoietic stem cell transplant recipients? A 23 year experience; Simoneau E et al.; Summary:Hematopoietic stem cell (HSC) transplantation is the most frequent underlying predisposing condition to invasive aspergillosis . However, the significance of positive blood culture with Aspergillus sp in this particular population remains uncertain . We retrospectively reviewed all blood cultures performed in 1453 patients who received HSC transplant at our institution between 1980 and 2002 . We identified 19 patients with positive blood cultures with Aspergillus sp . Only one of these patients had clinical, histologic or microbiologic evidence of invasive aspergillosis . Thus, even in a population at highest risk for invasive aspergillosis, positive blood cultures with Aspergillus sp remain unusual, and cannot be readily associated with invasive aspergillosis . A case by case assessment by treating physicians of the clinical and radiologic parameters should be systematically made to establish the significance of aspergillemia . Single bottle positivity, obtained with the lysis-centrifugation blood culture system, is a common indicator of pseudoaspergillemia.Bone Marrow Transplantation advance online publication, 6 December 2004; doi:10.1038/sj.bmt.1704793.

Med Oral Patol Oral Cir Bucal, 2004, 9 Suppl, 114 - 9; 108-14
Necrotising periodontal diseases; Bermejo-Fenoll A et al.; Necrotizing gingivitis (NG) or necrotising ulcerative gingivitis (NUG) is considered to be an acute opportunistic gingival infection caused by bacterial plaque . It appears more frequently in undernurished children and young adults as well as patients with immunodeficiency . In its pathogenesis, there are factors related to the oral microbiota with invasion processes on the one hand, and on the other hand, factors associated with the host, such as signs of capillary and immunological disorders as well as undernurishment . The disease is characterized by pain, bleeding and papillary necrosis with tendency to relapse . Diagnosis is made by a simple clinical examination . However, complementary tests ought to be performed in order to eliminate the possibility of illnesses systemic or immunodeficiency . Early and sustained treatment is strongly recommended . Lesions of the gums (craters in the interdental papillae) as an aftermath of the disease is a possibility, or if there is necrotizing periodontitis there will be loss of attachment tissue.

Med Oral Patol Oral Cir Bucal, 2004 Nov-Dec, 9(5), 467 - 70; 464-7
Fine-needle aspiration cytology in the diagnosis of cervicofacial actinomycosis: report of 15 cases; Custal-Teixidor M et al.; OBJECTIVES: Actinomycosis is quite an infrequent bacterial infection nowadays . However it can be considered in cases with a persistent cervicofacial disease . Although it is a bacterial infection, microbiologic cultures are frequently not diagnoses, therefore histopathologic studies and image studies are essential . Our interest is to explain our experience with cervicofacial actinomycosis; the clinical behaviour, evolution and treatment, always assisted by their elected diagnostic technique: the FNAC . STUDY DESIGN: In the last 16 years, 15 patients have been diagnosed with cervicofacial actinomycosis by FNAC, treated by Maxillofacial, Internal Medicine and Paediatrics units . Clinical course, evolution, anatomical space situation, antibiotic treatment, and surgical treatment have been studied . RESULTS AND CONCLUSIONS: The fine-needle aspiration cytology (FNAC) is an easy, safe and rapid method, with a high effect, that has made the final diagnosis in 15 cases in our Hospital . All the patients have had a good clinical evolution, only in one case did we need a new treatment for recidive . In all the cases treatment has been definitive . Our interest is to explain our experience in the treatment of cervicofacial actinomycosis, its clinical presentation and evolution, together with its elected method of diagnosis, FNAC.

J Bacteriol, 2004 Dec, 186(24), 8295 - 300
MazF-mediated cell death in Escherichia coli: a point of no return; Amitai S et al.; mazEF is a stress-induced toxin-antitoxin module, located on the chromosome of Escherichia coli, that we have previously described to be responsible for programmed cell death in E . coli . mazF specifies a stable toxin, and mazE specifies a labile antitoxin . Recently, it was reported that inhibition of translation and cell growth by ectopic overexpression of the toxin MazF can be reversed by the action of the antitoxin MazE ectopically overexpressed at a later time . Based on these results, it was suggested that rather than inducing cell death, mazF induces a state of reversible bacteriostasis (K . Pederson, S . K . Christensen, and K . Gerdes, Mol . Microbiol . 45:501-510, 2002) . Using a similar ectopic overexpression system, we show here that overexpression of MazE could reverse MazF lethality only over a short window of time . The size of that window depended on the nature of the medium in which MazF was overexpressed . Thus, we found "a point of no return," which occurred sooner in minimal M9 medium than it did in the rich Luria-Bertani medium . We also describe a state in which the effect of MazF on translation could be separated from its effect on cell death: MazE overproduction could completely reverse the inhibitory effect of MazF on translation, while not affecting the bacteriocidic effect of MazF at all . Our results reported here support our view that the mazEF module mediates cell death and is part of a programmed cell death network.

Biomaterials, 2005 May, 26(14), 2095 - 104
Efficacy of antibiotics-loaded interpenetrating network (IPNs) hydrogel based on poly(acrylic acid) and gelatin for treatment of experimental osteomyelitis: in vivo study; Changez M et al.; The safety and efficacy of gentamycin sulphate (GS)- or vancomycin hydrochloride (VCl)-loaded polymer devices based on poly(acrylic acid) and gelatin crosslinked selectively using 0.3mol % N,N'-methylene bisacrylamide and 1wt% glutaraldehyde were evaluated by varying the drug concentration onto the devices . The placebo and drug-loaded device of AxGx (acrylic acid:gelatin: 1:1 w/w) were employed for the treatment of experimental osteomyelitis in rabbit . Rabbits were categorized into four groups . Twelve rabbits in each group were treated with 12+/-1mg of AxGx-1a (22% w/w GS), 12+/-1mg of AxGx-1b (44% w/w GS), 16+/-1mg of AxGx-1b (44% w/w GS) and 16+/-1mg of AxGx-1c (44% w/w VCl) . The drug concentration was measured following implantation in the adjacent tissue of femoral cavity, and serum . In femoral cavity maximum drug concentration was found on the 7th day with all the four types of devices . No drug was found after 21 days, at the local site with devices AxGx-1a and AxGx-1b (12+/-1mg), whereas it was detected after 6 weeks with 16+/-1mg device (44% w/w GS or VCl) . Macroscopic evaluation after treatment revealed that swelling, redness, local warmth and drainage decreased depending upon the drug loading of the implants . Sequential radiographs, histology, microbiologic assay and scanning electron micrography demonstrated devices AxGx-1b and AxGx-1c (16+/-1mg of 44% w/w drug loading) to be the most suitable device, which heals the infection after 6 weeks of treatment . No significant difference (p>0.05) in the rate of healing was observed between GS- and VCl-loaded devices . None of the implant showed toxic level of drug in serum at any given time.

Am J Dent, 2004 Oct, 17(5), 351 - 3
In vivo enamel fluoride uptake after use of fluoride products; Campus G et al.; PURPOSE: To evaluate fluoride uptake in enamel after use of products containing different fluoride components of distinct oral hygiene regimen in vivo . METHODS: 84 healthy subjects (24 females and 60 males aged 19-26 years, mean 22.2) recruited from the students attending the courses of the Medical Faculty of the University of Sassari, Italy, were randomly assigned to one of four treatments groups: A: treated with a dentifrice containing sodium monofluorophosphate (1250 ppm F-); B: treated with dentifrice with 1250 ppm F- from AmF; C: treated with an AmF toothpaste (1250 ppm F-) and a mouth rinse with 250 ppm F- from AmF; or D: treated with a NaMFP toothpaste (1250 ppm F-) plus 1250 ppm NaMFP varnish . Acid-etched enamel microbiopsies on the vestibular surface of the first maxillary premolar were collected at baseline (to), after 20 days of test product use (t1) and after 24 hours without treatment (t2) . Fluoride concentration was measured using an ion-specific electrode . All measurements were made in triplicate and analyzed statistically using two-way factorial ANOVA for independent samples standard weighted-means analysis . RESULTS: The fluoride uptake in the enamel shows a considerable variation between the groups . Significant differences were found both within all products in terms of fluoride concentration in enamel for (P < 0.0001 for two-way ANOVA, and later Tukey HSD test) . After 20 days of treatment, the group using AmF toothpaste plus AmF mouthrinse had a higher fluoride uptake than the groups using either of the other products (P < 0.05) . In conclusion, all products lead to an appreciable amount of fluoride uptake in enamel; AmF products led to higher concentrations . The concentration of fluoride accumulated during treatment phase remained high after 24 hours following the cessation of use.

Appl Environ Microbiol, 2004 Dec, 70(12), 7355 - 64
PCR analysis of the distribution of unicellular cyanobacterial diazotrophs in the Arabian Sea; Mazard SL et al.; An oligonucleotide primer, NITRO821R, targeting the 16S rRNA gene of unicellular cyanobacterial N2 fixers was developed based on newly derived sequences from Crocosphaera sp . strain WH 8501 and Cyanothece sp . strains WH 8902 and WH 8904 as well as several previously described sequences of Cyanothece sp . and sequences of intracellular cyanobacterial symbionts of the marine diatom Climacodium frauenfeldianum . This oligonucleotide is specific for the targeted organisms, which represent a well-defined phylogenetic lineage, and can detect as few as 50 cells in a standard PCR when it is used as a reverse primer together with the cyanobacterium- and plastid-specific forward primer CYA359F (U . Nubel, F . Garcia-Pichel, and G . Muyzer, Appl . Environ . Microbiol . 63:3327-3332, 1997) . Use of this primer pair in the PCR allowed analysis of the distribution of marine unicellular cyanobacterial diazotrophs along a transect following the 67 degrees E meridian from Victoria, Seychelles, to Muscat, Oman (0.5 degrees S to 26 degrees N) in the Arabian Sea . These organisms were found to be preferentially located in warm (>29 degrees C) oligotrophic subsurface waters between 0 and 7 degrees N, but they were also found at a station north of Oman at 26 degrees N, 56 degrees 35'E, where similar water column conditions prevailed . Slightly cooler oligotrophic waters (<29 degrees C) did not contain these organisms or the numbers were considerably reduced, suggesting that temperature is a key factor in dictating the abundance of this unicellular cyanobacterial diazotroph lineage in marine environments.

Front Biosci, 2005 Jan 1, 10, 166 - 79 Print 2005 Jan 1.
Microsensors and microbiosensors for retinal implants; Zhou DD et al.; This paper concentrates on recent developments in microsensors and microbiosensors for the possible applications in visual prostheses, especially retinal prosthetic devices . A brief introduction on the developments of visual prosthesis will be presented . The importance for in-vivo pH measurements as well as the need for an implantable pH sensor will be demonstrated . Electrochemical biosensors developed for sensitive measurements of glucose and L-glutamate, a known neurotransmitter in the retina and brain will be reviewed . Novel electrode materials such as chemically modified thin-film diamond in applications for implantable biosensors will be shown . The challenges in the development of chronic implantable sensor systems, especially using MEMS technology for medical implants, will be discussed.

Environ Sci Technol, 2004 Nov 15, 38(22), 5939 - 49
Sugars--dominant water-soluble organic compounds in soils and characterization as tracers in atmospheric particulate matter; Simoneit BR et al.; The presence of saccharides is being reported for aerosols taken in urban, rural, and marine locales . The commonly found primary saccharides are alpha- and beta-glucose, alpha- and beta-fructose, sucrose, and mycose with lesser amounts of other monosaccharides . Saccharide polyols are also found in some airsheds and consist mainly of sorbitol, xylitol, mannitol, erythritol, and glycerol . In temperate climate areas these compounds increase from negligible concentrations in winter aerosols (usually dominated by levoglucosan and related anhydrosaccharides from biomass burning) to a maximum in late spring-summer, followed by a decrease to winter . The composition of the saccharide mixtures suggests soil and associated microbiota as the source . Saccharide analyses of soils confirmed these compositions . Therefore, we propose resuspension of soil (also unpaved road dust) from agricultural activities as a major component of aerosol particles and the saccharides are the source specific tracers . In addition, the saccharides as well as the anhydrosaccharide derivatives from biomass burning are completely water soluble and thus contribute significantly to the total water-soluble mass of aerosols.

J Microbiol Methods, 2005 Jan, 60(1), 107 - 13
McRAPD as a new approach to rapid and accurate identification of pathogenic yeasts; Plachy R et al.; Despite advances in antifungal prophylaxis and therapy, morbidity and mortality incurred by yeasts remain a significant burden . As pathogenic yeast species vary in their susceptibilities to antifungal agents, clinical microbiology laboratories face an important challenge to identify them rapidly and accurately . Although a vast array of phenotyping and genotyping methods has been developed, these are either unable to cover the whole spectrum of potential yeast pathogens or can do this only in a rather costly or laborious way . Random amplified polymorphic DNA (RAPD) fingerprinting was repeatedly demonstrated to be a convenient tool for species identification in pathogenic yeasts . However, its wider acceptance has been limited mainly due to special expertise and software needed for analysis and comparison of the resulting banding patterns . Based on a pilot study, we demonstrate here that a simple and rapid melting curve analysis of RAPD products can provide data for identification of five of the most medically important Candida species . We have termed this new approach melting curve of random amplified polymorphic DNA (McRAPD) to emphasize its rapidity and potential for automation, highly desirable features for a routine laboratory test.

Clin Diagn Virol, 1996 May, 5(2-3), 121 - 9
Laboratory diagnosis of intrauterine and perinatal virus infections; Best JM; Background: Intrauterine infection with rubella, cytomegalovirus (CMV), varicella zoster virus (VZV), parvovirus B19 and human immunodeficiency virus type 1 (HIV-1) may occur following maternal infection . Diagnosis of congenital infection in the neonate is dependant on the appropriate laboratory techniques being used . Prenatal diagnosis of intrauterine infection may also be indicated . Herpes simplex virus (HSV), HIV-1, VZV, enteroviruses, hepatitis B (HBV) and hepatitis C viruses (HCV), human T-cell lymphotropic viruses (HTLV-1 and 2) and genital papillomaviruses (PVs) may be acquired at delivery . Neonatal HSV, VZV and enterovirus infections may be severe or even fatal . Perinatally acquired HBV, HCV, HIV-1 and HTLVs are associated with persistent infection and chronic disease in later life . However, if the mother is identified as a carrier in the antenatal period, mother-infant transmission of HBV may be prevented by active/passive immunisation of the neonate, HIV-1 by caesarian section or antiviral therapy, and of HTLV-1 by avoiding breast feeding . Objectives and study design: To review the techniques available for the diagnosis of intrauterine infections, neonatal infections with HSV, HIV-1, VZV and enteroviruses, maternal infection with HBV, HCV and HIV-1 and prenatal diagnosis of intrauterine rubella, CMV and B19 . Results: Congenital rubella may be diagnosed by detection of specific IgM, but virus detection is the technique of choice for congenital cytomegalovirus . Congenital VZV may be diagnosed by serological techniques in up to 71% of cases . Detection of virus in vesicle scrapings or swabs from the oropharynx is the technique of choice for neonatal HSV, while enterovirus infections are best diagnosed by detection of viral RNA . A clinical diagnosis of congenital VZV is often possible . HIV-1 may be diagnosed within 3 months of birth by testing serial blood samples with a combination of techniques . Maternal infection with HBV, HCV, HIV and {Formula: see text} may be diagnosed by serological techniques and genital PVs by detection of viral DNA . Chorionic villus samples, amniotic fluid and fetal blood may be obtained for prenatal diagnosis of infection . Although detection of virus in amniotic fluid is the technique of choice for prenatal diagnosis of CMV, insufficient data is currently available to determine whether it may be used for intrauterine rubella . The most reliable technique for diagnosis of fetal B19 infection is detection of viral DNA in fetal blood . Conclusions: Close liaison between clinicians and microbiologists/virologists is required in order that appropriate specimens are collected from infant and/or mother and appropriate tests conducted . The use of TORCH screening should be discouraged.

Clin Diagn Virol, 1995 Jul, 4(1), 51 - 9
Laboratory acquired tick-borne meningoencephalitis: characterisation of virus strains; Avsic-Zupanc T et al.; Background: The handling of tick-borne encephalitis (TBE) virus is potentially hazardous, as indicated by a number of laboratory-acquired infections in the prevaccination era . Objectives: (1) To reemphasize the hazard of handling TBE virus without being vaccinated by describing the case of a laboratory-acquired full-blown TBE in a microbiologist who isolated the virus from a blood sample . (2) To molecularly characterize the causative virus strain isolated in Slovenia in comparison with the European prototype strain Neudoerfl . Study design: The virological diagnosis of the laboratory infection was established by serology and virus isolation . The virus was characterized by restriction fragment analysis of PCR products of amplified genomic sequences and a panel of monoclonal antibodies reacting with the major envelope protein . Results: The laboratory infection, most probably acquired by aerosol, resulted in a biphasic course of the disease with a severe meningoencephalitis in the second phase . Both by restriction fragment and monoclonal antibody analysis the Slovenian virus strains involved were indistinguishable from the European prototype strain . Conclusions: This report confirms the potential hazard of handling TBE virus in the laboratory without being vaccinated . The similarity of the virus isolates from Slovenia with the European prototype strain confirms the previously observed homogeneity of TBE virus strains from different European countries.

J Endod, 2004 Dec, 30(12), 851 - 4
Simultaneous detection of Dialister pneumosintes and Filifactor alocis in endodontic infections by 16S rDNA-directed multiplex PCR; Siqueira Junior JF et al.; Dialister pneumosintes and Filifactor alocis have been recently considered as candidate endodontic pathogens . In this study, we devised a 16S rDNA-directed multiplex PCR protocol for simultaneous detection of these two bacterial species in endodontic infections . Samples were taken from infected root canals associated with asymptomatic periradicular lesions as well as from cases of acute periradicular abscesses . DNA extracted from the samples was used as template for simultaneous detection of D . pneumosintes and F . alocis through a multiplex PCR assay . Two fragments of the expected sizes, one specific for D . pneumosintes and the other for F . alocis, were simultaneously amplified from a mixture of reference genomic DNA containing DNA from both species . Clinical samples that were positive for the target species showed a single band of the predicted size for each species . D . pneumosintes was detected by multiplex PCR in 11 samples (7 asymptomatic and 4 abscesses) and F . alocis was identified in 9 cases (6 asymptomatic and 3 abscesses) . Six samples (3 asymptomatic and 3 abscesses) shared the two species . Data from the present study confirmed that D . pneumosintes and F . alocis are common members of the microbiota present in primary endodontic infections and thereby may participate in the pathogenesis of periradicular lesions . The proposed multiplex PCR assay is a simple, rapid, and accurate method for the simultaneous detection of these two candidate endodontic pathogens.

An R Acad Nac Med (Madr), 2004, 121(2), 263 - 88
{Severe acute respiratory syndrome and aviar flu}; Eiros Bouza JM; Severe acute respiratoru syndrome (SARS) is a new disease that caused large ourbreaks in several countries in the first half of 2003, resulting in infection in more than 8.000 people and more than 900 deaths . The disease originated in southern China and a novel coronavirus (SARS CoV) has been implicated as the causative organism . We present an overview of the etiology, clinical presentation and diagnosis, based on the current state of knowledge derived from published studies and our experience in the National Microbiology Centre . Influenza is a zoonosis . This appreciation of influenza ecologyfacilitated recognition of the H5N1 'bird flu' incident in Hong Kong in 1997 in what was considered to be an incipient pandemic situation, the chicken being the source of virus for humans and . The current outbreak of avian influenza in South East Asia has resulted in a small number of human deaths . These findings highlight the importance of systematic virus surveillance of domestic poultry in recognizing changes in virus occurrence, host range and pathogenicity as signals at the avian level that could presage a pandemic.

J Surg Oncol, 2004 Dec 1, 88(3), 161 - 81
Errors in pathology and laboratory medicine: consequences and prevention; Hollensead SC et al.; Reducing errors and improving quality are an integral part of Pathology and Laboratory Medicine . The rate of errors is reviewed for the pre-analytical, analytical, and post-analytical phases for a specimen . The quality systems in place in pathology today are identified and compared with benchmarks for quality . The types and frequency of errors and quality systems are reviewed for surgical pathology, cytopathology, clinical chemistry, hematology, microbiology, molecular biology, and transfusion medicine . Seven recommendations are made to reduce errors in future for Pathology and Laboratory Medicine.

J Clin Periodontol, 2004 Dec, 31(12), 1117 - 21
In vitro studies on controlled-release cellulose acetate films for local delivery of chlorhexidine, indomethacin, and meloxicam; Cetin EO et al.; BACKGROUND: Delivery of medications into periodontal pockets to suppress or eradicate the pathogenic microbiota or modulate the inflammatory response, thereby limiting periodontal tissue destruction, has attracted significant interest with the purpose of effective periodontal treatment . However, no study has previously attempted to develop a controlled-release formulation of anti-inflammatory agents to be used in the field of periodontology . The aim of the present study was to examine the in vitro release profile of chlorhexidine gluconate, indomethacin, and meloxicam from cellulose acetate films . METHODS: Cellulose acetate films containing chlorhexidine gluconate, indomethacin, and meloxicam were prepared and cut in a form to fit to the periodontal pocket anatomy . The release of active agents was studied in 10 ml artificial saliva at 37 degrees C . Apparatus Vibrax was used at 150 r.p.m . Determinations were carried out spectrophotometrically and the release profiles were plotted as a function of time . RESULTS: The formulations showed two different release patterns for a total observation period of approximately 120 h . When the formulations of the three active agents were compared, the release patterns of meloxicam and chlorhexidine gluconate were found to be similar, while the indomethacin-containing formulation exhibited the fastest release rate . CONCLUSIONS: As a conclusion, cellulose acetate may be a suitable inert material for obtaining a prolonged local release of various anti-inflammatory agents like meloxicam . Further in vitro and in vivo studies are required before starting clinical applications of these controlled-release formulations of anti-inflammatory agents.

Clin Invest Med, 2004 Oct, 27(5), 253 - 8
Bloodstream infection complicating trauma; Laupland K et al.; BACKGROUND: Bloodstream infection (BSI) is recognized as an important infectious complication of major trauma . However, its occurrence, the risk factors contributing to its development, and its outcomes have not been well described . DESIGN: Cohort with linkage of regional trauma and microbiology databases . PATIENTS: Adult trauma patients with injury severity score (ISS) > or = 12 admitted to a regional trauma centre during a 33-month period . RESULTS: Of 1797 victims of acute trauma identified (median ISS 20; interquartile range {IQR} 16-25), 71 (4%) had 77 episodes of BSI, for an overall rate of 2.9 per 1000 hospital days . BSI in the majority of patients (37 of 72, or 52%) had onsets within the first week of hospitalization; 7 (10%) patients had community-acquired BSI (onset within 2 d) . Independently associated with the development of nosocomial BSI were higher ISSs (odds ratio {OR} 1.04, 95% confidence interval {CI} 1.02-1.07); requirement for ICU admission (OR 7.06, CI 3.38-14.75); and burns (OR 5.75, CI 2.16-15.30) . Although trauma-related BSI was a predictor of increased in-hospital case fatality (15/71 v . 208/1726; relative risk 1.75, CI 1.10-2.78), it was not an independent predictor of death . CONCLUSION: In our series, 1 in 25 major trauma cases was complicated by BSI . The infection occurred within the first week after injury in over half of our cases . Knowledge of the epidemiology of these infections will be important for planning preventive or early therapeutic efforts.

Pediatr Pulmonol, 2005 Jan, 39(1), 56 - 63
Use of laryngeal mask airway in flexible bronchoscopy in infants and children; Naguib ML et al.; In the past two decades, flexible bronchoscopy (FB) has gained increasing popularity among pediatric pulmonologists . The objective of this study was to review our experience with pediatric flexible bronchoscopy over the past 15 years, with special focus on route of bronchoscopy . This is a retrospective study . We reviewed our pediatric FB procedures performed at the University of Michigan, Mott Children's Hospital, from 1988-2003 . The study included 1,947 procedures in 1,548 patients, with a mean of 1.3 procedures/patient . The male:female ratio was 1.66:1, and age was 4.9 +/- 5.6 years (mean +/- SD) . Patients <2 years of age represented 46.6% of the study population . The laryngeal mask airway (LMA) was the most common route for flexible bronchoscopy in children 2 years of age and above . Complication rates were lower with the use of the LMA (1.9%) compared to the nasal route (3.5%) . Stridor was the most common indication in the age group <2 years (20.8%), while persistent pulmonary infiltrates were more common in the older age groups (32.2-37%) . Laryngomalacia was the most common finding in patients with stridor (31.5%), while inflammatory changes were more common with other indications . Procedure-related complications were reported in 2.3% of procedures . Bronchoalveolar lavage (BAL) samples were obtained and analyzed in 51.2% of FB procedures (n = 1,000), of which 19.4% yielded positive microbiology cultures . In conclusion, FB is a safe procedure in pediatrics . Children less than 7 years of age represent the majority of FB subjects . The LMA offered a lower rate of procedure-related complications when compared to the nasal route or endotracheal tube . It also reduced procedure time and anesthesia time . Pediatr Pulmonol . 2005; 39:56-63 . (c) 2005 Wiley-Liss, Inc.

Infect Dis Clin North Am, 2004 Dec, 18(4), 939 - 62
Healthcare-associated pneumonia in adults: management principles to improve outcomes; Craven DE et al.; Guidelines for Management of HAP were developed jointly by the ATS and IDSA in 2004 . These guidelines were designed to improve patient outcomes and to decrease the emergence of MDR pathogens (see Fig . 1).Principles include early initiation of appropriate and adequate antibiotic therapy after cultures of blood and sputum are obtained . Quantitative distal airway sampling by bronchoscopy provides greater diagnostic specificity for VAP: in one randomized study, improved outcomes were noted, compared with clinical diagnosis with qualitative endotracheal aspirates . Higher doses of initial, empiric antibiotics also are recommended . Assessment of the patient's clinical response to empiric antibiotics should be correlated with microbiologic results to streamline, de-escalate, or stop unnecessary anti-biotic treatment . Duration of therapy for uncomplicated HAP should be limited to 7 days followed by close monitoring for relapse after cessation of antibiotics . The authors suggest that prevention strategies target modifiable short- and long-term risk factors . They also advocate the use of a multidisciplinary team that is dedicated to the treatment and prevention of HCAP and the basic principle of the modern Hippocratic Oath: "I will prevent disease whenever I can, for prevention is preferable to cure."

Infect Dis Clin North Am, 2004 Dec, 18(4), 861 - 82, ix
Acute exacerbations of chronic bronchitis: new developments concerning microbiology and pathophysiology--impact on approaches to risk stratification and therapy; Sethi S et al.; Exacerbations are a characteristic feature of chronic obstructive pulmonary disease and contribute significantly to associated morbidity and mortality . Renewed interest in this common clinical problem and research using new investigative tools has enhanced substantially the understanding of the pathogenesis of exacerbations . Results of recent clinical trials and observational studies have allowed refinements in treatment of exacerbations that should im-prove patient outcomes . This article discusses a rational, stratified approach to the use of antibiotics for this condition based on these recent studies.

Rinsho Ketsueki, 2004 Oct, 45(10), 1100 - 4
{Methotrexate-induced interstitial pneumonitis in a child with acute lymphoblastic leukemia}; Tatsumoto C et al.; We report a case of 5-year-old boy with acute lymphoblastic leukemia who developed interstitial pneumonitis induced by methotrexate (MTX) . The patient was hospitalized with fever, cough, dyspnea and hypoxemia during maintenance treatment with low dose MTX and 6-mercaptopurine . A diagnosis of MTX pneumonitis was made based on the clinical findings, viral and serologic studies, negative microbiology and the radiological features . The patient recovered after cessation of the MTX treatment . Interstitial pneumonitis caused by MTX is well-recognized and the prevalence has been estimated to be 0.3-7.5% among patients with adult rheumatoid arthritis . However, there are few reports in the literature regarding this adverse effect in patients with leukemia . Furthermore, very few cases of childhood leukemia have been reported regarding MTX induced interstitial pneumonitis . Physicians should be aware of this rare complication during maintenance treatment with weekly low dose MTX for acute lymphoblastic leukemia in children.

Clin Orthop, 2004 Oct, (427), 132 - 7
Polymerase chain reaction can detect bacterial DNA in aseptically loose total hip arthroplasties; Clarke MT et al.; Identifying low-grade infection in failed total hip arthroplasties is an important but difficult task . This study investigated the ability of the polymerase chain reaction to identify low-grade infection during revision of total hip arthroplasties that failed from aseptic causes . One hundred thirteen specimens from 31 total hip arthroplasties revised for aseptic loosening were compared with 105 control specimens from 28 primary total hip arthroplasties . All surgeries were done in laminar flow operating rooms . No primary or revision specimen had positive microbiologic cultures . No revision specimen had histologic evidence suggestive of infection . Using the polymerase chain reaction with a detection threshold of 10 organisms per cubic centimeter of specimen, bacterial DNA was identified in 39 of 85 revision tissue specimens (46%) compared with 18 of 84 primary tissue specimens (21.4%) . Bacterial DNA was identified in the synovial fluid of three specimens taken from 28 revision total hip arthroplasties (10.7%) and in two specimens taken from 21 primary total hip arthroplasties (9.5%) . As multiple specimens were sent for each hip, a maximum of 16 of 31 revision total hip arthroplasties (52%) and eight of 28 primary total hip arthroplasties (29%) were considered to be infected . Bacterial DNA can be found in many specimens obtained from revised total hip arthroplasties considered to be aseptically loose . Because bacterial DNA identified at primary total hip arthroplasty was assumed to be attributable to contamination rather than present in healthy tissues, the overall specimen contamination rate of 19% and case contamination rate of 29% indicate that the polymerase chain reaction has poor specificity at this sensitivity level for diagnosing infection in revision total hip arthroplasty.

Biomed Microdevices, 2004 Dec, 6(4), 279 - 87
Membrane-based PDMS microbioreactor for perfused 3D primary rat hepatocyte cultures; Ostrovidov S et al.; To improve primary adult rat hepatocyte cultures, two types of PDMS microbioreactors containing a membrane, used as a scaffold for the attachment of cells, were built: one with a commercially-available polyester membrane, the other with a PDMS membrane (5 x 5 microm hole sizes) made in our laboratory . In that way, the relative surface area between blood perfusion and hepatocytes seen in vivo was mimicked and as cells were bathed in both sides by the culture medium they expressed much higher functions . A new technique to build such PDMS membrane was created . With this technique we could build various PDMS membranes with down to 5 x 5 microm holes and with thickness even below 20 microm . During the fifteen days of perfusion in these microbioreactors, good cell attachment then cell reorganization was observed . Moreover compared to static cultures in tissue-culture-treated dishes and in cultures in inserts with the same polyester membranes, seven and two times increases in the albumin secretion by the cultured primary rat adult hepatocyte were demonstrated, respectively . Ammonium removal also increased 7 times in perfused cultures compared to static cultures . These new microbioreactors, which closely mimic the in vivo liver architecture, revealed themselves to be very promising tools towards future applications in drug screening or liver tissue engineering.

Clin Orthop, 2004 Oct, 1(427), 132 - 137
Polymerase Chain Reaction Can Detect Bacterial DNA in Aseptically Loose Total Hip Arthroplasties; Clarke MT et al.; Identifying low-grade infection in failed total hip arthroplasties is an important but difficult task . This study investigated the ability of the polymerase chain reaction to identify low-grade infection during revision of total hip arthroplasties that failed from aseptic causes . One hundred thirteen specimens from 31 total hip arthroplasties revised for aseptic loosening were compared with 105 control specimens from 28 primary total hip arthroplasties . All surgeries were done in laminar flow operating rooms . No primary or revision specimen had positive microbiologic cultures . No revision specimen had histologic evidence suggestive of infection . Using the polymerase chain reaction with a detection threshold of 10 organisms per cubic centimeter of specimen, bacterial DNA was identified in 39 of 85 revision tissue specimens (46%) compared with 18 of 84 primary tissue specimens (21.4%) . Bacterial DNA was identified in the synovial fluid of three specimens taken from 28 revision total hip arthroplasties (10.7%) and in two specimens taken from 21 primary total hip arthroplasties (9.5%) . As multiple specimens were sent for each hip, a maximum of 16 of 31 revision total hip arthroplasties (52%) and eight of 28 primary total hip arthroplasties (29%) were considered to be infected . Bacterial DNA can be found in many specimens obtained from revised total hip arthroplasties considered to be aseptically loose . Because bacterial DNA identified at primary total hip arthroplasty was assumed to be attributable to contamination rather than present in healthy tissues, the overall specimen contamination rate of 19% and case contamination rate of 29% indicate that the polymerase chain reaction has poor specificity at this sensitivity level for diagnosing infection in revision total hip arthroplasty.

Cancer, 2004 Dec 15, 101(12), 2817 - 24
Rhabdomyosarcoma: value of myogenin expression analysis and molecular testing in diagnosing the alveolar subtype: an analysis of 109 paraffin-embedded specimens; Hostein I et al.; BACKGROUND: Identification of the alveolar subtype of rhabdomyosarcoma (ARMS) is important, because the poor prognosis associated with this subtype necessitates a modified therapeutic regimen . At present, ARMS diagnoses are made on the basis of histologic findings and the extent of myogenin immunopositivity . Nonetheless, the absence of an alveolar pattern in the solid variant, the low degree of differentiation in certain embryonal rhabdomyosarcomas (ERMS), and the increasing use of microbiopsy samples make the diagnosis of ARMS somewhat difficult . Two specific translocations have been found in ARMS, and fusion transcripts can be detected by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of paraffin-embedded tissue (PET) . METHODS: To assess the value of myogenin staining and molecular testing in the diagnosis of rhabdomyosarcoma, the authors examined 109 rhabdomyosarcoma samples (45 ARMS samples and 64 ERMS samples) . Real-time RT-PCR analysis of PET was performed in all 109 rhabdomyosarcomas, and RT-PCR analysis of frozen material was performed in 24 cases . RESULTS: PAX fusion transcripts were present in 44 cases (39 ARMS and 5 ERMS) and absent in 52 cases (2 ARMS and 50 ERMS) . In 13 cases (4 ARMS and 9 ERMS), the results were not interpretable . Results were concordant between paired frozen and fixed tumor samples . All 35 interpretable ERMS samples that contained < 50% myogenin-positive cells failed to yield detectable PAX fusion transcripts . Of the 61 interpretable tumor samples (41 ARMS and 20 ERMS) that contained > 50% myogenin-positive cells, 44 (39 ARMS and 5 ERMS) yielded detectable PAX fusion transcripts . CONCLUSIONS: The current study demonstrates that molecular detection of PAX fusion transcripts via real-time RT-PCR analysis of PET is a sensitive and specific method for the diagnosis of ARMS and that immunohistochemical analysis of myogenin expression can be used to select cases for such molecular testing . Although RT-PCR analysis appears not to possess diagnostic value in tumors with < 50% tumor cell immunopositivity, it is strongly recommended for the diagnosis of tumors containing > 50% myogenin-positive cells.

Pneumologie, 2004 Nov, 58(11), 777 - 90
{Current diagnostic approach to pleural effusion}; Frank W; Pleural effusion is a common pneumologic and interdisciplinary problem . Transudate/exsudate discrimination of the pleural fluid by thoracentesis remains the diagnostic basic algorithm . Regardless of a number of new markers, classical LIGHT's criteria comprising the pleural fluid protein- and LDH-values (or their serum ratio respectively) reveal the highest potency with an overall accuracy of 95 % . Expansion to cholesterol-determination (triplet test) may be helpful to identify transudates in indeterminate cases . The need for further local diagnostic evaluation is then usually restricted to exudates . Bacterial pleurisy, malignant and tuberculous effusion are the principal differential diagnoses . With the use of a variety of conventional biochemical, cytologic, immunologic and microbiologic investigations, thoracentesis will allow- or substantially narrow-diagnosis of exudates in about 70 %, with novel cell biological markers in some conditions up to 90 % . In bacterial pleurisy thoracentesis provides information directly relevant to management in terms of local interventions . It also constitutes a platform for more invasive imaging- or endoscopy-guided investigations with a focus on medical thoracoscopy (pleuroscopy) . Blind needle biopsy is diagnostic in a range of 40 - 70 % both in malignancy and inflammatory disease, thoracoscopy may clarify exudative conditions in about 95 % . Thus malignancy may be specifically diagnosed in 97 % of cases, tuberculous effusion in virtually 100 % . The value of thoracoscopy is augmented by interventional options including complete evacuation of the pleural cavity, eventually followed by talc pleurodesis ("poudrage") in recurrent effusions or adhesiolysis, irrigation and fibrinolysis protocols in certain inflammatory conditions . These combined features as accomplished in local anesthesia on a remarkably high safety level characterise medical thoracoscopy as a gold standard tool for the management of pleural disease even in comparison to more elaborate surgical procedures.

Proc Natl Acad Sci U S A, 2004 Nov 16, 101(46), 16322 - 7 Epub 2004 Nov 16.
Integrated regulatory responses of fimB to N-acetylneuraminic (sialic) acid and GlcNAc in Escherichia coli K-12; Sohanpal BK et al.; Bacterial-host attachment by means of bacterial adhesins is a key step in host colonization . Phase variation (reversible on-off switching) of the type 1 fimbrial adhesin of Escherichia coli involves a DNA inversion catalyzed by FimB (switching in either direction) or FimE (mainly on-to-off switching) . fimB is separated from the divergent yjhATS operon by a large (1.4 kbp) intergenic region . Short ( approximately 28 bp) cis-active elements (regions 1 and 2) close to yjhA stimulate fimB expression and are required for sialic acid (Neu(5)Ac) sensitivity of its expression {El-Labany, S., Sohanpal, B . K., Lahooti, M., Akerman, R . & Blomfield, I . C . (2003) Mol . Microbiol . 49, 1109-1118} . Here, we show that whereas NanR, a sialic acid-response regulator, binds to region 1, NagC, a GlcNAc-6P-responsive protein, binds to region 2 instead . The NanR- and NagC-binding sites lie adjacent to deoxyadenosine methylase (Dam) methylation sites (5'-GATC) that are protected from modification, and the two regulators are shown to be required for methylation protection at regions 1 and 2, respectively . Mutations in nanR and nagC diminish fimB expression, and both fimB expression and FimB recombination are inhibited by GlcNAc (3- and >35-fold, respectively) . Sialic acid catabolism generates GlcNAc-6-P, and whereas GlcNAc disrupts methylation protection by NagC alone, Neu(5)Ac inhibits the protection mediated by both NanR and NagC as expected . Type 1 fimbriae are proinflammatory, and host defenses enhance the release of both Neu(5)Ac and GlcNAc by a variety of mechanisms . Inhibition of type 1 fimbriation by these amino sugars may thus help balance the interaction between E . coli and its hosts.

J Nucl Med, 2004 Nov, 45(11), 1864 - 71
Diagnosing infection in the failed joint replacement: a comparison of coincidence detection 18F-FDG and 111In-labeled leukocyte/99mTc-sulfur colloid marrow imaging; Love C et al.; The objectives of this study were to investigate (18)F-FDG imaging, using a coincidence detection system, for diagnosing prosthetic joint infection and to compare it with combined (111)In-labeled leukocyte/(99m)Tc-sulfur colloid marrow imaging in patients with failed lower extremity joint replacements . METHODS: Fifty-nine patients--with painful, failed, lower extremity joint prostheses, 40 hip and 19 knee--who underwent (18)F-FDG, labeled leukocyte, and bone marrow imaging, and had histopathologic and microbiologic confirmation of the final diagnosis, formed the basis of this investigation . (18)F-FDG images were interpreted as positive for infection using 4 different criteria: criterion 1: any periprosthetic activity, regardless of location or intensity; criterion 2: periprosthetic activity on the (18)F-FDG image, without corresponding activity on the marrow image; criterion 3: only bone-prosthesis interface activity, regardless of intensity; criterion 4: semiquantitative analysis--a lesion-to-background ratio was generated, and the cutoff value yielding the highest accuracy for determining the presence of infection was determined . Labeled leukocyte/marrow images were interpreted as positive for infection when periprosthetic activity was present on the labeled leukocyte image without corresponding activity on the marrow image . RESULTS: Twenty-five (42%) prostheses, 14 hip and 11 knee, were infected . The sensitivity, specificity, and accuracy of (18)F-FDG, by criterion, were as follows: criterion 1: 100%, 9%, 47%; criterion 2: 96%, 35%, 61%; criterion 3: 52%, 44%, 47%; criterion 4: 36%, 97%, 71% . The sensitivity, specificity, and accuracy of labeled leukocyte/marrow imaging were 100%, 91%, and 95%, respectively . WBC/marrow imaging, which was more accurate than any of the (18)F-FDG criteria for all prostheses, as well as for hips and knees separately, was significantly more sensitive than criterion 3 (P < 0.001) and criterion 4 (P < 0.001) and was significantly more specific than criterion 1 (P < 0.001), criterion 2 (P < 0.001), and criterion 3 (P < 0.001) . CONCLUSION: Regardless of how the images are interpreted, coincidence detection-based (18)F-FDG imaging is less accurate than, and cannot replace, labeled leukocyte/marrow imaging for diagnosing infection of the failed prosthetic joint.

S Afr Med J, 2004 Oct, 94(10), 851 - 4
Serum procalcitonin as an early marker of neonatal sepsis; Ballot DE et al.; BACKGROUND: It has recently been suggested that procalcitonin (PCT) is of value in the diagnosis of neonatal sepsis, with varying results . This study was to evaluate the role of PCT as a single early marker of neonatal sepsis . SETTING: Neonatal Unit, Johannesburg Hospital, and Microbiology Laboratory, National Health Laboratory Service (NHLS), South Africa . SUBJECTS AND METHODS: Neonates undergoing evaluation for sepsis between April and August 2002 were eligible for inclusion . Patients were categorised into 'no infection', 'possible infection' and 'definite infection' on the basis of C-reactive protein (CRP), white cell count (WCC), platelet count and blood culture results . PCT was correlated with infection categories . RESULTS: One hundred and eighty-three neonates were enrolled . One hundred and eighteen had no infection, 52 possible infection and 13 definite infection . PCT differed significantly among infection categories (p < 0.0001) and correlated significantly with CRP at presentation (correlation coefficient 0.404, p < 0.001) and CRP at 24 hours (correlation coefficient 0.343, p < 0.001) . PCT predicted 89.5% of definite infection . Receiver operating characteristic (ROC) analysis for PCT to predict definite infection showed odds ratio (OR) 1.145 (95% confidence interval (CI): 1.05-1.25) with an area under the curve of 0.778 . PCT had a negative predictive value of 0.95 (95% CI: 0.915-0.988) for definite infection . CONCLUSIONS: Although PCT was significantly related to the category of infection, it is not sufficiently reliable to be the sole marker of neonatal sepsis . PCT would be useful as part of a full sepsis evaluation, but is relatively expensive . A negative PCT on presentation may rule out sepsis, but this needs to be evaluated further.

J Clin Microbiol, 2004 Nov, 42(11), 5065 - 9
Multilocus microsatellite typing system for Penicillium marneffei reveals spatially structured populations; Fisher MC et al.; For eukaryotic pathogens that have low levels of genetic variation, multilocus microsatellite typing (MLMT) offers an accurate and reproducible method of characterizing genetic diversity . Here, we describe the application of an MLMT system to the emerging pathogenic fungus Penicillium marneffei . Isolates used for this study were those held in the culture collections of the Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, and the Chiang Mai University Department of Microbiology, Chang Mai, Thailand . High genetic diversity and extensive spatial structure were observed among clinical isolates, with the geographical area of origin for each isolate strongly correlating with the occurrence of two deeply divided clades . Within each clade, multilocus linkage associations were highly significant and could be explained by genetically differentiated populations or by an exclusively clonal reproductive mode, or both . Our results show that southeast Asian penicilliosis is caused by a fungus with a complex population genetic structure . Furthermore, this MLMT system generates digital data that can be easily queried against a centrally held database via the internet ; this provides a powerful epidemiological tool for analyzing the underlying parameters that are responsible for the emergence of P . marneffei in human immunodeficiency virus-positive populations.

J Clin Microbiol, 2004 Nov, 42(11), 5058 - 64
Characterization of ancestral Mycobacterium tuberculosis by multiple genetic markers and proposal of genotyping strategy; Sun YJ et al.; Sixty-eight ancestral Mycobacterium tuberculosis isolates were previously identified by using the tuberculosis-specific deletion 1 (TbD1) PCR and mycobacterial interspersed-repetitive-unit-variable-number tandem repeat (MIRU-VNTR) typing (Y . J . Sun, R . Bellamy, A . S . G . Lee, S . T . Ng, S . Ravindran, S.-Y . Wong, C . Locht, P . Supply, and N . I . Paton, J . Clin . Microbiol . 42:1986-1993, 2004) . These TbD1(+) ancestral isolates were further characterized and typed in this study by IS6110 restriction fragment length polymorphism (RFLP) typing, VNTR typing using exact tandem repeats (VNTR-ETR), and spoligotyping of the direct-repeat region . To our knowledge, this is the first characterization of this genogroup by multiple genetic markers based on a fairly large sample size . In this genogroup, all spoligotypes were characterized by the absence of spacers 29 to 32 and 34 . In addition, VNTR-ETR typing could add further resolution to the clustered isolates identified by MIRU-VNTR, and the combination of MIRU-VNTR and VNTR-ETR, called MIRU-ETR, showed the highest discriminatory power for these strains compared to IS6110 RFLP typing and spoligotyping alone . However, MIRU-ETR appeared to still cluster some probably epidemiologically unrelated strains, as judged by IS6110 RFLP divergence . Therefore, a typing strategy based on stepwise combination of MIRU-ETR and IS6110 RFLP is proposed to achieve maximal discrimination for unrelated TbD1(+) strains . This typing strategy may be useful in areas where TbD1(+) ancestral strains are prevalent.

J Clin Microbiol, 2004 Nov, 42(11), 4937 - 46
Distribution of putative adhesins in different seropathotypes of Shiga toxin-producing Escherichia coli; Toma C et al.; The distribution of eight putative adhesins that are not encoded in the locus for enterocyte effacement (LEE) in 139 Shiga toxin-producing Escherichia coli (STEC) of different serotypes was investigated by PCR . Five of the adhesins (Iha, Efa1, LPF(O157/OI-141), LPF(O157/OI-154), and LPF(O113)) are encoded in regions corresponding to genomic O islands of E . coli EDL933, while the other three adhesins have been reported to be encoded in the STEC megaplasmid of various serotypes (ToxB {O157:H7}, Saa {O113:H21}, and Sfp {O157:NM}) . STEC strains were isolated from humans (n = 54), animals (n = 52), and food (n = 33) . They were classified into five seropathotypes (A through E) based on the reported occurrence of STEC serotypes in human disease, in outbreaks, and in the hemolytic-uremic syndrome (M . A . Karmali, M . Mascarenhas, S . Shen, K . Ziebell, S . Johnson, R . Reid-Smith, J . Isaac-Renton, C . Clark, K . Rahn, and J . B . Kaper, J . Clin . Microbiol . 41:4930-4940, 2003) . The most prevalent adhesin was that encoded by the iha gene (91%; 127 of 139 strains), which was distributed in all seropathotypes . toxB and efa1 were present mainly in strains of seropathotypes A and B, which were LEE positive . saa was present only in strains of seropathotypes C, D, and E, which were LEE negative . Two fimbrial genes, lpfA(O157/OI-141) and lpfA(O157/OI-154), were strongly associated with seropathotype A . The fimbrial gene lpfA(O113) was present in all seropathotypes except for seropathotype A, while sfpA was not present in any of the strains studied . The distribution of STEC adhesins depends mainly on serotypes and not on the source of isolation . Seropathotype A, which is associated with severe disease and frequently is involved in outbreaks, possesses a unique adhesin profile which is not present in the other seropathotypes . The wide distribution of iha in STEC strains suggested that it could be a candidate for vaccine development.

Radiol Med (Torino), 2004 Oct, 108(4), 335 - 44
Advantages and limits of percutaneous breast core biopsy with Mammotome and stereotactic equipment in upright seated patient; Della Sala SW et al.; PURPOSE: To confirm the diagnostic accuracy of percutaneous core breast biopsy with Mammotome and stereotactic equipment in upright seated patients, and to compare results from this method with those obtained using the dedicated unit with prone table . MATERIALS AND METHODS: We analysed 72 microbioptic samples from 69 upright seated patients . These samples were obtained in the course of mammographies assisted by digital stereotactic equipment (Diamond Delta32 General Electric Medical System/Instrumen-tarium), and further equipped with a lateral arm to support the Mammotone probe . We studied the following lesions: 57 cases of isolated microcalcification clusters, 12 masses with microcalcification and three cases of plain masses . We describe in detail the various phases of our 30-minute long microbiopsy procedure . All 72 samples were extracted without problems . Only in one case a large haematoma emerged, but quickly disappeared . RESULTS: We were able to characterise the histology of the lesions in all our samples: in 39/72 cases they were identified as benign, 28 were classified as malignant, and the remaining 5 as atypia . In the 39 lesions identified as benign, the microbiopsy procedure could be carried out without surgical biopsy - consequently, these patients were available for routine follow-up exams, which confirmed the benign nature of their lesions . Our identification of the 28 malignant lesions was confirmed by surgical biopsy . The microbiopsy finding and the histological result did not agree in only two cases . The 5 cases with atypical ductal hyperplasia showed no neoplastic alteration following surgical biopsy . In our personal experience, core biopsy with Mammotome achieved a high diagnostic accuracy . CONCLUSIONS: After careful analysis of the results obtained using Mammotome in upright seated patients with stereotactic equipment, we can confirm the diagnostic accuracy of this procedure in mammography alterations not visible in sonography . In agreement with previous studies, we also confirm that the diagnostic accuracy of this approach, as well as possible complications deriving from it, do not depend on the position of the patient.

Genetica, 2004 Jul, 121(3), 219 - 28
Number and nuclear localisation of nucleoli in mammalian spermatocytes; Berrios S et al.; In seven mammalian species, including man, the position and number of nucleoli in pachytene spermatocyte nuclei were studied from electron microscope (EM) nuclear sections or bivalent microspreads . The number and position of the nucleolar organiser regions (NORs) in mitotic and meiotic chromosomes were also analysed, using silver staining techniques and in situ hybridisation protocols . The general organisation of pachytene spermatocyte nucleoli was almost the same, with only minor morphological differences between species . The terminal NORs of Thylamys elegans (Didelphoidea, Marsupialia), Dromiciops gliroides (Microbiotheridae, Marsupialia), Phyllotys osgoodi (Rodentia, Muridae) and man, always gave rise to peripheral nucleoli in the spermatocyte nucleus . In turn, the intercalated NORs from Octodon degus, Ctenomys opimus (Rodentia, Octodontidae) and Chinchilla lanigera (Rodentia, Cavidae), gave rise to central nucleoli . In species with a single nucleolar bivalent, just one nucleolus is formed, while in those with multiple nucleolar bivalents a variable number of nucleoli are formed by association of different nucleolar bivalents or NORs that occupy the same nuclear peripheral space (Phyllotis and man) . It can be concluded that the position of each nucleolus within the spermatocyte nucleus is mainly dependent upon: (1) the position of the NOR in the nucleolar bivalent synaptonemal complex (SC), (2) the nuclear pathway of the nucleolar bivalent SC, being both telomeric ends attached to the nuclear envelope, and (3) the association between nucleolar bivalents by means of their NOR-nucleolar domains that occupy the same nuclear space . Thus, the distribution of nucleoli within the nuclear space of spermatocytes is non-random and it is consistent with the existence of a species-specific meiotic nuclear architecture.

Ann Allergy Asthma Immunol, 2004 Oct, 93(4), 381 - 9
The Copenhagen Prospective Study on Asthma in Childhood (COPSAC): design, rationale, and baseline data from a longitudinal birth cohort study; Bisgaard H; BACKGROUND: The atopic diseases asthma, atopic dermatitis, and allergic rhinitis are the most common chronic diseases in children, and their prevalence has increased recently in industrialized nations . Little is known about the genetic-environmental interaction factors driving such proliferation . OBJECTIVE: To investigate the relationships among genetic, environmental, and lifestyle factors in the development of atopic diseases in high-risk children with the aim of developing evidence-based prevention strategies . METHODS: The Copenhagen Prospective Study on Asthma in Childhood is a single-center, birth cohort study of children of asthmatic mothers . Objective assessments begin at birth, with scheduled visits every 6 months and when acute symptoms manifest . Clinical outcomes comprise preasthma, asthma, atopic dermatitis, allergic rhinitis, allergy, lung function, and bronchial responsiveness . Exposure assessments comprise respiratory, intestinal, and skin microbiology; the child's diet; indoor and outdoor air quality; allergens; and indicators of lifestyle . Genetic characteristics of probands and parents are evaluated . Quality assurance follows Good Clinical Practice guidelines . RESULTS: Four hundred eleven infants of asthmatic mothers were enrolled at the age of 1 month . The children were born between August 2, 1998, and December 28, 2001 . Compared with the Copenhagen population, mothers of the cohort population were less likely to have given natural childbirth . The households were slightly less affluent, with fewer children and fewer pets . Whites may be overrepresented . At age 2 years, 93% of the infants were still actively participating in the cohort . CONCLUSIONS: This longitudinal birth cohort study of high-risk Danish infants consists of objective phenotyping, detailed information on exposure, high data quality, and a high participant retention rate.

Nat Med, 2004 Nov, 10(11), 1177 - 85
From Pasteur to genomics: progress and challenges in infectious diseases; Rappuoli R; Over the past decade, microbiology and infectious disease research have undergone the most profound revolution since the times of Pasteur . Genomic sequencing has revealed the much-awaited blueprint of most pathogens . Screening blood for the nucleic acids of infectious agents has blunted the spread of pathogens by transfusion, the field of antiviral therapeutics has exploded and technologies for the development of novel and safer vaccines have become available . The quantum jump in our ability to detect, prevent and treat infectious diseases resulting from improved technologies and genomics was moderated during this period by the greatest emergence of new infectious agents ever recorded and a worrisome increase in resistance to existing therapies . Dozens of new infectious diseases are expected to emerge in the coming decades . Controlling these diseases will require a better understanding of the worldwide threat and economic burden of infectious diseases and a global agenda.

J Bacteriol, 2004 Nov, 186(22), 7726 - 35
Responses of the Rhodobacter sphaeroides transcriptome to blue light under semiaerobic conditions; Braatsch S et al.; Exposure to blue light of the facultative phototrophic proteobacterium Rhodobacter sphaeroides grown semiaerobically results in repression of the puc and puf operons involved in photosystem formation . To reveal the genome-wide effects of blue light on gene expression and the underlying photosensory mechanisms, transcriptome profiles of R . sphaeroides during blue-light irradiation (for 5 to 135 min) were analyzed . Expression of most photosystem genes was repressed upon irradiation . Downregulation of photosystem development may be used to prevent photooxidative damage occurring when the photosystem, oxygen, and high-intensity light are present simultaneously . The photoreceptor of the BLUF-domain family, AppA, which belongs to the AppA-PpsR antirepressor-repressor system, is essential for maintenance of repression upon prolonged irradiation (S . Braatsch et al., Mol . Microbiol . 45:827-836, 2002) . Transcriptome data suggest that the onset of repression is also mediated by the AppA-PpsR system, albeit via an apparently different sensory mechanism . Expression of several genes, whose products may participate in photooxidative damage defense, including deoxypyrimidine photolyase, glutathione peroxidase, and quinol oxidoreductases, was increased . Among the genes upregulated were genes encoding two sigma factors: sigmaE and sigma38 . The consensus promoter sequences for these sigma factors were predicted in the upstream sequences of numerous upregulated genes, suggesting that coordinated action of sigmaE and sigma38 is responsible for the upregulation . Based on the dynamics of upregulation, the anti-sigmaE factor ChrR or its putative upstream partner is proposed to be the primary sensor . The identified transcriptome responses provided a framework for deciphering blue-light-dependent signal transduction pathways in R . sphaeroides.

Laryngoscope, 2004 Nov, 114(11 Pt 3 Suppl 105), 1 - 26
Studies in otitis media: Children's Hospital of Pittsburgh-University of Pittsburgh progress report--2004; Bluestone CD; OBJECTIVES/HYPOTHESIS: The present Progress Report has summarized the key otitis media clinical trials and laboratory studies conducted since 1969 by investigators at the Children's Hospital of Pittsburgh-University of Pittsburgh (Pittsburgh, PA) . STUDY DESIGN: Review . METHODS: Included in the discussion are the following: 1) studies of the epidemiology and risk factors; 2) anatomy and pathology of the eustachian tube-middle ear from human temporal bone histopathological specimens; 3) physiology and pathophysiology of the eustachian tube-middle ear in humans and animal models; 4) pathogenesis; 5) otitis media in special populations (e.g., patients with cleft palate, Native Americans, patients with Down syndrome); 6) microbiology; 7) diagnosis; 8) outcomes of randomized clinical trials that evaluated efficacy of nonsurgical and surgical methods of treatment and prevention; 9) studies of certain complications and sequelae (e.g., effect of middle-ear effusion on hearing, early child development, and the vestibular system; chronic suppurative otitis media) . Also included are relevant summary tables and 256 references.

An Med Interna, 2004 Oct, 21(10), 495 - 7
{Nonbacterial thrombotic endocarditis as initial event of lung cancer}; Herrera de Pablo P et al.; Nonbacterial thrombotic endocarditis (NBTE) is a rare cause of peripheral embolism . NBTE is usually associated with malignant diseases and hypercoagulability states . Echocardiography is a very useful technique for the diagnosis . However valvular lesions in NBTE are similar to valvular vegetations observed from infectious endocarditis (IE), so it s necessary to establish a differential diagnosis . The treatment of thrombotic endocarditis is controversial, but the literature coincides in the use of intravenous heparin . We describe the case of a 42 years old woman with stroke in which transesophageal echocardiography (TEE) was useful in establishing the diagnosis of NBTE . The infectious origin was excluded with the realization of a exhaustive microbiologic study . In the search of causes for NBTE a lung adenocarcinoma was detected . In the present case the diagnosis of endocarditis preceded the neoplastic disease diagnosis.

J Laryngol Otol, 2004 Sep, 118(9), 706 - 9
Clinical evaluation of the electronic nose in the diagnosis of ear, nose and throat infection: a preliminary study; Shykhon ME et al.; The term electronic nose describes an electronic system that is able to mimic the human sense of smell . Electronic noses have been developed over the last 10 or more years to perform a variety of identification tasks in various industries . More recently electronic noses have attracted new interest in their application in the field of medical diagnosis . The aim of this study is to explore the use of an electronic nose to identify and classify pathogens associated with ear, nose and throat (ENT) infections . In this study 90 bacterial swab samples were collected from 90 patients with ENT infections . Some of these samples were analysed immediately with a commercial electronic nose (Cyranose C320) . Similar numbers of swabs were also taken from the same site of infection and were sent for microbiology culture and sensitivity . The electronic nose diagnosis was compared with the microbiology diagnosis and it was found that the electronic nose diagnosis was correct in 88.2 per cent of the cases, which is an encouraging result.

Am J Obstet Gynecol, 2004 Oct, 191(4), 1205 - 11
Development of a guinea pig model of chorioamnionitis and fetal brain injury; Patrick LA et al.; OBJECTIVE: The purpose of this study was to develop a guinea pig model of chorioamnionitis to study the mechanisms that lead to fetal brain injury.Study design Pregnant guinea pigs at 70% gestation were inoculated intracervically with 1000 to 2500 colony-forming units of Escherichia coli . Guinea pigs were killed 2 to 3 days after bacterial inoculation . Maternal blood and fetal amniotic fluid samples were analyzed for proinflammatory cytokine tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 levels with the use of enzyme-linked immunosorbent assay kits . Fetal brains were stained for evidence of cell death with NeuroTacs stain . RESULTS: Of 34 maternal guinea pigs that were given an intracervical inoculation of E coli, 8 guinea pigs showed microbiologic evidence of chorioamnionitis in the amniotic fluid . Tumor necrosis factor-alpha and interleukin-6 were significantly higher (P<.05) in amniotic fluid samples that were obtained from sows that were subjected to intracervical inoculation with bacteria as compared with control animals (n=6 control maternal animals) . These results were observed even if no bacteria were found subsequently on culture of the amniotic fluid from inoculated animals, which indicated that indirect exposure to infectious agents was sufficient to cause an elevated inflammatory response in the fetus . Levels of white matter injury were greater in fetuses that were exposed to bacterial infection in utero, as compared with control animals (P<.05) . This result was found in the staining of periventricular and cortical white matter for the immunolabeling of activated caspase 3 and NeuroTacs staining for cells that exhibited evidence of apoptotic cell death (positive stain with evidence of karyorrhexis) . CONCLUSION: Intracervical inoculation with E coli results in chorioamnionitis in guinea pigs that is associated with fetal brain injury.

Euro Surveill . 2004 Oct 01;9(10) {Epub ahead of print}
Preliminary results from the new HIV surveillance system in France; Lot F et al.; In addition to AIDS surveillance, data on HIV infection are necessary to better follow the dynamics of the epidemic . We report the first results of France's mandatory anonymous HIV notification system, which is linked to a virological surveillance of recent HIV infections and of circulating HIV types, groups and subtypes . HIV notifications are initiated by microbiologists who create an anonymous code of patient's identity . Clinicians complete the notification form with epidemiological and clinical data . Notifications are sent to the local health authorities and passed to the Institut de Veille Sanitaire (InVS).

J Biol Chem, 2005 Jan 7, 280(1), 564 - 570 Epub 2004 Oct 26.
Peroxiredoxin-linked Detoxification of Hydroperoxides in Toxoplasma gondii; Akerman SE et al.; The apicomplexan parasite Toxoplasma gondii is highly susceptible to oxidative stress caused by tert-butyl-hydroperoxide, juglone, and phenazine methylsulfate with IC(50) in the nanomolar range . Using dichlorofluorescein diacetate, a detector of endogenous oxidative stress, it was shown that juglone and phenazine methylsulfate are potentially toxic to the parasites without affecting the host cells . These results demonstrate that T . gondii is vulnerable to oxidative challenge that results from disruption of its redox balance and so this could be an effective approach to therapeutic intervention . This study has characterized redox active and antioxidant peroxidases belonging to the class of 1-Cys and 2-Cys peroxiredoxins that play crucial roles in maintaining redox balance . The tachyzoite stages of T . gondii express thioredoxin (TgTrx), 1-Cys peroxiredoxin (TgTrx-Px2), and a 2-Cys peroxiredoxin (TgTrx-Px1) and immunofluorescent studies revealed that all three proteins are located in the cytosol of the parasite confirming previous studies on TgTrx-Px1 (Kwok, L.Y., Schluter, D., Clayton, C., and Soldati, D . (2004) Mol . Microbiol . 51, 47-61) . TgTrx-Px1 showed K(m) values for H(2)O(2) and tert-butyl hydroperoxide in the nanomolar range, emphasizing the great affinity of the protein for theses substrates . Moreover, the catalytic efficiency of TgTrx-Px1 for these substrates at 10(6)-10(7) m(-1) s(-1) is unusually high, which qualifies the enzyme as an extremely potent antioxidant . Kinetic analyses revealed that TgTrx-Px1 is inhibited by tert-butyl hydroperoxide, and apparent inhibition constants were determined to be between 33 and 35.6 mum depending on the concentration of the non-inhibitory substrate thioredoxin . TgTrx-Px2 protected glutamine synthetase from inactivation by Fe(3+)/DTT, showing that it is an active peroxiredoxin.

J Endod, 2004 Nov, 30(11), 785 - 7
Soft tissue dissolution capacity of currently used and potential endodontic irrigants; Naenni N et al.; Necrotic soft-tissue remnants in root canals may provide a source of nutrition for surviving microbiota after root-canal therapy . This study assessed the necrotic tissue dissolution capacity of some popular and some potential root-canal irrigants: 1% (wt/vol) sodium hypochlorite (NaOCl), 10% chlorhexidine, 3% and 30% hydrogen peroxide, 10% peracetic acid, 5% dichloroisocyanurate (NaDCC), and 10% citric acid . Standardized necrotic tissue samples obtained from pig palates were incubated in these solutions, and their weight loss was measured over time . None of the test solutions except sodium hypochlorite had any substantial tissue dissolution capacity . It was concluded that this might be important when considering the use of irrigants other than NaOCl.

Cornea, 2004 Nov, 23(8), 819 - 22
Ocular cicatricial pemphigoid, keratomycosis, and intravenous immunoglobulin therapy; Iaccheri B et al.; PURPOSE: To report the case of a patient developing fungal keratitis in the context of uncontrolled ocular cicatricial pemphigoid (OCP), which, despite intravenous immunoglobulin (IVIg) and other immunomodulatory therapy, progressed to end-stage pemphigoid, with corneal opacification, ankyloblepharon, and xerosis . Keratoprosthesis (KPro) restored functional vision for the patient . METHODS: A 39-year-old man presented with uncontrolled CP and corneal ulcer in the left eye . Conjunctival biopsy diagnosed OCP; corneal scraping and biopsy diagnosed the cause of the corneal ulcer . OCP was treated with systemic steroids, immunosuppressive drugs, and IVIg . Visual rehabilitation was accomplished with Ahmed valve and a type II Dohlman KPro . RESULTS: Immunohistology of the biopsied conjunctiva showed IgG at the epithelial basement membrane zone, confirming the clinical diagnosis of OCP . Microbiologic studies of the corneal biopsy specimen were negative for Acanthamoeba and herpes but positive for Aspergillus niger . The patient's keratomycosis resolved with topical antifungal therapy . Treatment with Dapsone, intravenous-pulse steroid, oral cyclophosphamide, and intravenous immunoglobulin (IVIg) failed to control the OCP, with resultant complete conjunctivization of the cornea . Keratoprosthesis improved the patient's visual acuity from hand movements to 20/20 . CONCLUSIONS: Patients with uncontrolled OCP are at increased risk of corneal infection . The difficulty in diagnosing keratomycosis and the relatively rare occurrence of OCP explain the uniqueness of our reported case . OCP may progress to "end-stage" disease despite therapy . Keratoprosthesis can restore vision in selected otherwise seemingly hopeless cases.

J Hosp Infect, 2004 Nov, 58(3), 193 - 9
Professionals' awareness of operational antibiotic prescribing controls in UK NHS hospitals; Woodford EM et al.; In recent years, there have been increasing recommendations for multidisciplinary collaboration between clinical pharmacists and medical microbiologists in an attempt to control the quality (and quantity) of antibiotic prescribing . A questionnaire addressing the utilization of antibiotic prescribing controls was sent to the chief pharmacist and medical microbiologist in UK NHS hospitals . Responses were received from both the chief pharmacist and the medical microbiologist employed in the same hospital from 83 hospitals (a 30% response rate from two independent studies) . A high level of disagreement and poor awareness was identified between the interprofessional staff groups regarding the existence of antibiotic formulary (with disagreement between the two groups, or not known by one or both respondents, in 46% of the paired hospitals, N = 38) and guideline documents (13%, N = 11), performance of antibiotic prescribing audits (40%, N = 33), and whether pharmacists (52%, N = 43) and medical microbiologists (77%, N = 64) monitored physician compliance with antibiotic prescribing control documents . This study has identified poor knowledge of the existence of basic antibiotic prescribing control mechanisms and the role of professional colleagues . It is suggested that there is some way to go before 'Agenda for Change' principles of flexible and collaborative roles are met.

Emerg Infect Dis, 2004 Aug, 10(8), 1450 - 4
Human herpesviruses 6 and 7 and central nervous system infection in children; Ansari A et al.; The role and frequency of human herpesviruses (HHV)-6 and -7 in central nervous system (CNS) diseases of children are unclear . Cerebrospinal fluid samples from 245 pediatric patients (median age 43 days), submitted for evaluations of possible sepsis or of neurologic symptoms, were tested for HHV-6 and HHV-7 DNA by polymerase chain reaction . HHV-6 DNA was found in 3 of 245 samples, and HHV-7 was found in 0 of 245 samples . The three patients with HHV-6 DNA were <2 months of age . HHV-6 was likely pathogenic in two patients with meningitis who lacked evidence of another microbiologic cause . HHV-6 and HHV-7 are uncommon causes of CNS infection in children . HHV-6 may occasionally cause meningitis in young infants.

Mikrobiyol Bul, 2004 Jul, 38(3), 187 - 91
{Species and biovar distribution of Brucella isolates in Trakya University Hospital between 1997-2002}; Kuloglu F et al.; The aim of this study was to evaluate the species and biovar distribution of Brucella spp . isolated from blood cultures in Clinical Microbiology Laboratory of Trakya University Hospital, between 1997-2002 . A total of 48 Brucella spp . have been isolated from 14.815 patients (0.3%), and the strains were identified according to CO2 requirement, H2S production, basic fuchsin and thionin sensitivity, lysis by Tbilisi phages, and presence of agglutination with monospecific A and M antisera . As a result, 47 (97.9%) isolates were identified as B . melitensis, and one as B . abortus (2.1%) . Forty two (89.4%) of B . melitensis isolates were biovar 3, and five (10.6%) were biovar 1, while the single isolate of B . abortus was identified as an atypical strain.

Clin Microbiol Rev, 2004 Oct, 17(4), 840 - 62, table of contents
Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases; Clarridge JE 3rd; The traditional identification of bacteria on the basis of phenotypic characteristics is generally not as accurate as identification based on genotypic methods . Comparison of the bacterial 16S rRNA gene sequence has emerged as a preferred genetic technique . 16S rRNA gene sequence analysis can better identify poorly described, rarely isolated, or phenotypically aberrant strains, can be routinely used for identification of mycobacteria, and can lead to the recognition of novel pathogens and noncultured bacteria . Problems remain in that the sequences in some databases are not accurate, there is no consensus quantitative definition of genus or species based on 16S rRNA gene sequence data, the proliferation of species names based on minimal genetic and phenotypic differences raises communication difficulties, and microheterogeneity in 16S rRNA gene sequence within a species is common . Despite its accuracy, 16S rRNA gene sequence analysis lacks widespread use beyond the large and reference laboratories because of technical and cost considerations . Thus, a future challenge is to translate information from 16S rRNA gene sequencing into convenient biochemical testing schemes, making the accuracy of the genotypic identification available to the smaller and routine clinical microbiology laboratories.

Mol Cell Probes, 2004 Dec, 18(6), 437 - 41
Overcoming bacterial DNA contamination in real-time PCR and RT-PCR reactions for LacZ detection in cell therapy monitoring; Tondeur S et al.; Since the introduction of the polymerase chain reaction the presence of contaminating bacterial DNA in the Taq polymerase preparations has hampered the use of this technique in microbiology . Lately, this inconvenience has equally impeded gene quantification in the field of cell or gene therapy, where bacterial genes such as LacZ are often used as tags to detect vectors or cells after their injection in the recipient organism . Several means to overcome the DNA contamination of Taq Polymerase have been reported with variable degrees of decontamination efficiency and alteration of the PCR reaction . Here we propose two protocols to efficiently quantify DNA or RNA from the LacZ gene by real-time PCR using either decontamination by low concentrations of DNAse I prior to PCR amplification or a highly purified Taq Polymerase which is devoid of detectable contamination.

Mol Ecol, 2004 Nov, 13(11), 3329 - 44
Phylogeography recapitulates topography: very fine-scale local endemism of a saproxylic 'giant' springtail at Tallaganda in the Great Dividing Range of south-east Australia; Garrick RC et al.; Comparative phylogeography can reveal processes and historical events that shape the biodiversity of species and communities . As part of a comparative research program, the phylogeography of a new, endemic Australian genus and species of log-dependent (saproxylic) collembola was investigated using mitochondrial sequences, allozymes and anonymous single-copy nuclear markers . We found the genetic structure of the species corresponds with five a priori microbiogeographical regions, with population subdivision at various depths owing to palaeoclimatic influences . Closely related mtDNA haplotypes are codistributed within a single region or occur in adjacent regions, nuclear allele frequencies are more similar among more proximate populations, and interpopulation migration is rare . Based on mtDNA divergence, a late Miocene-late Pliocene coalescence is likely . The present-day distribution of genetic diversity seems to have been impacted by three major climatic events: Pliocene cooling and drying (2.5-7 million years before present, Mybp), early Pleistocene wet-dry oscillations (c . 1.2 Mybp) and the more recent glacial-interglacial cycles that have characterized the latter part of the Quaternary (<0.4 Mybp).

Ecotoxicol Environ Saf, 2005 Jan, 60(1), 73 - 80
Cyst-based toxicity tests XVII--prefeeding advantages in short-chronic rotifer microbiotests; Belgis C et al.; Attempts were made to overcome too-low reproductive output in the 48-h short-chronic microbiotest with the rotifer Brachionus calyciflorus fed with microalgae (Pseudokirchneriella subcapitata) from algal beads older than 4 months (Chemosphere 50 (2002a) 365) . Prefeeding of freshly hatched rotifers prior to the start of the actual growth experiments resulted in a satisfactory reproduction (i.e., an intrinsic growth rate r > 0.65), even with subsequent feeding of the rotifers with microalgae from beads stored up to 1 year . Rotirich is an attractive alternative inert food compared with the mixture of microalgae and Spirulina, originally used to prefeed the rotifers . When the rotifers were prefed for 2 h with 30 microg/mL Rotirich, it was then not necessary to add Spirulina to the microalgal food from beads . The above findings are a substantial simplification of the testing procedure of the new short-chronic "culture/maintenance-free" Rotoxkit microbiotest.

Fertil Steril, 2004 Oct, 82(4), 797 - 8
Microbiology of the endometrium and in vitro fertilization: do we yet understand the implications?
Barnhart KT.
The hypothesis that bacteria in the uterus may be detrimental to an embryo is very thought provoking . The data, however, do not yet support that this is a clinically relevant issue.

Mund Kiefer Gesichtschir, 2004 Sep, 8(5), 311 - 5 Epub 2004 Jul 29.
{Chronic cutaneous infiltration with abscess and fistula formation . A type of clinical course in atypical mycobacteriosis}; Moergel M et al.; BACKGROUND: Cutaneous lesions of atypical mycobacteriosis are very rare in Western Europe . Both the slow disease progression and the rather unspecific clinical appearance of the chronic infiltration render cutaneous mycobacteriosis a diagnostic challenge . In this communication, we present an uncommon, biphasic course of a cutaneous atypical mycobacteriosis . CASE REPORT: A 3-year-old child was referred to our institution due to submandibular cutaneous infiltration together with subcutaneous abscess formation . There was no carious lesion or other odontogenic potential source of infection . First-line therapy was incision of the abscess and local curettage to obtain a specimen for histologic and microbiologic evaluation, which revealed atypical mycobacteria . In spite of local curettage, only partial remission of the cutaneous infiltration was achieved . When the child was readmitted 9 months later, the lesion had progressed to a scrofulosis-like appearance even though specific antibiotic treatment had been administered . Complete surgical excision of the affected skin and extirpation of the respective lymph node and fistula was performed as second-line therapy . Complete remission was achieved without further adjuvant treatment . DISCUSSION: Our case demonstrates the problems that may arise with this rare type of mycobacterial infection . In all probability, scratch artifacts due to atopic dermatitis provided the source/entry of the infection in this specific case . Both histopathologic findings and clinical appearance were suggestive of classic cutaneous tuberculosis (tuberculosis cutis colliquativa), although it was in fact an atypical mycobacteriosis . Furthermore, obstinate persistence after minor surgical intervention and even after specific (triple) antibiotic treatment illustrates the difficulties of limited, minimally invasive treatment.

Nurs Times . 2004 Sep 7-13;100(36):56, 58.
Teaching nurses the importance of microbiology for infection control; Shuttleworth A; Ann Shuttleworth discusses whether nurses need an understanding of microbiology to ensure their infection control practice is effective, and whether they receive adequate education on the subject.

J Appl Toxicol, 2004 Sep-Oct, 24(5), 323 - 6
Sensitivity evaluation of the Daphtoxkit and Thamnotoxkit microbiotests on blind samples; Torokne A; At the Oregon State University Workshop we utilized two "culture/maintenance-free" microbiotests, the Thamnotoxkit F with the anostracan crustacean Thamnocephalus platyurus and the Daphtoxkit F magna with the cladoceran crustacean Daphnia magna, to determine the toxicity of water samples in a blind format . The Thamnotoxkit was applied to 7 samples and the Daphtoxkit to 12 samples . The chemical composition of the water samples to which the Toxkit microbiotests had been applied was disclosed a posteriori by the organizers and appeared to contain chlordimeform, colchicine, phosdrin, mercuric chloride, sodium arsenite, metham sodium, sodium cyanide, trimethylol propane phosphate, p-chlorophenol and a natural sediment sample containing mercury . Three of the water samples were blanks that had not been spiked with chemicals . No false positives were obtained with the two Toxkits and all the toxic waters were earmarked as such by the microbiotests, except trimethylol propane phosphate, which was not found to induce acute effects in Daphnia magna in the non-diluted water sample containing 100 mg l(-1) of this compound.

Braz J Infect Dis, 2004 Jun, 8(3), 259 - 62 Epub 2004 Sep 29.
Human ehrlichioses in Brazil: first suspect cases; Calic SB et al.; Brazilian spotted fever (BSF) rickettsiosis is the most common and recognized of the human rickettsioses in Brazil . It is difficult to establish the diagnosis of human rickettsiosis infection by routine microbiologic methods, creating a false idea that Rickettsia and Ehrlichia infections are rare and without importance . New tick-borne diseases, like human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME), have been described in many countries . These diseases can present symptoms similar to rickettsioses of the spotted fever group, and they are transmitted by ixodid ticks . The first two suspected cases of human ehrlichiosis in Brazil were first considered to be cases of BSF . The differential diagnosis was made at the Minas Gerais Rickettsiosis Public Health Laboratory . The clinical and laboratory findings, with positive serology for the HME agent, indicated suspected cases of human ehrlichioses in Brazil.

Laryngoscope, 2004 Sep, 114(9), 1533 - 5
Chronic rhizopus invasive fungal rhinosinusitis in an immunocompetent host; Scharf JL et al.; HYPOTHESIS: Rhizopus species may cause chronic invasive fungal rhinosinusitis in an immunocompetent host . STUDY DESIGN: Individual case report . METHODS: Review of inpatient and outpatient records, radiographic studies, pathologic examination, and microbiology of the presented case . Review of the literature . RESULTS: Rhizopus species may be the cause of chronic invasive fungal rhinosinusitis in an immunocompetent host and result in intracranial extension.

Gene, 2004 Oct 13, 340(2), 189 - 96
Marsupial relationships and a timeline for marsupial radiation in South Gondwana; Nilsson MA et al.; Recent marsupials include about 280 species divided into 18 families and seven orders . Approximately 200 species live in Australia/New Guinea . The remaining species inhabit South America with some of these secondarily ranging into North America . In this study, we examine marsupial relationships and estimate their divergences times using complete mitochondrial (mt) genomes . The sampling, which includes nine new mtDNAs and a total number of 19 marsupial genomes, encompasses all extant orders and 14 families . The analysis identified a basal split between Didelphimorphia and remaining orders about 69 million years before present (MYBP), while other ordinal divergences were placed in Tertiary times . The monotypic South American order Microbiotheria (Dromiciops gliroides, Monito del Monte) was solidly nested among its Australian counterparts . The results suggest that marsupials colonized Australia twice from Antarctica/South America and that the divergence between Microbiotheria and its Australian relatives coincided with the geological separation of Antarctica and Australia . Within Australia itself, several of the deepest divergences were estimated to have taken place close to the Eocene/Oligocene transition.

Recenti Prog Med, 2004 Sep, 95(9), 403 - 13
{Tick-borne infections}; Calza L et al.; Ticks are obligate, blood-sucking ectoparasites that are the most common agents of vector-borne infectious disease in the United States and European countries . Ticks play an important role in transmitting several infectious agents, such as viruses, bacteria, spirochetes, rickettsia, and parasites, and tick bites cause a variety of acute and chronic infectious diseases, including Lyme disease, tick-borne relapsing fever, Rocky Mountain and Mediterranean spotted fevers, ehrlichiosis, Q fever, tularemia, babesiosis, and tick-borne viral encephalitis . Since its identification nearly 30 years ago, Lyme disease has continued to spread, and it is now the most commonly reported arthropod-borne illness in American and European countries . Rickettsial infections are widely distributed throughout the world and have a remarkable influence on public health and military activities as a possible biological weapon . Tick-borne encephalitis virus is endemic in central, eastern and northern Europe and may cause a wide spectrum of clinical forms, ranging from asymptomatic infection to severe meningo-encephalitis . This article reviews the epidemiology, microbiology, clinical manifestation, diagnosis and treatment of the major tick-borne infectious diseases in the United States and Europe.

Lab Chip, 2004 Oct, 4(5), 446 - 52 Epub 2004 Sep 27.
Continuous cell washing and mixing driven by an ultrasound standing wave within a microfluidic channel; Hawkes JJ et al.; Ultrasound standing wave radiation force and laminar flow have been used to transfer yeast cells from one liquid medium to another (washing) by a continuous field-flow fractionation (FFF) approach . Two co-flowing streams, a cell-free suspending phase (flow rate > 50% of the total flow-through volume) and a yeast suspension, were introduced parallel to the nodal plane of a 3 MHz standing wave resonator . The resonator was fabricated to have a single pressure nodal plane at the centre line of the chamber . Laminar flow ensured a stable interface was maintained as the two suspending phases flowed through the sound field . Initiation of the ultrasound transferred cells to the cell-free phase within 0.5 s . This particle transfer procedure circumvents the pellet formation and re-suspension steps of centrifuge based washing procedures . In addition, fluid mixing was demonstrated in the same chamber at higher sound pressures . The channel operates under negligible back-pressure (cross-section, 0.25 {times} 10 mm) and with only one flow convergence and one flow division step, the channel cannot be easily blocked . The force acting on the cells is small; less than that experienced in a centrifuge generating 100g . The acoustically-driven cell transfer and mixing procedures described may be particularly appropriate for the increasingly complex operations required in molecular biology and microbiology and especially for their conversion to continuous flow processes.

J Clin Microbiol, 2004 Oct, 42(10), 4679 - 85
Evaluation and validation of real-time reverse transcription-pcr assay using the LightCycler system for detection and quantitation of norovirus; Pang X et al.; We developed an assay for the detection and quantitation of norovirus with the LightCycler SYBR Green-based real-time reverse transcription-PCR (real-time LC RT-PCR) and previously published primers in the capsid and the polymerase gene . One hundred thirty-two stool specimens from the Provincial Laboratory for Public Health (Microbiology), Alberta, Canada, and the Centers for Disease Control and Prevention, Atlanta, Ga., were used to validate the new assay . The samples were collected from patients involved in outbreaks of acute gastroenteritis or children who presented with sporadic gastroenteritis . The real-time LC RT-PCR assay detected norovirus strains from three genogroup I (G-I) clusters (G-I/1, G-I/2, and G-I/3) and 10 genogroup II (G-II) clusters (G-II/1, G-II/2, G-II/3, G-II/4, G-II/6, G-II/7, G-II/10, G-II/12, G-II/15, and G-II/16) . There was 100% concordance with the results from 58 stool specimens which tested positive by conventional RT-PCR assays . By dilution analysis, the real-time LC RT-PCR was 10,000 times more sensitive than the conventional RT-PCR . The new assay increased the number of samples in which noroviruses were detected by 19% . The real-time LC RT-PCR had a wide dynamic range, detecting from 5 to 5 x 10(6) copies of RNA per reaction, resulting in a theoretical lower limit of detection of 25,000 copies of RNA per g of stool . No cross-reactions were found with specimens containing sapovirus, rotavirus, astrovirus, and adenovirus . Because of the high sensitivity and specificity of the assay with a relatively rapid and simple procedure, the real-time LC RT-PCR will be useful as a routine assay for the clinical diagnosis of norovirus infection.

Nat Biotechnol, 2004 Oct, 22(10), 1268 - 74
The challenges of modeling mammalian biocomplexity; Nicholson JK et al.; Understanding the relationships between human genetic factors, the risks of developing major diseases and the molecular basis of drug efficacy and toxicity is a fundamental problem in modern biology . Predicting biological outcomes on the basis of genomic data is a major challenge because of the interactions of specific genetic profiles with numerous environmental factors that may conditionally influence disease risks in a nonlinear fashion . 'Global' systems biology attempts to integrate multivariate biological information to better understand the interaction of genes with the environment . The measurement and modeling of such diverse information sets is difficult at the analytical and bioinformatic modeling levels . Highly complex animals such as humans can be considered 'superorganisms' with an internal ecosystem of diverse symbiotic microbiota and parasites that have interactive metabolic processes . We now need novel approaches to measure and model metabolic compartments in interacting cell types and genomes that are connected by cometabolic processes in symbiotic mammalian systems.

Proc Natl Acad Sci U S A, 2004 Oct 19, 101(42), 15042 - 5 Epub 2004 Oct 06.
Wolbachia-induced cytoplasmic incompatibility as a means for insect pest population control; Zabalou S et al.; Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations . Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents . Wolbachia manipulate host reproduction by using several strategies, one of which is cytoplasmic incompatibility (CI) {Stouthamer, R., Breeuwer, J . A . J . & Hurst, G . D . D . (1999) Annu . Rev . Microbiol . 53, 71-102} . We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor . Wolbachia induced complete CI in the novel host . Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced CI could be used as a novel environmentally friendly tool for the control of medfly populations . The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations.

Cell Motil Cytoskeleton, 2004 Nov, 59(3), 201 - 14
Disruption of genes encoding predicted inner arm dynein heavy chains causes motility phenotypes in Tetrahymena; Liu S et al.; The multi-dynein hypothesis {Asai, 1995: Cell Motil Cytoskeleton 32:129-132} states: (1) there are many different dynein HC isoforms; (2) each isoform is encoded by a different gene; (3) different isoforms have different functions . Many studies provide evidence in support of the first two statements {Piperno et al., 1990: J Cell Biol 110:379-389; Kagami and Kamiya, 1992: J Cell Sci 103:653-664; Gibbons, 1995: Cell Motil Cytoskeleton 32:136-144; Porter et al., 1996: Genetics 144:569-585; Xu et al., 1999: J Eukaryot Microbiol 46:606-611} and there is evidence that outer arms and inner arms play different roles in flagellar beating {Brokaw and Kamiya, 1987: Cell Motil . Cytoskeleton 8:68-75} . However, there are few studies rigorously testing in vivo whether inner arm dyneins, especially the 1-headed inner arm dyneins, play unique roles . This study tested the third tenet of the multi-dynein hypothesis by introducing mutations into three inner arm dynein HC genes (DYH8, 9 and 12) that are thought to encode HCs associated with 1-headed inner arm dyneins . Southern blots, Northern blots, and RT-PCR analyses indicate that all three mutants (KO-8, 9, and 12) are complete knockouts . Each mutant swims slower than the wild-type cells . The beat frequency of KO-8 cells is lower than that of the wild-type cells while the beat frequencies of KO-9 and KO-12 are not different from that of wild-type cells . Our results suggest that each inner arm dynein HC is essential for normal cell motility and cannot be replaced functionally by other dynein HCs and that not all of the 1-headed inner arm dyneins play the same role in ciliary motility . Thus, the results of our study support the multi-dynein hypothesis {Asai, 1995: Cell Motil Cytoskeleton 32:129-132} .

J Wildl Dis, 2004 Jul, 40(3), 607 - 11
Foot infections associated with Arcanobacterium pyogenes in free-living fallow deer (Dama dama); Lavin S et al.; We describe foot infection associated with Arcanobacterium pyogenes in three adult male free-living fallow deer (Dama dama) from Sueve Regional Hunting Reserve (Principality of Asturias, Spain) . Affected fallow deer were culled in November 1997 and 1998 during the hunting season . Necropsy, radiography, and microbiologic analysis were carried out for each animal . Unilateral swelling of one extremity at the coronary band was observed in all three cases . Areas of bone loss, severe periosteal reaction, and soft tissue swelling were seen on radiography . Lead fragments were observed in one fallow deer . Seven bacterial species were isolated, but only Arcanobacterium pyogenes was routinely found . Weather conditions in the area (mild temperatures and high humidity), the land (alternating pasture land and rock), the animal population density (both fallow deer and domestic herds of cows, horses, sheep, and goats, live side by side in the same areas), and hunting activities could be related to the frequency of these infections.

J Cyst Fibros, 2004 Mar, 3(1), 8 - 14
Improved outcomes of patients with cystic fibrosis admitted to the intensive care unit; Vedam H et al.; BACKGROUND: Invasive mechanical ventilation (IMV) has been previously associated with a poor outcome for patients with cystic fibrosis (CF), but with improved survival and the availability of lung transplantation intensive care unit (ICU) admission is being increasingly considered . This study aimed to review the outcomes of adult CF patients admitted to ICU, and to identify factors that may have influenced outcomes . METHODS: A retrospective audit was conducted of CF patients admitted to ICU . Anthropometric data, spirometry, nutritional status, sputum microbiology, arterial blood gas tensions, and mode of ventilation used were recorded . Immediate and 1-year survival and lung transplantation utilisation were recorded . RESULTS: Twenty patients were admitted to ICU and nine (45%) survived to hospital discharge . Five of the nine survivors had potentially reversible conditions . Four patients admitted with respiratory exacerbations alone who survived, were maintained on non-invasive ventilation (NIV) . The relative risk of deaths for patients with a BMI < 18 and a FEV1 < 24% predicted were 3.25 (1.27-3.25), and 3.68 (1.11-16.33), respectively . CONCLUSIONS: The outcome of patients with CF admitted to ICU has improved, with 45% of the patients in this study surviving to hospital discharge . Five of these survivors underwent successful lung transplantation . Long-term use of NIV post discharge may have contributed to this favourable outcome . A BMI < 18 and FEV1 < 24% predicted were associated with a significantly higher relative risk of death.

Curr Allergy Asthma Rep, 2004 Nov, 4(6), 496 - 9
Animal models of sinusitis: relevance to human disease; Kara CO; Experimental animal models are necessary to assess the efficacy of both medical and surgical procedures . The researcher who wishes to confirm and publish his results must overcome controversies concerning the validity of his model and, therefore, needs guidelines for standardization . Various models are offered in the literature . In this article, the rabbit and murine models of experimental sinusitis and their relevance to human disease with anatomical, histopathologic, and microbiologic aspects are discussed.

Pediatr Dermatol, 2004 Sep-Oct, 21(5), 548 - 50
Infantile acute hemorrhagic edema and rotavirus infection; Di Lernia V et al.; Infantile acute hemorrhagic edema (AHE) is a benign condition characterized by a dramatic onset of inflammatory edema and ecchymotic purpura in a target or cockade pattern . It is considered an uncommon form of cutaneous vasculitis occurring in children younger than 2 years of age . The outbreak is frequently preceded by an immunization or various infections . We describe an 11-month-old girl with rosette-shaped purpuric plaques on the face and limbs, clinically consistent with a diagnosis of AHE of the skin, associated with fever and diarrhea . Laboratory investigations showed a rotavirus infection, which has not previously been reported in association with AHE of the skin . The disease had a benign course without relapses . Appropriate microbiologic investigations are advisable to confirm the possible etiologic role of rotavirus.

Rev Iberoam Micol, 2004 Mar, 21(1), 1 - 9
{Histopathological diagnosis of mycoses}; Mayayo Artal E; Infectious diseases emerge as a cause of pathology in our patients . Among the possible etiologies, mycoses have shown a considerable increase in the two last decades . In general, the clinical features of fungal diseases are not very distinctive . The morphology and the clinical aspects of the fungi serve as a protocol for their correct identification . Clinicians, microbiologists and pathologists are essential for the diagnosis . The pathologist using a simple and fast methodology can diagnose some types of mycosis, but they do not only identify the causal agent, but also the kind of injury that produces, the inflammatory response and the affected organ or organs . Moreover, they can classify the mycosis as superficial, cutaneous, subcutaneous, deep and systemic depending on the location . The present review paper describes study guidelines for the pathologist faced with a fungal infection and new technical advances that are established in pathology laboratories for a more precise identification of the mycoses.

Biotechnol Prog, 2004 Sep-Oct, 20(5), 1352 - 8
Improvement of the thermoregulated T7 expression system by using the heat-sensitive lacI; Wang ZW et al.; The thermoregulated T7 expression system was previously reported to be an effective way to produce massive amounts of recombinant proteins (Chao, Y . P.; Law, W . S.; Chen, P . T.; Hung, W . B . High production of heterologous proteins in Escherichia coli using the thermo-regulated T7 expression system . Appl . Microbiol . Biotechnol . 2002b, 58, 446-453) . To ensure its practical applicability, the system was improved for stringency with the construction of the T7lac-promoter-containing plasmid associated with the thermolabile lacI gene (lacIts) . Owing to the recessive feature of lacIts, the wild-type lacI was removed from the genome of the cell . Moreover, the cell was engineered to carry the chromosomal copy of the T7 gene 1 subject to the regulation of lambdaPL and lambdaPR promoters . To characterize the system, the lacZ gene was fused to the T7lac promoter, and subsequent experiments showed that various amounts of LacZ could be synthesized in the plasmid-bearing cell in response to heat . Among the producers, the cell with the plasmid containing lacIts (substitution of Gly265 with Asp in lacI) was able to produce the maximal LacZ, the production accounting for an amplification of more than 200-fold over the uninduced level . A further demonstration was carried out to illustrate the practical usefulness of the developed system by producing carbamoylase on a 4000 L scale . Cultured to reach high cell density, the carbamolyase-producing cell was shown to retain plasmids with 95% stability and to be capable of producing soluble protein equal to 13% of the total cell proteins . Overall, it illustrates the remarkable features of the developed system with tightness, high expression level, thermal inducibility, and high stability.

Acta Pharmacol Sin, 2004 Oct, 25(10), 1262 - 6
In vivo and in vitro chondrotoxicity of ciprofloxacin in juvenile rats; Li P et al.; AIM: To study the relationship between chondrotoxicity and toxicokinetics of ciprofloxacin (CPFX) . METHODS: Rats, 4-week old, were treated with CPFX 0, 400, 800, and 1200 mg/kg ig once daily on seven consecutive days . The knee joint cartilage was examined histopathologically . The concentration of CPFX in venous blood and knee joint cartilage samples were determined by a microbioassay using Escherichia coli 44102 . The effects of CPFX on proliferation of chondrocytes and secretion of soluble proteoglycans were determined with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and 1,9-dimethylmethylene blue (DMB) assay, respectively . RESULTS: Cartilage was severely lesioned after treatment with CPFX 800 or 1200 mg/kg for 7 d, such as matrix swelling and loss of chondrocytes . The thickness of cartilage was significantly decreased compared with the control group . The maximum serum concentration (Cmax), the area under the plasma concentration-time curve (AUC(0-infinity)), and concentration in cartilage was 16.3 +/- 2.1 mg/L, 97.2 +/- 12.3 mg x h x L(-1), and 13.4 +/- 2.8 microg x g(-1) and 21.8 +/- 2.5 mg/L, 143.1 +/- 22.3 mg x h x L(-1), and 20.3 +/- 3.5 microg x g(-1) after oral administration of CPFX 800 or 1200 mg/kg on d 1, respectively . The data on d 6 were similar with that on d 1 . CPFX inhibited proliferation of chondrocytes and the secretion of soluble proteoglycans . CONCLUSION: CPFX concentrations in serum and cartilage could provide a better basis for risk assessment .

Annu Rev Public Health . 2004 Sep 28; {Epub ahead of print}
New Microbiology Tools for Public Health and Their Implications; Robertson BH et al.; The realm of diagnostic assays for detection of acute infections is rapidly changing from antibody detection to pathogen detection, from clinical laboratory based to point-of-care based, from single analyte detection to multiple analyte detection, and is more focused on detection using less invasive approaches for collecting biological samples . New assays are typically more sensitive than are conventional assays and have the capability of providing more information that characterizes the pathogen or the host response to the pathogen . From a public health perspective, the advent of molecular epidemiology, which allows tracking of pathogens based on unique genetic sequences or antigenic properties, has revolutionized how epidemiologists investigate and evaluate epidemics and assess endemic diseases . In addition, the use of point-of-care (POC) devices can impact the detection and surveillance of infections and will enhance our ability to accurately identify the causes of illnesses . Expected online publication date for the Annual Review of Public Health Volume 26 is March 17, 2005 . Please see for revised estimates.

Indian J Pediatr, 2004 Sep, 71(9), 837 - 43
Treatment of invasive Aspergillosis in children with hematologic malignancies; Abuhammour W et al.; The respiratory tract is the most common organ involved with Aspergillosis in children with hematologic malignancies . Also Aspergillus species tend to invade blood vessels resulting in systemic dissemination to multiple organs . Early diagnosis and treatment are pivotal to the patient's outcome . A high index of suspicion should be maintained in children who have profound neutropenia and present with prolonged fever that is unresponsive to systemic antibiotics . Several diagnostic modalities should be used simultaneously in order to confirm the diagnosis in an expedited manner . Combination and sequential antifungal therapy have been shown to be of added benefit . Surgical intervention is associated with a high mortality rate, but may be indicated in children with a localized disease . In this article the authors review the epidemiology, microbiology, pathology, and clinical manifestations of invasive aspergillosis in children with hematologic malignancies . Current diagnostic approach, medical, and surgical treatment options are discussed.

Int J Syst Evol Microbiol, 2004 Sep, 54(Pt 5), 1649 - 52
Borrelia turcica sp . nov., isolated from the hard tick Hyalomma aegyptium in Turkey; Guner ES et al.; Previously, a novel, fast-growing spirochaete was isolated from the hard tick Hyalomma aegyptium, which infests tortoises (Testudo graeca), by using Barbour-Stoenner-Kelly (BSK) II medium; the tick samples were taken from the Istanbul area in northwestern Turkey {Guner et al . (2003) . Microbiology 149, 2539-2544} . Here is presented a detailed characterization of the spirochaete . Electron microscopy revealed that strain IST7T is morphologically similar to other spirochaetes of the genus Borrelia and possesses 15 to 16 flagellae that emerge from both polar regions . PFGE analysis revealed the genome to comprise a linear chromosome of approximately 1 Mb; two large linear plasmids of approximately 145 and 140 kb, and several small plasmids ranging from 50 to 20 kb in size were also found . The 16S rRNA gene sequence of this Borrelia isolate exhibited 99.4 to 99.8 % identity with other strains isolated from H . aegyptium and less than 99 % similarity with those of other Borrelia species . A phylogenetic tree, generated from 16S rRNA gene sequences, demonstrated that the spirochaete isolates from H . aegyptium clustered together and branched off from both Lyme-disease-related and relapsing-fever-associated Borrelia species . A single copy of the rrs gene was detected in the genome of strain IST7T by Southern hybridization . DNA-DNA hybridization results showed that strain IST7T was distinct from Lyme-disease-related Borrelia, Borrelia burgdorferi and the relapsing-fever-associated species Borrelia hermsii . The G+C content of strain IST7T is 30.0 mol% . From these genetic features, a novel Borrelia species, Borrelia turcica sp . nov., is proposed; the type strain is IST7T (= JCM 11958T = DSM 16138T).

Fed Regist, 2004 Sep 23, 69(184), 56934 - 6
Medical devices; immunology and microbiology devices; classification of the beta-glucan serological assay . Final rule; Food and Drug Administration et al.; The Food and Drug Administration (FDA) is classifying the beta-glucan serological reagent device into class II (special controls) . The special control that will apply to the device is the guidance document entitled "Class II Special Controls Guidance Document: Serological Assays for the Detection of Beta-Glucan." 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 1976 amendments), the Safe Medical Devices Act of 1990, the Food and Drug Administration Modernization Act of 1997, and the Medical Device User Fee and Modernization Act of 2002 . The agency is classifying the 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 publishing a notice of availability of a guidance document that is the special control for this device.

Rofo, 2004 Oct, 176(10), 1466 - 8
{Lymphadenitits caused by nontuberculous mycobacteria}; Mentzel HJ et al.; Infections caused by nontuberculous mycobacteria (NTM) commonly manifest as lymphadenitis in otherwise healthy children, as pneumonia in patients with chronic airway diseases, and as disseminated disease in patients with immunosuppression . The aim of this case report is to describe imaging findings (sonography, electrical impedance scanning, MRI) in children with NTM infection of the head and neck and to give some information about microbiologic findings and therapeutic options . In children presenting with enlarged neck masses and radiologically demonstrated nodal masses, the diagnosis of infections with NTM should be considered.

J R Army Med Corps, 2004 Jun, 150(2), 99 - 106
Surgical workload from an integrated UK field hospital during the 2003 Gulf conflict; Rew DA et al.; AIMS: This paper reports the surgical experience of a UK reserve field surgical hospital during military operations in Iraq during March and April 2003 . Particular reference is given to the integration of the surgical specialities, consultant led and multidisciplinary teamworking in the treatment of military and civilian casualties from all sources and of all causes . METHODS All surgical workload data was collected prospectively for the warfighting (Phase One) and specialist referral (Phase Two) phases of the operation . Standard military and Red Cross protocols were used for all injuries and microbiology studies were undertaken for penetrating ballistic injuries . Operations were classified by anatomical region; upper or lower limb; head and neck; thoracic; abdominal, including genitourinary; ophthalmic; and burns, and by whether they were primary; secondary planned or secondary unplanned . RESULTS: A wide range of patterns of wounding arising from combat and non-combat related military and civilian trauma and burns were seen and treated . 323 operations were performed . There were 116 operations in phase one and 207 operations in phase two . There were 160 primary procedures (new patients), 146 secondary planned and 17 unplanned procedures (range one to eight procedures per patient) . Close integration of general, orthopaedic, plastic, maxillofacial, ophthalmic and neurological surgeons and general and ITU anaesthetists allowed the delivery of a range of specialist treatment to a heterogenous patient population, including children as young as 6 months and a lady in the advanced stages of pregnancy . CONCLUSIONS: Patterns of wounding in casualties surviving to reach specialist field hospital care were similar to those reported in other high intensity conflicts . A consultant led, multidisciplinary approach with field intensive care facilities allowed combined, staged and safe surgery for complex, multi-organ and multi-limb trauma in an austere environment.

Cas Lek Cesk, 2004, 143(7), 485 - 9
{Problems of causality in medicine: The case of viruses and tumors--Part 2 . Viruses and human malignant tumors}; Vonka V; A special case in contemporary etiological studies is definition of the role of viruses in the pathogenesis of human cancer . Virus-associated cancer develops only in a small minority of infected subjects, which implies that, if the virus does play a role in the pathogenesis of the malignancy, other factors must be also involved . The author summarises the general properties of human tumour viruses, reviews the development of causal thinking in microbiology and proposes guidelines that might help to determine the role of viruses in human cancer.

J Microbiol Methods, 2004 Nov, 59(2), 253 - 62
Ergosterol as a measure of living fungal biomass: persistence in environmental samples after fungal death; Mille-Lindblom C et al.; The membrane lipid ergosterol is found almost exclusively in fungi, and is frequently used by environmental microbiologists as an indicator of living fungal biomass, based on the assumption that ergosterol is labile, and therefore rapidly degraded after the death of fungal hyphae . We studied the degradation of ergosterol in environmental samples without living fungi . Under the conditions used in this study, ergosterol was very stable both when added as a pure compound and when associated with dead fungi . The decrease of ergosterol was at most 34% during 2 months when protected from sunlight . Presence of a natural bacterial assemblage did not enhance degradation over this time period, as compared to sterile controls . However, photochemical degradation was significant, and led to a 43% decrease of in ergosterol content during 24 h . These results suggest that ergosterol should be used cautiously as a biomarker for living fungi.

Ceska Gynekol, 2004 Jul, 69(4), 253 - 7
{Evidence of antibodies against the chlamydial heat shock protein (cHSP60) in women from "in vitro" fertilization program}; Pospisil L et al.; OBJECTIVE: A contribution to the role of chlamydial heat shock protein in women from the IVF program . DESIGN: A serological study proving the antibodies against the chlamydial heat shock protein . SETTING: Veterinary Research Institute, Brno, Department of Clinical Microbiology, and Centrum of Reproductive Medicine, Faculty Hospital Brno . METHODS: The IgG antibodies against the chlamydial heat shock protein (cHSP60) and the IgA and IgG antibodies against species specific chlamydial major outer membrane protein (cMOMP) of Chlamydia trachomatis (C . trachomatis) and Chlamydophila pneumoniae (C . pneumoniae) in the blood serum of 70 females being in the fertilization program due to fertility disorders (Group 1) have been estimated and the results compared with those obtained in 50 females suffering from pelvic inflammation disease (PID) (Group 2) and in 51 female blood donors (Group 3) respectively . RESULTS: The anti-cHSP60 antibodies have been ascertained as follows: in 26 women from the first group (37.1%), in 16 of the second group (32.0%) and in 12 (23.5%) of female blood donors . The anti cHSP60-antibody-occurrence and mean index of positivity as well, found in the groups examined, was statistically insignificant . Similarly no difference has been found in the occurrence of the species specific anti-cMOMP antibodies C . trachomatis and C . pneumoniae in the groups examined . The anti-cMOMP antibodies against C . trachomatis and simultaneously against C . pneumoniae have been detected in 21 from 171 women of all groups (12.3%), only against C . trachomatis in 9 (5.3%) and against C . pneumoniae in 73 of them (42.7%) . There was considerably higher occurrence of the specific antibodies against C . trachomatis in women with a positive finding of antibodies against the cHSP60, especially in women suffering from gynecological disorders, than in women without such antibodies . CONCLUSION: C . trachomatis has a significant impact on the production of antibodies against the cHSP60 . This fact can be documented by considerably higher occurrence of the specific antibodies against C . trachomatis in women with a positive finding of antibodies against the cHSP60, than in women without such antibodies . Preceding infection C . trachomatis and following sensitization with chlamydial heat shock protein indicate an unfavourable prognosis of the reproductive outcome and impairs the perspective of a successful in vitro fertilization . The proof of antibodies against the chlamydial HSP60 can be recommended as a further auxiliary criterion in women suffering from fertility disorders.

J Clin Periodontol, 2004 Oct, 31(10), 894 - 902
Cytokine profile in gingival crevicular fluid of aggressive periodontitis: influence of smoking and stress; Kamma JJ et al.; BACKGROUND: Cigarette smoking and stress are considered risk factors that have been associated with periodontal disease progression . Conflicting results have been reported concerning the direct influence of smoking on the subgingival microbiota of periodontitis patients . Cytokine production may also be influenced by smoking and stress leading to an imbalance that disturbs the host-parasite relationship . AIM: The objective of the present study was to evaluate the influence of cigarette smoking on the gingival crevicular fluid (GCF) levels of interleukin (IL)-1beta, IL-4, IL-6 and IL-8 in aggressive or early onset periodontitis (EOP) patients and in healthy controls (H), psychosocial stress being considered as modifying factor . MATERIAL AND METHODS: Sixty-five EOP and 35 periodontally healthy individuals participated in this cross-sectional study . All the participants were interviewed about their smoking habits and their stressful social events . Clinical examination included the assessment of plaque index (PI), bleeding on probing (BOP), clinical attachment level (CAL) and probing pocket depth (PPD) . GCF was collected using durapore strips, from four sites per patient, randomly selected in each quadrant . The total amounts of IL-1beta, IL-4, IL-6 and IL-8 were measured in a total of 400 samples using commercially available enzyme-linked immunosorbent assays . RESULTS: All clinical parameters were significantly higher in the EOP group compared to the H group . There were no significant differences between EOP smokers and EOP non-smokers with regard to plaque accumulation, CAL and PPD of the sampling sites, whereas mean CAL and PPD of the diseased sites were greater in EOP smokers than in EOP non-smokers . In addition, EOP smokers seemed to have significantly less BOP and greater bone loss compared to EOP non-smokers . Significant interactions between "EOP" and "smoking" were present for total amounts of IL-1beta and IL-4 . IL-1beta, IL-6 and IL-8 showed significant main effects with healthy smokers and healthy non-smokers, respectively . For IL-8, stress presented a statistically significant interaction with smoking status and EOP (F=4.742, p=0.030) . More specifically EOP smokers were statistically affected by stress . CONCLUSIONS: Smoking influences host-related factors including cytokine network . The relative importance of smoking and stress-related alterations and their precise mode of action in increasing the risk of aggressive periodontitis remains to be elucidated . Copyright Blackwell Munksgaard, 2004

Dig Surg, 2004, 21(4), 297 - 304 Epub 2004 Sep 13.
Fungal infection but not type of bacterial infection is associated with a high mortality in primary and secondary infected pancreatic necrosis; Connor S et al.; INTRODUCTION: Knowledge of microbiology in the prognosis of patients with necrotizing pancreatitis is incomplete . AIM: This study compared outcomes based on primary and secondary infection after surgery for pancreatic necrosis . METHOD: From a limited prospective database of pancreatic necrosectomy, a retrospective case note review was performed (October 1996 to April 2003) . RESULTS: 55 of 73 patients had infected pancreatic necrosis at the first necrosectomy . 25 of 47 patients had resistant bacteria to prophylactic antibiotics (n = 21) or did not receive prophylactic antibiotics (n = 4), but this was not associated with a higher mortality (9 of 25) compared to those with sensitive organisms (4 of 22) . Patients with fungal infection (n = 6) had a higher initial median (95% CI) APACHE II score compared to those without (11 (9-13) verus 8.5 (7-10), p = 0.027) . Five of six patients with fungal infection died compared to 13 of 47 who did not (p = 0.014) . With the inclusion of secondary infections 21 (32%) of 66 patients had fungal infection with 10 (48%) deaths compared to 11 (24%) of 45 patients without fungal infection (p = 0.047) . CONCLUSION: Whether associated with primary or secondary infected pancreatic necrosis, fungal but not bacterial infection was associated with a high mortality .

J Clin Virol, 2004 Oct, 31(2), 96 - 9
Influenza A pneumonia presenting as progressive focal infiltrates in a stem cell transplant recipient; Scott JD et al.; BACKGROUND: Stem cell transplant recipients are susceptible to pulmonary infections, including influenza A . Typically, isolated influenza pneumonia has a diffuse, interstitial infiltrate pattern . OBJECTIVES: To describe the unusual clinical and radiographic course of influenza A pneumonia in a stem cell transplant recipient . STUDY DESIGN: Case report in which microbiologic assays, bronchoscopic and pathologic specimens are obtained . RESULTS: We describe a patient with influenza A pneumonia 8 months following a peripheral blood stem cell transplant who presented with minimal respiratory symptoms and rapidly progressing, focal pulmonary infiltrates . The large size and appearance of the masses have not been reported before in a patient with isolated influenza . CONCLUSION: This case highlights the differences of presentation and importance of early diagnosis and treatment of immunocompromised patients infected with influenza.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2004 Sep, 98(3), 321 - 3
Fusospirochetosis causing necrotic oral ulcers in patients with HIV infection; Salama C et al.; Under certain permissive circumstances, normally occurring fusiform bacteria and Borrelia spirochetes can result in a symbiotic overgrowth that leads to necrotic oral ulcers (stomatitis), gingivitis, and periodontitis . These lesions are collectively known as oral fusospirochetosis and may be under-appreciated in patients with HIV infection and AIDS . Fusospirochetal oral ulcers in patients with HIV are often large, necrotic, and malodorous; they respond completely to penicillin . We report 3 patients with HIV infection and fusospirochetal ulcerative stomatitis and review the clinical presentation, microbiologic diagnosis, potential pathogenesis, and treatment of these lesions.

J Immunol, 2004 Sep 15, 173(6), 4137 - 46
The essential role of the intestinal microbiota in facilitating acute inflammatory responses; Souza DG et al.; The restoration of blood flow, i.e., reperfusion, is the treatment of choice to save viable tissue following acute ischemia of a vascular territory . Nevertheless, reperfusion can be accompanied by significant inflammatory events that limit the beneficial effects of blood flow restoration . To evaluate the potential role of the intestinal microbiota in facilitating the development of tissue injury and systemic inflammation, germ-free and conventional mice were compared in their ability to respond to ischemia and reperfusion injury . In conventional mice, there was marked local (intestine) and remote (lung) edema formation, neutrophil influx, hemorrhage, and production of TNF-alpha, KC, MIP-2, and MCP-1 . Moreover, there was an increase in the concentration of serum TNF-alpha and 100% lethality . In germ-free mice, there was no local, remote, or systemic inflammatory response or lethality after intestinal ischemia and reperfusion and, in contrast to conventional mice, germ-free animals produced greater amounts of IL-10 . Similar results were obtained after administration of LPS, i.e., little production of TNF-alpha or lethality and production of IL-10 after LPS in germ-free mice . Blockade of IL-10 with Abs induced marked inflammation and lethality in germ-free mice after ischemia and reperfusion or LPS administration, demonstrating that the ability of these mice to produce IL-10 was largely responsible for their "no inflammation" phenotype . This was consistent with the prevention of reperfusion-associated injury by the exogenous administration of IL-10 to conventional mice . Thus, the lack of intestinal microbiota is accompanied by a state of active IL-10-mediated inflammatory hyporesponsiveness .

Annu Rev Microbiol . 2004 Jun 16; {Epub ahead of print}
The Small RNA Regulators of Escherichia coli: Roles and Mechanisms; Gottesman S; Small noncoding RNAs have been found in all organisms, primarily as regulators of translation and message stability . The most exhaustive searches have taken place in E . coli, resulting in identification of more than 50 small RNAs, or 1%-2% of the number of protein-coding genes . One large class of these small RNAs uses the RNA chaperone Hfq; members of this class act by pairing to target messenger RNAs . Among the members of this class are DsrA and RprA, which positively regulate rpoS translation, OxyS, which negatively regulates rpoS translation and fhlA translation, RyhB, which reapportions iron use in the cell by down-regulating translation of many genes that encode Fe-containing proteins, and Spot 42, which changes the polarity of translation in the gal operon . The promoters of these small RNAs are tightly regulated, frequently as part of well-understood regulons . Lessons learned from the study of small RNAs in E . coli can be applied to finding these important regulators in other organisms . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 Jun 11; {Epub ahead of print}
The Cellulosomes: Multienzyme Machines for Degradation of Plant Cell Wall Polysaccharides; Bayer EA et al.; The discrete multi-component, multi-enzyme cellulosome complex of anaerobic cellulolytic bacteria provides enhanced synergistic activity among the different resident enzymes to efficiently hydrolyze intractable cellulosic and hemicellulosic substrates of the plant cell wall . A pivotal noncatalytic subunit called scaffoldin secures the various enzymatic subunits into the complex, via the cohesin-dockerin interaction . The specificity characteristics and tenacious binding between the scaffoldin-based cohesin modules and the enzyme-borne dockerin domains dictate the supramolecular architecture of the cellulosome . The diversity in cellulosome architecture among the known cellulosome-producing bacteria is manifest in the arrangement of their genes in either multiple-scaffoldin or enzyme-linked clusters on the genome . The recently described three-dimensional crystal structure of the cohesin-dockerin heterodimer sheds light on the critical amino acids that contribute to this high-affinity protein-protein interaction . In addition, new information regarding the regulation of cellulosome-related genes, budding genetic tools and emerging genomics of cellulosome-producing bacteria promises new insight into the assembly and consequences of the multi-enzyme complex . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 Jun 10; {Epub ahead of print}
Biosynthesis of Nonribosomal Peptides; Finking R et al.; Bacteria and fungi utilize large multifunctional enzymes, the so called nonribosomal peptide synthetases (NRPSs), to produce peptides of broad structural and biological activity . Biochemical studies have contributed substantially to the understanding of the key principles of these modular enzymes that can draw on a much larger number of catalytic tools for the incorporation of unusual features compared with the ribosomal system . Several crystal structures of NRPS domains have yielded deep insight into the catalytic mechanisms involved, led to a better prediction of the products assembled and to the construction of hybrid enzymes . In addition to the structure-function relationship of the core and tailoring domains of NRPSs, which is the main focus of this review, different biosynthetic strategies and essential enzymes for posttranslational modification and editing are discussed . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 Jun 10; {Epub ahead of print}
Cell-Mediated Immunity and the Outcome of Hepatitis C Virus Infection; Shoukry NH et al.; The hepatitis C virus (HCV) infects approximately three percent of the world's population . Some individuals resolve the infection spontaneously, but the majority develop persistent viremia that often causes progressive liver disease . There is an emerging consensus that cellular immune responses are essential for spontaneous resolution of acute hepatitis C and long-term protection from persistent infection . This review focuses on the recent advances in understanding mechanisms of protective immunity and why they fail in most infected individuals . The distinct yet complementary role of CD4+ and CD8+ T lymphocytes in this process is highlighted . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 May 21; {Epub ahead of print}
Early Molecular Investigations of Lichen-Forming Symbionts: 19862001; DePriest PT; From the mid-1980s the symbionts in lichen associations, heterotrophic fungi and photosynthetic algae or cyanobacteria, were the subject of increasing numbers of molecular investigations . Many of the studies examined the phylogenetic placement of the individual symbiotic partners with their free-living relatives, refining their nomenclature and classification . Resulting phylogenies permitted the mapping of transitions to and from lichenization and stimulated discussion of the relative ease of gaining and losing symbiotic lifestyles . Comparing symbiont phylogenies both rejected strict cospeciation and mirrored phylogenies and hinted at more complex forces of coevolution, including symbiont switching and selection . Studies at the species and population levels examined patterns of species delimitation and geographic dispersion and processes such as gene flow, self-fertilization, and founder effect . Significant genetic variation often was associated with mobile elements, group I and spliceosomal introns . This review examines the influence of molecular investigation on lichenology during this first 15 years . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 May 21; {Epub ahead of print}
Herpes Vector-Mediated Gene Transfer in Treatment of Diseases of the Nervous System; Glorioso JC et al.; Vectors constructed from recombinant herpes simplex virus (HSV) have special utility for gene transfer to the nervous system . Nonreplicating vectors created by deletion of essential immediate early genes can be propagated to high titers on complementing cell lines that provide the missing gene product(s) in trans . Direct inoculation of these vectors into neural parenchyma has been demonstrated to be effective in rodent models of brain tumor, Parkinson disease, spinal cord injury and spinal root trauma . Subcutaneous inoculation of the HSV vectors can be used to transduce neurons of the dorsal root ganglion to provide a therapeutic effect in models of polyneuropathy and chronic regional pain . In human trials, direct injection of replication competent HSV into brain tumors has proven safe . Human trials of nonreplicating HSV gene transfer by direct inoculation for treatment of glioblastoma, and HSV gene transfer by subcutaneous inoculation for the treatment of chronic intractable pain, should commence soon . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 May 14; {Epub ahead of print}
Ribosomal Crystallography: Initiation, Peptide Bond Formation, and Amino Acid Polymerization are Hampered by Antibiotics; Yonath A et al.; High-resolution structures of ribosomal complexes revealed that minute amounts of clinically relevant antibiotics hamper protein biosynthesis by limiting ribosomal mobility or perturbing its elaborate architecture, designed for navigating and controlling peptide bond formation and continuous amino acid polymerization . To accomplish this, the ribosome contributes positional rather than chemical catalysis, provides remote interactions governing accurate substrate alignment within the flexible transferase-center (PTC) pocket, and ensures nascent-protein chirality through spatial limitations ensuring nascent protein chirality . Peptide bond formation is concurrent with aminoacylated-tRNA 3' end translocation, and is performed by a rotary motion around the axis of a sizable ribosomal symmetry-related region, which is identified around the PTC in all known crystal structures . Guided by ribosomal-RNA scaffold along an exact pattern, the rotary motion results in stereochemistry that is optimal for peptide bond formation and for nascent proteins entrance into their exit tunnel, the main target of antibiotics targeting ribosomes . By connecting between the PTC, the decoding center, and the tRNA entrance and exit regions, the symmetry-related region can transfer intraribosomal signals, guaranteeing smooth processivity of amino acids polymerization . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 Apr 16; {Epub ahead of print}
Viral Error Catastrophe by Mutagenic Nucleosides; Anderson JP et al.; Riboviruses and retroviruses have the highest rates of mutations of any known organisms . Increasing the mutation rate of these viruses could exceed the error threshold for viability of a viral population within a host . Recent experiments with mutagenic nucleoside analogs validate this new approach to treating infection of RNA viruses . Lethal mutagenesis has been documented with HIV infected cells in culture has been documented and has been postulated to be the mechanism for treatment of hepatitis C with ribavirin . We consider the viral dynamics involved in the formation of a quasispecies, the choice of mutagenic nucleoside analogs, and the studies that have demonstrated the feasibility of lethal mutagenesis . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 Apr 16; {Epub ahead of print}
Stationary-Phase Physiology; Nystrom T; Bacteria enjoy an infinite capacity for reproduction as long as they reside in an environment supporting growth . However, their rapid growth and efficient metabolism ultimately results in depletion of growth-supporting substrates and the population of cells enters a phase defined as the stationary phase of growth . In this phase, their reproductive ability is gradually lost . The molecular mechanism underlying this cellular degeneration has not been fully deciphered . Still, recent analysis of the physiology and molecular biology of stationary-phase E . coli cells has revealed interesting similarities to the aging process of higher organisms . The similarities include increased oxidation of cellular constituents and its target specificity, the role of antioxidants and oxygen tension in determining life span, and an apparent trade-off between activities related to reproduction and survival . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Annu Rev Microbiol . 2004 Mar 26; {Epub ahead of print}
Signaling in Myxobacteria; Kaiser D; Myxobacteria use soluble and cell-contact signals during their starvation-induced formation of fruiting bodies . These signals coordinate developmental gene expression with the cell movements that build fruiting bodies . Early in development, the quorum-sensing A-signal in Myxococcus xanthus helps to assess starvation and induce the first stage of aggregation . Later, the morphogenetic C-signal helps to pattern cell movement and shape the fruiting body . C-signal is a 17-kDa cell surface protein that signals by contact between the ends of two cells . The number of C-signal molecules per cell rises 100-fold from the beginning of fruiting body development to the end, when spores are formed . Traveling waves, streams, and sporulation have increasing thresholds for C-signal activity, and this progression ensures that spores form inside fruiting bodies . Expected online publication date for the Annual Review of Microbiology Volume 58 is September 8, 2004 . Please see for revised estimates.

Mol Gen Mikrobiol Virusol, 2004, (3), 30 - 40
{Fluorescence detection methods and their use in microbiology}; A model for consolidation of clinical microbiology laboratory services within a multihospital health-care system; Department of Pathology, University of South Alabama, Mobile, AL, USATo determine the cost-effectiveness of consolidating clinical microbiology services in a three-hospital health-care network while maintaining high-quality laboratory services, a retrospective review of the total costs of maintaining separate clinical microbiology laboratories within our health-care system was compared to the cost of providing these services after consolidation . Turnaround times before and after consolidation were compared to assess efficiency of the consolidated services . Input of clinicians was also solicited to ensure that quality of services and customer satisfaction remained high . The results of the consolidation project show that the net fiscal saving because of consolidation of clinical microbiology services within our health-care system will be approximately 100,000 dollars per fiscal year . This value includes increased courier charges as well as personnel savings . Although fiscal savings are an integral part of any laboratory consolidation plan, the financial considerations must be balanced by quality of service . The response to consolidation from clinicians was decidedly mixed before implementation of the plan because of fear of increased turnaround times and limited access to laboratory information . The consolidation process, however, was smooth with few physician complaints . The consolidation of our clinical microbiology services illustrates that significant financial savings can be achieved without compromise of efficiency or quality of service.

J Biol Chem, 2004 Nov 12, 279(46), 47543 - 54 Epub 2004 Aug 30.
Novel phenotypes of Escherichia coli tat mutants revealed by global gene expression and phenotypic analysis; Ize B et al.; The Tat protein export system serves to export folded proteins harboring an N-terminal twin arginine signal peptide across the cytoplasmic membrane . In this study, we have used gene expression profiling of Escherichia coli supported by phenotypic analysis to investigate how cells respond to a defect in the Tat pathway . Previous work has demonstrated that strains mutated in genes encoding essential Tat pathway components are defective in the integrity of their cell envelope because of the mislocalization of two amidases involved in cell wall metabolism (Ize, B., Stanley, N . R., Buchanan, G., and Palmer, T . (2003) Mol . Microbiol . 48, 1183-1193) . To distinguish between genes that are differentially expressed specifically because of the cell envelope defect and those that result from other effects of the tatC deletion, we also analyzed two different transposon mutants of the DeltatatC strain that have their outer membrane integrity restored . Approximately 50% of the genes that were differentially expressed in the tatC mutant are linked to the envelope defect, with the products of many of these genes involved in self-defense or protection mechanisms, including the production of exopolysaccharide . Among the changes that were not explicitly linked to envelope integrity, we characterized a role for the Tat system in iron acquisition and copper homeostasis . Finally, we have demonstrated that overproduction of the Tat substrate SufI saturates the Tat translocon and produces effects on global gene expression that are similar to those resulting from the DeltatatC mutation.

Am J Respir Cell Mol Biol, 2004 Dec, 31(6), 657 - 62 Epub 2004 Dec.
Immune response to Mycoplasma pulmonis in nasal mucosa is modulated by the normal microbiota; Henriksson G et al.; The impact of commensal bacteria on lymphocyte responses in the upper airways was studied in rat nasal mucosa after infection with the pathogen Mycoplasma pulmonis . Phenotyping was performed in situ by paired immunofluorescence staining in germ-free (GF) and conventional (CV) rats before and 3 wk after the monoinfection . Intraepithelial lymphocytes had expanded significantly in GF (P = 0.02) but not in CV rats . Furthermore, a striking proportional increase of T-cell receptor (TCR)alphabeta(+)CD4(+) cells was observed both in the lamina propria and epithelium of GF (P < 0.01) but not of CV rats . Notably, in contrast to the pre-infection state, both mucosal compartments showed a percentage of TCRalphabeta(+)CD4(+) cells that was significantly higher in GF (P = 0.03-P < 0.01) than in CV rats after the monoinfection . In parallel, both compartments displayed a percentage of TCRalphabeta(+) CD8(+) cells that was decreased in GF (P < 0.01) but not in CV rats . The small fraction of TCRgammadelta(+) T cells observed (< 5%) did not change quantitatively or phenotypically after infection . The size of organized nose-associated lymphoid tissue was, on average, increased 5.2-fold in GF rats versus 2.6-fold in CV rats . Collectively, our results demonstrated that the normal microbiota modulated markedly the nasal immune response elicited by monoinfection with M . pulmonis.

Appl Environ Microbiol, 2004 Sep, 70(9), 5138 - 44
Surveillance of bulk raw and commercially pasteurized cows' milk from approved Irish liquid-milk pasteurization plants to determine the incidence of Mycobacterium paratuberculosis; O'Reilly CE et al.; Over the 13-month period from October 2000 to November 2001 (inclusive), the Food Safety Authority of Ireland (FSAI) carried out surveillance of Irish bulk raw (n = 389) and commercially pasteurized (n = 357) liquid-milk supplies to determine the incidence of Mycobacterium paratuberculosis . The pasteurization time-temperature conditions were recorded for all pasteurized samples . Overall, 56% of whole-milk pasteurized samples had been heat treated at or above a time-temperature combination of 75 degrees C for 25 s . All analyses were undertaken at the Department of Food Science (Food Microbiology) laboratory at Queen's University Belfast . Each milk sample was subjected to two tests for M . paratuberculosis: immunomagnetic separation-PCR (IMS-PCR; to detect the presence of M . paratuberculosis cells, live or dead) and chemical decontamination and culture (to confirm the presence of viable M . paratuberculosis) . Overall, M . paratuberculosis DNA was detected by IMS-PCR in 50 (12.9%; 95% confidence interval, 9.9 to 16.5%) raw-milk samples and 35 (9.8%; 95% confidence interval, 7.1 to 13.3%) pasteurized-milk samples . Confirmed M . paratuberculosis was cultured from one raw-milk sample and no pasteurized-milk samples . It is concluded that M . paratuberculosis DNA is occasionally present at low levels in both raw and commercially pasteurized cows' milk . However, since no viable M . paratuberculosis was isolated from commercially pasteurized cows' milk on retail sale in the Republic of Ireland, current pasteurization procedures are considered to be effective.

Environ Microbiol, 2004 Oct, 6(10), 1081 - 5
Estimating bacterial diversity from clone libraries with flat rank abundance distributions; Lunn M et al.; There are a number of parametric and non-parametric methods for estimating diversity . However all such methods employ either the proportional abundance of the most abundant taxon in a sample or require that a specific taxon is sampled more than once . Consequently, the available methods for estimating diversity cannot be applied to samples consisting entirely of singletons, which might be characteristic of some hyperdiverse communities . Here we present a non-parametric method that estimates the probability that a given number of unique taxa would be sampled from a community with a particular diversity . We have applied this approach to a well known data set of 100 unique clones from a sample of Amazonian soil (Borneman and Triplett (1997) Appl Environ Microbiol 63: 2647-2653) and determine the probability that this observation would be made from an environment of a given diversity . On this basis we can state this observation would be very unlikely (P = 0.006) if the soil diversity was less than 10(3), and quite unlikely (P = 0.6) if the diversity was less than 10(4), and probable (P = 0.95) if the diversity was about 10(5) . There are essentially no contestable assumptions in our method . Thus we are able to offer almost unequivocal evidence that the bacterial diversity, of at least soils, is very large and a method that may be used to interpret samples consisting entirely of singletons from other hyperdiverse communities.

Acta Pharmacol Sin, 2004 Sep, 25(9), 1186 - 94
Learning of medical pharmacology via innovation: a personal experience at McMaster and in Asia; Kwan CY; Pharmacology, a discrete preclinical discipline of the traditional medical curriculum, identifies itself distinctly different from the other preclinical or clinical subjects in knowledge base as well as learning/teaching instructions . It exists in series with other pre-clinical courses (e.g., anatomy, biochemistry and physiology), and in parallel with other paraclinical courses such as pathology, microbiology and community medicine . Such arrangement makes learning of pharmacology rather difficult and deficient with regard to its therapeutic relevance and clinical application . In recent years, medical curricula based on clinical cases have emerged as a platform in which pharmacology is one integrated component in a holistic approach to medical education . In this problem-based learning (PBL) model, students learn, with teachers' facilitation, in a student-centered environment, based on self-directed, clinically relevant and case-oriented approach, usually in a small-group tutorial format . In PBL, pharmacology is learned in concert with other subject issues relevant to the case-problem in question, such as anatomy, physiology, pathology, microbiology, population health, behavior science, etc . Students learn via problem-evoked curiosity and motivation, in an environment which encourages free inquiries and intensive discussions in a cooperative rather than competitive atmosphere . Teachers facilitate students' learning objectives rather than deliver pre-packed knowledge and dictate what they think students should learn . A change towards PBL curriculum appears to be beneficial in better preparing the medical students as life-long learners capable of coping with changes in knowledge and skills associated with the progressive and dynamic social/economic transformation in the Asia-Pacific regions . Evidence is presented that this is indeed happening.

Int J Food Microbiol, 2004 Sep 15, 95(3), 321 - 31
IGS-RFLP analysis and development of molecular markers for identification of Fusarium poae, Fusarium langsethiae, Fusarium sporotrichioides and Fusarium kyushuense; Konstantinova P et al.; The intergenic spacer (IGS) regions of the rDNA of several Fusarium spp . strains obtained from the collaborative researchers (Int . J . Food Microbiol . (2003)) were amplified by polymerase chain reaction (PCR), and an IGS-RFLP analysis was performed . Restriction digestion with AluI, MspI and PstI allowed differentiation between the related Fusarium poae and Fusarium kyushuense species . Fusarium langsethiae was also separated from Fusarium sporotrichioides (including var . minus) on the basis of the banding patterns after MspI digestion, while specific XhoI, AluI and MspI restriction patterns were found in the IGS amplicons of F . sporotrichioides var . minus . According to the phylogenetic analysis of IGS-RFLP patterns, F . langsethiae (except for one strain), F . sporotrichioides, F . poae and F . kyushuense strains formed four well-supported clades with high-bootstrap values . Based on the sequence differences in the IGS region, species-specific primers were designed for the F . langsethiae/F . sporotrichioides group and for F . poae . The specificity and sensitivity of the primers were tested on various Fusarium species and isolates, and on several other important fungal genera associated with cereals . The F . poae-specific primers, designed in this study, showed the same specificity as primers Fp82f/Fp82r developed previously . The two phylogenetic subgroups of F . langsethiae, found by IGS sequencing analysis, were separated on the basis of size differences of the amplification products with primers CNL12/PulvIGSr specific for the F . langsethiae/F . sporotrichioides group . RFLP analysis of the amplified IGS region is a useful molecular assay for characterisation and a phylogenetic study of several related Fusarium species-F . langsethiae, F . sporotrichioides, F . sporotrichioides var . minus, F . poae and F . kyushuense . The primers designed in this study were highly specific and allowed identification of F . poae and the F . langsethiae/F . sporotrichioides group.

Saudi Med J, 2004 Aug, 25(8), 1066 - 9
Plasma copper and zinc levels in chronic viral hepatitis; Meram I et al.; OBJECTIVE: The relationship between chronic hepatitis and trace metals has not been understood clearly . Copper (Cu) and zinc (Zn) are essential trace elements for several metabolic processes . Overload or deficiency of these elements can lead to metabolic disorders and some other diseases . In this study, we aimed to examine the relationship between chronic hepatitis and plasma Cu and Zn levels . METHODS: Forty-three patients with chronic viral hepatitis (CVH) and 30 healthy controls were included in this study . The patients were consecutively admitted to the Department of Infectious Disease and Clinical Microbiology, Faculty of Medicine, Gaziantep University, Turkey, between January 2000 and November 2000 . Plasma Cu and Zn levels, and hepatic function test results of the patients and controls were compared . Serologic and virologic markers, and histopathologic assessments were performed for confirmation of CVH . Plasma Cu and Zn concentrations were determined with Bathocuproin, using deproteinization method and 5-Br-PAPS methods . RESULTS: Patients plasma Cu level was 16.0 +/- 2.8 and plasma Zn level was 26.0 +/- 7.3 . The corresponding values were 12.2 +/- 5.4 and 26.6 +/- 5.6 in the healthy controls . The patients with CVH had a higher plasma Cu level than the controls (p<0.05) while the Zn levels were similar in both groups (p>0.08) . CONCLUSION: Although there is an increased plasma Cu level in CVH, its mechanism is unclear . However, this condition may have clinical importance as Cu is a hepatotoxic element.

Shock, 2004 Sep, 22(3), 197 - 203
Ethnic disparities in outcome from posttransplant infections; Pelletier SJ et al.; Black transplant recipients have decreased graft survival and increased rejection rates compared with whites . Because increased rejection rates may lead to more immunosuppression in black recipients, ethnic differences may exist for outcomes of posttransplant infectious complications . All episodes of infection between December 1996 and October 1998 on the transplant services at the University of Virginia Health Sciences Center were prospectively evaluated . Parameters recorded included self-designated ethnicity, demographics, APACHE II scores, laboratory and microbiologic data, immunosuppression, episodes of rejection, and outcome measures . Evaluation of 303 episodes of infection demonstrated an increased mortality rate for white compared with black recipients (19% vs . 3%, P = 0.0006) despite having a similar severity of illness (APACHE II score) . Among renal transplant recipients, episodes of infection occurring in black recipients (n = 46) were also associated with a decreased mortality rate versus whites (n = 89) (0% vs . 15%, P = 0.006) and shorter mean length of stay (12 +/- 2 vs . 25 +/- 4 days, P = 0.002) despite similar severity of illness and rejection rates . For posttransplant infections in liver transplant recipients, blacks (n = 23) demonstrated a trend toward decreased mortality (9% vs . 26%, P = 0.07) but equal lengths of stay despite similar APACHE II scores, rejection rates, and age . White liver transplant recipients had an increased incidence of viral infections (15% vs . 0%, P = 0.03) . All other infecting organisms were similar . The unexpected finding of a significantly decreased rate of mortality associated with posttransplant infections in black recipients remains largely unexplained but may be related to subtle differences in immune response between racial or ethnic groups.

Infez Med, 2004 Jun, 12(2), 118 - 25
The impact of automation on organizational changes in a community hospital clinical microbiology laboratory; Camporese A; The diagnosis of infectious diseases and the role of the microbiology laboratory are currently undergoing a process of change . The need for overall efficiency in providing results is now given the same importance as accuracy . This means that laboratories must be able to produce quality results in less time with the capacity to interpret the results clinically . To improve the clinical impact of microbiology results, the new challenge facing the microbiologist has become one of process management instead of pure analysis . A proper project management process designed to improve workflow, reduce analytical time, and provide the same high quality results without losing valuable time treating the patient, has become essential . Our objective was to study the impact of introducing automation and computerization into the microbiology laboratory, and the reorganization of the laboratory workflow, i.e . scheduling personnel to work shifts covering both the entire day and the entire week . In our laboratory, the introduction of automation and computerization, as well as the reorganization of personnel, thus the workflow itself, has resulted in an improvement in response time and greater efficiency in diagnostic procedures.

Res Microbiol, 2004 Sep, 155(7), 571 - 4
Candida pararugosa isolation from the oral cavity of an Italian denture wearer; Giammanco GM et al.; Candida pararugosa was first isolated from human feces, but after the initial description no further recovery from humans has been reported . During a study on oral Candida colonization in denture wearers living in Palermo (Italy), we isolated C . pararugosa from a 61-year-old woman without signs of oral candidosis . This constitutes, to the authors' knowledge, the first isolation of C . pararugosa from the oral cavity . After six months, colonization by C . pararugosa persisted, suggesting that this species could be a component of the normal oral microbiota . The identification procedure we used could be useful in elucidating the epidemiology of C . pararugosa and for establishing its clinical significance.

Biochem Biophys Res Commun, 2004 Sep 10, 322(1), 133 - 46
Molecular characterization of a phenanthrene degradation pathway in Mycobacterium vanbaalenii PYR-1; Stingley RL et al.; Mycobacterium vanbaalenii PYR-1 is capable of degrading a number of polycyclic aromatic hydrocarbons (PAHs) to ring cleavage metabolites via multiple pathways . Genes for the large and small subunits of a pyrene dioxygenase, nidA and nidB, respectively, were previously identified in M . vanbaalenii PYR-1 {Appl . Environ . Microbiol . 67 (2001) 3577} . A library of the M . vanbaalenii PYR-1 genome was constructed in a fosmid vector to identify additional genes involved in PAH degradation . Twelve fosmid clones containing nidA were identified by Southern hybridization . Sequence analysis of one nidA-positive clone, pFOS608, revealed a number of additional genes involved in PAH degradation . At this locus, one putative operon contained genes involved in phthalate degradation, and another contained genes encoding a putative ABC transporter(s) . A number of the genes found in this region are homologous to those involved in phenanthrene degradation via the phthalic acid pathway . The majority of phenanthrene degradation genes were located between putative transposase genes . In Escherichia coli, pFOS608 converted phenanthrene into phenanthrene cis-3,4-dihydrodiol, and converted 1-hydroxy-2-naphthoic acid into 2'-carboxybenzalpyruvate, 2-carboxybenzaldehyde, and phthalic acid . A subclone containing nidA and nidB converted phenanthrene into phenanthrene cis-3,4-dihydrodiol, suggesting that the NidAB dioxygenase is responsible for an initial attack on phenanthrene . This study is the first to identify genes responsible for the degradation of phenanthrene via the phthalic acid pathway in Mycobacterium species.

World J Gastroenterol, 2004 Sep 15, 10(18), 2769 - 71
Intestinal perforation after combined liver-kidney transplantation for a case of congenital polycystic disease; Peng T et al.; AIM: To highlight the intestinal perforation (IP), an uncommon and catastrophic complication after combined liver-kidney transplantation . METHODS: Combined liver-kidney transplantation (LKTx) with left kidney excision and a cyst fenestration procedure on the right kidney were performed on a case of 46-year-old female with congenital polycystic disease (CPCD) . RESULTS: Two sites of IP were noted 40-50 cm proximal to ileocecal area during emergent laparotomy 10 d postoperatively . Despite aggressive surgical and medical management, disease progressed toward a fatal outcome due to sepsis and multiple organ failure 11 d later . CONCLUSION: Long duration of operation without venovenous bypass, overdose of steroid together with postoperative volume excess may all contribute to the risk of idiopathic multiple IPs . Microbiology and pathology inspections suggested that the infected cyst of the fenestrated kidney might be one reason for the fatal intra-peritoneal infection . Thus for the CPCD patients who seem to be very susceptible to infectious complications, any sign of suspected renal-infection found before or during LKTx is indication for the excision of original kidney . And the intensity of immunosuppression therapy should be controlled cautiously.

Am J Transplant, 2004 Sep, 4(9), 1383 - 90
Xenotransplantation: infectious risk revisited; Fishman JA et al.; Xenotransplantation is a possible solution for the shortage of tissues for human transplantation . Multiple hurdles exist to clinical xenotransplantation, including immunologic barriers, metabolic differences between pigs--the source species most commonly considered--and humans, and ethical concerns . Since clinical trials were first proposed almost 10 years ago, the degree of risk for infection transmitted from the xenograft donor to the recipient has been extensively investigated . A number of potential viral pathogens have been identified including porcine endogenous retrovirus (PERV), porcine cytomegalovirus (PCMV), and porcine lymphotropic herpesvirus (PLHV) . Sensitive diagnostic assays have been developed for each virus . Human-tropic PERV are exogenous recombinants between PERV-A and PERV-C sequences and are present in only a subset of swine . Porcine cytomegalovirus can be excluded from herds of source animals by early weaning of piglets . In contrast, the risks associated with PLHV remain undefined . Microbiologic studies and assays for potential xenogeneic pathogens have furthered understanding of risks associated with xenotransplantation . Thus far, clinical xenotransplantation of pig tissues has not resulted in transmission of viral infection to humans; significant risks for disease transmission from swine to humans have not been confirmed . If immunologic hurdles can be overcome, it is reasonable to initiate carefully monitored clinical trials .

Clin Infect Dis, 2004 Aug 1, 39 Suppl 2, S115 - 22
Controversies in diagnosing and managing osteomyelitis of the foot in diabetes; Jeffcoate WJ et al.; The optimal approach to diagnosing and managing osteomyelitis of the foot in diabetes is unclear . Diagnosis is based on clinical signs, supplemented by a variety of imaging tests . Bone biopsy is the accepted criterion standard for diagnosis but is not used by many . Management traditionally involves surgical removal of infected bone, combined with antibiotic therapy . However, recent studies have shown that antibiotics alone may apparently eliminate bone infection in many cases . There is also evidence that early amputation of infected digits is frequently noncurative . Agreement on criteria for diagnosing osteomyelitis is required, and randomized trials are urgently needed, to determine the relative benefits of various surgical interventions and the optimal deployment of antibiotics . We review the microbiology of osteomyelitis of the foot in diabetes, the benefits and limitations of various diagnostic procedures, and the evidence for the effectiveness of both surgical and nonsurgical approaches to management.

South Med J, 2004 Jul, 97(7), 702 - 4
Blastoschizomyces capitatus pneumonia in an immunocompetent male; Wills TS et al.; Blastoschizomyces capitatus is an emerging fungal pathogen . It has been well characterized as a cause of local and disseminated disease in immunocompromised hosts, especially in the setting of neutropenia . We describe a case of B . capitatus pneumonia in an otherwise healthy man and review the clinical presentation, microbiologic characteristics, and treatment strategies for B . capitatus infections.

Am J Primatol, 2004 Aug, 63(4), 225 - 37
Use of primates in research: a global overview; Carlsson HE et al.; We assessed the use of nonhuman primates and nonhuman primate biological material in research by reviewing studies published in 2001 in peer-reviewed journals . The number and species of primates used, the origin of the animals, the type of study, the area of research of the investigation, and the location at which the research was performed were tabulated . Additionally, factors related to the animals that may have affected the outcome of the experiments were recorded . A total of 2,937 articles involving 4,411 studies that employed nonhuman primates or nonhuman primate biological material were identified and analyzed . More than 41,000 animals were represented in the studies published in 2001 . In the 14% of studies for which re-use could be determined, 69% involved animals that had been used in previous experiments . Published studies most commonly used nonhuman primates or nonhuman primate biological material from the species Chlorocebus aethiops (19%), Macaca mulatta (18%), M . fascicularis (9%), and Papio spp . (6%) . Of these studies, 54% were classified as in vitro studies, 14% as noninvasive, 30% as chronic, and 1% were considered acute . Nonhuman primates were primarily used in research areas in which they appear to be the most appropriate models for humans . The most common areas of research were microbiology (including HIV/AIDS (26%)), neuroscience (19%), and biochemistry/chemistry (12%) . Most (84%) of the primate research published in 2001 was conducted in North America, Europe, and Japan . The animals and conditions under which they were housed and used were rarely described . Although it is estimated that nonhuman primates account for an extremely small fraction of all animals used in research, their special status makes it important to report the many husbandry and environmental factors that influence the research results generated . This analysis has identified that editors rarely require authors to provide comprehensive information concerning the subjects (e.g., their origin), treatment conditions, and experimental procedures utilized in the studies they publish . The present analysis addresses the use of primates for research, including the effects of a shortage of suitable nonhuman primate subjects in many research areas .

Hist Philos Life Sci, 2003, 25(2), 167 - 91
Mesosomes and scientific methodology; Hudson RG; In his recent article, Nicolas Rasmussen (2001) is harshly critical of what he terms 'empirical philosophy of science', a philosophy that takes seriously the history of science in advancing philosophical pronouncements about science . He motivates his criticism by reflecting on recent history in microbiology involving the 'discovery' of a new bacterial organelle, the mesosome, during the 1950's and 1960's, and the subsequent retraction of this discovery by experimental microbiologists during the late 1970's and early 1980's . In particular, he argues that there was a lack of constancy in the methods microbiologists used in approaching the issue of the existence of mesosomes, and that in fact a similar sort of 'methodological flux' pervades all experimental work . My goal here is to refute Rasmussen's doctrine of flux, and in turn to re-establish order in our understanding of the methods and strategies of experimenters . My strategy in achieving this goal is to re-visit the same crucial research articles in the history of the mesosome episode that Rasmussen (2001) visits; and what I find upon returning to this literature is not flux, as Rasmussen seems to find, but a constancy of method in experimental reasoning, a constancy codified by what I call 'reliable process reasoning'.

Pediatr Infect Dis J, 2004 Aug, 23(8), 786 - 8
Cerebrospinal latex agglutination fails to contribute to the microbiologic diagnosis of pretreated children with meningitis; Nigrovic LE et al.; We conducted a 10-year retrospective study of all children who had cerebrospinal fluid latex agglutination testing for bacterial antigens performed at 1 tertiary care urban children's hospital . Of the 176 patients with culture-negative meningitis who were pretreated with antibiotics before lumbar puncture, none had a positive latex agglutination study (0 of 176; 95% confidence interval, 0-2%) . Latex agglutination studies identified no additional cases of bacterial meningitis beyond those identified by culture in pretreated patients . Clinical decision-making algorithms for the management of pretreated patients at risk for bacterial meningitis should not include latex agglutination testing.

Eur Respir J, 2004 Jul, 24(1), 171 - 81
Atypical pathogens and respiratory tract infections; Blasi F; The atypical respiratory pathogens Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila are now recognised as a significant cause of acute respiratory-tract infections, implicated in community-acquired pneumonia, acute exacerbations of chronic bronchitis, asthma, and less frequently, upper respiratory-tract infections . Chronic infection with C . pneumoniae is common among patients with chronic obstructive pulmonary disease and may also play a role in the natural history of asthma, including exacerbations . The lack of a gold standard for diagnosis of these pathogens still handicaps the current understanding of their true prevalence and role in the pathogenesis of acute and chronic respiratory infections . While molecular diagnostic techniques, such as polymerase chain reaction, offer improvements in sensitivity, specificity and rapidity over culture and serology, the need remains for a consistent and reproducible diagnostic technique, available to all microbiology laboratories . Current treatment guidelines for community-acquired pneumonia recognise the importance of atypical respiratory pathogens in its aetiology, for which macrolides are considered suitable first-line agents . The value of atypical coverage in antibiotic therapy for acute exacerbations of chronic bronchitis and exacerbations of asthma is less clear, while there is no evidence to suggest that atypical pathogens should be covered in antibiotic treatment of upper respiratory-tract infections.

Med Oral, 2004 Aug-Oct, 9(4), 280 - 7
Orofacial infections of odontogenic origin; Gutierrez-Perez JL et al.; The polymicrobial nature of the odontogenic infections as well as the variety of associated conditions are a consequence of the diversity of the buccal microbiota and the anatomical and functional complexity of the oral cavity . In addition to this, all these processes can give way to multiple complications which range from the local to the systemic level . The appropriate choice of antibiotic and posology is crucial in the successful management of these infections . Pharmacodynamics provides those parameters that make it possible to assess how antibiotics activity varies in time . As a general rule, the first step in the initial management of orofacial infections in adults, included odontogenic infections, will be the administration of 875 mg of amoxicillin and 125 mg of clavulanic every 8 hours . Therapeutic compliance is paramount to avoid resistance, therefore patient acceptance must be sought . In this sense, it has been proved that Augmentine Plus (2000/125) every twelve hours both as profylaxis and as treatment significantly decreases the rate of infective complications associated to extraction of the third molar.

J Bacteriol, 2004 Aug, 186(16), 5480 - 5
Tra proteins characteristic of F-like type IV secretion systems constitute an interaction group by yeast two-hybrid analysis; Harris RL et al.; Using yeast two-hybrid screens, we have defined an interaction group of six Tra proteins encoded by the F plasmid and required by F(+) cells to elaborate F pili . The six proteins are TraH, TraF, TraW, TraU, TrbI, and TrbB . Except for TrbI, these proteins were all identified as hallmarks of F-like type IV secretion systems (TFSSs), with no homologues among TFSS genes of P-type or I-type systems (T . Lawley, W . Klimke, M . Gubbins, and L . Frost, FEMS Microbiol . Lett . 224:1-15, 2003) . Also with the exception of TrbI, which is an inner membrane protein, the remaining proteins are or are predicted to be periplasmic . TrbI consists of one membrane-spanning segment near its N terminus and an 88-residue, hydrophilic domain that extends into the periplasm . Hence, the proteins of this group probably form a periplasmic cluster in Escherichia coli . The interaction network identifies TraH as the most highly connected node, with two-hybrid links to TrbI, TraU, and TraF . As measured by transcriptional activation of lacZ, the TrbI-TraH interaction in Saccharomyces cerevisiae requires the TraH amino acid segment from residues 193 to 225 . The TraU and TraF interactions are localized to C-terminal segments of TraH (amino acids 315 to 458 for TraF and amino acids 341 to 458 for TraU) . The TrbI-TraH interaction with full-length (less the signal peptide) TraH is weak but increases 40-fold with N-terminal TraH deletions; the first 50 amino acids appear to be critical for inhibiting TrbI binding in yeast . Previous studies by others have shown that, with the exception of trbB mutations, which do not affect the elaboration or function of F pili under laboratory conditions, a mutation in any of the other genes in this interaction group alters the number or length distribution of F pili . We propose a model whereby one function of the TraH interaction group is to control F-pilus extension and retraction.

Mund Kiefer Gesichtschir . 2004 Jul 29; {Epub ahead of print}
{Chronic cutaneous infiltration with abscess and fistula formationA type of clinical course in atypical mycobacteriosis}; Moergel M et al.; BACKGROUND . Cutaneous lesions of atypical mycobacteriosis are very rare in Western Europe . Both the slow disease progression and the rather unspecific clinical appearance of the chronic infiltration render cutaneous mycobacteriosis a diagnostic challenge . In this communication, we present an uncommon, biphasic course of a cutaneous atypical mycobacteriosis . CASE REPORT . A 3-year-old child was referred to our institution due to submandibular cutaneous infiltration together with subcutaneous abscess formation . There was no carious lesion or other odontogenic potential source of infection . First-line therapy was incision of the abscess and local curettage to obtain a specimen for histologic and microbiologic evaluation, which revealed atypical mycobacteria . In spite of local curettage, only partial remission of the cutaneous infiltration was achieved . When the child was readmitted 9 months later, the lesion had progressed to a scrofulosis-like appearance even though specific antibiotic treatment had been administered . Complete surgical excision of the affected skin and extirpation of the respective lymph node and fistula was performed as second-line therapy . Complete remission was achieved without further adjuvant treatment . DISCUSSION . Our case demonstrates the problems that may arise with this rare type of mycobacterial infection . In all probability, scratch artifacts due to atopic dermatitis provided the source/entry of the infection in this specific case . Both histopathologic findings and clinical appearance were suggestive of classic cutaneous tuberculosis (tuberculosis cutis colliquativa), although it was in fact an atypical mycobacteriosis . Furthermore, obstinate persistence after minor surgical intervention and even after specific (triple) antibiotic treatment illustrates the difficulties of limited, minimally invasive treatment.

Surg Laparosc Endosc Percutan Tech, 2004 Apr, 14(2), 49 - 52
Management of splenic abscess in a critically ill patient; Farres H et al.; Because of the increased number of immunocompromised patients within the general population, the incidence of splenic abscesses has increased over the last decade . This cohort of immune-deficient patients with splenic abscesses engenders a distinct evolution in the pathogenesis and microbiology of the disease process . Moreover, the morbidity and mortality rates for splenic abscesses are increased in this unique population . Clinically, these patients do not have a characteristic presentation . Diagnostically, computed tomography of the abdomen is the test of choice . Antibiotics and splenectomy remain the standard of care in most clinical settings . However, percutaneous drainage is reported with solitary and unilocular abscesses and in poor operative candidates . An unusual case of a patient with a splenic abscess awaiting heart transplantation is presented . This patient was successfully treated with percutaneous drainage and antibiotics . The literature regarding the presentation, diagnosis, pathogenesis, and treatment of splenic abscesses is reviewed as well.

Biotechnol Bioeng, 2004 Aug 20, 87(4), 485 - 94
Growth-rate recovery of Escherichia coli cultures carrying a multicopy plasmid, by engineering of the pentose-phosphate pathway; Flores S et al.; Expression of plasmid-encoded genes in bacteria is the most common strategy for the production of specific proteins in biotechnological processes . However, the synthesis of plasmid-encoded proteins and plasmid-DNA replication often places a metabolic load (metabolic burden) into the cell's biochemical capacities that usually reduces the growth rate of the producing culture (Glick BR . Biotechnol Adv 1995;13:247-261) . This metabolic burden may be related to a limited capacity of the cell to supply the extra demand of building blocks and energy required to replicate plasmid DNA and express foreign multicopy genes . Some of these required blocks are intermediaries of the pentose phosphate (PP) pathway, e.g., ribose-5-phosphate, erythrose-4-phosphate . Due to the important impact of metabolic burden on biotechnological processes, several groups have worked on developing strategies to overcome this problem, like reduction of plasmid copy number (Seo JH, Bailey JE . Biotechnol Bioeng 1985;27:1668-1674; Jones KL, Kim S, Keasling JD . Metab Eng 2000;3:328-338), chromosomal insertion of the gene which product is desired, or changing the plasmid-coded antibiotic resistance gene (Hong Y, Pasternak JJ, Glick BR . Can J Microbiol 1995;41:624-628) . However, few efforts have been attempted to overcome the reduction of growth rate due to protein over-expression, by modifying central metabolic pathways (Chou C-H, Bennett GN, San KY . Biotechnol Bioeng 1994;44:952-960) . We constructed a high-copy number plasmid carrying the gene for glucose-6-phosphate dehydrogenase, zwf, under the control of an inducible trc promoter (pTRzwf04 plasmid) . By transforming a wild-type strain and inducing with IPTG, it was possible to recover growth-rate from 0.46 h(-1) (uninduced) to 0.64 h(-1) (induced) . The same transformation in an Escherichia coli zwf(-), allows a growth-rate recovery from 0.43 h(-1) (uninduced) to 0.62 h(-1) (induced) . We also studied this effect as part of a laboratory-scale biotechnology process: production of a recombinant insulin peptide by co-transforming E . coli JM101 strain with pTRzwf07, a low-copy-number plasmid that carries the same inducible construction as pTRzwf04, and with the pTEXP-MMRPI vector that carries a TrpLE-proinsulin hybrid gene . In this system, production of TrpLE-proinsulin strongly reduces growth rate; however, overexpression of zwf gene recovers with a growth rate from 0.1 h(-1) in the TrpLE-proinsulin induced strain, to 0.37 h(-1) when both zwf and TrpLE-proinsulin genes were induced . In this paper, we show that the engineering of the pentose phosphate pathway by modulation of the zwf gene expression level partially overcomes the possible bottleneck for the supply of building blocks and reducing power synthesized through the PP pathway, that are required for plasmid replication and plasmid-encoded protein expression.

Int J Food Microbiol, 2004 Sep 1, 95(2), 231 - 4
Estimation of accordance and concordance in inter-laboratory trials of analytical methods with qualitative results; van der Voet H et al.; In this short communication, we discuss alternative ways to estimate accordance and concordance, which are, for qualitative methods, the analogues of the well-known concepts repeatability and reproducibility . We argue that estimators in a random framework appear to be more appropriate than the estimators in a fixed framework as they were originally introduced by Langton et al . {International Journal of Food Microbiology 79 (2002) 171} . A simple example of their application is given.

Appl Microbiol Biotechnol . 2004 Jul 23; {Epub ahead of print}
Deficiency in the glycerol channel Fps1p confers increased freeze tolerance to yeast cells: application of the fps1Delta mutant to frozen dough technology; Izawa S et al.; Intracellular glycerol content affects the freeze-thaw stress tolerance of Saccharomyces cerevisiae . We have recently reported that intracellular-glycerol-enriched cells cultured in glycerol medium acquire tolerance to freeze stress and retain high leavening ability even in dough after frozen storage {Izawa et al . (2004) Appl Microbiol Biotechnol A deletion mutant of the FPS1 gene, which encodes a glycerol channel, accumulates glycerol inside the cell without an exogenous supply of glycerol into the medium . We found that the fps1Delta cells acquired tolerance to freeze stress and retained high leavening ability in dough after frozen storage for 7 days . These results suggest that the fps1Delta mutant is a useful strain for developing better frozen-dough with a commercial advantage.

J Med Microbiol, 2004 Aug, 53(Pt 8), 741 - 8
Influence of normal microbiota on some aspects of the immune response during experimental infection with Trypanosoma cruzi in mice; Duarte R et al.; To study the influence of normal associated microbiota on systemic immunological responses during experimental Chagas' disease, germ-free and conventional NIH Swiss mice were infected with Y strain of Trypanosoma cruzi . Although no statistical differences in mortality and parasitaemia were found, conventional mice showed IFN-gamma, TNF-alpha and NO production (P < 0.05) by spleen cell cultures and higher blood levels of immunoglobulins of the IgG2a isotype (P < 0.05) when compared to their germ-free counterparts . Moreover, higher levels of IgG1 were also found in conventional animals . On the other hand, no differences in IL10 production were found between germ-free and conventional mice after infection (P < 0.05) . Interestingly, spleen cell cultures from non-infected germ-free mice spontaneously produced higher levels of IL10 than cultures from conventional mice . Moreover, cultures from non-infected germ-free mice responded to T . cruzi antigens with IFN-gamma production, contrary to cultures from conventional animals . In conclusion, the presence of the normal microbiota skews the immune response towards production of inflammatory cytokines during experimental infection with T . cruzi in mice . However, the increase in production of cytokines that is linked to resistance to this parasite did not alter the outcome of infection significantly, probably due to high virulence of the Y strain .

Environ Toxicol, 2004 Aug, 19(4), 416 - 20
Increase of crustacean sensitivity to purified hepatotoxic cyanobacterial extracts by manipulation of experimental conditions; Drobniewska A et al.; Toxic cyanobacterial blooms are one of the most common consequences of water eutrophication . Microbiotests with crustaceans are not expensive and are easy to prepare for screening tests . They can be applied in the determination of bioactivity and interaction between toxic substances in water, including hepatotoxins . The principal aim of this study was to modify the standard conditions in the Thamnotoxkit F trade mark and Artoxkit M in order to increase crustacean sensitivity to purified cyanobacterial extracts containing microcystins . The results reported show that exposure time, higher temperature, and presence of DMSO can increase the sensitivity of microbiotests to microcystins . The best sensitivity with the Artemia salina test was achieved after a 48-h exposure at 25 degrees C . The tests using a 24-h exposure at 27 degrees C were the most sensitive for Thamnocephalus platyurus . The test without preincubation with DMSO provided the best correlation of microcystin concentration and LC(50) for Thamnocephalus platyurus and is recommended .

Environ Toxicol, 2004 Aug, 19(4), 296 - 301
Effect of deicing salts on urban soils and health status of roadside trees in the Opole region; Czerniawska-Kusza I et al.; This article reports on a study whose aim was to evaluate the impact of snow removal salts on urban soil properties and the health of roadside trees . The evaluation was done by chemical analyses of soil samples and plant matter combined with toxicity testing, performed with a Protoxkit F, a protozoan microbiotest . Samples were collected at 45 locations on three main roads in the town of Opole (Poland) . The roads differed in the snow removal technology and amount of chemical substances (mostly NaCl) used on them during the winter . The study showed that when soil was exposed to a high level of NaCl, it tended to be more alkaline and also exhibited increased content of Na(+) and Cl(-) . The toxic effects of the soil extract on protozoa appeared at 26.0 mg Na(+)/100 g soil dry mass (s.d.m.) and 12.0 mg Cl(-)/100 g s.d.m., whereas salt injury symptoms (chlorosis and necrosis of the edge of leaf blades) appeared at 13.2 mg Na(+)/100 g s.d.m . and 3.9 mg Cl(-)/100 g s.d.m., becoming more severe at 26.0 mg Na(+)/100 g s.d.m . and 12.0 mg Cl(-)/100 g s.d.m . because of extensive necrosis and defoliation . The lysimetric experiment, which was used to test soil samples collected from the city park area, indicated that salt plays a significant role in the pollution of soil in urban areas, with the least toxic salt being CaCl(2) .

Am J Transplant, 2004 Aug, 4(8), 1366 - 71
Invasive pneumococcal infections in adult lung transplant recipients; de Bruyn G et al.; An increased risk of invasive pneumococcal infection (IPI) has been described among kidney or heart transplant recipients, but the epidemiology of IPI among lung transplant recipients has not been previously reported . We undertook a single center, retrospective cohort study to define the incidence, timing, clinical, and microbiologic features of IPI in lung transplant patients . Fourteen out of 220 recipients (6.4%) developed IPI at a median of 1.3 years after transplantation (incidence rate: 22.7 cases per 1000 person-years) . All patients were receiving trimethoprim-sulfamethoxazole (TMP-SMX) prophylaxis at the time of diagnosis, and 10 (71%) had TMP-SMX-resistant isolates . All isolates were from the 23 valent polysaccharide vaccine-associated serogroups . The high incidence of IPI in lung transplant recipients is similar to that reported in kidney and heart recipients . Alternative prevention strategies, including use of the conjugated pneumococcal vaccine, should be explored in future studies.

J Small Anim Pract, 2004 Jul, 45(7), 351 - 6
Complications of exploratory coeliotomy in 70 cats; Lester S et al.; Records of all cats that had undergone exploratory coeliotomy at the University of Edinburgh during the period November 1995 to July 2002 were reviewed . Seventy records were retrieved . There were 30 cats in which infection or inflammatory disorders predominated, 17 cats with neoplasia, three cats with trauma and 20 cats with other disorders . Exploratory coeliotomy was performed for diagnostic purposes in 28 cats (40 per cent), treatment in 34 cats (49 per cent) and for diagnosis and treatment in eight cats (11 per cent) . Methods of intraoperative diagnosis included incisional biopsy of abdominal organs (52 cats), cytology (two cats), microbiology (17 cats) and gross appearance (17 cats) . Fifty-eight cats (83 per cent) survived the hospitalisation period . Complications occurred in 18 cats (26 per cent) and were related to anaesthesia (four cats), the underlying disease process (15 cats), surgery (five cats) and were undetermined in one cat.

Minerva Stomatol, 2004 Jun, 53(6), 345 - 53
{Genetic and microbiologic tests in periodontal disease}; Checchi L et al.; AIM: The presence of genetic polymorphisms IL-1A and IL-1B was demonstrated to be associated to an increased clinical severity of Periodontitis . The aim of our study was to verify the existence of a relationship between such genetic polymorphisms and pathogenic variations of Actinobacillus actinomycetemcomitans (Aa), Prevotella intermedia (Pi) and Porphyromonans gingivalis (Pg), in patients with periodontitis before and after active therapy . METHODS: Twenty-five patients were randomly selected among those attending the Department of Periodontology, University of Bologna, on the basis of radiographic and clinical data . At the end of active therapy, subjects had to undergo "PST" test . RESULTS: The "PST" was positive in 32% (8 out of 25) of patients; the majority of them (4 out of 8) were classified ADA IV, 3 were ADA III and 1 was ADA I (p > 0.05) . All subjects with good hygiene therapy (13 out of 24 patients) have shown a reduced bacterial count after active clinical therapy (Fisher test p = 0.03) . CONCLUSION: No statistically significant association was observed between positivity to "PST" and severity of periodontal disease.

Curr Infect Dis Rep, 2004 Aug, 6(4), 276 - 282
Fungal Endocarditis; Nadir E et al.; Recent advances in medicine have caused fungal endocarditis (FE) to be a more common disease entity . Many fungi are potential pathogens in FE, although Candida species and Aspergillus species are the most common . Valvular heart disease is the necessary underlying condition for FE, with intravenous devices and antibiotic use being the predisposing factors for yeast endocarditis, whereas immunosuppression in patients with valvulopathy predisposes for mold endocarditis . Better prognosis of FE depends on fast and accurate diagnosis and subsequent treatment . Echocardiography was the most valuable recent technique in the past two decades that allowed early diagnosis of FE and is probably responsible for the improved prognosis of patients with FE . In the future, development of nonculture-based diagnostic tests may further improve the sensitivity, specificity, and rapidity of microbiologic diagnosis of FE . Novel approaches in treatment, such as new antifungal drugs, also may assist in achieving cure and further improving the prognosis of this disease entity.

Minerva Stomatol, 2004 May, 53(5), 215 - 30
Peri-implant diseases . A systematic review of the literature; Romeo E et al.; Peri-implantitis surrounding oral implants is an inflammatory process affecting the soft and hard tissues resulting in rapid loss of supporting bone associated with bleeding and suppuration . The etiopathogenesis of early and late peri-implantitis is less well understood and seems to be related to the peri-implant environment and to the soft tissues/implant interface, to patient-related factors (smoke, systemic diseases, plaque control) and to host-parasite equilibrium . However, there is wide scientific evidence that demonstrates the direct correlation between oral microbiota and peri-implant mucositis or peri-implantitis . In particular, adherence and colonization of microbiota on plaque-exposed biomaterials, like c.p . titanium, are discriminant factors for the development of infection . Regenerative or resective surgical approaches are proposed for the treatment of peri-implantitis depending on the morphology and the shape of bone defects . The therapeutical approach of peri-implantitis surrounding dental implants comprises several aspects such as the removal of supra-gingival bacterial plaque, an appropriate surgical approach, the removal of granulation tissue and detoxification of the exposed implant surface, the elimination of the anaerobic ecosystem by the removal of peri-implant pocket (gingivectomy or apically repositioned flap) or by the regeneration of the peri-implant hard tissues and, finally, the establishment of an efficient plaque control regimen.

Vet Microbiol, 2004 Aug 6, 101(4), 225 - 34
Paratuberculosis in farmed and free-living wild ruminants in the Czech Republic (1999-2001); Machackova M et al.; Due to the occurrence of the infection of Mycobacterium avium subspecies paratuberculosis among domestic ruminants and the rapid development of farmed deer industry and the market of cloven-hoofed game we have carried surveys of paratuberculosis, beginning in 1997, in the most common four species of wild ruminants in the Czech Republic {Pavlik et al., Vet . Microbiol . 77 (2000) 231-251} . From 1999 the prevalence of paratuberculosis has been slightly reduced in all three types of husbandry of wild ruminants . Nevertheless paratuberculosis has been diagnosed in wild ruminants in three districts, in four game parks and in five farms . M . a . paratuberculosis was isolated from 128 (5.3%) out of 2,403 wild ruminants of four animal species: 106 red deer, 2 roe deer, 4 fallow deer and 16 mouflons . In red deer farms, the highest number of clinical paratuberculosis cases was in yearling deer . RFLP type B-C1 of M . a . paratuberculosis predominated during the second period (1999-2001) in all types of husbandry with no relationship to wild ruminant species . New "cattle" RFLP types B-C5 and B-C16 of M . a . paratuberculosis were described in infected farmed red deer and one "intermediate" RFLP type R-I4 in fallow deer from one game park . The survival of M . a . paratuberculosis was found to be 4 months during winter in the pasture after destocking of all cattle infected with paratuberculosis . We found that non-vertebrates, wild ruminants or non-ruminant wildlife can be vectors and potentially become a risk factor in the spread of M . a . paratuberculosis infection.

Clin Microbiol Rev, 2004 Jul, 17(3), 681 - 94, table of contents
Competency assessment in the clinical microbiology laboratory; Sharp SE et al.; The laboratory comprises an invaluable part of the total health care provided to patients . Competency assessment is one method by which we can verify that our employees are competent to perform laboratory testing and report accurate and timely results . To derive the greatest benefit from the inclusion of competency assessment in the laboratory, we must be sure that we are addressing areas where our efforts can be best utilized to optimize patient care . To be competent, an employee must know how to perform a test, must have the ability to perform the test, must be able to perform the test properly without supervision, and know when there is a problem with the test that must be solved . In some cases, competency assessment protocols may demonstrate areas of competence but can fail to disclose incompetence . For example, challenges of low-complexity tasks (such as reading the technical procedure manual) are inferior to challenges that measure understanding and execution of a protocol, and poorly designed competency challenges will probably not detect substandard laboratory performance . Thus, if we are to receive the greatest benefit from our competency assessment programs, which may be time-consuming for the supervisors and the staff as well, we must not only meet the letter of the law but also find a way to make these assessments meaningful, instructive, and able to detect areas of concern . As we address competency assessment in our laboratories, we must understand that when done properly, competency assessment will reward our organizations and assist us in providing the best possible care to our patients.

Presse Med, 2004 Jun 19, 33(11), 707 - 9
{Evolution in the prevalence of intestinal parasitosis in the Fort de France University Hospital (Martinique)}; Edouard A et al.; OBJECTIVE: Determine the prevalence of intestinal parasitosis between January 1, 1997 and December 31, 1999 in the microbiology laboratory of the Fort de France University Hospital . METHOD: Retrospective study of the results of 4684 parasitological examinations of stools performed in 2704 patients between January 1, 1997 and December 31, 1999 in this laboratory . RESULTS: This survey showed the high prevalence of anguillulosis, found in 51.69% of infested patients and in 4.56% of the population studied, the ever high prevalence of non or scarcely pathogenic amoebas (Endolimax nanus, Dientamoeba fragilis, Entamoeba coli) found in 27.19% of infested patients and 1.88% of the population studied, together with that of hookworms (12.80% of infested patients and 1.13% of the population), and the persistence of lambliasis . This study also revealed the presence of cryptosporidies (7 cases) and microsporidies (4 cases) in the patients infected by the human immunodeficiency virus . DISCUSSION: These results confirm the trend of the past twenty Years and the results of surveys initiated by the national statistics board (INSERM) in 1978, 1988 and 1995-1995, with the regression of bilharziosis and the persistence of anguillulosis and hookworms . CONCLUSION: The improvement in living conditions and hygiene, the combined efforts of the health care workers and Authorities in Martinique over the past 30 Years in the fight against parasites have led to a great reduction in the prevalence of classical intestinal parasitosis . However, new parasites associated with HIV infection have appeared.

Cell Tissue Bank, 2001, 2(3), 179 - 83
Bone bank service in Odense, Denmark . Audit of the first ten years with bone banking at the Department of Orthopaedics, Odense University Hospital; Nielsen HT et al.; There has been an increase in the demand for allograft bone in recent years . The Odense University Hospital bone bank has been in function since 1990, and this paper outlines our results during the 10 year period 1990-1999 . Potential donors were screened by contemporary banking techniques which include a social history, donor serum tests for HIV, hepatitis B and C, and graft microbiology . The bones were stored at -80 degrees C . No type of secondary sterilisation was made . 423 femoral heads were approved and donated to 300 patients,1-6 heads/operation . The allografts have been used mainly to reconstruct defects at revision hip arthroplasty (34%), and for fracture surgery (24%) . 7 % of all transplanted patients were reoperated because of infection . In the hip revision group the infection rate was 4 % . There were no cases of disease transmission . During the 10 year period there was a change in the clinical use of the allografts . In the first years the allografts were mainly used for spinal fusion surgery, but today the majority are used in hip revision and fracture surgery . The clinical results correspond to those reported in larger international series.

Mol Microbiol, 2004 Aug, 53(3), 985 - 1000
Transcription of the sporulation gene ssgA is activated by the IclR-type regulator SsgR in a whi-independent manner in Streptomyces coelicolor A3(2); Traag BA et al.; SsgA plays an important role in the control of sporulation-specific cell division and morphogenesis of streptomycetes, and ssgA null mutants have a rare conditionally non-sporulating phenotype . In this paper we show that transcription of ssgA and of the upstream-located ssgR, an iclR-type regulatory gene, is developmentally regulated in Streptomyces coelicolor and activated towards the onset of sporulation . A constructed ssgR null mutant was phenotypically very similar to the ssgA mutant . The absence of ssgA transcription in this mutant is probably the sole cause of its sporulation deficiency, as wild-type levels of sporulation could be restored by the SsgR-independent expression of ssgA from the ermE promoter . Binding of a truncated version of SsgR to the ssgA promoter region showed that ssgA transcription is directly activated by SsgR; such a dependence of ssgA on SsgR in S . coelicolor is in clear contrast to the situation in S . griseus, where ssgA transcription is activated by A-factor, and its control by the SsgR orthologue, SsfR, is far less important . Our failure to complement the ssgR mutant with S . griseus ssfR suggests functional differences between the genes . These observations may explain some of the major differences in developmental control between the phylogenetically divergent species S . coelicolor and S . griseus, highlighted in a recent microreview (Chater and Horinouchi (2003) Mol Microbiol 48: 9-15) . Surprisingly, transcription of ssgA and ssgR is not dependent on the early whi genes (whiA, whiB, whiG, whiH, whiI and whiJ ) .

J Perinatol, 2004 Nov, 24(11), 679 - 85
The possible role of intrauterine infections in unexplained second trimester abortions and macerated stillbirths: a study from a single center; Atay GA et al.; OBJECTIVE: To investigate the role of intrauterine infections in unexplained second trimester abortions and stillbirths . STUDY DESIGN: Histopathologic and microbiologic evidence of intrauterine infection in the placentas, fetal membranes and fetal lung tissues of 18 unexplained second trimester abortions and macerated stillbirth cases as well as the placentas and fetal membranes of 10 healthy term neonates were investigated in a prospective study conducted in Ankara University School of Medicine, Turkey . RESULTS: Histopathologic chorioamnionitis and placental culture positivity rates in the study and control groups were 64.7 vs 0% . Bacteria were recovered from 90.9% of placentas and 36.4% of fetal lungs of the cases with histopathologic chorioamnionitis . Intrauterine infection was found in 66.7% of the whole study group, in 85.7% of the unexplained second trimester abortions, and in 54.5% of the macerated stillbirths . CONCLUSION: Intrauterine infection may be an important factor in unexplained stillbirths and second trimester abortions in centers where pregnancy follow-ups lack evaluation for asymptomatic infection.

Z Orthop Ihre Grenzgeb, 2004 May-Jun, 142(3), 333 - 6
{Soft tissue reconstruction following total knee arthroplasty: treatment possibilities and retrospective analysis of plastic surgery therapy treatment possibilities}; Unglaub F et al.; AIM: We performed a retrospective analysis of patients with soft tissue defects following total knee arthroplasty and therapy . Furthermore, we described the possibilities of covering soft tissue defects following knee arthroplasty . METHOD: In 5 patients, soft tissue defects following knee arthroplasty were covered with medial M . gastrocneminus flaps . Localisation and size of the defect, microbiology, risk factors, and interval between arthroplasty and the occurrence of the soft tissue defect were retrospectively analysed . RESULTS: On average, defects occurred 9 weeks after prosthesis implantation . In all cases, the soft tissue defect developed in the area of the incision . In 3 cases, wound infection was diagnosed . Four patients exhibited factors associated with wound-healing failure . No prosthesis was lost . CONCLUSION: The gastrocneminus muscle flap provides good quality coverage with small donor site defect, permits early mobilisation, and allows for fast rehabilitation . The risk of flap loss is minimal . Early and adequate defect coverage can reduce both prosthesis loss and amputation rates.

Chest, 2004 Jul, 126(1), 268 - 72
The clinical utility of flexible bronchoscopy in the evaluation of chronic cough; Barnes TW et al.; STUDY OBJECTIVES: To assess the clinical utility of flexible bronchoscopy (FB) in the evaluation of patients with chronic cough and normal or nonlocalizing chest radiographic findings . DESIGN: Retrospective chart review . SETTING: Tertiary referral center . PATIENTS: Forty-eight patients with chronic cough and no other clinical or radiographic indications for FB who underwent the procedure from 1996 to 2001 . RESULTS: Before FB, 45 patients had a chest radiograph and 21 patients had a chest CT performed . On visual inspection, 37 of 48 of the FB findings (82%) were normal, 9 were consistent with "bronchitis," one patient had a "small tracheal plaque," and one patient had "minimal arytentoid redundancy." Three of the 27 patients who had a microbiologic assessment performed during their FB demonstrated potentially pathogenic organisms . Antibiotic treatment based on culture data did not result in improvement in cough . Cytologic examination was performed in specimens from 33 patients; 1 patient was noted to have reactive squamous atypia, and the remainder were normal . The patient noted to have minimal arytenoid redundancy and the patient with a tracheal plaque each had another etiology for their cough identified, the treatment of which resulted in cough improvement . CONCLUSIONS: FB adds little to the diagnosis of chronic cough in the context of normal or nonlocalizing chest radiographic or CT findings . FB did not result in successful treatment alteration, nor did it contribute to the identification of the cough etiology.

Pediatr Infect Dis J, 2004 Jul, 23(7), 642 - 9
Invasive pneumococcal infections among hospitalized children in Bamako, Mali; Campbell JD et al.; BACKGROUND: Prevention of invasive pneumococcal disease (IPD) in children is a global public health priority, and determination of the most common serotypes is crucial for vaccine development and implementation . METHODS: We performed prospective surveillance for IPD in hospitalized children in Bamako, Mali . All febrile children and others suspected to have invasive bacterial disease had an admission blood culture and cultures of additional anatomic sites when indicated . Standard microbiologic methods were used to identify, serotype and determine antibiograms for pneumococcal isolates . RESULTS: Of 2,049 children enrolled, 106 (5%) had an IPD, including 47 cases of meningitis and 44 bacteremic pneumonias . The incidence was highest in infants (84/100,000/year) . The overall IPD case fatality rate was 24% . Only 2 of 96 isolates were nonsusceptible to penicillin . The serotypes isolated were 5 (54%), 2 (14%), 7F (10%), 19F (8%), 6A/B (3%), 9V (3%), 1 (2%) and 14 (1%) . CONCLUSIONS: IPD is common and frequently fatal among hospitalized children in Mali, but surprisingly little resistance has occurred . Notably, 91% of the serotypes causing IPD in Bamako children are found in the 11-valent pneumococcal conjugate vaccine.

Curr Gastroenterol Rep, 2004 Aug, 6(4), 273 - 9
Pyogenic and amebic liver abscesses; Kurland JE et al.; Pyogenic and amebic liver abscesses are the two most common hepatic abscesses . Amebic abscesses are more common in areas where Entamoeba histolytica is endemic, whereas pyogenic abscesses are more common in developed countries . Pyogenic abscess severity is dependent on the bacterial source and the underlying condition of the patient . Amebic liver abscess is more prevalent in individuals with suppressed cell-mediated immunity, men, and younger people . The right lobe of the liver is the most likely site of infection in both types of hepatic abscess . Patients usually present with a combination of fever, right-upper-quadrant abdominal pain, and hepatomegaly . Jaundice is more common in the pyogenic abscess . The diagnosis is often delayed and is usually made through a combination of radiologic imaging and microbiologic, serologic, and percutaneous techniques . Treatment involves antibiotics along with percutaneous drainage or surgery.

J Clin Microbiol, 2004 Jul, 42(7), 2988 - 95
Coexistence of multiple genotypes, including newly identified genotypes, in outbreaks of gastroenteritis due to Norovirus in Japan; Kageyama T et al.; Norovirus (NV) (formerly called Norwalk-like virus) is the most common cause of acute nonbacterial gastroenteritis in humans . Recently, we reported an NV genotyping scheme based on variability in the capsid N-terminal/shell (N/S) domain gene (Katayama et al., Virology 299:225-239, 2002) . We found 19 genotypes, including nine of genogroup I and 10 of genogroup II . In the present study, we investigated the molecular epidemiology of NV from 66 outbreaks that occurred in Saitama Prefecture, Japan, from 1997 to 2002 . We screened 416 stool specimens by a real-time reverse transcription (RT)-PCR method (Kageyama et al., J . Clin . Microbiol . 41:1548-1557, 2003) and detected 156 NV-positive specimens, from which we amplified the capsid N/S domain gene by RT-PCR and then cloned the PCR products . After sequencing these clones, we obtained 368 sequence variants (strains) . By applying our classification scheme to the strains from Saitama and other published strains, we identified a total of 31 genotypes, including an additional five genotypes for genogroup I and seven for genogroup II . Of the 31 genotypes, 26 were present in the Saitama area during that time period . These results provide additional evidence for the great diversity of human NV genotypes . Specimens from all shellfish-related infections contained multiple genotypes, including several new genotypes . On the other hand, single genotypes were observed mostly in outbreaks that originated in semiclosed communities . Thus, the number of NV genotypes in each outbreak depended on the route of transmission.

Indian J Lepr, 2003 Oct-Dec, 75(4), 317 - 25
Modular teaching programme on leprosy; Karthikeyan K et al.; Though repeated attention has been drawn to a lack of proper teaching-learning modules in leprosy endemic countries, no satisfactory module exists . Keeping in view this fact, we attempted to draft a suitable module on leprosy that could be used to teach leprosy to undergraduate medical students in a simple and comprehensive manner . We used two different modules, Module A and Module B, to teach two different batches of students of the pre-final year (VI and VII semesters) of the MBBS course . Both these modules were conducted by the Department of Dermatology and STD, with participation by the Departments of Microbiology, Pathology and Preventive and Social Medicine . The drafts of the modules were discussed before hand in the Department, keeping in mind the number of days allotted to us . Both the modules were different in certain aspects, but the basic concept was the same . Because Module A had more time, certain practical aspects were also discussed . It was interesting to note that the percentage of increase in the post-test score was 17 for Module A and 15 for Module B, thus proving that both the modules were effective in conveying the core message about leprosy.

Appl Environ Microbiol, 2004 Jul, 70(7), 3979 - 87
Toxic and nontoxic microcystis colonies in natural populations can be differentiated on the basis of rRNA gene internal transcribed spacer diversity; Janse I et al.; Assessing and predicting bloom dynamics and toxin production by Microcystis requires analysis of toxic and nontoxic Microcystis genotypes in natural communities . We show that genetic differentiation of Microcystis colonies based on rRNA internal transcribed spacer (ITS) sequences provides an adequate basis for recognition of microcystin producers . Consequently, ecological studies of toxic and nontoxic cyanobacteria are now possible through studies of rRNA ITS genotypic diversity in isolated cultures or colonies and in natural communities . A total of 107 Microcystis colonies were isolated from 15 lakes in Europe and Morocco, the presence of microcystins in each colony was examined by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and they were grouped by rRNA ITS denaturing gradient gel electrophoresis (DGGE) typing . Based on DGGE analysis of amplified ITSa and ITSc fragments, yielding supplementary resolution (I . Janse et al., Appl . Environ . Microbiol . 69:6634-6643, 2003), the colonies could be differentiated into 59 classes . Microcystin-producing and non-microcystin-producing colonies ended up in different classes . Sequences from the rRNA ITS of representative strains were congruent with the classification based on DGGE and confirmed the recognition of microcystin producers on the basis of rRNA ITS . The rRNA ITS sequences also confirmed inconsistencies reported for Microcystis identification based on morphology . There was no indication for geographical restriction of strains, since identical sequences originated from geographically distant lakes . About 28% of the analyzed colonies gave rise to multiple bands in DGGE profiles, indicating either aggregation of different colonies, or the occurrence of sequence differences between multiple operons . Cyanobacterial community profiles from two Dutch lakes from which colonies had been isolated showed different relative abundances of genotypes between bloom stages and between the water column and surface scum . Although not all bands in the community profiles could be matched with isolated colonies, the profiles suggest a dominance of nontoxic colonies, mainly later in the season and in scums.

Appl Environ Microbiol, 2004 Jul, 70(7), 3910 - 7
Serotypes and virulence gene profiles of shiga toxin-producing Escherichia coli strains isolated from feces of pasture-fed and lot-fed sheep; Djordjevic SP et al.; Shiga toxin-producing Escherichia coli (STEC) strains possessing genes for enterohemolysin (ehxA) and/or intimin (eae), referred to here as complex STEC (cSTEC), are more commonly recovered from the feces of humans with hemolytic uremic syndrome and hemorrhagic colitis than STEC strains that do not possess these accessory virulence genes . Ruminants, particularly cattle and sheep, are recognized reservoirs of STEC populations that may contaminate foods destined for human consumption . We isolated cSTEC strains from the feces of longitudinally sampled pasture-fed sheep, lot-fed sheep maintained on diets comprising various combinations of silage and grain, and sheep simultaneously grazing pastures with cattle to explore the diversity of cSTEC serotypes capable of colonizing healthy sheep . A total of 67 cSTEC serotypes were isolated, of which 21 (31.3%), mainly isolated from lambs, have not been reported . Of the total isolations, 58 (86.6%) were different from cSTEC serotypes isolated from a recent study of longitudinally sampled healthy Australian cattle (M . Hornitzky, B . A . Vanselow, K . Walker, K . A . Bettelheim, B . Corney, P . Gill, G . Bailey, and S . P . Djordjevic, Appl . Environ . Microbiol . 68:6439-6445, 2002) . Our data suggest that cSTEC serotypes O5:H(-), O75:H8, O91:H(-), O123:H(-), and O128:H2 are well adapted to colonizing the ovine gastrointestinal tract, since they were the most prevalent serotypes isolated from both pasture-fed and lot-fed sheep . Collectively, our data show that Australian sheep are colonized by diverse cSTEC serotypes that are rarely isolated from healthy Australian cattle.

Int J Infect Dis, 2004 Jul, 8(4), 210 - 6
Nosocomial infective endocarditis in critically ill patients: a report of three cases and review of the literature; Haddad SH et al.; Nosocomial infective endocarditis (NIE) is a relatively uncommon but nevertheless a serious complication affecting critically ill hospitalized patients who are frequently exposed to life-saving invasive procedures . We report three cases of NIE in a tertiary-care hospital encountered during a period of two years . The first case developed in a 50% burn-injured patient; the second in a liver transplant recipient; and the third in a renal transplant recipient . All patients met indications for cardiac surgical intervention, however, the patient who had received a liver transplant (case 2) was considered a poor candidate and unfit for surgery; she subsequently died . The other two patients underwent open-heart surgery . The burns patient (case 1) survived; conversely, the renal transplant recipient (case 3) died postoperatively . We have reviewed the literature concerning NIE in critically ill patients and describe the epidemiology, microbiology and clinical features of this uncommon infection and comment on its diagnosis and management.

Nature, 2004 Jul 1, 430(6995), 68 - 71
Role of metal-reducing bacteria in arsenic release from Bengal delta sediments; Islam FS et al.; The contamination of ground waters, abstracted for drinking and irrigation, by sediment-derived arsenic threatens the health of tens of millions of people worldwide, most notably in Bangladesh and West Bengal . Despite the calamitous effects on human health arising from the extensive use of arsenic-enriched ground waters in these regions, the mechanisms of arsenic release from sediments remain poorly characterized and are topics of intense international debate . We use a microscosm-based approach to investigate these mechanisms: techniques of microbiology and molecular ecology are used in combination with aqueous and solid phase speciation analysis of arsenic . Here we show that anaerobic metal-reducing bacteria can play a key role in the mobilization of arsenic in sediments collected from a contaminated aquifer in West Bengal . We also show that, for the sediments in this study, arsenic release took place after Fe(III) reduction, rather than occurring simultaneously . Identification of the critical factors controlling the biogeochemical cycling of arsenic is one important contribution to fully informing the development of effective strategies to manage these and other similar arsenic-rich ground waters worldwide.

Clin Infect Dis, 2004 Jun 15, 38(12), e119 - 22 Epub 2004 May 24.
Prevention of laboratory-acquired brucellosis; Robichaud S et al.; Brucellosis is one of the most common laboratory-acquired infections, mostly because aerosolization is a mechanism of transmission in this setting . We report an exposure to Brucella melitensis that occurred in a large microbiology laboratory and describe the strategy chosen for antibiotic prophylaxis and serological follow-up of exposed workers.

Acta Clin Belg, 2004 Mar-Apr, 59(2), 79 - 83
The one fixed point in a changing age . An analysis of half-secular trends among original papers published in The Lancet 1945-95; Selvais P et al.; We performed a systematic survey of original papers published in The Lancet throughout the years 1945, 1946, 1970 and 1995 . The Lancet was chosen due to a conservative layout through these years, and the year 1946 added to the analysis to ponder for any war-induced bias . The general layout and structure of the journal were indeed maintained throughout these years . The number of authors per paper increased steadily (from 1.7 +/- 1.3 in 1945 to 6.4 +/- 3.4 in 1995), as well as the number of papers written on behalf of investigation groups (from 1 in 1945 to 37 in 1995) . Major changes were observed in the area of origin of the papers, from an overwhelming British Isles' preponderance (87% of papers published in 1945) to a partake with Continental Europe, America and international groups (27, 33, 15 and 12%, respectively of the 1995 papers) . The most often addressed fields of medicine shifted from war-, surgery- and microbiology-related subjects in 1945 towards cardiology-, oncology-, and virus-related works in 1995 . The most impressive and enduring change was the increase in the use of statistical tools for data analysis and presentation (0.3% of papers in 1945 vs . 60% in 1995).

J Med Assoc Thai, 2004 May, 87(5), 486 - 91
Bacterial isolation with on-site inoculation of ascites fluid into hemoculture bottle in spontaneous bacterial peritonitis; Charoenhirunyingyos S et al.; OBJECTIVE: To compare the on-site bacterial inoculation of ascites fluid into hemoculture bottle with routine method in the patients who were preliminary diagnosed of SBP . MATERIAL AND METHOD: A retrospective analysis of case records during January-December 2001 . RESULTS: A total of 673 specimens from 325 patients were retrieved from the data records at the Department of Microbiology, Siriraj Hospital in 2001 . The neutrocytic ascites were found in 163 specimens (94 patients) . The routine method and on-site inoculation into the hemoculture bottle were employed in 107 and 56 specimens respectively . Culture-positive neutrocytic ascites was found in the routine method 16 (14.9%) specimens and in the on-site inoculation 26 (46.4%) specimens (p < 0.0001) . Among these samples, the two methods were simultaneously performed in 42 specimens of which 18 paired specimens were eligible for analysis . Positive culture was found in 2 samples in which the routine method and in an additional 5 samples in which on-site inoculation into hemoculture bottle method . Using Kappa analysis (e score = 0.328, 95% CI = -0.172 to 0.829) that can be interpreted the on-site inoculation method had a higher yield than the routine technique . Moreover, 21 cases also had their blood and ascites samples simultaneously collected and cultured . 4 of ten (19%) and 5 of eleven (23.8%) cases were found in the routine and on-site and direct inoculation groups respectively . These finding suggested that the severity of infection in among two groups were similar and unlikely to be the cause of the difference of the positive isolation rate in both groups . CONCLUSION: The on-site and direct inoculation of ascites into hemoculture bottle method had a significantly higher isolation rate than routine method (i.e . 46.4% versus 14.9% p < 0.0001) either with separated or paired samples of ascites.

J Am Soc Echocardiogr, 2004 Jul, 17(7), 766 - 8
Mortality in patients with paravalvular abscess diagnosed by transesophageal echocardiography; Cosmi JE et al.; BACKGROUND: Paravalvular abscess is a complication of endocarditis that may lead to persistent infection, conduction abnormalities, fistula formation, worsening congestive heart failure, and death . METHODS: Between 1991 and 2001, paravalvular abscess was identified on transesophageal echocardiography in 24 patients who subsequently underwent surgical treatment . Echocardiographic findings were reviewed for location of abscess, presence of a valvular prosthesis, valvular function, and presence of vegetations . Information gathered included the time interval between transesophageal echocardiography diagnosis and operation, inhospital mortality, and microbiologic data . RESULTS: Of 24 patients, 9 died, for a mortality of 38% . Of the patients who died, the average survival after operation was 43 days, with a range of 1 to 238 days . Of the 14 patients with significant valvular or paravalvular regurgitation, 8 died (57%) . However, of the 10 patients with mild or no regurgitation, only 1 died (10%) . This difference was statistically significant (P =.02) . CONCLUSION: Patients without preserved valve function (those with moderate or severe regurgitation) have a significantly worse outcome (57% mortality) than do those with normal valvular function (10% mortality).

Infect Immun, 2004 Jul, 72(7), 4151 - 8
Tritrichomonas foetus induces apoptotic cell death in bovine vaginal epithelial cells; Singh BN et al.; Tritrichomonas foetus is a serious veterinary pathogen, causing bovine trichomoniasis, a sexually transmitted disease leading to infertility and abortion . T . foetus infects the mucosal surfaces of the reproductive tract . Infection with T . foetus leads to apoptotic cell death of bovine vaginal epithelial cells (BVECs) in culture . An affinity-purified cysteine protease (CP) fraction yielding on sodium dodecyl sulfate-polyacrylamide gel electrophoresis a single band with an apparent molecular mass of 30 kDa (CP30) also induces BVEC apoptosis . Treatment of CP30 with the protease inhibitors TLCK (Nalpha-p-tosyl-l-lysine chloromethyl ketone) and E-64 {l-trans-epoxysuccinyl-leucylamide-(4-guanido)-butane} greatly reduces induction of BVEC apoptosis . Matrix-assisted laser desorption ionization-time-of-flight MALDI-TOF mass spectrometry analysis of CP30 reveals a single peak with a molecular mass of 23.7 kDa . Mass spectral peptide sequence analysis of proteolytically digested CP30 reveals homologies to a previously reported cDNA clone, CP8 (D . J . Mallinson, J . Livingstone, K . M . Appleton, S . J . Lees, G . H . Coombs, and M . J . North, Microbiology 141:3077-3085, 1995) . Induction of apoptosis is highly species specific, since the related human parasite Trichomonas vaginalis and associated purified CPs did not induce BVEC death . Fluorescence microscopy along with the Cell Death Detection ELISA(PLUS) assay and flow cytometry analyses were used to detect apoptotic nuclear condensation, DNA fragmentation, and changes in plasma membrane asymmetry in host cells undergoing apoptosis in response to T . foetus infection or incubation with CP30 . Additionally, the activation of caspase-3 and inhibition of cell death by caspase inhibitors indicates that caspases are involved in BVEC apoptosis . These results imply that apoptosis is involved in the pathogenesis of T . foetus infection in vivo, which may have important implications for therapeutic interference with host cell death that could alter the course of the pathology in vivo.

Am J Trop Med Hyg, 2004 Jun, 70(6), 651 - 7
Reducing diarrhea through the use of household-based ceramic water filters: a randomized, controlled trial in rural Bolivia; Clasen TF et al.; Ceramic water filters have been identified as one of the most promising and accessible technologies for treating water at the household level . In a six-month trial, water filters were distributed randomly to half of the 50 participating households in a rural community in Bolivia; the remaining households continued to use customary water handling practices and served as controls . In four rounds of sampling following distribution of the filters, 100% of the 96 water samples from the filter households were free of thermotolerant coliforms compared with 15.5% of the control household samples . Diarrheal disease risk for individuals in intervention households was 70% lower than for controls (95% confidence interval {CI} = 53-80%; P < 0.001) . For children less than five years old, the reduction in risk was 83% (95% CI = 51-94%; P < 0.001) . These results show that affordable ceramic water filters enable low-income households to treat and maintain the microbiologic quality of their drinking water.

Enferm Infecc Microbiol Clin, 2004 May, 22(5), 299 - 304; quiz 305
{Real-time PCR}; Costa J; PCR based assays are currently used routinely in most microbiology laboratories . But, with few exceptions, they are restricted to the field of virology, especially to a limited number of viral targets with important economical interest for which commercially well standardized assays are available . For several reasons, it has had a poor implementation of the PCR assays into routine diagnostics for other infectious diseases despite they are advantageous . Combined with automated sample isolation of nucleic acids, real-time PCR gives an ideal platform for the development of molecular assays for a wide range of infectious agents with clinical interest . Because of its advantages, as simplicity, rapidity and minor risk of contamination, real-time PCR will go replace conventional PCR assays and its use will extend to a wide range of applications in clinical microbiology.

Enferm Infecc Microbiol Clin, 2004 May, 22(5), 272 - 8
{Epidemiology of human immunodeficiency virus (HIV) infection in the province of Salamanca (1985-2002)}; Gonzalez-Celador R et al.; INTRODUCTION: Since effective treatment against HIV has become available, HIV infection surveillance is focussed not only on AIDS cases, but also on new cases of HIV infection . One of the methods used for this task is compilation of case records . We report the results of 18 years of recording information on new diagnoses of HIV (1985-2002) in the area of Salamanca (Spain) . METHODS: Our report system is based on active monitoring of results from the microbiology laboratory of the Salamanca Health Area . All patients testing positive in HIV analytical studies and who had not been previously diagnosed as HIV-infected are included in the report system . RESULTS: A total of 188 579 analytical results were studied . We found 16,772 (9.23%) HIV-positive results, 1984 of which were new HIV infection diagnoses . The incidence for the entire period was 556 cases/100,000 inhabitants . The highest incidence was found in 1992 (62.2 cases/100,000), with a progressive decrease thereafter to 2002 (5.18/100,000) . Among the total, 75.3% of patients were men between 20 and 39 years old and more than half (62.2%) were intravenous drug users . Over the period studied, there has been an increase in the age of patients at the time of diagnosis, a trend to increases in the number of cases acquired through heterosexual transmission and a reduction in the number of drug users . During the study period, 312 AIDS cases were declared . CONCLUSIONS: Epidemiological information systems for HIV surveillance are essential to know the features, magnitude and evolution of the epidemic . Creation of a national HIV infection surveillance system to compile the information from smaller regional or local organisms would contribute substantially to our understanding of the epidemiology and evolution of HIV infection.

J Biomed Mater Res, 2004 Jul 15, 70B(1), 139 - 45
Diagnosis of infection after total hip replacement; Savarino L et al.; Subclinical infection in patients with pain following total hip replacement (THR) is an underestimated condition that needs consideration because it mimics aseptic loosening, contributes to periprosthetic osteolysis, and necessitates proper treatment . We aimed to define the reliability of diagnostic parameters that are routinely used before revision surgery for the assessment of infection . A continuous series of 26 subjects who underwent THR revision surgery was considered, including 21 cases diagnosed as aseptic loosening (group A) and 5 hip revisions with a clinical diagnosis for infection (group B) . Seven subjects at the time of the primary arthroplasty were used as negative controls (group C) . Technetium-99m labeled hydroxymethylene diphosphonate {(99m)Tc-HDP}- and technetium-99m hexamethylpropyleneamine oxide {(99m)Tc-HMPAO)}-labeled granulocyte scintigraphy, histology of peri-implant tissues, laboratory tests for inflammation, and microbiology were performed . Scintigraphy was positive for loosening {positive (99m)Tc-HDP scan} but negative for infection {negative (99m)Tc-HMPAO-labeled granulocyte scan} in all group A patients, whereas in 11 cases (52%) a positive culture was unexpectedly obtained . Histology showed conflicting results: Polymorphonuclear cells (PMNs) were found only in 5 of 11 culture-positive patients, whereas in 2 cases the presence of PMNs did not correspond to a positive culture . In group B patients, both isotope scans and microbiology were found to be positive . All control subjects (group C) had negative cultures . In our opinion, smoldering infection could be present in a significant proportion of cases of failed hip implants currently diagnosed as "nonseptic." The inflammatory response to wear debris and the presence of superimposed, slowly growing bacteria could act synergically, both contributing to the pathogenesis of periprosthetic osteolysis .

Environ Microbiol, 2004 Jul, 6(7), 678 - 85
Design and application of two oligonucleotide probes for the identification of Geodermatophilaceae strains using fluorescence in situ hybridization (FISH); Urzi C et al.; Bacteria of the family of Geodermatophilaceae are actively involved in the decay processes {Urzi, C . and Realini, M . (1998) Int Biodeterior Biodegrad 42: 45-54; Urzi, C., Salamone, P., Schumann, P., and Stackebrandt, E . (2000) Int J Syst Evol Microbiol 50: 529-536} of stone monuments . Characterization of isolates includes phenotypic, chemotaxonomic and genetic analysis often requiring long-term procedures . The use of specific probes for members of Geodermatophilaceae family could be useful for the easy detection of those strains colonizing rock surfaces and involved in the biodeterioration . Two 16S rRNA-targeted oligonucleotide probes were designed for the specific detection of members of the family Geodermatophilaceae using fluorescence in situ hybridization (FISH); one probe specific for members of the two genera Geodermatophilus/Blastococcus and the second for members of the genus Modestobacter.

J Clin Microbiol, 2004 Jun, 42(6), 2438 - 44
Analysis of the allelic diversity of the mycobacterial interspersed repetitive units in Mycobacterium tuberculosis strains of the Beijing family: practical implications and evolutionary considerations; Mokrousov I et al.; A study set comprised 44 Mycobacterium tuberculosis strains of the Beijing family selected for their representativeness among those previously characterized by IS6110-RFLP and spoligotyping (Northwest Russia, 1997 to 2003) . In the present study, these strains were subjected to mycobacterial interspersed repetitive units (MIRU) typing to assess a discriminatory power of the 12-MIRU-loci scheme (P . Supply et al., J . Clin . Microbiol . 39:3563-3571, 2001) . The 44 Russian Beijing strains were subdivided into 12 MIRU types with identical profiles: 10 unique strains and two major types shared by 10 and 24 strains . Thus, basically, two distinct sublineages appear to shape the evolution of the Beijing strains in Russia . Most of the MIRU loci were found to be (almost) monomorphic in the Russian Beijing strains; the Hunter-Gaston discriminatory index (HGDI) for all 12 loci taken together was 0.65, whereas MIRU26 (the most variable in our study) showed a moderate level of discrimination (0.49) . The results were compared against all available published MIRU profiles of Beijing strains from Russia (3 strains) and other geographic areas (51 strains in total), including South Africa (38 strains), East Asia (7 strains), and the United States (4 strains) . A UPGMA (unweighted pair-group method with arithmetic averages)-based tree was constructed . Interestingly, no MIRU types were shared by Russian and South African strains (the two largest samples in this analysis), whereas both major Russian types included also isolates from other locations (United States and/or East Asia) . This implies the evolution of the Beijing genotype to be generally strictly clonal, although a possibility of a convergent evolution of the MIRU loci cannot be excluded . We propose a dissemination of the prevailing local Beijing clones to have started earlier in South Africa rather than in Russia since more monomorphic loci were identified in Russian samples than in South African samples (mean HGDI scores, 0.08 versus 0.17) . To conclude, we suggest to use a limited number of MIRUs for preliminary subdivision of Beijing strains in Russian (loci 26 + 31), South African (10 + 26 + 39), and global settings (10 + 26 + 39).

Int J Tuberc Lung Dis, 2004 Jun, 8(6), 772 - 7
Evaluation of a new automated, rapid, colorimetric culture system using solid medium for laboratory diagnosis of tuberculosis and determination of anti-tuberculosis drug susceptibility; Baylan O et al.; SETTING: Department of Microbiology and Clinical Microbiology, Gulhane Military Medical Academy, Ankara, Turkey, a tertiary referral hospital in a region endemic for tuberculosis . OBJECTIVE: To evaluate the performance of the Dio-TK Culture System (CS), a new, rapid, automated colorimetric culture system . DESIGN: Dio-TK CS results were compared with routinely used Lowenstein Jensen (LJ) medium and Bactec 460 TB . RESULTS: In this study, 449 specimens, mostly sputum samples obtained from 348 patients, were evaluated . Mycobacteria were isolated from 31 (6.9%), 23 (5.1%), 18 (4.0%) and 21 (4.7%) of the specimens using Bactec 12B, LJ, Dio-TK Medium and Dio-TK SLC (selective), respectively . The mean time to detection of growth of 13 isolates by Bactec 12B, Dio-TK Medium, Dio-TK SLC and LJ medium was respectively 8.9, 15.1, 17.0 and 26.1 days . CONCLUSION: Dio-TK may be a practical and rapid culture system for daily use . However, the manufacturer should improve the system to minimise the effects of manipulation errors . Comparative studies with a larger number of isolates are needed to standardise drug concentrations used in anti-tuberculosis drug susceptibility testing.

DNA Repair (Amst), 2004 Mar 4, 3(3), 323 - 34
A role for DNA polymerase V in G --> T mutations from the major benzo{a}pyrene N2-dG adduct when studied in a 5'-TGT sequence in E . coli; Yin J et al.; Benzo{a}pyrene (B{a}P), a potent mutagen/carcinogen, is metabolically activated to (+)-anti-B{a}PDE, which induces a full spectrum of mutations (e.g . GC --> TA, GC --> AT, etc.) principally via its major adduct {+ta}-B{a}P-N2-dG . Recent findings suggest that different lesion bypass DNA polymerases may be involved in different mutagenic pathways, which is the subject of this report . {+ta}-B{a}P-N2-dG built into a plasmid in a 5'-TGT sequence gives approximately equal numbers of G --> T and G --> A mutations when host E . coli are UV irradiated prior to transformation, so this sequence context was chosen to investigate what DNA polymerases are involved in G --> T versus G --> A mutations . G --> T mutations decline (>10-fold) if E . coli either are not UV-irradiated or are deficient in DNA polymerase V ((delta)umuD/C), demonstrating a role for damage-inducible DNA Pol V in a G --> T pathway . G --> T mutations are not affected by transformation into E . coli deficient in either DNA polymerases II or IV . While the work herein was in progress, Lenne-Samuel et al . {Mol . Microbiol . 38 (2000) 299} built the same adduct into a plasmid in a 5'-GGA sequence, and showed that the frequency of G --> T mutations was similar in UV-irradiated and unirradiated host E . coli cells, suggesting no involvement by damage-inducible, lesion bypass DNA polymerases (i.e., not II, IV or V); furthermore, a role for DNA Pol V was explicitly ruled out . The easiest way to reconcile the findings of Lenne-Samuel et al . with the findings herein is if two G --> T mutagenic pathways exist for {+ta}-B{a}P-N2-dG, where sequence context dictates which pathway is followed . In contrast to the G --> T mutations, herein G --> A mutations from {+ta}-B{a}P-N2-dG in the 5'-TGT sequence context are shown not to be affected by UV-irradiation of host E . coli, and are not dependent on DNA Pol V, or Pol II, Pol IV, or the damage-inducible, but SOS-independent UVM system . Published studies, however, have shown that G --> A mutations are usually enhanced by UV-irradiation of host E . coli prior to the introduction of plasmids either site-specifically modified with {+ta}-B{a}P-N2-dG or randomly adducted with (+)-anti-B{a}PDE; both findings imply the involvement of a lesion-bypass DNA polymerase . These disparate results suggest the existence of two G --> A mutagenic pathways for {+ta}-B{a}P-N2-dG as well, although confirmation of this awaits further study . In conclusion, a comparison between the evidence presented herein and published findings suggests the existence of two distinct mutagenic pathways for both G --> T and G --> A mutations from {+ta}-B{a}P-N2-dG, where in each case one pathway is not damage-inducible and not dependent on a lesion-bypass DNA polymerase, while the second pathway is damage-inducible and dependent on a lesion-bypass DNA polymerase . Furthermore, DNA sequence context appears to dictate which pathway (as defined by the involvement of different DNA polymerases) is followed in each case .

Am J Infect Control, 2004 Jun, 32(4), 185 - 8
Antibiotic susceptibility of glutaraldehyde-tolerant Mycobacterium chelonae from bronchoscope washing machines; Nomura K et al.; BACKGROUND: Contamination of bronchoalveolar lavage fluid is a major problem in the world . Although 2% glutaraldehyde (GA) is widely used as a disinfectant for bronchoscope cleaning, recently, GA-tolerant mycobacteria have been isolated, which makes this problem more complicated . METHODS: We studied the susceptibility to GA and antibiotics of mycobacteria isolated from bronchoscope washing machines in our hospital . We also studied the minimum inhibitory concentrations of GA and antibiotics with pump inhibitors . RESULTS: Twenty-nine mycobacteria were isolated, of which 26 were Mycobacterium chelonae . Among 18 isolates of M chelonae, excluding 8 isolates in which some results were not reproducible, 50% (9 of 18) were 2% GA-tolerant . One hundred percent (9 of 9) of the GA-tolerant isolates and 11% (1 of 9) of the GA-sensitive isolates were either resistant or intermediately resistant to 2 or 3 classes of antibiotics . Efflux pump inhibitors did not influence the susceptibility to GA and antibiotics . CONCLUSIONS: It was suggested that there might be an association of GA tolerance with antibiotic resistance in M chelonae . There may a different mechanism(s) other than that involving efflux pumps with regard to GA tolerance and antibiotic resistance in M chelonae . When bronchoscopy-related mycobacterial infections are suspected, physicians and clinical microbiologists should exercise care in handling GA-tolerant mycobacteria, which may be resistant to multiple antibiotics.

Curr Allergy Asthma Rep, 2004 Jul, 4(4), 270 - 5
Dietary modification of atopic disease: Use of probiotics in the prevention of atopic dermatitis; Isolauri E; The increased prevalence of atopic diseases, atopic dermatitis, allergic rhinitis, and asthma has been described as an epidemic . New approaches in the fight against allergic diseases are called for, the target being the persistence of the atopic T helper 2-skewed immune responder pattern beyond infancy . Atopic dermatitis, the earliest of these conditions, might act as a portal for the development of IgE-mediated atopic manifestations . Abundant evidence implies that specific strains selected from the healthy gut microbiota exhibit powerful antipathogenic and anti-inflammatory capabilities, and several targets for the probiotic approach have emerged in atopic dermatitis: degradation/structural modification of enteral antigens, normalization of the properties of aberrant indigenous microbiota and of gut barrier functions, regulation of the secretion of inflammatory mediators, and promotion of the development of the immune system . Better understanding of the effects of different probiotic strains and deeper insight into the mechanisms of the heterogeneous manifestations of atopic disease are needed for the validation of specific strains carrying anti-allergic potential.

Ter Arkh, 2004, 76(4), 25 - 30
{In Process Citation}; Kingella kingae: from medical rarity to an emerging paediatric pathogen; Clinical Microbiology Laboratories, Soroka University Medical Center, Ben-Gurion University of the Negev, Beer-Sheva, Israel . yagupsky@bgumail.bgu.ac.il

In recent years, Kingella kingae has emerged as an important cause of invasive infections in young children, especially septic arthritis, osteomyelitis, spondylodiscitis, bacteraemia, and endocarditis, and less frequently lower respiratory tract infections and meningitis . The organism is part of the pharyngeal flora of young children and is transmitted from child-to-child . The clinical presentation of invasive K kingae disease is often subtle and laboratory tests are frequently normal . A substantial fraction of children with invasive K kingae infections have a recent history of stomatitis or symptoms of upper-respiratory-tract infection . The organism is susceptible to a wide array of antibiotics that are usually given empirically to young children including beta lactams, and with the exception of cases of endocarditis, the disease runs a benign clinical course . Although isolation and recognition of the organism is not difficult, clinicians and microbiologists should be aware of its fastidious nature . To optimise the recovery of K kingae, inoculation of synovial fluid specimens into blood culture vials is strongly recommended.

Sex Transm Infect, 2004 Jun, 80(3), 207 - 11
Diagnosis of genital chlamydia in primary care: an explanation of reasons for variation in chlamydia testing; McNulty CA et al.; OBJECTIVES: To explore the reasons for the 40-fold variation in diagnostic testing for genital Chlamydia trachomatis by general practices . METHODS: A qualitative study with focus groups . We randomly selected urban and rural high and low testing practices served by Bristol, Hereford, and Gloucester microbiology laboratories . Open questions were asked about the investigation of C trachomatis in men and women in different clinical contexts . RESULTS: The high and low testing practices did not differ in their age/sex make-up or by deprivation indices . There were major differences between high and low chlamydia testing practices . Low testing practices knew very little about the epidemiology and presentation of genital chlamydia infection and did not consider it in their differential diagnosis of genitourinary symptoms until patients had consulted several times . Low testers were less aware that chlamydia was usually asymptomatic, thought it was an inner city problem, and had poor knowledge of how to take diagnostic specimens . High testing practices either had a general practitioner with an interest in sexual health or a practice nurse who had completed specialist training in family planning . High testing practices were more cognizant of the symptoms and signs of chlamydia and always considered it in their differential diagnosis of genitourinary symptoms, including patients attending family planning clinics . CONCLUSIONS: Any programme to increase chlamydia testing in primary care must be accompanied by an education and awareness programme especially targeted at low testing practices . This will need to include information about the benefits of testing and who, when, and how to test.

Sex Transm Infect, 2004 Jun, 80(3), 204 - 6
How is the high vaginal swab used to investigate vaginal discharge in primary care and how do GPs' expectations of the test match the tests performed by their microbiology services?
Noble H, Estcourt C, Ison C, Goold P, Tite L, Carter YH.
OBJECTIVES: To describe the management of vaginal discharge in general practice, with particular regard to the use of the high vaginal swab (HVS), and to compare GPs' expectations of this test with the processing and reporting undertaken by different laboratories . METHODS: A postal questionnaire survey of 2146 GPs in the North Thames area and postal questionnaire study of the 22 laboratories serving the same GPs were carried out . GPs were asked how they would manage a young woman with vaginal discharge and what information they would like on an HVS report . Laboratories were asked how they would process and report on the HVS sample from the same patient . RESULTS: Response rate was 26% . 72% of GPs would take an HVS and 62% would refer on to a genitourinary medicine (GUM) clinic . 45% would offer empirical therapy and 47% of these would treat for candida initially . 75% of GPs routinely request "M,C&S" on HVS samples but 55% only want to be informed about specific pathogens . Routine processing of HVS samples varies widely between laboratories and 86% only report specific pathogens . 78% of GPs would like to be offered a suggested diagnosis on HVS reports, and 74% would like a suggested treatment . 43% of laboratories ever provide a diagnosis, and 14% provide a suggested treatment . CONCLUSIONS: GPs frequently manage vaginal discharge and most of them utilise the HVS . GPs' expectations of the test are not well matched to laboratory processing or reporting of the samples.

Rinsho Byori, 2004 Apr, 52(4), 307 - 11
Postgraduate education in laboratory medicine and certification/re-certification of clinical pathologists in Taiwan; Sun CF; The Taiwan Society of Clinical Pathologists (TSCP) plays a central role in postgraduate education of laboratory medicine and the certification/re-certification of clinical pathologists in Taiwan . For the certification of clinical pathologists, TSCP establishes "Guidelines and Scope of Resident Training" and "Standards for Training Hospitals in Clinical Pathology(CP)", administers board examinations, and issues board certifications/re-certifications . There are two types of CP resident training programs, including a straight CP program with 3 years of CP training for a CP certificate and a combined program with 3 years of Anatomic Pathology training and 2 years of CP training for both the CP and AP certificates . The core curriculum for CP training includes: (1) Clinical Chemistry (at least 4 months), (2) Clinical Microscope with Parasitology (at least 3 months), (3) Clinical Hematology (at least 4 months), and (4) Clinical Microbiology with Clinical Virology (at least 4 months), (5) Immunohematology and Blood Banking (Transfusion Medicine) (at least 3 months), (6) Clinical Serology and Immunology(at least 4 months), and (7) Laboratory Management (at least 2 months) . The curriculum for third-year training is not specified and may be in any field . In recent years, the board examination has emphasized the topics of Molecular Biology and Laboratory Informatics . The TSCP has also established an accreditation and inspection program for the CP resident raining hospitals . Each accredited CP training hospital is required to have a detailed teaching protocol of CP training . Quotas are assigned according to the available CPs of the accredited hospitals . The accreditation period is 3 years . Through sponsoring scientific and educational programs, the TSCP offers credit hours of education in laboratory medicine, which are required for re-certification of CPs in Taiwan . The members of the TSCP meet at least twice a year for scientific presentations and seminars . In addition, two to four symposia, offering 8 credit hours each, are held each year in various subspecialties of CP . In 2003, 22 hospitals were accredited as CP training hospitals for a total quota of 26 . Until 2003, the TSCP had certified 116 CPs . At the present time, only 103 certified CPs are actively practicing laboratory medicine . Re-certification requires 100 credit hours of continuing education . The requirements for board certification and re-certification are the two main driving forces for CPs in Taiwan to seek continuing education . Our model of education for CPs has proven to be effective . The number of practicing CPs increased from 21 (one per 3,083 beds) in 1991 to 103 (one per 929 beds) in 2002 . Most of the CPs are associated with medical centers(62/103, 60.2%) and regional hospitals(38/103, 36.9%).

J Clin Virol, 2004 Aug, 30(4), 313 - 9
Inter-laboratory comparison of qualitative and quantitative detection of hepatitis C (HCV) virus RNA in diagnostic virology: a multicentre study (MS) in Italy; Mancini C et al.; BACKGROUND: The importance of the standardisation of nucleic acid amplification technology (NAT) assays for the detection of hepatitis C virus RNA is well known today, as many studies carried out in different European countries attest . The results of a previous study performed in Italy (J . Clin . Virol . 1 (2003) 83) by the Italian Society of Clinical Microbiology (AMCLI) showed that the use of external reference standards and of multicentre collaborative studies significantly improves laboratory performance for the qualitative evaluation of HCV RNA . OBJECTIVES: the AMCLI organised a new study on the standardisation of both the qualitative and the quantitative evaluation of HCV RNA with NAT in order to improve the implementation of the diagnostic methods for HCV RNA detection . STUDY DESIGN: seventeen diagnostic centres of major Italian Hospitals participated in this quality control study . The study consisted of testing three panels, each made up of 10 coded samples including negative and positive samples . Positive samples contained four levels of HCV RNA (genotype 1) . RESULTS AND CONCLUSION: Seven out of 510 qualitative results obtained were incorrect (1.4%), two false negative and five false positive . The results gave a sensitivity of 99.5% and a specificity of 95.8% . Regarding quantitative tests, the geometric mean (GM) and standard deviation (S.D.) could be calculated only for the three highest HCV RNA levels . The percentage of results within the range of GM +/- 0.5 log(10) varied from 91% to 100% . Some laboratories had some difficulty in the exact quantification of the lowest (3.00 log IU/ml) as well as of the highest viral levels (6.35 log IU/ml) values, very near to the limits of the dynamic range of the assays . The comparison of the results of this study with that previously carried out one confirms that a regular participation in external quality assessment (EQA) assures the achievement of a high proficiency level in the diagnosis of HCV infection.

J Basic Microbiol, 2004, 44(3), 197 - 202
Evaluation of three new culture media for the cultivation and isolation of Leishmania parasites; Limoncu ME et al.; The aim of this study was to establish novel culture media for Leishmania parasites with a potential of obtaining high amounts of promastigotes with long-term viability, and consisting of ingredients that were available in microbiology or parasitology laboratories . Other features of these media included no requirement for blood, FCS (Fetal calf serum) or erythrocyte lysate, inexpensiveness and easiness in preparation . In addition, aspiration samples obtained from cutaneous leishmaniasis (CL) suspected patients were cultivated in these media . Three culture media were prepared; trypticase beef extract hemoglobine (TBH) medium, including trypticase, beef extract and yeast extract as the protein source, glucose as the carbohydrate source, FeNH4 and bovine hemoglobine; Peptone-Yeast extract medium (PY), found to be effective in our previous studies for cultuvation of on Leishmania parasites, with bovine hemoglobine (PYH) and Brain Heart medium, containing bovine hemoglobine (BKH) . The number of promastigotes were the highest on day 8 and 13 in RPMI 1640 and BKH medium, respectively . In TBH and PYH, the peak level of reproduction was between day 16 and 19, and it was found to be higher in TBH medium after the day 20 . The number of promastigotes were found to be close in BKH, TBH and RPMI-1640 media and lower in PYH medium . Examination of the cultivation of the aqueous lesion specimens of the 10 CL-suspected cases in media revealed reproduction in 9 flasks of RPMI-1640 containing 10% FCS, 7 TBH, 6 BKH and 4 PYH . The differences between the culture media were not found to be statistically significant . These results suggested that, three liquid culture media, assessed in this study, with no requirement for FCS or erythrocyte lysate, were effective in the reproduction of promastigotes, and could be used effectively in the patient isolation and field studies, as well .

Clin Infect Dis, 2004 May 15, 38(10), 1428 - 39 Epub 2004 Apr 30.
Nontuberculous mycobacterial infection in hematopoietic stem cell and solid organ transplant recipients; Doucette K et al.; Nontuberculous mycobacteria (NTM) are ubiquitous environmental organisms . In immunocompetent hosts, they are a rare cause of disease . In immunocompromised hosts, disease due to NTM is well documented . Reports of NTM disease have increased in hematopoietic stem cell transplant (HSCT) and solid organ transplant (SOT) recipients . This increase may reflect increased numbers of transplants, intensification of immune suppressive regimens, prolonged survival of transplant recipients, and/or improved diagnostic techniques . The difficulty of diagnosis and the impact associated with infections due to NTM in HSCT and SOT recipients necessitates that, to ensure prompt diagnosis and early initiation of therapy, a high level of suspicion for NTM disease be maintained . The most common manifestations of NTM infection in SOT recipients include cutaneous and pleuropulmonary disease, and, in HSCT recipients, catheter-related infection . Skin and pulmonary lesions should be biopsied for histologic examination, special staining, and microbiologic cultures, including cultures for bacteria, Nocardia species, fungi, and mycobacteria . Mycobacterial infections associated with catheters may be documented by tunnel or blood (isolator) cultures . Susceptibility testing of mycobacterial isolates is an essential component of optimal care . The frequent isolation of NTM other than Mycobacterium avium complex (MAC) from transplant recipients limits the extrapolation of therapeutic data from human immunodeficiency virus-infected individuals to the population of transplant recipients . Issues involved in the management of NTM disease in transplant recipients are characterized by a case of disseminated infection due to Mycobacterium avium complex in a lung transplant recipient, with a review of the relevant literature.

J Immunol, 2004 Jun 1, 172(11), 6894 - 901
Production of IFN-gamma by CD4 T cells is essential for resolving ehrlichia infection; Bitsaktsis C et al.; To address the role of cellular immunity during ehrlichia infection, we have used a newly described model of monocytic ehrlichiosis that results from infection of mice by an ehrlichia that was isolated from an Ixodes ovatus tick (Ixodes ovatus ehrlichia, IOE) . Immunocompetent C57BL/6 and BALB/c mice exhibited a dose-dependent susceptibility to IOE infection . Mice infected with a high dose inoculum ( approximately 1000 organisms) exhibited pronounced thrombocytopenia, lymphopenia, anemia, and morbidity within 12 days postinfection . Infection was associated with bacterial colonization of a number of tissues . In contrast, mice infected with a low dose inoculum ( approximately 100 organisms) exhibited only transient disease and were able to resolve the infection . SCID mice were highly susceptible to low-dose infection, indicating that adaptive immunity was required . Resistance to sublethal challenge in both C57BL/6 and BALB/c mice was CD4-, but not CD8-, dependent and required IL-12p40-dependent cytokines, IFN-gamma, and TNF-alpha, but not IL-4 . CD4 T cells purified from infected mice proliferated in vitro in response to IOE Ags . T cell proliferation was associated with production of IFN-gamma, and the production of this cytokine by CD4 T cells rescued IFN-gamma-deficient mice from fatal infection . Exogenous IFN-gamma was capable of inducing microbiocidal activity in infected macrophages . The data suggest that classical immune mechanisms involving CD4 cells and type 1 cytokines are responsible for macrophage activation and for elimination of this intracellular bacterial pathogen.

Methods Mol Biol, 2004, 266, 213 - 27
Microarrays for bacterial typing: realistic hope or holy grail?
Van Ijperen C, Saunders NA.
Microbiology has entered the postgenomic era and it is clear that bacterial typing should aim to be based on analysis of complete genomes . Although complete genome sequencing for epidemiological typing remains unrealistic for the present, microarrays that provide information on gene content are now becoming available . Microarrays comprised of several thousand probes on glass slides can now be manufactured in the laboratory using robotic arrayers . The gene probes are either PCR products or synthetic oligonucleotides that can be irreversibly attached to a reactive glass surface . The target nucleic acids to be hybridized to the probe array are tagged with fluorescent dyes . Relative probe hybridization signals can be measured when two or more different preparations are labeled with distinguishable fluorophores . Microarrays that include probes for every gene within a genome provide excellent comparative data, although a focus on variable genes may be more useful for typing purposes . Composite arrays of variable genes are under development.

Methods Mol Biol, 2004, 266, 167 - 89
Molecular diagnostics: future probe-based strategies; Marsh P et al.; Nucleic acid amplification technologies (NAATs) represent powerful tools in clinical microbiology, particularly in areas where traditional culture-based methods alone prove insufficient . A notable advantage is in reducing the time from taking samples to reporting results . This, and the specificity and sensitivity imparted by NAATs, can help to improve patient care . Both thermal and isothermal NAATs have been adapted to aid diagnosis in clinical laboratories . Current molecular diagnostic assays are generally high-tech, and are expensive to buy and perform . Easy-to-use NAATs are beginning to appear, not only facilitating acceptable throughput in clinical laboratories, but also allowing tests to move out of the laboratory, closer to the point of care . Demand for simpler, miniaturized equipment and assays, and the trend toward personalized medicine, is leading towards the development of fully integrated automation and home-use kits . The integration of diverse disciplines, such as genomics, molecular biology, microelectromechanical systems, microfluidics, microfabrication, and organic chemistry, is behind the emerging DNA microarray technology . Development of DNA microchips allows the simultaneous detection of potentially thousands of target sequences, not only favoring high throughput, but also the potential for genotyping patient subsets with respect to their response to particular drug types (pharmakogenomics) . It is envisaged that the future of probe-based technologies will see the development of fully integrated assays and devices suitable for nonskilled users.

Klin Mikrobiol Infekc Lek, 2004 Apr, 10(2), 80 - 7
{Comparison of the results of in vitro antifungal susceptibility testing using the Fungitest kit and the disc diffusion test}; Buchta V et al.; AIM OF STUDY: Substantial progress has been achieved in the field of in vitro antifungal susceptibility testing, including the interpretation of the test results towards the clinical practice . Reference methods for antifungal testing of yeasts (M27-A) and filamentous fungi (M38-P) are available now . There are several alternatives to these standard protocols as the potential candidates for a routine laboratory testing of antifungal drugs . The aim of the study is to compare the results of antifungal susceptibility testing obtained by Fungitest and disk diffusion method . MATERIALS AND METHODS: From 1997 to 2000, 134 strains of 16 yeast species from the patients with suspected or proven mycosis were evaluated in the mycology laboratory of the Department of Clinical Microbiology, Teaching Hospital in Hradec Krlove . All these strains were tested by both disc test on Sabouraud agar (pH = 5.6) and Fungitest . The agreement of the results of the both methods was determined for all antimycotics and fungal species tested in terms of the categories susceptible, intermediate and resistant strain . RESULTS: Overall, there was only 60.3 % agreement between these methods for all antifungal drugs with the best results for flucytosine (71.4 % agreement), further for ketoconazole (66.2 %), amphotericin B (60.2 %), fluconazole (57.9 %), itraconazole (52.6 %), and miconazole (50.5 %) . The best overall agreement between the results for individual species was found with C . lusitaniae (73.9 %), C . pelliculosa (73.5 %), C . parapsilosis (71.7 %), and partly with C . albicans (67.5 %) . The poorest results were associated with fluconazole-resistant yeasts Saccharomyces cerevisiae (30.0 %), C . krusei (32.5 %) and C . glabrata (47.7 %) . Regressive analysis revealed a relative close relationship between the results of these methods for miconazole (R = 0.49, P < 0.001), fluconazole (R = 0.45, P < 0.001), itraconazole (R = 0.42, P < 0.001) and flucytosine (R= 0.37, P < 0.001), but not for amphotericin B (R = 0.10) and ketoconazole (R = 0.16) . CONCLUSION: The comparison of the results of the disk test and Fungitest showed some discrepancies which were dependent on an antifungal drug, a yeast species and the test conditions . Sabouraud agar and the criteria for interpretation of the results of the disk test seemed to be main sources of the disagreement . A modified Mueller-Hinton agar (2 % glucose, methylene blue) as a recommended test medium in a proposed agar diffusion method M44-P is the way to the improvement of standardization and reproducibility of the test.

Clin Rev Allergy Immunol, 2004 Apr, 26(2), 75 - 83
The role of parasites in genetic susceptibility to allergy: IgE, helminthic infection and allergy, and the evolution of the human immune system; Hagel I et al.; There have been numerous studies in the mouse illustrating the dichotomy of T cell responses, with the common classification orchestrated around Th1 vs . Th2 responses . This classification is now widely applied to human disease as well and the generic conclusion is that the Th1 responses are more likely to occur secondary to specific microbiologic insult but also inflammatory responses . In contrast, the Th2 response is the prevalent response in subjects with atopy and allergic disease but is also the mechanism for protection against helminthic infections . Unfortunately, the paradigm of Th1 vs . Th2 is not as clear in the human as it is in mouse models . Even so, the immunological mechanisms responsible for IgE production that are protective in helminthic infections, i.e . Schistosoma, are similar to those for the production of specific IgE against allergens . In fact, there also appear to be associations in the memory T cell subpopulation CD4+CD45RO+ and the elicitation of IgE against both parasites and allergens . In this review, we present the overall contemporary scheme on the role of parasites in genetic susceptibility to allergic IgE, helminthic infections with specific discussion of its implications for the evolution of the human immune system.

Int J Food Microbiol, 2004 May 1, 92(3), 249 - 54
Rare occurrence of heterotrophic bacteria with pathogenic potential in potable water; Stelma GN Jr et al.; Since the discovery of Legionella pneumophila, an opportunistic pathogen that is indigenous to water, microbiologists have speculated that there may be other opportunistic pathogens among the numerous heterotrophic bacteria found in potable water . The US Environmental Protection Agency (USEPA) developed a series of rapid in vitro assays to assess the virulence potential of large numbers of bacteria from potable water to possibly identify currently unknown pathogens . Results of surveys of potable water from several distribution systems using these tests showed that only 50 of the approximately 10,000 bacterial colonies expressed one or more virulence characteristics . In another study, 45 potable water isolates that expressed multiple virulence factors were tested for pathogenicity in immunocompromised mice . None of the isolates infected mice that were compromised either by treatment with carrageenan (CG), to induce susceptibility to facultative intracellular pathogens, or by cyclophosphamide (CY), to induce susceptibility to extracellular pathogens . These results indicate that there are very few potential pathogens in potable water and that the currently developed in vitro virulence screening tests give an overestimation of the numbers of heterotrophic bacteria that may be pathogens . Current efforts are focused on using the animal models to screen concentrated samples of waters known to contain large numbers of heterotrophic bacteria and newly discovered Legionella-like organisms that parasitize amoebae .

Int J Food Microbiol, 2004 May 1, 92(3), 241 - 7
Heterotrophic plate count measurement in drinking water safety management: report of an Expert Meeting Geneva, 24-25 April 2002; Bartram J et al.; A group of microbiology and public health experts including regulatory and medical expertise was convened in Geneva, Switzerland, 25-26 April 2002 to consider the utility of heterotrophic plate count (HPC) measurements in addressing drinking water quality and safety . The group was convened following the NSF International/World Health Organization Symposium on HPC Bacteria in Drinking Water--Public Health Implications? The Expert Meeting was attended by 31 participants from Australia, Canada, France, Germany, Italy, Japan, the Netherlands, South Africa, Switzerland, UK and USA .

J Hosp Infect, 2004 May, 57(1), 1 - 7
Stenotrophomonas maltophilia: the significance and role as a nosocomial pathogen; Senol E; Stenotrophomonas maltophilia is a significant pathogen that primarily affects patients with co-morbid illnesses, usually malignancies, who have been hospitalized for prolonged periods and received broad-spectrum antibiotic therapy . The organism has been associated with an expanding spectrum of clinical manifestations associated with high case to fatality ratios . Therapy of these infections presents a significant challenge both for the clinician and the microbiologists because of its high-level antibiotic resistance to most of the currently used agents and methodological difficulties in susceptibility testing with this organism.

Bioorg Med Chem, 2004 Jun 1, 12(11), 2937 - 50
Modeling and interactions of Aspergillus fumigatus lanosterol 14-alpha demethylase 'A' with azole antifungals; Gollapudy R et al.; Recent identification of the sterol 14-alpha demethylase genes (CYP51 A and B) from Aspergillus fumigatus and other species by Mellado et al . (J . Clin . Microbiol . 2001, 39(7), 2431-2438), has opened up possibilities of investigating the interactions of azole antifungals with the enzyme(s) from fungi . This study describes for the first time, a model of the three-dimensional structure of A . fumigatus 14-alpha demethylase (AF-CYP51A), using the crystal structure of Mycobacterium tuberculosis 14-alpha demethylase (PDB code:1EA1) as a template . The paper also describes the various interactions between azole antifungals and the target from A . fumigatus (AF-CYP51A) . Quantitative evaluation of these interactions is done using COMBINE analysis to understand contributions of active site residues to ligand activity . It also provides explanation for the activity/inactivity of different ligands for AF-CYP51A.

Med Clin (Barc), 2004 May 1, 122(16), 610 - 6
{Meta-analysis on safety and efficacy of liposomal amphotericin B in the empirical treatment of febrile neutropenia}; Badia X et al.; BACKGROUND AND OBJECTIVE: We compared the safety and efficacy of liposomal amphotericin B with other formulations of amphotericin B and voriconazol as an empirical treatment of febrile neutropenia . MATERIAL AND METHOD: Several randomized controlled clinical trials, designed to evaluate the safety and/or efficacy of liposomal amphotericin B in the empirical treatment of febrile neutropenia, in comparison with other amphotericin B formulations or voriconazol, were identified by means of a search in MEDLINE, EMBASE and Cochrane Controlled Trials Register's data bases . American Society of Microbiology and American Society of Clinical Oncology abstracts, presented between 1999 and 2002, were also included in the search . Prior to data extraction, concepts like nephrotoxicity, infusion-related adverse events and efficacy (success rate and mortality) were defined . RESULTS: Two studies comparing liposomal amphotericin B with conventional amphotericin B, one comparing liposomal amphotericin B with amphotericin B lipid complex and one comparing liposomal amphotericin B with voriconazol were included in the analysis . Patients treated with liposomal amphotericin B had lower nephrotoxicity than patients treated with other amphotericin B formulations or with conventional amphotericin B (RR = 0.49); conversely, no statistically significant differences with regard to voriconazol were observed . In terms of efficacy, mortality rate and therapeutic failure, patients treated with limposomal amphotericin B showed a slightly higher efficacy . CONCLUSIONS: Liposomal amphotericin B is a lipidic formulation with a slightly higher efficacy than other amphotericin B formulations and voriconazol . In terms of nephrotoxicity, liposomal amphotericin B showed lower nephrotoxicity than other amphotericines while its safety rates were similar to those of voriconazol.

Orthopade, 2004 Apr, 33(4), 439 - 54
{Principles of surgical therapy concepts for postoperative and chronic osteomyelitis}; Kutscha-Lissberg F et al.; Infection of the bone is one of the most serious complications in the field of orthopedic and trauma orthopedic surgery . Sufficient treatment protocols not only contain complex surgical procedures but also sophisticated diagnostic tools, proper use of antibiotics, and intensive physical therapy right from the beginning . Even in light of these advanced treatment protocols, which have great impact on both patients and health care systems, persisting infection and residual functional deficits of the extremities are not rare . In cases of early (acute) infection, the main objective is to avoid chronification by diligent surgical interventions . The surgical principle is the meticulous debridement and lavage of the situs . Revision of only the epifascial layers is as inadequate as the simple reopening of the wound without excision of the whole wound including all tissue layers . In cases of chronic soft tissue and bone infection, radical debridement of all infected and scar tissue is also the basic requirement of treatment . Reconstruction of the soft tissue envelope is done by local or free flap surgery . Because of they are better resistant to infection, musculo(cutaneous)flaps are preferred . Bony reconstruction is done by autologous cancellous bone grafting (partial defects), segment transport (full thickness defects), or freely transplanted vascularized bone grafts (large partial defects) . Both soft tissue and osseous reconstruction take a relatively long period of time requiring several operations and periods of hospitalization . These have to be discussed and explained to the patients extensively . If the required amount of resection and the capability of reconstruction do not coincide, the surgeon and the patient have to decide whether restoration of function without definitive infection care, symptomatic infection therapy, or amputation is the most proper treatment option according to the patient's everyday needs and lifestyle . Because each treatment protocol is a composition of orthopedic trauma surgeons, plastic surgeons, radiologists, microbiologists, and physical therapists, reliable cooperation and communication is essential.

Ceska Gynekol, 2004 Mar, 69(2), 133 - 40
{Antifungal immunity and its mechanism in female genitalia}; Spacek J et al.; OBJECTIVE: A review of immunological aspects of fungal infections with focus on vulvovaginal candidiasis . DESIGN: Review . SETTING: Department of Obstetrics and Gynaecology, Medical Faculty, Hradec Kralove, Charles University in Prague, Department of Biological and Medical sciences, Faculty of Pharmacy, Hradec Kralove, Charles University in Prague, Department of Clinical Immunology and Allergology, University Hospital, Hradec Kralove, Department of Clinical Microbiology, University Hospital, Hradec Kralove . SUBJECT AND METHOD: Survey and discussion of current immunological aspects of vulvovaginal candidias based on literature review and personal experience . CONCLUSION: The current knowledge of immunity to Candida vulvovaginitis, particularly from experimental studies on animal models, support a more relevant role of local protection mechanisms than systemic immunity . As a consequence of this compartmentalization of the immune response, the role of innate and adaptive immunity remains unclear . The development of vulvovaginal candidiasis depends on a number of factors, including both the individual predisposition of a macroorganism and the properties of a fungal strain, and the environmental conditions influencing a delicate balance between the host and the yeast.

Oecologia, 2004 Oct, 141(2), 269 - 81 Epub 2004 May 08.
A multi-scale perspective of water pulses in dryland ecosystems: climatology and ecohydrology of the western USA; Loik ME et al.; In dryland ecosystems, the timing and magnitude of precipitation pulses drive many key ecological processes, notably soil water availability for plants and soil microbiota . Plant available water has frequently been viewed simply as incoming precipitation, yet processes at larger scales drive precipitation pulses, and the subsequent transformation of precipitation pulses to plant available water are complex . We provide an overview of the factors that influence the spatial and temporal availability of water to plants and soil biota using examples from western USA drylands . Large spatial- and temporal-scale drivers of regional precipitation patterns include the position of the jet streams and frontal boundaries, the North American Monsoon, El Nino Southern Oscillation events, and the Pacific Decadal Oscillation . Topography and orography modify the patterns set up by the larger-scale drivers, resulting in regional patterns (10(2)-10(6) km2) of precipitation magnitude, timing, and variation . Together, the large-scale and regional drivers impose important pulsed patterns on long-term precipitation trends at landscape scales, in which most site precipitation is received as small events (< 5 mm) and with most of the intervals between events being short (< 10 days) . The drivers also influence the translation of precipitation events into available water via linkages between soil water content and components of the water budget, including interception, infiltration and runoff, soil evaporation, plant water use and hydraulic redistribution, and seepage below the rooting zone . Soil water content varies not only vertically with depth but also horizontally beneath versus between plants and/or soil crusts in ways that are ecologically important to different plant and crust types . We highlight the importance of considering larger-scale drivers, and their effects on regional patterns; small, frequent precipitation events; and spatio-temporal heterogeneity in soil water content in translating from climatology to precipitation pulses to the dryland ecohydrology of water availability for plants and soil biota.

Fed Regist, 2004 May 11, 69(91), 26036 - 8
Medical devices; immunology and microbiology devices; classification of the immunomagnetic circulating cancer cell selection and enumeration system . Final rule; Food and Drug Administration et al.; The Food and Drug Administration (FDA) is classifying the Immunomagnetic Circulating Cancer Cell Selection and Enumeration System device into class II (special controls) . The special control that will apply to the device is the guidance document entitled "Class II Special Controls Guidance Document: Immunomagnetic Circulating Cancer Cell Selection and Enumeration System." 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 (the 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 the 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 publishing a notice of availability of a guidance document that is the special control for this device.

J Heart Lung Transplant, 2004 May, 23(5), 570 - 6
Clinical usefulness of bronchoalveolar lavage in heart transplant recipients with suspected lower respiratory tract infection; Lehto JT et al.; BACKGROUND: Bronchoscopy with bronchoalveolar lavage (BAL) is the recommended initial invasive diagnostic procedure when lower respiratory tract infection is suspected in solid-organ transplant recipients . In this study, we evaluated the clinical impact and safety of bronchoscopy with BAL in heart transplant recipients . METHODS: We reviewed all 44 consecutive diagnostic bronchoscopies with BAL that were performed in 35 heart transplant recipients at Helsinki University Central Hospital between May 1988 and December 2001 . RESULTS: Bronchoscopy findings established specific microbiologic diagnoses in 18 of 44 (41%) cases, and 14 of 44 (32%) bronchoscopic findings led to changes in therapy . The diagnostic yield of bronchoscopy from 1 to 6 months after transplantation was 73%, significantly better (p = 0.002) than diagnostic yield during the first month (18%) and after 6 months (28%) . Pneumocystis carinii and cytomegalovirus were the most frequently detected pathogens in the BAL fluid . Cytomegalovirus pneumonia carried a high mortality rate (44%), whereas all patients with P carinii pneumonia recovered . Fourteen episodes were diagnosed as bacterial pneumonia, but because of empiric antibiotic therapy that was started widely before bronchoscopy, a microbiologic diagnosis was established in only 1 case . However, all patients with community-acquired pneumonia responded to empiric therapy . Four cases of major complications occurred after bronchoscopy, all cardiovascular but none fatal . CONCLUSIONS: Bronchoscopy with BAL is a useful diagnostic tool in heart transplant recipients, especially between 1 and 6 months after transplantation.

Appl Environ Microbiol, 2004 May, 70(5), 3055 - 63
Comparative analysis of the conventional and novel pmo (particulate methane monooxygenase) operons from methylocystis strain SC2; Ricke P et al.; In addition to the conventional pmoA gene (pmoA1) encoding the active site polypeptide of particulate methane monooxygenase, a novel pmoA gene copy (pmoA2) is widely distributed among type II methanotrophs (methane-oxidizing bacteria {MOB}) (M . Tchawa Yimga, P . F . Dunfield, P . Ricke, J . Heyer, and W . Liesack, Appl . Environ . Microbiol . 69:5593-5602, 2003) . Here we report that the pmoA1 and pmoA2 gene copies in the type II MOB Methylocystis strain SC2 are each part of a complete pmoCAB gene cluster (pmoCAB1, pmoCAB2) . A bacterial artificial chromosome (BAC) library of strain SC2 genomic DNA was constructed, and BAC clones carrying either pmoCAB1 or pmoCAB2 were identified . Comparative sequence analysis showed that these two gene clusters exhibit low levels of identity at both the DNA level (67.4 to 70.9%) and the derived protein level (59.3 to 65.6%) . In contrast, the secondary structures predicted for PmoCAB1 and PmoCAB2, as well as the derived transmembrane-spanning regions, are nearly identical . This suggests that PmoCAB2 is, like PmoCAB1, a highly hydrophobic, membrane-associated protein . A total of 190 of the 203 amino acid residues representing a highly conserved consensus sequence of the currently known PmoCAB1 and AmoCAB sequence types could be identified in PmoCAB2 . The amoCAB gene cluster encodes ammonia monooxygenase and is evolutionarily related to pmoCAB . Analysis of a set of amino acid residues that allowed differentiation between conventional PmoA and AmoA provided further support for the hypothesis that pmoCAB2 encodes a functional equivalent of PmoCAB1 . In experiments in which we used 5' rapid amplification of cDNA ends we identified transcriptional start sites 320 and 177 bp upstream of pmoC1 and pmoC2, respectively . Immediately upstream of the transcriptional start sites of both pmoCAB1 and pmoCAB2, sequence motifs similar to Escherichia coli sigma(70) promoters were identified.

J Med Assoc Thai, 2004 Mar, 87(3), 246 - 54
Comparison of topical lomefloxacin 0.3 per cent versus topical ciprofloxacin 0.3 per cent for the treatment of presumed bacterial corneal ulcers; Booranapong W et al.; PURPOSE: To compare the efficacy and safety of topical lomefloxacin 0.3 per cent with topical ciprofloxacin 0.3 per cent for treating mildly severe suspected bacterial corneal ulcers . METHOD: This prospective, randomized, double-masked controlled clinical trial was conducted on 41 patients (41 eyes) with suspected bacterial corneal ulcers who were randomized into 2 groups: 23 patients were in the lomefloxacin group and 18 patients in the ciprofloxacin group . All of these corneal ulcers were scraped for gram's stain, KOH preparation and microbiologic cultures before starting treatment . The clinical success rate, the time to cure, the rates of treatment failures, ocular signs and symptoms and the adverse effects of the study medication were evaluated . RESULTS: Topical lomefloxacin is equivalent clinically and statistically to topical ciprofloxacin . No statistically significant treatment differences were found between lomefloxacin (100%) and ciprofloxacin (100%) in terms of success rate . Similarly, no differences were noted in the time to cure (p > 0.05), the treatment failure, or the resolution of the clinical signs and symptoms (p > 0.05) . The adverse effects of lomefloxacin were superficial punctate keratitis (26.1%) and irritation (8.7%), whereas those of ciprofloxacin were superficial punctate keratitis (22.2%), white precipitate (11.1%) and irritation (11.1%) . However, no statistically significant differences of these adverse effects were found between the two groups (p > 0.05) . CONCLUSION: Lomefloxacin ophthalmic solution (0.3%) is equivalent clinically and statistically to ciprofloxacin ophthalmic solution (0.3%) for the treatment of mildly severe presumed bacterial corneal ulcers without statistically significant differences in the adverse effects and discomfort.

Zentralbl Gynakol, 2004 Apr, 126(2), 73 - 6
{Mastitis puerperalis - causes and therapy}; Peters J; Between 1995 and 2003 a total of 1 827 women suffering from puerperal mastitis was studied . Etiology of the disease was defined, white cell counts and microbiologic cultures were obtained . According to the results the illness was classified and treated . Since in all cases we found either simple congestion or non-infectious inflammation of the breast treatment consisted of intensified emptying of the breast by breast feeding every 2 hours and in some cases additional manual expression of milk . Antibiotics and bromocriptine were not used . In 1 826 cases fever and flue symptoms had resolved within 25-36 hours . Redness and soreness had resolved within 2-3 days . Relapse did not occur and none of the women developed an abscess . One woman had to be treated with antibiotics because the baby refused to nurse at the diseased breast and emptying had to be done by hand.

J Am Coll Surg, 2004 May, 198(5), 748 - 57
Experimental obstructive jaundice disrupts intestinal mucosal barrier by altering occludin expression: beneficial effect of bombesin and neurotensin; Assimakopoulos SF et al.; BACKGROUND: Little is known of the molecular events leading to increased intestinal permeability in obstructive jaundice . This study was undertaken to investigate the influence of experimental obstructive jaundice on the expression of the tight junction-associated protein occludin in the intestinal epithelium . STUDY DESIGN: Seventy male Wistar rats were randomly divided into five groups: I, controls; II, sham-operated; III, bile duct ligation (BDL); IV, BDL+Bombesin (BBS) (30 microg/kg/d); and V, BDL+Neurotensin (NT) (300 microg/kg/d) . At the end of the experiment, on day 10, endotoxin was measured in portal and aortic blood . Tissue sections of the terminal ileum were examined histologically and immunohistochemically for evaluation of occludin expression in the intestinal epithelium . Lipid peroxidation and protein oxidation were determined on tissue homogenates from terminal ileum and microbiologic analysis was performed in cecal contents . RESULTS: Obstructive jaundice resulted in portal and aortic endotoxemia, which was significantly reduced after BBS or NT administration . In the BDL group, there was total loss of occludin expression in numerous enterocytes mainly at the upper third of the villi, while a gradient of positivity existed from crypt to tip . Occludin expression was restored to control state after treatment with BBS or NT . In addition, both peptides reduced intestinal lipid peroxidation, while BBS reduced protein oxidation as well . CONCLUSIONS: Experimental obstructive jaundice induces regional loss of occludin expression in the intestinal epithelium, which may be a key factor contributing to the disruption of the mucosal barrier . Gut regulatory peptides BBS and NT prevent this alteration, leading to lower portal and systemic endotoxemia.

South Med J, 2004 Apr, 97(4), 398 - 400
Histoplasmosis in two human immunodeficiency virus-positive immigrants to Italy: clinical features and management in the highly active antiretroviral therapy era; Faggian F et al.; We report two cases of histoplasmosis occurring in human immunodeficiency virus-positive patients who immigrated to Italy, and focus our attention on the clinical features and therapeutic aspects, with particular emphasis on secondary prophylaxis . The patients had comparable human immunodeficiency virus baseline parameters, but had a completely different compliance over therapeutic regimens . The two patients were followed in two different city hospitals of our region, Padua and Verona, and the diagnosis was made on the basis of instrumental, histologic, and microbiologic findings . One of them was treated with corticosteroids because of nephrotic syndrome.

Swiss Med Wkly, 2004 Feb 21, 134(7-8), 97 - 102
Burden of illness imposed by severe sepsis in Switzerland; Schmid A et al.; OBJECTIVE: This study aims to determine the burden of illness imposed by severe sepsis in Switzerland by evaluating the direct and indirect patient-related costs for critically ill patients with severe sepsis . METHODS: In order to estimate the direct costs a retrospective analysis was undertaken using records from 61 adult patients treated in three intensive care units (ICUs) in three different University hospitals in Switzerland, in 2001 . Resource use was determined by a bottom up approach and valued using centre-specific unit costs for medication, nutrition, blood products, disposables and official tariffs for laboratory and microbiology analysis, diagnostic services, and clinical procedures . By adding centre-specific personnel and basic bed (hotel) costs total direct costs in the ICU were calculated . Indirect costs resulting from unfitness for work, early retirement, and premature death were calculated using official Swiss statistics for the years 1998-2000 . RESULTS: The mean total direct costs for a severely septic patient are CHF 41,790 (+/- 33,222 CHF) or CHF 3244 (+/- 757 CHF) per day . Nonsurvivors cause significantly higher costs than survivors (CHF 45,956 vs . CHF 37,759, p <0.001) . The total intensive care costs in Switzerland due to severe sepsis amount to CHF 146-355 million . Indirect costs were estimated to range from CHF 347 to 844 million (predominantly due to premature death) . Consequently the burden of illness of severe sepsis can be estimated to range from CHF 493 to 1199 million per year in Switzerland (1 CHF = 0.662 Euro in 2001) . CONCLUSION: Patients suffering from severe sepsis in Switzerland have a high mortality rate and spend a prolonged time in the ICU, leading to high direct and indirect costs . Particularly productivity losses due to premature death represent a considerable burden to the Swiss society.

Klin Mikrobiol Infekc Lek, 2004 Feb, 10(1), 30 - 2
{Skin infection caused by Mycobacterium marinum}; Loveckova Y et al.; Mycobacterium (M.) marinum is a human opportunistic pathogen, capable of causing skin infections and possibly also affecting deeper tissues . The authors describe a case of skin infection caused by M . marinum in a female patient with an appropriate case history . They emphasize the need of close cooperation of the clinician with a microbiologist when establishing the diagnosis.

J Assist Reprod Genet, 2004 Jan, 21(1), 11 - 3
What regulations for preimplantation genetic diagnosis?
Hill DL, Li M.
PURPOSE: To discuss the qualifications of personnel involved in either (a) embryo biopsy/slide fixation, (b) performance of preimplantation genetic diagnosis-specific assays, or (c) interpretation of findings and results from such studies . METHODS: Illustrate similarities and differences between the qualifications and licensing requirements for personnel involved in high complexity testing in other laboratory specialities, such as microbiology or endocrinology . RESULTS: Parallels can in fact be drawn between specific personnel certification and licensing requirements for reporting test results following preimplantation genetic testing and those for other clinical specimens submitted for testing . CONCLUSIONS: Assisted reproductive technology programs generally do not have personnel certified in cytotechnology or clinical genetics . Programs that perform the actual interpretation of specimens submitted for evaluation by fluorescent in situ hybridization will have to have supervisory personnel certified to perform such interpretations to ensure the level of quality control and assurance now afforded in centers for assisted reproductive technology.

Laryngoscope, 2004 Mar, 114(3), 557 - 60
Epiglottitis in the Hemophilus influenzae type B vaccine era: changing trends; Shah RK et al.; OBJECTIVE: To describe the epidemiology, natural history, and treatment of epiglottitis in the Hemophilus influenzae type B (Hib) vaccine era . DESIGN: Ten-year retrospective study . SETTING: Tertiary-care children's hospital . PATIENTS: Nineteen patients with a discharge diagnosis of epiglottitis . INTERVENTION: As indicated by the presentation of the patient, including direct laryngoscopy, intubation, intravenous antibiotics, and steroids . MAIN OUTCOME MEASURES: Presentation, management, and microbiology . RESULTS: Presenting symptoms included fever, drooling, and hot potato voice . Patients' ages ranged from 15.8 months to 17.5 (mean 8.9) years . From 1992 to 1997, the mean age was 5.8 years; from 1998 to 2002, the mean age was 11.6 years . Sixty-eight percent of patients were transferred from an outside hospital . A lateral neck radiograph was recorded in 84% of patients . Mean leukocyte count was 16600 . Direct laryngoscopy with intubation was performed in 79%; intubation was for an average of 3.5 days . H . influenzae was cultured in six patients, five of which had up-to-date immunizations . The most commonly administered antibiotics were ampicillin/sulbactam and ceftriaxone . Steroids were administered to 63% of patients . Complications included deep neck space infection (2), seizure (1), recurrent illness (1), and vocal granuloma (1) . There were no deaths . CONCLUSION: The demographics, causative organisms, and natural history of epiglottitis have changed substantially in the Hib vaccination era . The clinician evaluating and treating patients with epiglottitis should be aware of current trends.

Crit Care Med, 2004 Mar, 32(3), 858 - 73
Surviving Sepsis Campaign guidelines for management of severe sepsis and septic shock; Dellinger RP et al.; OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for severe sepsis and septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis . DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee . METHODS: We used a modified Delphi methodology for grading recommendations, built on a 2001 publication sponsored by the International Sepsis Forum . We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade . Pediatric considerations were provided to contrast adult and pediatric management . RESULTS: Key recommendations, listed by category and not by hierarchy, include early goal-directed resuscitation of the septic patient during the first 6 hrs after recognition; appropriate diagnostic studies to ascertain causative organisms before starting antibiotics; early administration of broad-spectrum antibiotic therapy; reassessment of antibiotic therapy with microbiology and clinical data to narrow coverage, when appropriate; a usual 7-10 days of antibiotic therapy guided by clinical response; source control with attention to the method that balances risks and benefits; equivalence of crystalloid and colloid resuscitation; aggressive fluid challenge to restore mean circulating filling pressure; vasopressor preference for norepinephrine and dopamine; cautious use of vasopressin pending further studies; avoiding low-dose dopamine administration for renal protection; consideration of dobutamine inotropic therapy in some clinical situations; avoidance of supranormal oxygen delivery as a goal of therapy; stress-dose steroid therapy for septic shock; use of recombinant activated protein C in patients with severe sepsis and high risk for death; with resolution of tissue hypoperfusion and in the absence of coronary artery disease or acute hemorrhage, targeting a hemoglobin of 7-9 g/dL; appropriate use of fresh frozen plasma and platelets; a low tidal volume and limitation of inspiratory plateau pressure strategy for acute lung injury and acute respiratory distress syndrome; application of a minimal amount of positive end-expiratory pressure in acute lung injury/acute respiratory distress syndrome; a semirecumbent bed position unless contraindicated; protocols for weaning and sedation/analgesia, using either intermittent bolus sedation or continuous infusion sedation with daily interruptions/lightening; avoidance of neuromuscular blockers, if at all possible; maintenance of blood glucose <150 mg/dL after initial stabilization; equivalence of continuous veno-veno hemofiltration and intermittent hemodialysis; lack of utility of bicarbonate use for pH > or =7.15; use of deep vein thrombosis/stress ulcer prophylaxis; and consideration of limitation of support where appropriate . Pediatric considerations included a more likely need for intubation due to low functional residual capacity; more difficult intravenous access; fluid resuscitation based on weight with 40-60 mL/kg or higher needed; decreased cardiac output and increased systemic vascular resistance as the most common hemodynamic profile; greater use of physical examination therapeutic end points; unsettled issue of high-dose steroids for therapy of septic shock; and greater risk of hypoglycemia with aggressive glucose control . CONCLUSION: Evidence-based recommendations can be made regarding many aspects of the acute management of sepsis and septic shock that are hoped to translate into improved outcomes for the critically ill patient . The impact of these guidelines will be formally tested and guidelines updated annually and even more rapidly as some important new knowledge becomes as available.

J Bacteriol, 2004 May, 186(9), 2757 - 65
A novel tetrahydrofolate-dependent O-demethylase gene is essential for growth of Sphingomonas paucimobilis SYK-6 with syringate; Masai E et al.; Sphingomonas paucimobilis SYK-6 degrades syringate to 3-O-methylgallate (3MGA), which is finally converted to pyruvate and oxaloacetate via multiple pathways in which protocatechuate 4,5-dioxygenase, 3MGA dioxygenase, and gallate dioxygenase are involved . Here we isolated the syringate O-demethylase gene (desA), which complemented the growth deficiency on syringate of a Tn5 mutant of the SYK-6 derivative strain . The desA gene is located 929 bp downstream of ferA, encoding feruloyl-coenzyme A synthetase, and consists of a 1,386-bp open reading frame encoding a polypeptide with a molecular mass of 50,721 Da . The deduced amino acid sequence of desA showed 26% identity in a 325-amino-acid overlap with that of gcvT of Escherichia coli, which encodes the tetrahydrofolate (H(4)folate)-dependent aminomethyltransferase involved in glycine cleavage . The cell extract of E . coli carrying desA converted syringate to 3MGA only when H(4)folate was added to the reaction mixture . DesA catalyzes the transfer of the methyl moiety of syringate to H(4)folate, forming 5-methyl-H(4)folate . Vanillate and 3MGA were also used as substrates for DesA; however, the relative activities toward them were 3 and 0.4% of that toward syringate, respectively . Disruption of desA in SYK-6 resulted in a growth defect on syringate but did not affect growth on vanillate, indicating that desA is essential to syringate degradation . In a previous study the ligH gene, which complements the growth deficiency on vanillate and syringate of a chemical-induced mutant of SYK-6, DC-49, was isolated (S . Nishikawa, T . Sonoki, T . Kasahara, T . Obi, S . Kubota, S . Kawai, N . Morohoshi, and Y . Katayama, Appl . Environ . Microbiol . 64:836-842, 1998) . Disruption of ligH resulted in the same phenotype as DC-49; its cell extract, however, was found to be able to convert vanillate and syringate in the presence of H(4)folate . The possible role of ligH is discussed.

J Dent Hyg, 2004 Winter, 78(1), 39 - 45
Predicting academic and National Board Dental Hygiene Examination performance based on Academic Factors; Bauchmoyer SM et al.; PURPOSE: Numerous studies have explored reliable variables that predict student success in dental hygiene programs and on the National Board Dental Hygiene Examination (NBDHE) . However, no studies were found using data collected since the NBDHE format changed in 1998 to investigate if traditional predictors hold true . The objective of this study was to examine the relationship between pre-admission requirements, basic college science requirements, site of academic preparation, cumulative dental hygiene grade point average (CDHYGPA) and the NBDHE score . METHODS: Data from the academic records of 173 graduates of the dental hygiene program at The Ohio State University from 1998 through 2002 were entered into an Excel spreadsheet using identification numbers . Demographic information for the description of the subjects, course transfer data, course grades in program prerequisites and basic science requirements, CDHYGPA, and NBDHE scores were entered . Data were analyzed using the Statistical Package for the Social sciences (SPSS-version 10), Pearson's r correlations, regression analysis, and ANOVA with a predetermined level of significance at .05 . RESULTS: Of the 173 records entered, 132 had complete data (76.3%) . Results indicate the existing prerequisites for the dental hygiene program remain strong predictors for success . A strong correlation was noted between human nutrition courses and the CDHYGPA . Other core science courses completed while in the program-anatomy, physiology and microbiology--also rendered a moderately strong correlation to the CDHYGPA . The greatest predictors for success on the NBDHE were the student's CDHYGPA and the prerequisite three science GPA . Consistency in site of science preparation also revealed a positive correlation to the CDHYGPA . CONCLUSIONS: This study confirmed the continued use of the three science GPA pre-requisite and entering GPA for predicting success in this dental hygiene program and on the NBDHE even after the format changed to include case-based items . Other predictors for success that were identified in the study may aide dental hygiene program admission committees in their selection process . These committees may also consider the site of science preparation noting that institution consistency played a role in academic performance.

AIDS, 2004 Jan 23, 18(2), 257 - 64
A study of the safety, immunology, virology, and microbiology of adjunctive etanercept in HIV-1-associated tuberculosis; Wallis RS et al.; OBJECTIVE: Tumor necrosis factor (TNF), an important inflammatory mediator in tuberculosis, has been implicated in causing accelerated HIV disease progression in HIV-associated tuberculosis . However, TNF blockade, particularly by monoclonal antibody, has been associated with the reactivation of latent Mycobacterium tuberculosis infection by the impairment of mycobacterial immunity . This phase 1 study examined the safety, microbiology, immunology, and virology of TNF blockade using etanercept (soluble TNF receptor, Enbrel) during the initial treatment of HIV-associated tuberculosis . DESIGN: A single-arm trial, with key endpoints compared with historical controls, conducted in Mulago Hospital, Kampala, Uganda . SUBJECTS:: Sixteen HIV-1-infected patients and 42 CD4-frequency-matched controls with sputum smear-positive tuberculosis and CD4 cell counts > 200 cells/microl . INTERVENTION: Etanercept 25 mg, eight doses administered subcutaneously twice weekly beginning on day 4 of tuberculosis therapy . MAIN OUTCOME MEASURES: Serial examination, radiography, sputum culture, CD4 T-cell counts, plasma log10 HIV-RNA copy numbers . RESULTS: Trends towards superior responses to tuberculosis treatment were evident in etanercept-treated subjects in body mass, performance score, number of involved lung zones, cavitary closure, and time to sputum culture conversion . Etanercept treatment resulted in a 25% increase in CD4 cells by week 4 (P = 0.1 compared with controls) . The change in CD4 cell count was inversely related to the change in serum neopterin, a marker of macrophage activation . There was no effect on plasma HIV RNA . CONCLUSION: Etanercept can be safely administered during the initial treatment of pulmonary tuberculosis . Further studies are warranted to examine the effects of etanercept on T-cell numbers, activation and apoptosis in AIDS and tuberculosis.

J Intensive Care Med, 2004 Mar-Apr, 19(2), 83 - 104
Critical care aspects of lung transplantation; Lau CL et al.; Lung transplantation currently is the preferred treatment option for a variety of end-stage pulmonary diseases . Remarkable progress has occurred through refinements in technique and improved understanding of transplant immunology and microbiology . As a result, recipients are surviving longer after their transplant . Despite improvements in short- and intermediate-term survival, long-term success with lung transplantation remains limited by chronic allograft rejection, also known as bronchiolitis obliterans syndrome . Despite its long-term limitations, lung transplantation remains the only hope for many with end-stage pulmonary disease, and during the past 20 years, it has become increasingly accepted and used . As a result, clinicians working in an intensive care unit (ICU) are more likely to be exposed to these patients both in the immediate postoperative period as well as throughout their remaining lives . It is thus important that the ICU team have a working knowledge of the common complications, when these complications are most likely to occur, and how best to treat them when they do arise . The main focus of this review is to address the variety of potential graft and life-threatening problems that may occur in lung transplant recipients . Because the ICU is also the most common setting where a potential donor is identified, donor issues will briefly be addressed.

Radiology, 2004 Apr, 231(1), 250 - 7
Infected aortic aneurysms: imaging findings; Macedo TA et al.; PURPOSE: To determine the imaging characteristics of infected aortic aneurysms . MATERIALS AND METHODS: Review of records of patients with surgical and/or microbiologic proof of infected aortic aneurysm obtained over a 25-year period revealed 31 aneurysms in 29 patients . This study included 21 men and eight women (mean age, 70 years) . One radiologist reviewed 28 computed tomographic (CT) studies (22 patients underwent CT once and three patients underwent CT twice), 12 arteriograms (12 patients underwent arteriography once), eight nuclear medicine studies (six patients underwent nuclear medicine imaging once and one patient underwent nuclear medicine imaging twice), and three magnetic resonance (MR) studies (three patients underwent MR imaging once) . Features evaluated included aneurysm size, shape, and location; branch involvement; aortic wall calcification; gas; radiotracer uptake on nuclear medicine studies; and periaortic and associated findings . The location of infected aortic aneurysms was compared with that of arteriosclerotic aneurysms . RESULTS: Aneurysms were located in the ascending aorta (n = 2, 6%), descending thoracic aorta (n = 7, 23%), thoracoabdominal aorta (n = 6, 19%), paravisceral aorta (n = 2, 6%), juxtarenal aorta (n = 3, 10%), infrarenal aorta (n = 10, 32%), and renal artery (n = 1, 3%) . Two patients had two infected aortic aneurysms . CT revealed 25 saccular (93%) and two fusiform (7%) aneurysms with a mean diameter at initial discovery of 5.4 cm (range, 1-11 cm) . Paraaortic soft-tissue mass, stranding, and/or fluid was present in 13 (48%) of 27 aneurysms, and early periaortic edema with rapid aneurysm progression and development was present in three (100%) patients with sequential studies . Other findings included adjacent vertebral body destruction with psoas muscle abscess (n = 1, 4%), kidney infarct (n = 1, 4%), absence of calcification in the aortic wall (n = 2, 7%), and periaortic gas (n = 2, 7%) . Angiography showed 13 saccular aneurysms with lobulated contour in 10 (77%) . Nuclear medicine imaging showed increased activity consistent with infection in six (86%) of seven aneurysms . MR imaging showed three saccular aneurysms . Adjacent abnormal vertebral body marrow signal intensity was seen in one (33%) of three patients . CONCLUSION: Saccular aneurysms (especially those with lobulated contour) with rapid expansion or development and adjacent mass, stranding, and/or fluid in an unusual location are highly suspicious for an infected aneurysm . Copyright RSNA, 2004

Commun Dis Public Health, 2003 Dec, 6(4), 317 - 9
Chest physicians' and microbiologists' awareness and demand for drug monitoring in the treatment of TB; Pooransingh S et al.; There is a role for therapeutic drug monitoring (TDM) to achieve the optimum therapeutic concentration of anti-tuberculous drugs . This work aimed to determine the current awareness of TDM in TB control among chest physicians and to estimate the demand for this service . Responses from a sample of chest physicians in the West Midlands revealed that 60% were aware of TDM and 33% had used it . Responses were received from half of a UK group of microbiologists who reported a median of nine requests in the past year . It appeared that more was known about services for rifampicin and streptomycin than other first-line drugs . There appears to be a need for both increased awareness among potential service users and for coordination of assay services.

J Chromatogr A, 2004 Apr 2, 1032(1-2), 1 - 6
Determination of vitamin B5 in a range of fortified food products by reversed-phase liquid chromatography-mass spectrometry with electrospray ionisation; Mittermayr R et al.; Methods for Vitamin B5 determination in food products remain limited by their low sensitivity and poor selectivity . Here, we have developed a liquid chromatography-mass spectrometry (LC-MS) method for Vitamin B5 determination in wide range of fortified food products . Vitamin B5 was extracted from food samples by heat treatment and analysed by LC-MS in the positive mode using electrospray ionisation (ESI) . Vitamin B5 was quantified using hopantenic acid (HOPA) as internal standard after their separation on a C18 narrow-bore column with a gradient of mobile phase made of water/acetonitrile and trifluoroacetic acid (TFA) 0.025% . MS with single ion monitoring mode at mass m/z 220 was used for Vitamin B5 quantification . Calibration curve between 0.5 and 10 microg/ml of Vitamin B5 was linear (r2=0.9993) and the detection limit was determined to be 800 pg . The overall quantitative efficiency of the method was evaluated using Nestle reference sample (infant formula) . The intra-assay RSD was 4.8% (n=8), the inter-assay RSD 6.4% (n=4) and the recoveries of the spiked samples were above 95% . Application of the LC-MS method to Vitamin B5 determination in wide range of fortified food products including three US National Institute of Standards and Technology (NIST) reference samples (RM 8435, RM 8415 and SRM 1546) shows consistent results with those obtained by microbiology and recoveries of Vitamin B5 between 93 and 104% for the spiked samples.

Genome Biol . 2004;5(4):R27 . Epub 2004 Mar 18.
Comparative genomics of gene-family size in closely related bacteria; Pushker R et al.; BACKGROUND: The wealth of genomic data in bacteria is helping microbiologists understand the factors involved in gene innovation . Among these, the expansion and reduction of gene families appears to have a fundamental role in this, but the factors influencing gene family size are unclear . RESULTS: The relative content of paralogous genes in bacterial genomes increases with genome size, largely due to the expansion of gene family size in large genomes . Bacteria undergoing genome reduction display a parallel process of redundancy elimination, by which gene families are reduced to one or a few members . Gene family size is also influenced by sequence divergence and physiological function . Large gene families show wider sequence divergence, suggesting they are probably older, and certain functions (such as metabolite transport mechanisms) are overrepresented in large families . The size of a given gene family is remarkably similar in strains of the same species and in closely related species, suggesting that homologous gene families are vertically transmitted and depend little on horizontal gene transfer (HGT) . CONCLUSIONS: The remarkable preservation of copy numbers in widely different ecotypes indicates a functional role for the different copies rather than simply a back-up role . When different genera are compared, the increase in phylogenetic distance and/or ecological specialization disrupts this preservation, albeit in a gradual manner and maintaining an overall similarity, which also supports this view . HGT can have an important role, however, in nonhomologous gene families, as exemplified by a comparison between saprophytic and enterohemorrhagic strains of Escherichia coli.

J Endod, 2004 Mar, 30(3), 135 - 7
Nested PCR detection of Centipeda periodontii in primary endodontic infections; Siqueira Junior JF et al.; In recent years, molecular genetic methodologies have provided significant additional knowledge about components of the microbiota associated with infections of endodontic origin . Following this research line, the purpose of the present study was to investigate the prevalence of Centipeda periodontii in primary endodontic infections using a species-specific nested PCR assay . Samples were collected from fifty teeth having carious lesions, necrotic pulps, and different forms of periradicular diseases . DNA extracted from the samples was initially amplified using universal 16S rDNA primers, and a second round of amplification used the first PCR products to detect a specific fragment of C . periodontii 16S rDNA . This species was detected in 3 (13%) of 23 asymptomatic cases, in 1 (14%) of 7 cases diagnosed as acute apical periodontitis, and in 3 (15%) of 20 pus samples aspirated from acute periradicular abscesses . There was no significant association between C . periodontii and the presence of clinical symptoms . Overall, C . periodontii was detected in 14% of the cases of endodontic infections . This is probably the hitherto first study to detect C . periodontii in primary endodontic infections . The specific role played by this bacterial species in infections of endodontic origin awaits further clarification.

Int J Parasitol, 2004 Mar 9, 34(3), 393 - 400
The NF-kappaB signaling pathway: immune evasion and immunoregulation during toxoplasmosis; Shapira S et al.; The NF-kappaB family of transcription factors is part of an evolutionarily conserved system that plays an important role in the regulation of genes associated with the development of innate and adaptive responses required for the recognition and immunologic control of pathogens {Clin . Microbiol . Rev . 15 (2002) 414; Annu . Rev . Immunol . 16 (1998) 225; Infect . Immun . 70 (2002) 3311} . In addition, NF-kappaB regulates other cellular processes required for a coordinated immune response, such as cellular growth and differentiation, cell adhesion, survival and apoptosis . Recent studies have highlighted the prominent role played by the NF-kappaB system in resistance to Toxoplasma gondii but it is becoming apparent that this intracellular parasite can directly modulate this signalling pathway . This article briefly reviews the biology of NF-kappaB, examines the complex interaction that takes place between T . gondii and these transcription factors in infected cells, and highlights the role of different NF-kappaB family members during the development of a protective immune response to this pathogen.

Orthopade, 2004 Apr, 33(4), 389 - 96
{From "wet caries" to controllable complication . History of post-traumatic and postoperative osteitis}; Rauschmann MA et al.; In former times treatment of osteomyelitis was based on very few surgical principles such as incision of the infected region, debridement with sequestrectomy, and continuous drainage to prevent superficial closing of the wound . There were no rules for asepsis, anesthesia, and microbiology in the treatment of bone infection . Therapy improved dramatically when J . Lister introduced and performed antisepsis.With the beginning of the twentieth century it became much more standardized and differentiated . In the 4th decade of the twentieth century, the development of antibiotics changed the regime of osteomyelitis therapy again . Especially systemic intravenous antibiotic therapy was very successful in the treatment of the hematogeneous osteomyelitis . Treatment failed in post-traumatic and postoperative osteitis . Local antibiotic therapy on a delivery system was the breakthrough for these cases . The application of PMMA cement as an antibiotic delivery system led to this new possibility for treating bone infections and is still in use nowadays.

Lancet Infect Dis, 2004 Apr, 4(4), 201 - 12
Infections related to the ingestion of seafood Part I: Viral and bacterial infections; Butt AA et al.; Foodborne diseases cause an estimated 76 million illnesses in the USA each year . Seafood is implicated in 10-19% of these illnesses . A causative agent can be traced in about 44% of seafood-related outbreaks, viruses accounting for around half of these illnesses . Although viruses are the most common cause of seafood-related infections, most hospitalisations and deaths are due to bacterial agents . A wide variety of viruses, bacteria, and parasites have been implicated in seafood-related outbreaks, which are reported worldwide . The factor most commonly associated with infection is consumption of raw or undercooked seafood . People with underlying disorders, particularly liver disease, are more susceptible to infection . The first part of this two-part review summarises the general incidence of seafood-related infections and discusses the common viral and bacterial causes of these infections . For each agent, the microbiology, epidemiology, mode of transmission, and treatment are discussed . In the May issue of the journal we will discuss parasites associated with seafood consumption, the safety of seafood, and the measures put in place in the USA to increase its safety.

Ann Ig, 2003 Nov-Dec, 15(6), 945 - 52
{Quality evaluation multiapproach of sediments in water bodies in Southern Italy}; Isidori M et al.; This study reports the toxicity of sediments from rivers and canals collected at two six-month intervals from 19 sampling points of the province of Naples in Southern Italy . The toxicity of the pore-water from sediments was determined with a battery composed of different freshwater microbiotests . The toxicity varied substantially from station to station, but none of the Toxkit tests was either the most or the least sensitive for all stations . Chemical analyses were accomplished on some heavy metals . Furthermore, the in situ biological communities of macroinvertebrates were assessed . Most of tested sediments were found to be very toxic for several if not all the microbiotests as well as the biological communities (EBI) . Comparison between toxicological data and heavy metals revealed a positive correlation for zinc and copper . This study confirms that only an integrated approach is suitable for the evaluation of contaminated sediments and that single parameters are not always able to define a situation of environmental deterioration.

Ann Ig, 2003 Nov-Dec, 15(6), 851 - 61
{Molecular diagnosis of Chlamydia pneumoniae diseases}; Visca P et al.; Chlamydia pneumoniae (C . p.) is an intracellular parasite directly involved in respiratory disease and more recently in chronic degenerative pathologies as atherosclerosis and asthma . Its peculiar life cycle makes cultural isolation difficult, thus, troublesome the diagnosis of the disease . Serology is so far the most common method of diagnosis of the, although the indirect based evidence of the serology may give clinically misleading results . Nucleic acid amplification methods offer indeed rapid, reliable and low cost assessment pathogen bacteria isolation, with relevant benefits for the patient's management . These molecular methods are nowadays essential in presence of bacteria of difficult cultivation or method inconsistent with temporal clinical needs, for they allow to rapidly detect even nucleic acid traces of the infectious agent, providing direct evidence of its presence in the biological samples and hence the relevant therapy . Nucleic acid methods are extensively applied in laboratory diagnosis of Chlamydia trachomatis bringing about the development of sensitive and reliable commercial kits . This review analyses the literature of the genic amplification methods in the search of C . p . in clinical samples highlighting methodological and diagnostic aspects . Although genic amplification methods have been implemented presently by the clinical research labs only, it is anticipated that through their standardisation they could be used by most clinical microbiology laboratories.

Neurol Neurochir Pol, 2004 Jan-Feb, 38(1), 31 - 6
{Tuberculous meningitis in 8-year observation of the Department of Infectious Diseases and Neuroinfections of the Medical Academy in Białystok}; Grygorczuk S et al.; BACKGROUND AND PURPOSE: Tuberculous meningitis (tbm), although comparatively rare, remains a serious clinical problem . MATERIAL AND METHODS: We have analyzed medical records of patients with tbm diagnosis hospitalized in our Department in the years 1995-2002 . RESULTS: During that period 11 cases of tbm were diagnosed in the Department . Family contact with a person with diagnosed tuberculosis was revealed in 2 of them, previously treated pulmonary tuberculosis in 2 and active tuberculosis outside cns was diagnosed in 4 . On admission 9 of the patients were unconscious . Neurologic abnormalities were present in all 11 patients, most commonly they were pathologic reflexes and cranial nerve abnormalities . Cerebrospinal fluid (csf) showed abnormalities in all patients, however, with a high variability of its parameters (cytosis: 12-4400 cells per microliters, protein concentration 52-482 mg/dL, low glucose concentration in 2 patients) . CT scan revealed cns abnormalities (most typically hypodense foci and/or ventricular enlargement) in 5 patients . Microbiologic confirmation (M . tuberculosis growth from csf) was obtained in only 1 patient, despite intensive diagnostic efforts . The mean time from admission to proper diagnosis and start of antituberculous treatment was 9.9 days . Treatment effects were favorable (healing without serious neurologic sequelae) in 7 patients, permanent sequelae were observed in 3 and one patient died while in hospital . CONCLUSIONS: Our data confirm the presence of serious difficulties in the diagnosis and treatment of tbm . A prompt diagnosis and quick start of empirical treatment remain the most important means of reducing the mortality and frequency of permanent sequelae associated with this form of meningitis.

Radiology, 2004 May, 231(2), 333 - 41 Epub 2004 Mar 24.
FDG PET for differentiation of infection and aseptic loosening in total hip replacements: comparison with conventional radiography and three-phase bone scintigraphy; Stumpe KD et al.; PURPOSE: To compare the diagnostic efficacy of fluorine 18 fluorodeoxyglucose (FDG) positron emission tomography (PET) with that of conventional radiography and three-phase bone scintigraphy in patients suspected of having infection in their total hip replacements . MATERIALS AND METHODS: Thirty-five patients with painful total hip replacements and possible septic prosthetic loosening were examined with FDG PET, conventional radiography, and three-phase bone scintigraphy . PET, radiographic, and scintigraphic images were each evaluated by two independent observers in a blinded fashion . For 32 of 35 patients, serial conventional radiographs were available . Results of microbiologic examinations of surgical specimens represented the standard of reference in 26 patients, and results of joint aspiration plus clinical follow-up of at least 6 months represented the standard of reference in the remaining nine patients . Sensitivity, specificity, accuracy, and interobserver variability (kappa) values were calculated . The imaging modalities were compared in terms of diagnostic confidence by using the sign test . RESULTS: Nine patients had septic and 21 patients had aseptic loosening . In five patients, neither loosening nor infection was confirmed . For diagnosing infection with FDG PET, conventional radiography, and bone scintigraphy, respectively, sensitivity values for reader 1 and reader 2 were 33% and 22%, 89% and 78%, and 56% and 44%, while specificity values were 81% and 85%, 50% and 65%, and 88% and 92% and accuracy values were 69% for both readers, 60% and 69%, and 80% for both readers . PET was significantly more specific (P =.035) but less sensitive (P =.016) than conventional radiography for the diagnosis of infection . CONCLUSION: In a study population of patients suspected of having infected total hip replacements, FDG PET performed similarly to three-phase bone scintigraphy . FDG PET was more specific but less sensitive than conventional radiography for the diagnosis of infection.

Obes Res, 2004 Mar, 12(3), 547 - 55
Alteration of amine oxidase activity in the adipose tissue of obese subjects; Visentin V et al.; OBJECTIVE: To explore the activity of monoamine oxidases (MAOs) and semicarbazide-sensitive amine oxidases (SSAOs) in adipose tissue and blood of lean and moderately obese subjects and to study whether there is a link between these hydrogen peroxide-generating enzymes and blood markers of oxidative stress . RESEARCH METHODS AND PROCEDURES: Nine obese male subjects (BMI 32.6 +/- 0.4 kg/m(2)) and nine controls (BMI 23.4 +/- 0.5) of 24- to 40-year-old subjects were included in the study . MAO and SSAO activities were measured on microbiopsies of abdominal subcutaneous adipose tissue by quantifying (14)C-tyramine and (14)C-benzylamine oxidation . Levels of soluble SSAO, lipid peroxidation products, and antioxidant agents were measured in plasma, whereas cytoprotective enzymes were determined in blood lysates . RESULTS: The high MAO activity found in adipose tissue was diminished by one-half in obese subjects (maximum initial velocity of 1.2 vs . 2.3 nmol tyramine oxidized/mg protein/min) . There was no change in SSAO activity, either under its adipose tissue-bound or plasma-soluble form . Plasma levels of lipid peroxidation products and antioxidant vitamins remained unmodified, as well as erythrocyte antioxidant enzymes, whereas circulating triglycerides, insulin, and leptin were increased . DISCUSSION: Although they already exhibited several signs of endocrino-metabolic disorders, the obese men did not exhibit the increase in blood markers of oxidative stress or the decrease in antioxidant defenses reported to occur in very obese or diabetic subjects . The reduced MAO and the unchanged SSAO activities found in obesity suggest that these hydrogen peroxide-generating enzymes expressed in adipocytes are probably not involved in the onset of the oxidative stress found in severe obesity and/or in its complications.

Acad Med, 2004 Apr, 79(4), 366 - 75
Guidelines for preclerkship bioterrorism curricula; Coico R et al.; PURPOSE: To develop medical school curriculum guidelines related to bioterrorism to ensure that future medical graduates are armed with the critical knowledge, skills, and attitudes to face this emerging threat . METHOD: An Internet-based Delphi survey was performed in 2002 under the auspices of the Association of Medical School Microbiology and Immunology Chairs involving 64 medical educators in microbiology, immunology, and infectious diseases representing 54 U.S . medical schools . A 12-member bioterrorism expert reference panel participated in the final phase of the survey . RESULTS: Study participants identified the top educational objectives for the following bioterrorism-related curriculum categories: general issues, biodefense, public health, infection control, infectious diseases, and toxins . CONCLUSION: The study focused on preparedness training through the development of curricular guidelines in predominantly preclerkship medical education by identifying basic science and clinical laboratory aspects of putative biologic weapons organisms and toxins, clinical manifestations of bioterrorist attacks, treatment strategies, epidemiology, and prophylaxis.

Ceska Gynekol, 2003 Nov, 68(6), 432 - 9
{Host-yeast interactions in relation to the urogenital tract, vulvovaginal candidiasis and urologic aspects of mycotic diseases}; Spacek J et al.; OBJECTIVE: A review of the role of yeasts in indigenous microbiota with the focus on urogenital tract . The yeast in vaginal environment and their relation to acute and recurrent form of vulvovaginal candidiasis . DESIGN: A review . SETTING: Department of Obstetrics and Gynecology, Department of Urology, Medical Faculty, Charles University, Hradec Kralove . SUBJECT AND METHODS: Review of current literature data with the contribution on own experience . CONCLUSIONS: Vulvovaginal candidiasis is one of the most frequent complaints in patients attending gynecologist . Recurrent form of this disease is still an unsolved problem because the etiopathogenesis is unexplained and successful therapeutic management from the point of view of next attacks is very difficulty.

Int J Legal Med, 2004 Jun, 118(3), 163 - 9 Epub 2004 Mar 24.
German study on sudden infant death (GeSID): design, epidemiological and pathological profile; Findeisen M et al.; The German study on sudden infant death (GeSID) is a multi-centre case-control study aiming at the assessment of etiological factors and risk factors of SIDS . This report describes the study design and the methods applied and presents some general findings . Between 1998 and 2001, 455 cases of sudden and unexpected death of infants aged between 8 and 365 days were recruited into the study . The study comprised at least 11 out of the 16 German states with 18 centres involved . In 1999 and 2000, 75% of all SIDS cases registered with the Federal Office of Statistics (ICD 10/R95, n=384) in the study area were recruited into the study (n=286) . A standardised autopsy including extended histology, microbiology, virology, toxicology and neuropathology investigations was carried out . Of the parents 82% (n=373) agreed to fill in an extensive questionnaire containing 120 questions reflecting all important aspects of the infant's development . For each SIDS case, the parents of three living control infants were interviewed . These controls were matched for age, gender and region (n=1,118) . The response rate of the controls was 58.7% . Data were linked with medical records obtained from obstetrics departments, the children's hospitals, and general practitioners . Death scene investigation was performed in 4 study areas (cases: n=64, controls: n=191) . All cases were classified into one of 4 categories using defined criteria: 7.3% of the children were assigned to category 1 (no pathological findings: SIDS), 61.1% to category 2 (minor findings: SIDS+), 20.4% to category 3 (severe findings: SIDS+) and 11.2% to category 4 (findings which explained the death: non-SIDS) . In case conferences the previous history and circumstantial factors were included and an extended category (E-cat.) was defined . The consideration of these factors for the final classification is of great importance in the causal explanation of some cases . An analysis of 18 main variables in cases of categories 1-3 (SIDS) compared to the cases of category 4 (non-SIDS) showed significant differences for the sleeping position, coughing the day before death and breast-feeding indicating that the cases of both groups should be separated for further analyses.

Nat Rev Microbiol . 2003 Oct;1(1):3.
A new beginning...; Management of community-acquired pneumonia in persons with spinal cord injury; Spinal Cord Injury Service, VA Puget Sound Health Care System, and the Department of Rehabilitation Medicine and Harborview Injury Prevention and Research Center, University of Washington, Seattle, WA, USASTUDY DESIGN: Retrospective case series . OBJECTIVES: Respiratory disorders are the leading cause of death in persons with spinal cord injury (SCI), but the epidemiology and medical management of pneumonia in persons with chronic SCI is not well characterized . We describe the clinical presentation of persons with SCI with community-acquired pneumonia (CAP), characterize its management and compare practice to recommendations for CAP in the general population . SETTING: Three United States Veterans Affairs Medical Centers with specialized SCI services . METHODS: Chart abstraction was performed for all persons with chronic SCI seen at participating centers for treatment of CAP during a 2-year period . Collected data included presenting signs and symptoms, laboratory and imaging results, initial antibiotic therapy, secretion mobilization techniques, in-patient vs outpatient management, length of stay, and mortality . RESULTS: In all, 41 persons with SCI received treatment for CAP during the study period . A total of 32 (78.0%) patients were admitted for treatment; two (4.8%) required intubation and mechanical ventilation . Initial antibiotic coverage met guideline recommendations for only half of inpatients and infrequently provided adequate antipseudomonal coverage . Microbiologic testing was performed on 26 cases (63.4%) and demonstrated a specific pathogen in only five cases (12.2% of total) . Three cases (7.3%) died during treatment for CAP, and 16 (42.1%) of 38 CAP survivors died within a median follow-up of 3 years . CONCLUSION: The majority of chronic SCI patients who present to specialized SCI centers with CAP are admitted for treatment . Short-term mortality is comparable to CAP in the general population.

Curr Opin Microbiol, 2004 Feb, 7(1), 4 - 10
Bacterial subversion of lipid rafts; Lafont F et al.; Bacteria rely on numerous basic cellular functions of their target cells to reach successful infection . The recent discovery that the plasma membrane contains specialized microdomains, called lipid rafts, with many specific functions but in particular with the ability to concentrate signaling molecules, has therefore attracted the attention of cellular microbiologists . Since then an increasing number of bacteria and their products have been shown to interact with lipid rafts to promote infection or intoxication . Here we review why certain bacteria and/or their products are attracted toward these lipid microdomains.

Enferm Infecc Microbiol Clin, 2003 May, 21 Suppl 2, 81 - 3
{The present and future of clinical virology}; Telenti A; Clinical virology faces challenges of opposing sign . On one hand, there will be an increasing demand for identification of novel pathogenic viruses, for resistance testing in a new era of antiviral therapy, and for full participation in the molecular revolution in diagnostic technologies . On the other hand, the responsibilities of clinical virology are being transferred to the general diagnostic microbiology laboratory, that risks limiting the numbers of specialists in virology available for meeting future challenges and demands.

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