J Am Osteopath Assoc, 2004 Dec, 104(12), 527 - 35 Sexually Transmitted Infections and Increased Risk of Co-infection with Human Immunodeficiency Virus; Nusbaum MR et al.; The incidence of trichomoniasis (Trichomonas vaginalis) in the United States is estimated at 5 million cases annually; chlamydia (Chlamydia trachomatis) at 3 million; gonorrhea (Neisseria gonorrhoeae), 650,000; and syphilis (Treponema pallidum), 70,000 . However, most sexually transmitted infections (STIs) are asymptomatic-contributing to underdiagnosis estimated at 50% or more . Diagnosis of an STI signals sexual health risk because an STI facilitates the transmission and acquisition of other STIs, including human immunodeficiency virus (HIV) . In fact, comorbid STIs increase patients' susceptibility of acquiring and transmitting HIV by two- to fivefold . Several studies have shown that aggressive STI prevention, testing, and treatment reduces the transmission of HIV . The authors discuss common clinical presentations, screening, diagnosis, and treatment for trichomoniasis, chlamydia, gonorrhea, syphilis, and herpes simplex virus. Vaccine, 2005 Jan 26, 23(10), 1312 - 21 Interactions of proteoliposomes from serogroup B Neisseria meningitidis with bone marrow-derived dendritic cells and macrophages: adjuvant effects and antigen delivery; Rodriguez T et al.; Exposure to proteoliposomes from serogroup B Neisseria meningitidis (PL) induced up-regulation of MHC-II, MHC-I, CD40, CD80 and CD86 expression on the surface of murine bone marrow-derived dendritic cells (DC) . CD40, CD80 and CD86 were up-regulated on bone marrow-derived macrophages (MPhi) upon stimulation with PL . Both DC and MPhi released TNFalpha, but only DC produced IL12(p70) in response to PL . A small increase in the expression of MHC-II, CD40 and CD86, as well as production of IL12(p70), was observed on the cell surface of DC, but not MPhi from LPS-non-responder C3H/HeJ after exposure to PL . DC, but not MPhi, incubated with PL containing ovalbumin (PL-OVA) presented OVA-specific peptides to CD4(+) and CD8(+) OVA-specific T-cell hybridomas . These data clearly indicate that PL exert an immunomodulatory effect on DC and MPhi, with some contribution of non-LPS components besides the main role of LPS . The work also shows the potential of PL as a general system to deliver antigens to DC for presentation to CD4(+) and CD8(+) T-cells. Br J Biomed Sci, 2004, 61(4), 175 - 8 Aetiological agents of vaginitis in Nigerian women; Otuonye NM et al.; This study focuses on the identification of aetiological agents of vaginitis in Nigerian women . Study subjects are drawn from patients presenting with lower abdominal pain, vaginal discharge and itching at the gynaecology clinic of Lagos University Teaching Hospital and at the Clinical Centre of the Nigerian Institute of Medical Research, Yaba, Lagos, between January 2001 and July 2002 . A total of 250 patients gave informed consent to participate in the study . The patients also had pre- and post-test human immunodeficiency virus (HIV) counselling . Each patient completed a questionnaire in order to provide biographical data, past clinical history and socio-economic background information . A cervical swab (CS) and a high-vaginal swab (HVS) were obtained from each patient . Swab samples were examined for pH and under light microscopy by Gram's stain and as wet preparations in 10% potassium hydroxide . Subsequently, samples were cultured on appropriate media at optimal conditions and a drug sensitivity profile for all isolates was determined by standard methods . Blood samples were screened and confirmed for HIV antibodies . Bacterial, fungal and parasitic pathogens were identified or isolated in samples from 241 (96.4%) of the women . Bacterial agents (Neisseria, Streptococcus and Staphylococcus species) were predominant in 128 (51.2%) patients, followed by fungi in 108 (43.2%) and parasites (Trichomonas vaginalis) in five (2.0%) . Sensitivity to ciprofloxacin was seen in 40% of Staphylococcus species and in 90% of Neisseria species . Positive HIV serology was seen in 25 (10%) of the 250 women studied, 20 (80%) of which had concurrent microbial infections . Overall, a broad spectrum of microbial agents were shown to be responsible for vaginitis in the group of patients studied. Gynakol Geburtshilfliche Rundsch, 2005 Jan, 45(1), 14 - 8 {Adnexitis and pelvic inflammatory disease}; Lauper U et al.; Pelvic inflammatory disease and upper genital tract infection describe inflammatory changes in the upper female genital tract of any combination: endometritis, salpingitis, tubo-ovarian abscess, peritonitis in the small pelvis . The International Infectious Disease Society for Obstetrics and Gynecology recommends a revision of the CDC guidelines taking into account the type of germ or the triggering agent and the seriousness of the disease . Infections with Chlamydia trachomatis and Neisseria gonorrhoeae are increasing worldwide . They are one of the main causes of tubal sterility, chronic abdominal pain and ectopic pregnancies . More than 30% of the infections are subclinical and asymptomatic . Therefore it is most recommendable to generally screen young, sexually active women with any of the risks mentioned above . Antibiotic therapy should be started as early as possible, in case of doubt even probatively, and should cover a broad spectrum of germs . C . trachomatis and N . gonorrhoeae should be treated according to resistance testing . In uncomplicated cases, hospitalization is unnecessary, ambulant therapy is sufficient. J Huazhong Univ Sci Technolog Med Sci, 2004, 24(5), 417 - 20 Construction of prokaryotic expression plasmid of fusion protein including porin A and porin B of Neisseria gonorrhoeae and its expression in E . coli; Liao F et al.; In order to provide a rational research basis for clinical detection and genetic engineering vaccine, plasmid pET-28a (+) encoding both Porin gene PIA and PIB of Neisseria gonorrhoeae was constructed and a fusion protein in E . coli DE3 expressed . The fragments of PIA and PIB gene of Neisseria gonorrhoeae were amplified and cloned into prokaryotic expression plasmid pET-28a (+) with double restriction endonuclease cut to construct recombinant pET-PIB-PIA . The recombinant was verified with restriction endonuclease and sequenced and transformed into E . coli DE3 to express the fusion protein PIB-PIA after induced with IPTG . The results showed PIA-PIB fusion DNA fragment was proved correct through sequencing . A 67 kD (1 kD= 0.992 1 ku) fusion protein had been detected by SDS-PAGE . It was concluded that the fusion protein was successively expressed. Carbohydr Res, 2005 Feb 7, 340(2), 221 - 33 Synthesis of deoxy and acylamino derivatives of lactose and use of these for probing the active site of Neisseria meningitidis N-acetylglucosaminyltransferase; Westerlind U et al.; Derivatives of lactose with the galactose ring substituents replaced by deoxy or acylamino functions were prepared . The 2'-, 3'-, 4'- and 6'-deoxy, 3'-acetamido and 3'-benzamido derivatives of phenyl 4-O-(beta-d-galactopyranosyl)-beta-d-glucopyranoside (phenyl beta-lactoside) were synthesized from disaccharide or monosaccharide precursors . The derivatives were tested as substrates for the N-acetylglucosaminyltransferase from Neisseria meningitidis, which uses lactosyl derivatives as acceptors and UDP-GlcNAc as the donor in a beta-(1-->3) glycosylation reaction . The 6'-deoxy derivative was nearly threefold as active as phenyl beta-lactoside, whereas the 2'- and 4'-deoxy derivatives were less active . The other derivatives were inactive, as expected. APMIS, 2004 Dec, 112(11-12), 771 - 84 Molecular genetic methods for diagnosis and characterisation of Chlamydia trachomatis and Neisseria gonorrhoeae: impact on epidemiological surveillance and interventions; Fredlund H et al.; Fredlund H, Falk L, Jurstrand M, Unemo M . Molecular genetic methods for diagnosis and characterisation of Chlamydia trachomatis and Neisseria gonorrhoeae: impact on epidemiological surveillance and interventions . APMIS 2004;112:771-84.One of the mainstays in the prevention of Chlamydia trachomatis and Neisseria gonorrhoeae infections is the availability of laboratory diagnostics with high sensitivity and specificity . Assays for diagnosis of C . trachomatis include cell culture and nucleic acid amplification tests (NAATs) . The major target sequences for C . trachomatis diagnosis by NAATs are located at the cryptic plasmid and the major target used for characterisation is the omp1 gene . The gold standard for diagnosis of N . gonorrhoeae is culture . However, numerous NAATs for identification of N . gonorrhoeae and a number of molecular genetic methods for characterisation of N . gonorrhoeae have been developed . Probably no routine laboratory can attain as high sensitivity by culturing C . trachomatis or N . gonorrhoeae as by using NAATs . For that reason NAATs can be recommended for diagnosing C . trachomatis, but not as the only diagnostic assay for N . gonorrhoeae, due to lack of antibiotic susceptibility testing and specificity problems, most pronounced for pharyngeal and rectal samples . Genotyping of C . trachomatis or N . gonorrhoeae provides additional information for contact tracing . It is recommended for N . gonorrhoeae, at least in low prevalence geographic areas, but cannot today be recommended for C . trachomatis . This is due to the low genetic variability and hence the limited benefits for partner notification . However, genotyping of C . trachomatis may play an important role under special circumstances. Res Microbiol, 2005 Jan-Feb, 156(1), 17 - 29 A conserved polar region in the cell division site determinant MinD is required for responding to MinE-induced oscillation but not for localization within coiled arrays; Szeto J et al.; A region in the cell division site determinant MinD required for stimulation by MinE and which determines MinD topological specificity along coil-like structures has been identified . Structural modeling of dimeric MinD and sequence alignment of 24 MinD proteins revealed a conserved polar region in Gram-negative bacterial MinD proteins, corresponding to residues 92-94 of Neisseria gonorrhoeae MinD (MinD(Ng)) . Using MinD(Ng) as a paradigm for MinD functionality in Gram-negative organisms, mutation of these conserved residues did not abrogate MinD(Ng) self-association, nor its interaction with MinE(Ng) and the cell division inhibitor MinC . Although the MinD(Ng) mutant dimerized in the presence of ATP, its ATPase activity was not stimulated by MinE(Ng), unlike wild-type MinD(Ng) . GFP fusions to either MinD(Ng) or to Escherichia coli MinD bearing simultaneous or individual mutations to residues 92-94 localized within coiled arrays along the E . coli inner cell periphery, similar to wild-type GFP-MinD . However, unlike wild-type GFP-fusions, the mutant proteins were distributed uniformly throughout the array, despite the presence of MinE, which normally imparts topological specificity to MinD by inducing the latter to oscillate from pole-to-pole and away from midcell . Hence, despite localizing along the inner cell periphery as a polymeric structure, the mutant MinD proteins in this study have lost the ability to be efficiently stimulated by MinE(Ng), resulting in a loss of distinct pole-to-pole oscillation. J Clin Microbiol, 2005 Jan, 43(1), 368 - 75 Genetic typing of the porin protein of Neisseria gonorrhoeae from clinical noncultured samples for strain characterization and identification of mixed gonococcal infections; Lynn F et al.; Molecular methods that characterize the Neisseria gonorrhoeae porin protein Por are needed to study gonococcal pathogenesis in the natural host and to classify strains from direct clinical samples used with nucleic acid amplification-based tests . We have defined the capabilities of por variable region (VR) typing and determined suitable conditions to apply the method to direct clinical specimens . Nested PCR from spiked urine samples detected 1 to 10 copies of template DNA; freezing spiked whole urine greatly reduced the ability to amplify porB . In a laboratory model of mixed gonococcal infections, the por type of one strain could be determined in the presence of a 100-fold excess of another . por VR typing was used to examine clinical samples from women enrolled in studies conducted in Baltimore, Md., and Madagascar . por type was determined from 100% of paired cervical swab and wick samples from 20 culture-positive women from Baltimore; results for eight individuals (40%) suggested infection with more than one strain . In frozen urine samples from Madagascar, porB was amplified and typed from 60 of 126 samples from ligase chain reaction (LCR)-positive women and 3 samples from LCR-negative women . The por VR types of 13 samples (21%) suggested the presence of more than one gonococcal strain . Five por types, identified in >45% of women with typed samples, were common to both geographic areas . Molecular typing is an important adjunct to nucleic acid amplification-based diagnostics . Methods that utilize direct clinical samples and can identify mixed infections may contribute significantly to studies of host immunity, gonococcal epidemiology, and pathogenesis. J Clin Microbiol, 2005 Jan, 43(1), 199 - 207 Microbial DNA typing by automated repetitive-sequence-based PCR; Healy M et al.; Repetitive sequence-based PCR (rep-PCR) has been recognized as an effective method for bacterial strain typing . Recently, rep-PCR has been commercially adapted to an automated format known as the DiversiLab system to provide a reliable PCR-based typing system for clinical laboratories . We describe the adaptations made to automate rep-PCR and explore the performance and reproducibility of the system as a molecular genotyping tool for bacterial strain typing . The modifications for automation included changes in rep-PCR chemistry and thermal cycling parameters, incorporation of microfluidics-based DNA amplicon fractionation and detection, and Internet-based computer-assisted analysis, reporting, and data storage . The performance and reproducibility of the automated rep-PCR were examined by performing DNA typing and replicate testing with multiple laboratories, personnel, instruments, DNA template concentrations, and culture conditions prior to DNA isolation . Finally, we demonstrated the use of automated rep-PCR for clinical laboratory applications by using isolates from an outbreak of Neisseria meningitidis infections . N . meningitidis outbreak-related strains were distinguished from other isolates . The DiversiLab system is a highly integrated, convenient, and rapid testing platform that may allow clinical laboratories to realize the potential of microbial DNA typing. J Clin Microbiol, 2005 Jan, 43(1), 127 - 31 Ability of new APTIMA CT and APTIMA GC assays to detect Chlamydia trachomatis and Neisseria gonorrhoeae in male urine and urethral swabs; Chernesky MA et al.; A clinical evaluation was conducted in six North American centers to determine the ability of APTIMA CT (ACT) and APTIMA GC (AGC) nucleic acid amplification assays to detect Chlamydia trachomatis and Neisseria gonorrhoeae infections in 1,322 men by testing their urethral swabs and first-catch urine (FCU) . The results obtained with ACT and AGC assays were compared to an infected patient status determined by testing the specimens with the APTIMA Combo 2 and the BD ProbeTec energy transfer multiplex assays . Symptoms did not influence the values . Positive and negative agreements of the ACT and AGC assays for individual specimens were high, with each comparator assay ranging between 94.3 and 100% for positives and 93.9 and 99.4% for negatives . The ACT and AGC assays performed on noninvasive specimens such as FCU effectively identified C . trachomatis or N . gonorrhoeae infections in symptomatic and asymptomatic men and should be suitable for screening male populations. J Infect Dis, 2005 Feb 1, 191(3), 333 - 8 Epub 2004 Dec 22. Contribution of HIV-1 Infection to Acquisition of Sexually Transmitted Disease: A 10-Year Prospective Study; McClelland RS et al.; Background . Sexually transmitted diseases (STDs) enhance human immunodeficiency virus (HIV)-1 susceptibility, but few studies have examined the reciprocal effect of HIV-1 on STD acquisition.Methods . Data from a prospective cohort study conducted among female sex workers in Mombasa, Kenya between 1993 and 2003 were used to determine the effect of HIV-1 infection on STD susceptibility . The cohort included 1215 HIV-1-seronegative women who underwent monthly HIV-1 and STD screening, of whom 238 experienced seroconversion to HIV-1 during follow-up . Andersen-Gill proportional-hazards models were used to compare the incidence rates for genital-tract infections (syphilis, genital ulcer disease {GUD}, Neisseria gonorrhoeae infection, Chlamydia trachomatis infection, Trichomonas vaginalis infection, vulvovaginal candidiasis, and bacterial vaginosis) in HIV-1-seropositive versus HIV-1-seronegative women, after controlling for sexual behavior and other potential confounding factors.Results . HIV-1 infection was associated with a significantly higher incidence of GUD (hazard ratio {HR}, 2.8; 95% confidence interval {CI}, 2.0-3.9), gonorrhea (HR, 1.6; 95% CI, 1.1-2.2), and vulvovaginal candidiasis (HR, 1.5; 95% CI, 1.3-1.8) . The risks of GUD and vulvovaginal candidiasis increased with progressive levels of immunosuppression.Conclusions . The increased incidence of genital-tract infections among HIV-1-seropositive women could promote the spread of both HIV-1 and other STDs, particularly in areas where these conditions are highly prevalent. Am J Epidemiol, 2005 Jan 15, 161(2), 186 - 95 Douching, pelvic inflammatory disease, and incident gonococcal and chlamydial genital infection in a cohort of high-risk women; Ness RB et al.; Douching has been linked to gonococcal or chlamydial cervicitis and pelvic inflammatory disease (PID) in retrospective studies . The authors conducted a 1999-2004 prospective observational study of 1,199 US women who were at high risk of acquiring chlamydia and were followed for up to 4 years . Cervical Neisseria gonorrhoeae and Chlamydia trachomatis were detected from vaginal swabs by nucleic acid amplification . PID was characterized by histologic endometritis or pelvic pain and tenderness plus one of the following: oral temperature >38.3 degrees C, leukorrhea or mucopus, erythrocyte sedimentation rate >15 mm/hour, white blood cell count >10,000, or gonococcal/chlamydial lower genital tract infection . Associations between douching and PID or gonococcal/chlamydial genital infections were assessed by proportional hazards models . The 4-year incidence rate of PID was 10.9% and of gonococcal and/or chlamydial cervicitis was 21.9% . After adjustment for confounding factors, douching two or more times per month at baseline was associated with neither PID (adjusted hazard ratio = 0.76, 95% confidence interval: 0.42, 1.38) nor gonococcal/chlamydial genital infection (adjusted hazard ratio = 1.16, 95% confidence interval: 0.76, 1.78) . Frequency of douching immediately preceding PID or gonococcal/chlamydial genital infection was not different between women who developed versus did not develop outcomes . These data do not support an association between douching and development of PID or gonococcal/chlamydial genital infection among predominantly young, African-American women. Jpn J Infect Dis, 2004 Dec, 57(6), 253 - 6 Neisseria gonorrhoea, Chlamydia trachomatis, and Treponema pallidum Infection in Antenatal and Gynecological Patients at Korle-Bu Teaching Hospital, Ghana; Apea-Kubi KA et al.; Five hundred and seventeen women attending the gynecology and obstetrics clinics of the Korle-Bu Teaching Hospital were examined for sexually transmitted infections (STIs) . Vaginal swabs were examined for Trichomonas vaginalis, Candida albicans, and Gardnerella vaginalis infection . Endocervical swabs were examined for Neisseria gonorrhoea and Chlamydia trachomatis using a recently developed RNA detection kit . Strain typing was performed to identify serovars of C . trachomatis . Sera were analyzed for Treponema pallidum with a passive-particle agglutination assay kit . The prevalence of infection with N . gonorrhoea was 0.6%, C . trachomatis 3.0%, and T . pallidum 5.6% . Eight samples were PCR-positive for C . trachomatis . Five of these were serovar G, and the rest were serovar E . All cases of mixed infections occurred in pregnant women . In conclusion, a high transmissible risk of T . pallidum infection was observed among our study population and in particular among our pregnant women . The absence of association between the presenting symptoms, clinical findings, and specific pathogens has implications for the syndromic approach to STI case management . The low prevalence of C . trachomatis and N . gonorrhoea may be due to self medication and requires further research in primary health institutions in rural areas to compare rates. Int J Antimicrob Agents, 2005 Jan, 25(1), 68 - 74 Clarithromycin treatment selects for persistent macrolide-resistant bacteria in throat commensal flora; Jonsson M et al.; The aim of the study was to determine the effect of clarithromycin treatment on resistance development in the commensal throat flora . Alpha-haemolytic streptococci and Neisseria spp . were isolated from patients receiving clarithromycin for eradication of Helicobacter pylori . The treatment resulted in an immediate increase in the number of macrolide-resistant streptococci, which remained for one year after treatment, but declined to background level three years later . The most prevalent resistance gene was mef(A) . Neisseria spp . were less affected by the treatment: the number of resistant isolates increased in only in one case during treatment . In conclusion, a one-week standard therapy with clarithromycin selects for an increased prevalence of macrolide-resistant streptococci that persisted for more than one year. Antimicrob Agents Chemother, 2005 Jan, 49(1), 137 - 43 Emergence and spread of Neisseria gonorrhoeae clinical isolates harboring mosaic-like structure of penicillin-binding protein 2 in Central Japan; Ito M et al.; Of 150 clinical isolates of Neisseria gonorrhoeae recovered in 2001, we examined 55 clinical isolates of N . gonorrhoeae for which cefixime MICs were > or=0.125 microg/ml and randomly selected 15 isolates for which cefixime MICs were < or =0.06 microg/ml for analysis of alterations in the penicillin-binding protein 2 (PBP 2) gene . We found insertion of an extra codon (Asp-345a) in the transpeptidase domain of PBP 2, and this insertion occurred alone or in conjunction with other amino acid substitutions . We also found a mosaic PBP 2 that was composed of fragments of the PBP 2 proteins from Neisseria cinera and Neisseria perflava . This mosaic PBP 2 was significantly associated with decreased susceptibilities to penicillin and cephalosporins, especially oral cephalosporins . For most of the isolates with a mosaic PBP 2, the cefixime MICs were > or =0.5 microg/ml and the cefdinir MICs were > or =1 microg/ml . Analysis of chromosomal DNA restriction patterns by pulsed-field gel electrophoresis revealed that most isolates with the mosaic PBP 2 were genetically similar . The recombination events that generated the mosaic PBP 2 would likely have contributed to the decreased sensitivities to cephalosporins . Isolates with the mosaic PBP 2 appear to threaten the efficacy of the currently recommended regimen with cefixime . The emergence of such strains may be the result of the in vivo generation of clones in which interspecies recombination occurred between the penA genes of N . gonorrhoeae and commensal Neisseria species. Clin Infect Dis, 2005 Jan 1, 40(1), 188 - 92 Epub 2004 Dec 01. High prevalence of ciprofloxacin-resistant Neisseria gonorrhoeae in Northern Taiwan; Hsueh PR et al.; Among 55 preserved isolates collected in northern Taiwan from 1999 through 2003, ciprofloxacin resistance (minimum inhibitory concentration, >or=1 microg/mL) was found in 1 (25%) of 4 isolates obtained in 1999-2000 and in 27 (93.1%) of 29 isolates obtained in 2003 . Pulsed-field gel electrophoresis analysis indicated that several clones predominated among the ciprofloxacin-resistant isolates. Sex Transm Dis, 2005 Jan, 32(1), 13 - 19 Sexually Transmitted Infections Among Brothel-Based Sex Workers in Bangladesh: High Prevalence of Asymptomatic Infection; Nessa K et al.; OBJECTIVE AND GOAL:: The goal of this study was to study the prevalence of sexually transmitted infections (STIs)/reproductive tract infections (RTIs) among brothel-based sex workers (SWs) in Bangladesh . STUDY:: A cross-sectional study was conducted among SWs in 4 randomly selected brothels . A sociodemographic and behavioral survey and pelvic examination was conducted . Specimens including endocervical swab, high vaginal swab, and blood were collected and were examined for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, bacterial vaginosis candidiasis, and syphilis . RESULTS:: A total of 439 SWs were enrolled and 49.6% had genital symptoms . Among all SWs, 17.5% were positive for N . gonorrhoeae; 15.5% for C . trachomatis; 7.5% for T . vaginalis, and 6.6% had active syphilis . A total of 67.4% SWs were positive for at least 1 cervical and/or vaginal infection . CONCLUSION:: The prevalence of STIs/RTIs among SWs in brothels in Bangladesh is high . An intervention strategy addressing both symptomatic and asymptomatic infections and periodic screening of SWs for RTIs/STIs is essential for successful HIV and STIs prevention programs. Mol Microbiol, 2005 Jan, 55(1), 65 - 77 PilX, a pilus-associated protein essential for bacterial aggregation, is a key to pilus-facilitated attachment of Neisseria meningitidis to human cells; Helaine S et al.; Summary The attachment of pathogenic Neisseria species to human cells, in which type IV pili (Tfp) play a key but incompletely defined role, depends on the ability of these bacteria to establish contacts with the target cells but also interbacterial interactions . In an effort to improve our understanding of the molecular mechanisms of N . meningitidis adherence to human cells, we screened a collection of defined mutants for those presenting reduced attachment to a human cell line . Besides underscoring the central role of Tfp in this process, this analysis led to the identification of mutants interrupted in a novel gene termed pilX, that displayed an adherence as impaired as that of a non-piliated mutant but quantitatively and qualitatively unaltered fibres . Moreover, the pilX gene, which encodes a pilin-like protein that copurifies with Tfp fibres, was also found to be essential for bacterial aggregation . We provide here several piece of evidence suggesting that PilX has intrinsic aggregative but no adhesive properties and that the reduced numbers of adherent bacteria seen with a pilX mutant result from the absence of interbacterial interactions . These data extend the current model for Tfp-facilitated adherence of N . meningitidis to human cells by suggesting that the pili lead to an increase in net initial adherence primarily by mediating a cooperation between the bacteria, which is supported by the finding that a major effect on initial adherence could be observed in a wild-type (WT) genetic background after a mechanical removal of the bacterial aggregates. Mol Microbiol, 2005 Jan, 55(1), 54 - 64 Type IV pilus biogenesis in Neisseria meningitidis: PilW is involved in a step occurring after pilus assembly, essential for fibre stability and function; Carbonnelle E et al.; Summary Type IV pili (Tfp) play a critical role in the pathogenic lifestyle of Neisseria meningitidis and N . gonorrhoeae, notably by facilitating bacterial attachment to human cells, but our understanding of their biogenesis, during which the fibres are assembled in the periplasm, then emerge onto the cell surface and are stabilized, remains fragmentary . We therefore sought to identify the genes required for Tfp formation in N . meningitidis by screening a genome-wide collection of mutants for those that were unable to form aggregates, another phenotype mediated by these organelles . Fifteen proteins, of which only seven were previously characterized, were found to be essential for Tfp biogenesis . One novel component, named PilW, was studied in more detail . We found that PilW is an outer-membrane protein necessary for the stabilization of the fibres but not for their assembly or surface localization, because Tfp could be restored on the surface in a pilW mutant by a mutation in the twitching motility gene pilT . However, Tfp-linked properties, including adherence to human cells, were not restored in a pilW/T mutant, which suggests that PilW is also essential for the functionality of the fibres . Together with the finding that PilW is important for the stability of PilQ multimers, our results extend the current model for Tfp biogenesis by suggesting that a multiprotein machinery in the outer-membrane is involved in the terminal stage of Tfp biogenesis during which growing fibres are not only stabilized, but also become perfectly functional. Sex Transm Dis, 2004 Dec, 31(12), 734 - 9 Sex partner concurrency, geographic context, and adolescent sexually transmitted infections; Jennings J et al.; BACKGROUND: Geographic areas characterized by a high prevalence of sexually transmitted infections (STIs) are critical to the maintenance and persistence of STIs within populations . Sex partner concurrency has been shown to be associated with increased risk for individual-level STIs . OBJECTIVES: The objectives of this study were to determine whether gonorrhea rate per census block group and sex partner concurrency independently and interactively are associated with a current bacterial STI among adolescents . STUDY: Face-to-face interviews and urine testing for Chlamydia trachomatis and Neisseria gonorrhoeae were conducted among female, sexually active, 14- to 19-year-olds presenting for reproductive clinic care between August 2000 and June 2002 . RESULTS: Gonorrhea rate per census block group and sex partner concurrency were not independently but were interactively associated with a current bacterial STI . Among participants with a main sex partner who practiced concurrency, living in high-prevalence geographic areas was significantly associated with a current bacterial STI . CONCLUSIONS: The results suggest that geographic context may moderate an adolescent sex partner's behaviors . The research adds to the basic understanding of sexually transmitted disease transmission and acquisition in a high-prevalence inner-city setting. Sex Transm Dis, 2004 Dec, 31(12), 702 - 8 Fluoroquinolone resistance among Neisseria gonorrhoeae isolates in Hawaii, 1990-2000: role of foreign importation and increasing endemic spread; Iverson CJ et al.; OBJECTIVES: In 1999, an increase in ciprofloxacin-resistant Neisseria gonorrhoeae isolates was identified in Hawaii, prompting initiation of investigative studies . GOALS: The goal of this study was epidemiologic evaluation of this increase . STUDY: The authors conducted a review of laboratory data; case-series and case-control studies based on medical record review; and a prospective case-control study based on patient interviews . RESULTS: A total of 10.4% (21 of 201) of gonococcal isolates from Hawaii in 2000 were ciprofloxacin-resistant compared with <1.5% per year from 1990 to 1997 . From medical record review for patients diagnosed with ciprofloxacin-resistant N . gonorrhoeae infection from 1990 to 1999, 59% were Asian/Pacific Islanders and 91% were heterosexual . From review of 1998 and 1999 sexually transmitted disease (STD) clinic medical records, patients with ciprofloxacin-resistant N . gonorrhoeae were more likely to report recent foreign travel or a sex partner with recent foreign travel than patients with ciprofloxacin-susceptible N . gonorrhoeae (6 of 12 vs . 10 of 117, P <0.001), but 50% (6 of 12) acquired a ciprofloxacin-resistant strain locally from a partner with no recent travel . In 2000, 70% (7 of 10) of STD clinic patients with ciprofloxacin-resistant N . gonorrhoeae acquired their infection locally from partners with no reported recent travel . CONCLUSIONS: Infections with ciprofloxacin-resistant N . gonorrhoeae are increasing and evolving in Hawaii. Urol Int, 2004, 73(4), 358 - 60 Periurethral abscess formation due to Neisseria gonorrhoeae; Kraus S et al.; Abscess formation secondary to chronic urethritis of the posterior male urethra caused by Neisseria gonorrhoeae has become an atypical and rare urological complication . Periurethral abscess formation has been reported to be associated with gonococcal infections, but in such rare cases a delay in the diagnosis of gonococcal infection seems to have been the cause for this periurethral complication . A case with a primary diagnosis of periurethral abscess with N . gonorrhoeae infection is reported--successful treatment included transurethral incision and antibiotic therapy . 2004 S . Karger AG, Basel. J Bacteriol, 2005 Jan, 187(1), 392 - 5 Proteomics analysis by two-dimensional differential gel electrophoresis reveals the lack of a broad response of Neisseria meningitidis to in vitro-produced AI-2; Schauder S et al.; To investigate the effect of the autoinducer AI-2 on protein expression in Neisseria meningitidis, a luxS mutant of strain MC58 was grown in the presence and absence of in vitro-produced AI-2, and differential protein expression was assessed by two-dimensional differential gel electrophoresis . N . meningitidis did not show a global response to AI-2 signaling activity. Int J STD AIDS, 2004 Dec, 15(12), 822 - 8 Screening of males for Chlamydia trachomatis and Neisseria gonorrhoeae infections at STD clinics in three US cities -- Indianapolis, New Orleans, Seattle; Kohl KS et al.; We assessed prevalence and risk factor data for men routinely screened for Chlamydia trachomatis and Neisseria gonorrhoeae in STD clinics in four US cities from May 1995-March 1999 . Data were analysed separately for 'test-visits' (self-reported symptoms, clinical signs or sexual contact to an STD) and 'screen-visits' (STD screen only) for 32,595 men with 45,390 visits . Among test-visits in Seattle, Indianapolis and New Orleans, 8.7% (807/9285), 15.3% (1305/8519), and 10.1% (1551/15,296) of men were positive for C . trachomatis, and 10.2% (773/7543), 24.9% (2108/8478), and 30.4% (4746/ 15,629) for N . gonorrhoeae . Among screen-visits, 2.1% (88/4103), 7.3% (130/1790), and 5.6% (292/5183) of men were positive for C . trachomatis, and 1.8% (46/2576), 1.7% (31/ 1786), and 5.2% (274/5235) for N . gonorrhoeae . Positivity for screen-visits was particularly high among young men (15-24 years), and those reporting > 1 sex partner in the past 60 days . Substantial variation among sites in positivity warrants local determination of prevalence and risk factors to inform screening strategies. Eur J Clin Microbiol Infect Dis . 2004 Dec 15; {Epub ahead of print} Orogenital transmission of Neisseria meningitidis serogroup C confirmed by genotyping techniques; Urra E et al.; Urethritis caused by Neisseria meningitidis in heterosexual patients is presumed to occur via orogenital contact, but confirmation has not been possible in most cases . Presented here is a case of urethritis caused by N . meningitidis, serogroup C, and the isolation of the same microorganism from the nasopharynx and endocervix of the patient's sexual partner . The similarity of the urethral and nasopharyngeal isolates' electrophoretic patterns, obtained using pulsed-field gel electrophoresis, proves the infection was transmitted via orogenital contact. Zhonghua Nan Ke Xue, 2004 Nov, 10(11), 815 - 8 {Analysis of the infection of sexually transmitted diseases among high-risk population in Nanjing}; Huang Y et al.; OBJECIVE: To survey the prevalence of sexually transmitted diseases among high-risk population in Nanjing . METHODS: A high-risk population of 1539 underwent physical examinations from September 2001 to May 2004 . The results were recorded, the urogenital swabs collected to further detect Neisseria gonorrhoeae (GN), Chlamydia trachomatis (Ct), Ureaplasma urealyticum (Uu), and the blood specimens tested for syphilis and HIV antibodies . RESULTS: The infection rates were 13.5% for GN, 10.3% for syphilis, 2.3% for condyloma acuminata, 35.9% for Ct and 22.4% for Uu respectively . Nongonococcal urethritis (NUG) was shown to be the most prevalent STD in the group . No subject was found to be positive with HIV antibody . CONCLUSION: Screening strategy in the highrisk population is useful and efficient in controlling the spread of STDs and HIV. Biosens Bioelectron, 2005 Jan 15, 20(7), 1341 - 8 Micromachined polymerase chain reaction system for multiple DNA amplification of upper respiratory tract infectious diseases; Liao CS et al.; This paper presents a micro polymerase chain reaction (PCR) chip for the DNA-based diagnosis of microorganism genes and the detection of their corresponding antibiotic-resistant genes . The micro PCR chip comprises cheap biocompatible soda-lime glass substrates with integrated thin-film platinum resistors as heating/sensing elements, and is fabricated using micro-electro-mechanical-system (MEMS) techniques in a reliable batch-fabrication process . The heating and temperature sensing elements are made of the same material and are located inside the reaction chamber in order to ensure a uniform temperature distribution . This study performs the detection of several genes associated with upper respiratory tract infection microorganisms, i.e . Streptococcus pneumoniae, Haemopilus influenze, Staphylococcu aureus, Streptococcus pyogenes, and Neisseria meningitides, together with their corresponding antibiotic-resistant genes . The lower thermal inertia of the proposed micro PCR chip relative to conventional bench-top PCR systems enables a more rapid detection operation with reduced sample and reagent consumption . The experimental data reveal that the high heating and cooling rates of the system (20 and 10 degrees C/s, respectively) permit successful DNA amplification within 15min . The micro PCR chip is also capable of performing multiple DNA amplification, i.e . the simultaneous duplication of multiple genes under different conditions in separate reaction wells . Compared with the large-scale PCR system, it is greatly advantageous for fast diagnosis of multiple infectious diseases . Multiplex PCR amplification of two DNA segments in the same well is also feasible using the proposed micro device . The developed micro PCR chip provides a crucial tool for genetic analysis, molecular biology, infectious disease detection, and many other biomedical applications. Mol Immunol, 2005 Feb, 42(3), 335 - 344 Molecular analysis of monoclonal antibodies to group variant capsular polysaccharide of Neisseria meningitidis: recurrent heavy chains and alternative light chain partners; Berry JD et al.; We determined the molecular sequence of monoclonal antibodies (mAbs) to serogroups B and C capsular polysaccharides (PS) of Neisseria meningitidis . N . meningitidis infections are a leading cause of bacterial septicemia and meningitis in humans . Antibodies to PS are fundamental to host defense and diagnostics . The polysaccharide capsule of group B N . meningitidis is poorly immunogenic and thus is an important model for studying pathogen-host co-evolution through understanding the molecular basis of the host immune response . We used a modified reverse-transcriptase PCR to amplify and sequence the V-genes of murine hybridomas produced against types B and C capsular PS . Databank analysis of the sequences encoding the V-genes of type C capsular PS mAb, 4-2-C, reveal that heavy chain alleles are recurrently used to encode this specificity in mice . Interestingly, a V-gene from the same germline family also encodes the V-domain of mAbs 2-2-B, which targets the antigenically distinct serogroup B capsular PS . Somatic mutation, junctional diversity and alternative light chains collectively impart the specificity for these serologically distinct epitopes . Knowledge of the specific immunoglobulin genes used to target common bacterial virulence factors may lead to insights on pathogen-host co-evolution, and the potential use of this information in pre-symptomatic diagnosis is discussed. West Afr J Med, 2004 Jul-Sep, 23(3), 215 - 20 A case control study of ophthalmia Neonatorum in Kaduna II: causative agents and their antibiotic sensitivity; Olatunji FO; AIM: To identify the causative agents of ophthalmia neonatorum and their antibiotic sensitivity in the hospitals of study . PATIENTS AND METHODS: A case-control study of ophthalmia neonatorum was carried out in 6 hospitals in Kaduna metropolis in northern Nigeria . Two hundred cases were matched with two hundred controls . RESULTS: The male to female ratio was 1.1:1 . The mean age of the patients was 8.9 days, and range was birth to 28 days . The age at the onset of the symptoms was birth to 27 days with a mean of 3.9 days . The most common pathogen isolated in this study was Staphylococcus aureus, both in cases (69.7%) and in controls (48%) . Only one case of Neisseria gonorrhoea was found . Thirteen and four E . coli were isolated from cases and control while 7 and 31 Staphylococcus epidermidis were isolated from cases and controls respectively . Giemsa staining carried out in only 3 out of the 6 hospitals yielded 9 and 3 isolates of Chlamydia trachomatis in cases and controls respectively . The percentage sensitivity of Staphylococcus aureus to penicillin, chloramphenicol, tetracycline, erythromycin and gentamicin were 3, 73, 37, 59 and 77 respectively . The corresponding sensitivity of E . coli to the above antibiotics was 0, 69, 46, 10 and 62 respectively . CONCLUSION: Most of the cases of ophthalmia neonatorum are mild. J Clin Microbiol, 2004 Dec, 42(12), 5819 - 24 Detection of gyrA and parC mutations associated with ciprofloxacin resistance in Neisseria gonorrhoeae by use of oligonucleotide biochip technology; Zhou W et al.; An oligonucleotide biochip that specifically detects point mutations in the gyrA and parC genes of Neisseria gonorrhoeae was designed and subsequently evaluated with 87 untreated clinical specimens . The susceptibilities of the N . gonorrhoeae strains were tested to determine the prevalence of ciprofloxacin-resistant strains in Anhui Province, People's Republic of China . Conventional DNA sequencing was also performed to identify mutations in gyrA and parC and to confirm the biochip data . The study demonstrates that all of the point mutations in the gyrA and parC genes of N . gonorrhoeae were easily discriminated by use of the oligonucleotide biochip . Fifteen different alteration patterns involved in the formation of ciprofloxacin resistance were identified by the biochip assay . Double mutations in both Ser91 and Asp95 of the GyrA protein were seen in all nonsensitive isolates . Double mutations in Ser91 and Asp95 of GyrA plus mutation of Glu91 or Ser87 of the ParC protein lead to significant high-level resistance to ciprofloxacin in N . gonorrhoeae isolates . The results obtained by use of the oligonucleotide biochip were identical to those obtained by use of DNA sequencing . In conclusion, the oligonucleotide biochip technology has potential utility for the rapid and reliable identification of point mutations in the drug resistance genes of N . gonorrhoeae. J Clin Microbiol, 2004 Dec, 42(12), 5751 - 6 Direct comparison of the BD ProbeTec ET system with in-house LightCycler PCR assays for detection of Chlamydia trachomatis and Neisseria gonorrhoeae from clinical specimens; Koenig MG et al.; The commercial BD ProbeTec ET (BDPT) system for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis from clinical specimens was compared with our in-house LightCycler real-time PCR (LC-PCR) assays . Specimens initially positive by the BDPT system were retested by our LC-PCR assays . Our results for C . trachomatis testing indicate a 91.2% agreement when the results of 114 clinical specimens, initially positive by BDPT over a wide range of method-other-than-acceleration (MOTA) scores, were retested by our LC-PCR assay . The agreement between the two systems improved to 96% when only MOTA scores of >30,000 were retested by the LC-PCR assay . The overall agreement between the two systems for Neisseria gonorrhoeae detection from 155 clinical specimens was only 77.4%, with agreement particularly low (24.1%) for MOTA scores ranging from 2,000 to 19,999 . Repeat testing of specimens with the BDPT only closely correlated with that seen by others demonstrating that reproducibility of the BDPT system for specimens initially within the MOTA score range from 2,000 to 9,999 is problematic, especially for Neisseria gonorrhoeae testing . With our study, we proposed an algorithm for C . trachomatis and N . gonorrhoeae testing which involves screening with the BDPT system followed by selective use of our in-house LC-PCR assays. J Pediatr Adolesc Gynecol, 2004 Oct, 17(5), 331 - 9 Comparison of nucleic acid amplification tests and culture techniques in the detection of Neisseria gonorrhoeae and Chlamydia trachomatis in victims of suspected child sexual abuse; Kellogg ND et al.; STUDY OBJECTIVES: (1) To identify factors predictive for gonorrhea and chlamydia positivity by LCR testing based on history and physical findings encountered during the sexual abuse evaluations . (2) To compare Ligase Chain Reaction (LCR), Polymerase Chain Reaction (PCR), and culture methods in the detection of chlamydia and gonorrhea infection among prepubertal and adolescent girls referred for sexual abuse evaluations . DESIGN: Prevalence odds ratios and logistic regression analysis were used to identify factors among patients' physical symptoms and signs, history of sexual activity, and abuse characteristics that were associated with positive test results for gonorrhea and chlamydia . The Kappa statistic was used to perform pairwise comparisons of LCR, PCR, and culture identification of gonorrhea and chlamydia infection . SETTING: A specialized sexual abuse clinic in San Antonio, Texas . PARTICIPANTS: A consecutive sample of 229 girls between the ages of 6 and 20 who reported, or had indicators of, abusive genital-genital or genital-anal contact . MAIN OUTCOME MEASURES: Patients' history and physical findings predicting positive test results for gonorrhea and chlamydia infection; and relative sensitivity of testing sites (vaginal swab and urine) and methodologies (LCR, PCR, and culture) in identifying gonorrhea and chlamydia infection . RESULTS: (1) Gonorrhea infection: 3.2% of subjects were positive for gonorrhea by LCR at one or more sites; 2.4% had positive gonorrhea cultures . There was excellent agreement between vaginal swab LCR and PCR; agreement between urine samples was limited by the small number of positive tests . The sole factor that predicted gonorrhea positivity was increased number of white blood cells seen on wet mount . (2) Chlamydia infection: 11.1% of subjects were positive for chlamydia by at least one LCR test; only 0.8% had positive chlamydia cultures . Both urine and vaginal swab testing showed good agreement between PCR and LCR but not between culture and either of the newer methodologies . Factors that predicted chlamydia positivity were: patient history of consensual sexual contact, patient history of vaginal discharge, and the presence of concerning or definitive findings of genital trauma . CONCLUSIONS: While LCR, PCR, and culture techniques appeared comparable for detecting gonorrhea, LCR techniques detected significantly more patients with chlamydia infection when compared with the culture technique . PCR was comparable to LCR in detecting chlamydia infection . The LCR vaginal swab detected more patients with chlamydia and gonorrhea than the LCR urine sample . Risk factors for chlamydia and gonorrhea infection were present in most, but not all, of the children with positive LCR findings . LCR and PCR appear to detect more chlamydial and gonorrheal infections than do cultures. AIDS, 2004 Nov 5, 18(16), 2179 - 84 Hormonal contraception and risk of cervical infections among HIV-1-seropositive Kenyan women; Lavreys L et al.; OBJECTIVE: To evaluate the relationship between hormonal contraceptive use and the acquisition of cervical sexually transmitted infections (STI) among HIV-1-infected women . DESIGN: A prospective cohort study of 242 commercial sex workers in Mombasa, Kenya, followed from the time of HIV-1 infection . METHODS: At monthly follow-up visits, sexual behavior and contraceptive use were recorded, and laboratory screening for STI was performed . Multivariate Andersen-Gill proportional hazards models were constructed to examine the association between the use of hormonal contraception and the occurrence of cervical STI . RESULTS: The median duration of follow-up after HIV-1 acquisition was 35 months, and 799 person-years of follow-up were accrued . After adjustment for demographic factors and sexual behavior, women using the injectable contraceptive depot medroxyprogesterone acetate were at increased risk of Chlamydia trachomatis infection {hazard ratio (HR) 3.1, 95% confidence interval (CI) 1.0-9.4, P = 0.05} and cervicitis (HR 1.6, 95% CI 1.0-2.3, P = 0.03) compared with women using no contraception . The use of oral contraceptive pills was associated with an increased risk of cervicitis (HR 2.3, 95% CI 1.4-3.8, P = 0.001) . Hormonal contraception was not associated with an increased risk of infection with Neisseria gonorrhoeae . CONCLUSION: The use of hormonal contraception by HIV-1-infected women was associated with an increased risk of cervicitis and cervical chlamydia infection . HIV-1-seropositive women using hormonal contraception should be counseled about the importance of consistent condom use to prevent both STI and HIV-1 transmission. J Bacteriol, 2004 Dec, 186(24), 8443 - 52 The RpoH-mediated stress response in Neisseria gonorrhoeae is regulated at the level of activity; Laskos L et al.; The general stress response in Neisseria gonorrhoeae was investigated . Transcriptional analyses of the genes encoding the molecular chaperones DnaK, DnaJ, and GrpE suggested that they are transcribed from sigma32 (RpoH)-dependent promoters upon exposure to stress . This was confirmed by mutational analysis of the sigma32 promoter of dnaK . The gene encoding the gonococcal RpoH sigma factor appears to be essential, as we could not isolate viable mutants . Deletion of an unusually long rpoH leader sequence resulted in elevated levels of transcription, suggesting that this region is involved in negative regulation of RpoH expression during normal growth . Transcriptional analyses and protein studies determined that regulation of the RpoH-mediated stress response is different from that observed in most other species, in which regulation occurs predominantly at the transcriptional and translational levels . We suggest that an increase in the activity of preformed RpoH is primarily responsible for induction of the stress response in N . gonorrhoeae. Glycobiology . 2004 Dec 1; {Epub ahead of print} Inner core assembly and structure of the lipooligosaccharide of Neisseria meningitidis: capacity of strain NMB to express all known immunotype epitopes; Kahler CM et al.; Neisseria meningitidis expresses a heterogeneous population of lipooligosaccharide 3 (LOS) inner cores variously substituted with alpha1-3 linked glucose and O-3, O-6, and O-7 linked phosphoethanolamine (PEA), as well as glycine, attached to HepII . Combinations of these attachments to the LOS inner core represent immunodominant epitopes that are being exploited as future vaccine candidates . Historically, each LOS immunotype was structurally assessed and prescribed a certain unique inner core epitope . We report that a single isolate, strain NMB, possesses the capacity to produce all of the known neisserial LOS inner core immunotype structures . Analysis of the inner cores from parental LOS revealed the presence or absence of alpha1,3 linked glucose, O-6 and/or O-7 linked PEA, in addition to glycine attached at the 7 position of the HepII inner core . Identification and inactivation of lpt-6 in strain NMB resulted in the loss of both O-6 and O-7 linked PEA groups from the LOS inner core, suggesting that Lpt-6 of strain NMB may have bifunctional transferase activities or that the O-6 linked PEA groups once attached to the inner core undergo non-enzymatic transfer to the O-7 position of HepII . Although O-3 linked PEA was not detected in parental LOS inner cores devoid of alpha1-3 linked glucose residues, LOS glycoforms bearing O-3 PEA groups accumulated in a truncated mutant, NMBlgtK (Hep2Kdo2-lipidA) . Since these structures disappeared upon inactivation of the lpt-3 locus, strain NMB expresses a functional O-3 PEA transferase . The LOS glycoforms expressed by NMBlgtK were also devoid of glycine attachments, indicating that glycine was added to the inner core after the completion of the gamma-chain by LgtK . In conclusion, strain NMB has the capability to express all known inner core structures, but in in vitro culture L2 and L4 immunotype structures are predominantly expressed. East Mediterr Health J, 2002 Nov, 8(6), 794 - 804 {Urethral discharge in Morocco: prevalence of microorganisms and susceptibility of gonococcos}; Alami K et al.; We studied 422 patients with urethral discharge recruited from 4 sentinel sites in Morocco to determine sociodemographic characteristics, history of STI infection, infecting organism and antibiotic susceptibility of Neisseria gonorrhoeae . The mean age of the sample was 28 years (range 16-67 years), and most were single, had multiple sex partners without taking protective measures and came from all social backgrounds; 59.9% had a history of a previous STI . The majority (87%) of the infections were the acute form . By polymerase chain reaction of urine samples of 399 patients, 41.6% had N . gonorrhoeae infection, 6.3% Chlamidia trachomatis and 10.8% both organisms; in 41.4% no organism was identified . N . gonorrhoeae was strongly susceptible to ciprofloxacin. Pharmacoepidemiol Drug Saf . 2004 Nov 25; {Epub ahead of print} From suspicion to action: the chemical conjunctivitis and silver nitrate connexion example in Brazilian hospitals; Napchan BM et al.; BACKGROUND: During the 20th century, silver nitrate (SN) eye drops instillation to newborns had been the best prophylaxis against ophthalmia neonatorum (ON) caused by Neisseria gonorrheae, the most frequent cause of ocular infections leading to blindness . At present, this treatment has been questioned because there is a growing prevalence of other pathogens such as Chlamydia trachomatis, and SN is associated with chemical conjunctivitis (ChC) . In addition, SN could present some conservation problems in tropical climates . Among other alternative drugs, 2.5% povidone-iodine has a proven efficacy, not only against Neisseria, but also against Chlamydia, has no conservation problems, has not been associated with ChC and is cheap . METHODS: The recently created Setor de Farmacovigilancia (SF) of the Sao Paulo State in Brazil has created a pharmacovigilance network of 11 big public hospitals . During a periodical signal searching process, the SF found a cluster of 33 ChC reports from one of the network hospitals . RESULTS: The problem was discussed with the remaining participant hospitals and this signal was used as a way to strengthen the network . Thirteen months later, 622 ChC reports were received from six hospitals . DISCUSSION: Thus, this well-known side-effect in the literature was highlighted as a 'real' problem in Brazil, and some participant hospitals began a discussion period together with the delivery and newborn care professionals in order to switch SN by povidone-iodine . This is an example of both, how a simple pharmacovigilance exercice could improve the implication of health professionals with their own therapeutic problems and how a pharmacovigilance network could be strengthened . Copyright (c) 2004 John Wiley & Sons, Ltd. Br J Anaesth, 2005 Feb, 94(2), 222 - 228 Epub 2004 Nov 19. Release of tumour necrosis factor {alpha} (TNF{alpha}) by TNF{alpha} cleaving enzyme (TACE) in response to septic stimuli in vitro; Robertshaw HJ et al.; BACKGROUND: Tumour necrosis factor alpha (TNFalpha), in its soluble form (solTNF), has been well described as an important cytokine in inflammatory states including sepsis . The transmembrane precursor of solTNF, membrane-bound TNFalpha (memTNF), is cleaved by TNFalpha cleaving enzyme (TACE), the regulation of which is poorly understood . We hypothesized that the diversity of clinical features seen with sepsis caused by different organisms may be a result of differential regulation of TACE . Therefore, we measured these proteins in models of sepsis using flow cytometric methods that we have developed . METHODS: Surface protein expression of memTNF and TACE, and TACE catalytic activity were measured in human monocytes by flow cytometry following cell stimulation by lipopolysaccharide (LPS), zymosan (a yeast cell wall product) or heat-inactivated Neisseria meninigitidis . RESULTS: Unstimulated human monocytes express memTNF on the cell surface (mean fluorescence intensity, MFI 131) and this is down-regulated initially in response to LPS (MFI 57) but then recovers to exceed the resting protein expression (MFI 614) . TACE protein is also present on the surface of resting cells (MFI 389) but is catalytically inactive until cell stimulation . Stimulation of monocytes with LPS, zymosan and Neisseria meningitidis produced different patterns of TACE activation with time . CONCLUSIONS: The regulation of memTNF by TACE on monocytes is an important regulatory event in the pro-inflammatory cytokine cascade . As monocytes are important in the inflammatory cascade, we suggest that the control of memTNF and TACE activity on monocytes may play a role in the pathophysiology of sepsis. Bioorg Med Chem, 2004 Dec 15, 12(24), 6427 - 35 Chemoenzymatic synthesis of CMP-sialic acid derivatives by a one-pot two-enzyme system: comparison of substrate flexibility of three microbial CMP-sialic acid synthetases; Yu H et al.; Three C terminal His6-tagged recombinant microbial CMP-sialic acid synthetases {EC 2.7.7.43} cloned from Neisseria meningitidis group B, Streptococcus agalactiae serotype V, and Escherichia coli K1, respectively, were evaluated for their ability in the synthesis of CMP-sialic acid derivatives in a one-pot two-enzyme system . In this system, N-acetylmannosamine or mannose analogs were condensed with pyruvate, catalyzed by a recombinant sialic acid aldolase {EC 4.1.3.3} cloned from E . coli K12 to provide sialic acid analogs as substrates for the CMP-sialic acid synthetases . The substrate flexibility and the reaction efficiency of the three recombinant CMP-sialic acid synthetases were compared, first by qualitative screening using thin layer chromatography, and then by quantitative analysis using high performance liquid chromatography . The N . meningitidis synthetase was shown to have the highest expression level, the most flexible substrate specificity, and the highest catalytic efficiency among the three synthetases . Finally, eight sugar nucleotides, including cytidine 5'-monophosphate N-acetylneuraminic acid (CMP-Neu5Ac) and its derivatives with substitutions at carbon-5, carbon-8, or carbon-9 of Neu5Ac, were synthesized in a preparative (100-200 mg) scale from their 5- or 6-carbon sugar precursors using the N . meningitidis synthetase and the aldolase. Am J Prev Med, 2004 Dec, 27(5), 404 - 10 Health risk factors among detained adolescent females; Crosby R et al.; OBJECTIVES: To identify the prevalence of health risk factors among a sample of detained adolescent females and determine whether there are racial/ethnic differences . DESIGN AND SETTING: A cross-sectional survey of 197 adolescent females (aged 14 to 18 years) recruited within eight detention facilities . OUTCOME MEASURES: Thirty-five measures, comprising four domains, were assessed . Domains were sex-related risk factors, violence-related risk factors, selected mental health issues, and substance abuse behaviors . Measures were collected using audio-computer-assisted self-interviewing . A biological assessment for the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis was also conducted . RESULTS: Several health risk factors were especially prominent . Mean age of sexual debut was 13 years . The mean number of sex partners (lifetime) was 8.8 . Twenty percent tested positive for an STD, and 32.2% had ever been pregnant . Of those reporting sexual activity, 33.9% had not used any form of contraception in the past 2 months and about 40% reported having recent sex with a casual partner . More than 40% reported that a friend had been beaten, attacked, or hurt by others in the past year . More than one half had witnessed violence (past year) and nearly 30% had ever belonged to a gang . Forty percent had thought about committing suicide in the past 2 months, with 35% informing someone of their intent, and 25% attempting suicide . Recent use of illegal substances was common . Correlations between the four domains were significant (p < 0.03), but weak . With some notable exceptions (STD prevalence, trading sex for money, witnessing violence), no difference was found between minority and nonminority adolescents with respect to risk factors . After creating an index of all the risk factors, a normal distribution was obtained (mean number of factors, 10.3; standard deviation, 5.0) . CONCLUSIONS: Preventive medicine programs for adolescent females within detention facilities are warranted . Our evidence suggests that health risk behaviors in this population may be quite diverse; therefore, these programs should be broad in scope and that depth of the programs should vary as a function of risk level. Curr Opin Microbiol, 2004 Dec, 7(6), 610 - 6 Biogenesis of the Gram-negative bacterial outer membrane; Bos MP et al.; Gram-negative bacteria are bounded by two membranes . The outer membrane consists of phospholipids, lipopolysaccharides, lipoproteins and integral outer membrane proteins, all of which are synthesized in the cytoplasm . Recently, much progress has been made in the elucidation of the mechanisms of transport of these molecules over the inner membrane, through the periplasm and into the outer membrane, in part by exploiting the extraordinary capacity of Neisseria meningitidis to survive without lipopolysaccharide. Zhonghua Liu Xing Bing Xue Za Zhi, 2004 Sep, 25(9), 779 - 82 {Application of random amplification polymorphic DNA in the genotyping of Neisseria gonorrhoeae.}; Zhang TJ et al.; OBJECTIVE: To set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection . METHODS: Four different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared . Arbitrary sequence was then used to amplify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains . Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between patient and his/her sexual partner was compared . RESULTS: Cetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the fingerprint of different Neisseria gonorrhoeae were distinctive . However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar . CONCLUSION: Based on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes . RAPD fingerprint maps could be used to trace the source of infection. Mikrobiol Z, 2004 Sep-Oct, 66(5), 23 - 9 {Convertible energy sources in Neisseria gonorrhoeae}; {The rise in fluoroquinolone-resistant Neisseria gonorrhoeae among people attending the Municipal Health Service's clinic for sexually transmitted diseases in Amsterdam et al.; Kolader M, Peerbooms PG, Vader PC, van Bergen JE, Fennema JS, de Vries HJ. GG&GD, Nieuwe Achtergracht 100, 1018 WT AmsterdamOBJECTIVE: To report the incidence of gonorrhoea and the development of resistance to Neisseria gonorrhoeae among attendees at the Municipal Health Service's STD-clinic in Amsterdam, The Netherlands, 2000-2003 . DESIGN: Descriptive . METHOD: Urethral or cervical swabs for culture for N . gonorrhoea were taken from attendees at the STD-clinic . Depending on reported sexual techniques throat and rectal swabs were also taken . The disk diffusion technique in combination with a beta-lactamase test were used for sensitivity testing . RESULTS . The number of Neisseria gonorrhoeae (NG) isolates collected at the Amsterdam Municipal Health Service's STD-clinic decreased from 1047 in 2002 to 772 in 2003 . The number of fluoroquinolone-resistant NG (FRNG) isolates rose from 3 in 2000 to 56 in 2003 (p < 0.001) . FRNG isolates amongst men who have sex with men increased from 1 in 568 isolates (0.2%) in 2000 to 50 in 478 isolates (10.5%) in 2003 (p < 0.001) . Amongst heterosexual men, FRNG rose from 2 per 275 (0.7%) in 2000 to 16 per 297 (5.4%) in 2002 and dropped to 6 per 190 (3.4%) in 2003 (p = 0.146) . No FRNG isolates were found in women in 2003 (2000: 0/180 (0.0%); 2001: 2/160 (1.3%); 2002: 4/183 (2.2%) . CONCLUSION: The recent incidence of FRNG among men who have sex with men to over 5% makes ciprofloxacin and other fluoroquinolones obsolete as the first-choice treatment option for uncomplicated gonorrhoea if no antibiogram is available . It is advised to use cefotaxim when an antibiogram is not available (yet). J Infect Dis, 2004 Dec 15, 190(12), 2109 - 20 Epub 2004 Dec 15. Detection of novel organisms associated with salpingitis, by use of 16S rDNA polymerase chain reaction; Hebb JK et al.; Although Chlamydia trachomatis and Neisseria gonorrhoeae are established causes of salpingitis, the majority of cases have no known etiology . We used broad-range 16S rDNA polymerase chain reaction to identify novel, possibly uncultivable, bacteria associated with salpingitis and identified bacterial 16S sequences in Fallopian-tube specimens from 11 (24%) of 45 consecutive women with laparoscopically confirmed acute salpingitis (the case patients) and from 0 of 44 women seeking tubal ligations (the control subjects) at Kenyatta National Hospital, Nairobi, Kenya . Bacterial phylotypes most closely related to Leptotrichia spp . were detected as the sole phylotypes in 1, and mixed with other bacterial phylotypes in 2, specimens . Novel bacterial phylotypes and those associated with bacterial vaginosis, including Atopobium vaginae, were identified in 3 specimens . N . gonorrhoeae and Streptococcus pyogenes were identified in 2 and 1 specimens, respectively . The finding of novel phylotypes associated with salpingitis has important implications for the etiology, pathogenesis, and treatment of this important reproductive-tract disease syndrome. Immunol Cell Biol, 2004 Dec, 82(6), 603 - 10 Novel adjuvant based on a proteoliposome-derived cochleate structure containing native lipopolysaccharide as a pathogen-associated molecular pattern; Perez O et al.; Proteoliposomes (PL) from Neisseria meningitidis B have been widely used as a core antigen for antimeningococcal vaccination . PL contain major outer membrane proteins, LPS and phospholipids, and they induce a strong Th1 immune response, but they have low stability in solution . Attending to the need for new vaccine adjuvants, we developed a highly stable cochleate structure (CS) from PL using a technology that allows easy incorporation of new antigens . We explored the ability of PLCS to activate the immune system and its possible application as an adjuvant for parenteral and mucosal routes . Our results showed that PLCS were able to upregulate the expression of MHC class II and costimulatory molecules on human dendritic cells, as well as being able to stimulate the production of soluble mediators of a Th1 response, such as IL-12 and nitric oxide . High levels of anti-PL IgG were detected in serum after i.m . or mucosal (oral and nasal) administration, but also anti-PL secretory IgA was produced in saliva following nasal delivery . The immune response polarization to a Th1 pattern was confirmed by the induction of IgG2a antibodies, positive delayed type hypersensitivity reactions, and IFN-gamma production by splenocytes from immunized mice . The adjuvant potential was explored using PLCS containing ovalbumin (Ova) . PLCS-Ova was able to elicit a substantial increase in anti-Ova IgG compared with Ova alone . In addition, a significant reduction in lesion size was observed in mice immunized with Leishmania major antigens in PLCS after challenge with virulent protozoa, suggesting at least partial modulation of the Th2 environment induced by this parasite . In conclusion, our results support the use of PLCS as a potent Th1 adjuvant for parenteral and mucosal vaccines. J Formos Med Assoc, 2004 Nov, 103(11), 858 - 9 Acute urethritis and arthritis-dermatitis syndrome due to Neisseria meningitidis; Sun HY et al.; Acute urethritis and arthritis-dermatitis syndrome after sexual contact are often assumed to be caused by Neisseria gonorrhoeae . We report a case of arthritis-dermatitis syndrome in a 32-year-old man who presented with generalized maculopapular and petechial skin lesions and polyarthritis . Acute urethritis developed 1 week after oro-genital sexual contact with a sex worker about 3 weeks before admission . No pathogen was found on smear of urethral discharge and skin lesions, but Gram-negative diplococci were noted in joint fluid, and blood culture yielded N . meningitidis . His condition improved gradually after repeated arthrocentesis and antibiotic therapy with ceftriaxone followed by ciprofloxacin . Oro-genital contact is a transmission route for N . meningitidis infection manifesting as acute urethritis and arthritis-dermatitis syndrome. J Muscle Res Cell Motil, 2004, 25(4-5), 343 - 50 Neisseria gonorrhoeae porin, P.IB, causes release of ATP from yeast actin; Wen KK et al.; Neisserial porins may play a role in the invasion of the host cell by the bacterium . The protein translocates to the host cell membrane and then to the cytosol during the invasive process, and we have shown it interacts with actin in vitro . Here, we have examined the nucleotide-dependence of the interaction of Neisseria porin, P.IB, with fluorescently labeled yeast G actin . Increasing free ATP between 0 to 0.5 mM retards complex formation between the two proteins . The ATP effect probably results from binding of the nucleotide to actin rather than to porin . Complex formation results in a biphasic release of bound nucleoside triphosphate from actin in the absence of free nucleotide at a rate slower than that of complex formation, but it does not induce hydrolysis of the actin-bound nucleotide . ATP prevents the porin-induced distortion of F-actin structure, and addition of ATP to the complex formed in the absence of free nucleotide induces actin polymerization indicating that P.IB stabilizes nucleotide-free G-actin . Our results suggest that P.IB causes an actin conformation change leading to the production of a polymerization-competent nucleotide-free protein. An Med Interna, 2004 Nov, 21(11), 551 - 3 {Anticoagulation with bemiparina after intracerebral hemorrhage as complication of bacterial endocarditis on metallic prosthetic valve}; Munoz Morente A et al.; We present the case of a patient with an intracerebral hemorrhage as complication of an infectious endocarditis (EI) produced by Neisseria sicca on a prosthetic mitral valve . The patient was anticoagulated previously with Acenocumarol as prophylaxis of thrombosis of the prosthetic valve . He was diagnosed as having IE and later he presented neurological symptoms as consequence of several intracerebral hemorrhagic foci . We decided to continue the anticoagulation with sodium heparin followed of Bemiparina and no new hemorrhagic complications nor thrombosis of the valve were observed after three months of the event . We have not found in the literature any case where low molecular weight heparin has been used as method of anticoagulation in these cases beyond two weeks. J Immunother, 2004 Nov-Dec, 27(6), 442 - 51 Active specific immunotherapy of melanoma with a GM3 ganglioside-based vaccine: a report on safety and immunogenicity; Guthmann MD et al.; A novel cancer vaccine was obtained by combining GM3 ganglioside with Neisseria meningitidis outer membrane protein complex to obtain very-small-size proteoliposomes (GM3/VSSP) . The authors report the results of a phase 1 study of intramuscular administration of GM3/VSSP/Montanide ISA 51 to patients with metastatic melanoma . Twenty-six patients were included in three dose-level cohorts of 120, 240, and 360 mug . The first five doses (induction phase) were given at 2-week intervals, and the remaining four doses were given monthly . Patients were evaluated for dose-related toxicities and antitumor effects . In addition, serum and peripheral blood mononuclear cells were obtained at baseline and throughout treatment to evaluate humoral and cellular immune responses . One episode of severe hypotension and fever was observed in a patient included at the highest dose level . Other toxicities consisted of local reactions at the site of injection and mild fever and chills . Five doses of GM3/VSSP induced an anti-GM3 IgM response in 44% of patients . Serum reactivity was also observed against melanoma cell lines and tumor biopsies . GM3/VSSP was shown to induce very strong in vitro IFNgamma secretion in all evaluated melanoma patients . Furthermore, in one patient IFNgamma secretion was shown to be GM3-specific . A 62% reduction of a mediastinal mass was documented in one patient (partial response), while a second patient benefited from initial disease stabilization followed by tumor reduction in nonmeasurable soft tissue lesions accompanied by vitiligo. Int J Med Microbiol, 2004 Oct, 294(5), 295 - 301 Genome-based vaccines; Knaust A et al.; Vaccination is an effective possibility to prevent many bacterial or viral infections, but for several important pathogens still no vaccines are available . The sequences of complete genomes are now decoded for an increasing number of bacterial pathogens and offer the possibility for comprehensive screenings to identify targets for vaccine development . In this article current genomic approaches to identify antigenic proteins of Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, and Chlamydia pneumoniae are summarized. Prev Med, 2005 Apr, 40(4), 420 - 31 Evaluation of a cognitive-behavioral, group, randomized controlled intervention trial to prevent sexually transmitted infections and unintended pregnancies in young women; Boyer CB et al.; BACKGROUND: Few cognitive-behavioral interventions have focused on preventing sexually transmitted infections (STIs) and unintended pregnancies (UPs) in young, sexually active women in a single study . Military recruit training provides a well-defined, national, nonclinic sample in which to evaluate such an intervention . METHODS: All female Marine recruits (N = 2,288) in training were approached . Of these, 2,157 (94.3%) voluntarily agreed and were randomly assigned, by platoons, to participate in cognitive-behavioral interventions to prevent STIs or UPs or to prevent physical training injuries and cancer . Participants completed self-administered questionnaires and were screened for pregnancy, Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis at baseline and, on average, 1 and 14 months postintervention . RESULTS: A higher proportion of the control group had a postintervention STI or UP {odds ratio (OR) = 1.41, 95% confidence interval (CI) = 1.01-1.98} . Among participants who had no history of STIs or pregnancy, but who engaged in risky sexual behaviors just before recruit training, the control group was more likely to acquire a postintervention STI (OR = 3.24, CI = 1.74-6.03) . Among participants who were not sexually experienced at baseline, the control group was more likely to have casual (OR = 2.05, 95% CI = 1.04-4.08) and multiple (OR = 1.87, 95% CI = 1.01-3.47) sexual partners postintervention . CONCLUSIONS: This randomized controlled trial indicates that cognitive-behavioral interventions are effective for reducing behavioral risk and preventing STIs and UPs in young, sexually active women who are not seeking health care. Harefuah, 2004 Aug, 143(8), 576 - 7, 622 {Vaccines for all occasions}; Passwell JH; The strategy of immunizing a population at risk of infectious disease has been enormously successful medically and has also proven to be cost effective . Development of effective immunogens, that induce active immunization, is a long process that requires careful monitoring and assurance of short and long term safety, induction of protective immunity and proven efficacy in preventing the disease . A successful immunization program is also dependent on delivery of the vaccine to as many susceptible individuals as possible, so as to attain herd immunity . Passive immunization with antibodies, usually used prior to the development of active vaccines has also been remarkably effective . The special circumstances of the field and crowded conditions have demanded that the Medical Corps of the Israeli army cater for the needs of our soldiers . In this issue, the past achievements and current immunization policy are outlined for the first time . Their contribution to the health of our soldiers is commendable . Close monitoring of the epidemiology of infectious disease in the special circumstances of field conditions has prompted successful programs to markedly reduce infectious hepatitis A by passive immunization with gamma globulin in the past and, nowadays, with the killed active viral vaccine . In addition, prevention of influenza by killed viral vaccine and invasive bacterial disease by Neisseria meningitidis with multivalent polysaccharide vaccines are being used . This group has also improved hygienic conditions in the field to cope with shigellosis and salmonella infections . Research in the development of effective vaccines for protection of shigellosis has also been addressed by this group . New challenges posed by the emerging infectious diseases and the possible effects of bioterrorism are certain to keep this group on their toes. Mol Microbiol, 2004 Nov, 54(4), 962 - 79 Characterization of a novel Neisseria meningitidis Fur and iron-regulated operon required for protection from oxidative stress: utility of DNA microarray in the assignment of the biological role of hypothetical genes; Grifantini R et al.; We have previously shown that in the human pathogen Neisseria meningitidis group B (MenB) more than 200 genes are regulated in response to growth with iron . Among the Fur-dependent, upregulated genes identified by microarray analysis was a putative operon constituted by three genes, annotated as NMB1436, NMB1437 and NMB1438 and encoding proteins with so far unknown function . The operon was remarkably upregulated in the presence of iron and, on the basis of gel retardation analysis, its regulation was Fur dependent . In this study, we have further characterized the role of iron and Fur in the regulation of the NMB1436-38 operon and we have mapped the promoter and the Fur binding site . We also demonstrate by mutant analysis that the NMB1436-38 operon is required for protection of MenB to hydrogen peroxide-mediated killing . By using both microarray analysis and S1 mapping, we demonstrate that the operon is not regulated by oxidative stress signals . We also show that the deletion of the NMB1436-38 operon results in an impaired capacity of MenB to survive in the blood of mice using an adult mouse model of MenB infection . Finally, we show that the NMB1436-38 deletion mutant exhibits increased susceptibility to the killing activity of polymorphonuclears (PMNs), suggesting that the 'attenuated' phenotype is mediated in part by the increased sensitivity to reactive oxygen species-producing cells . This study represents one of the first examples of the use of DNA microarray to assign a biological role to hypothetical genes in bacteria. Carbohydr Res, 2004 Nov 15, 339(16), 2641 - 9 Synthesis of unnatural sugar nucleotides and their evaluation as donor substrates in glycosyltransferase-catalyzed reactions; Khaled A et al.; New unnatural sugar nucleotides, UDP-Fuc and CDP-Fuc were synthesized from fucose-beta-1-phosphate and nucleotide monophosphates activated as morpholidates . Furthermore, a nucleotide analogue was prepared by phosphorylation of 1-(beta-D-ribofuranosyl)cyanuric acid, itself obtained as a protected derivative by condensation of the persilylated derivative of cyanuric acid with 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose in 74% yield . This phosphate activated according to the same procedure was condensed with fucose-beta-1-phosphate, affording a new sugar nucleotide conjugate (NDP-Fuc) which was evaluated together with UDP-Fuc, CDP-Fuc and ADP-Fuc, as fucose donors in alpha-(1-->4/3)-fucosyltransferase (FucT-III) catalyzed reaction . Fucose transfer could be observed with each of the donors and kinetic parameters were determined using a fluorescent acceptor substrate . Efficiency of the four analogues towards FucT-III was in the following order: UDP-Fuc=ADP-Fuc>NDP-Fuc>CDP-Fuc . According to the same strategy ADP-GlcNAc was prepared from AMP-morpholidate and N-acetylglucosamine-alpha-1-phosphate; tested as a glucosaminyl donor towards Neisseria meningitidis N-acetylglucosaminyl transferase (LgtA), ADP-GlcNAc was recognized with 0.1% efficiency as compared with UDP-GlcNAc, the natural donor substrate. Biochemistry, 2004 Nov 9, 43(44), 14290 - 8 Identification and mechanism of a bacterial hydrolyzing UDP-N-acetylglucosamine 2-epimerase; Murkin AS et al.; This paper reports the first identification of a fully functional hydrolyzing UDP-N-acetylglucosamine 2-epimerase from a bacterial source . The epimerase (known as SiaA or NeuC) from Neisseria meningitidis MC58 group B is shown to catalyze the conversion of UDP-GlcNAc into ManNAc and UDP in the first step of sialic acid (N-acetylneuraminic acid) biosynthesis . The mechanism is proposed to involve an anti elimination of UDP to form 2-acetamidoglucal as an intermediate, followed by the syn addition of water . The observation that the alpha-anomer of ManNAc is the true product and that solvent deuterium is incorporated at C-2 is consistent with this mechanism . The use of the (18)O-labeled substrate confirms that the overall hydrolysis reaction proceeds via cleavage of the C-O bond . Furthermore, the putative intermediate 2-acetamidoglucal is shown to serve as a catalytically competent substrate and is enzymatically hydrated to give ManNAc exclusively . Isotope effect studies show that cleavage of the C-H bond is not rate limiting during catalysis . Mutagenesis studies show that three active site carboxylate residues are crucial for catalysis . In two of the mutants that were studied (E122Q and D131N), 2-acetamidoglucal was released from the active site during catalysis, providing direct evidence that the enzyme is capable of catalyzing the anti elimination of UDP from UDP-GlcNAc. J Bacteriol, 2004 Nov, 186(22), 7811 - 4 Mutation of a single lytic transglycosylase causes aberrant septation and inhibits cell separation of Neisseria gonorrhoeae; Cloud KA et al.; The function of lytic peptidoglycan transglycosylases is poorly understood . Single lytic transglycosylase mutants of Escherichia coli have no growth phenotype . By contrast, mutation of Neisseria gonorrhoeae ltgC inhibited cell separation without affecting peptidoglycan monomer production . Thus, LtgC has a dedicated function in gonococcal cell division. J Biol Chem . 2004 Oct 29; {Epub ahead of print} Structural and mechanistic analysis of sialic acid synthase NeuB from neisseria meningitidis in complex with Mn2+, phosphoenolpyruvate, and N-acetylmannosaminitol; Gunawan J et al.; Sialic acid synthase catalyze the condensation of phosphoenolpyruvate (PEP) and N-acetyl mannosamine (ManNAc), directly forming N-acetylneuraminic acid (NeuNAc or sialic acid) . The importance of understanding the mechanism of sialic acid production is made evident by the vital role this sugar plays in a number of biological processes.We have determined the high resolution crystal structures of a sialic acid synthase, that of the enzyme NeuB from the bacterial pathogen Neisseria meningitidis, alone and in complex with Mn(2+) its natural substrate PEP, and substrate analog rManNAc . The structure reveals a unique domain-swapped homodimer architecture consisting of a (ss/a)(8) barrel type fold at the C-terminal end, and a novel domain with high sequence identity and structural similarity to the ice binding type III antifreeze proteins at the N-terminal end of the enzyme . The domain-swapped dimer appears to be mechanistically important for the creation of the NeuB active site, as both the TIM barrel and antifreeze-like domain provide contacts for enzyme:sugar substrate interactions.In addition to the identification of potential catalytic residues, our structural data shows that a bound Mn(2+) ion is positioned correctly relative to substrate to play a direct catalytic role in the production of sialic acid, an observation that contrasts against recent enzymatic research on other PEP sugar synthases including DAH7PS and KDO8PS . Using isotopic (18)O-labelling and a newly developed continous assay we have also further characterized the catalytic mechanism of sialic acid synthase by confirming a C-O bond cleavage process occurs during the formation of sialic acid. J Obstet Gynaecol, 1998, 18(6), 556 - 60 Gynaecological and microbiological findings in women attending for a general health check-up; Tchoudomirova K; Two hundred apparently healthy sexually active women, 17-34 years of age, who had presented for a general health check-up at the Clinic of Dermatology and Venereology, Medical University, Plovdiv, Bulgaria, were asked about genital symptoms, sexual behaviour, contraceptive use and smoking habits, and examined for signs of genital infections . They were searched for genital chlamydial infection, gonorrhoea, trichomoniasis, bacterial vaginosis (BV) and vulvovaginal candidosis, syphilis and HIV . Polymerase chain reaction (PCR) was used for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine samples and the results were compared with direct immunofluorescence (DFA) and enzyme immunoassays (EIA) for C . trachomatis in urethral, cervical and urine samples . In 56 (28%) women, an STD and/or an STD-related condition were diagnosed . The prevalence of genital chlamydial infection, trichomoniasis, BV and vulvovaginal candidosis was 4.5%, 0.5%, 17.5% and 7.5% respectively . On direct questioning 39 (19.5%) women reported symptoms suggestive of an infection, while 58 (29%) had signs that may have been caused by genital infection . In urine the PCR tests detected more (3.5%) chlamydia-positive women than the DFA (2.5%) and EIA tests (1.5%) . The urine PCR test was as sensitive as the DFA when testing cervical samples . The chlamydia-positive women and women with BV were less likely to have a steady partner than the controls . No woman had syphilis or HIV infection . The women with BV were more frequent users of an intrauterine device and were more likely to smoke heavily compared with other women . STDs and STD-related conditions are common among adult women who consider themselves gynaecologically healthy . Screening for genital infections among women in reproductive age attending for health check-up could improve women's reproductive health. J Obstet Gynaecol, 1998, 18(2), 164 - 8 Laparoscopic and microbiological features of acute salpingitis in developing countries; O Arowojolu R A Bakare A A Oni A O Ilesanmi A; Summary One hundred and twenty-four out of 198 consecutive women who underwent diagnostic lasparoscopy for clinical symptoms and signs of acute salpingitis at the University College Hospital, Ibadan, Nigeria had acute salpingitis . These were slightly younger than those without acute salpingitis, otherwise there were no differences in the sociodemographic characteristics of the two groups . Urinary and gastrointestinal symptoms, abnormal vaginal discharge, fever ( 38 C) and sexually transmitted organisms were significantly more in women with acute salpingitis . Ninetyfive per cent of the Neisseria gonorrhoea cultured were of the PPNG strain . Pelvic adhesions were present in 69.4% of the women with acute salpingitis . Forty-one per cent of the women had tubal occlusion . It was concluded that laparoscopy rather than clinical findings alone would determine the severity of acute salpingitis . This should be performed along with culture of genital discharges and peritoneal fluid for optimum management . The use of a single dose broad spectrum antibiotics active against both PPNG and non-PPNG strains, and chlamydial infections is advocated for treatment in developing countries. Commun Dis Intell, 2003, 27(4), 488 - 91 Surveillance of antibiotic resistance in Neisseria gonorrhoeae in the WHO Western Pacific Region, 2002; Reproductive tract infections in rural women from the highlands et al.; Center for AIDS and STDs, University of Washington, Harborview Medical Center Box 359931, Seattle, WA 98104, USAOBJECTIVE: To define the prevalences and manifestations of reproductive tract infections (RTIs) in rural Peruvian women . METHODS: During 1997-98, we visited 18 rural districts in coastal, highlands, and jungle regions of Peru . We administered standardized questionnaires and pelvic examinations to members of women's community-based organizations; and collected vaginal fluid for pH, amine odour, Gram stain, microscopy, and culture for Trichomonas vaginalis; cervical specimens for Chlamydia trachomatis, Neisseria gonorrhoeae; human papilloma virus (HPV) by polymerase chain reaction (PCR) assays, and blood for syphilis serology . FINDINGS: The 754 participants averaged 36.9 years of age and 1.7 sex partners ever; 77% reported symptoms indicative of RTIs; 51% and 26% reported their symptoms spontaneously or only with specific questioning, respectively . Symptoms reported spontaneously included abnormal vaginal discharge (29.3% and 22.9%, respectively) . One or more RTIs, found in 70.4% of participants, included bacterial vaginosis (43.7%), trichomoniasis (16.5%), vulvovaginal candidiasis (4.5%), chlamydial infection (6.8%), gonorrhoea (1.2%), syphilis seropositivity (1.7%), cervical HPV infection (4.9%), and genital warts or ulcers (2.8%) . Of 715 adequate Pap smears, 7 revealed cancer, 4 high-grade squamous intra-epithelial lesions (SIL) and 15 low-grade SIL . Clinical algorithms had very low sensitivity and predictive values for cervical infection, but over half the women with symptoms of malodorous vaginal discharge, signs of abnormal vaginal discharge, or both, had bacterial vaginosis or trichomoniasis . CONCLUSION: Overall, 77% of women had symptoms indicative of RTIs, and 70% had objective evidence of one or more RTIs . Women with selected symptoms and signs of vaginal infection could benefit from standard metronidazole therapy. Infect Immun, 2004 Nov, 72(11), 6743 - 7 Identification of Neisseria meningitidis genetic loci involved in the modulation of phase variation frequencies; Alexander HL et al.; It has been proposed that increased phase variation frequencies in Neisseria meningitidis augment transmissibility and invasiveness . A Himar1 mariner transposon mutant library was constructed in serogroup A N . meningitidis and screened for clones with increased phase variation frequencies . Insertions increasing the frequency of slippage events within mononucleotide repeat tracts were identified in three known phase variation-modulating genes (mutS, mutL, and uvrD), as well as six additional loci (pilP, fbpA, fbpB, NMA1233, and two intergenic regions) . The implications of these insertion mutations are discussed. Infect Immun, 2004 Nov, 72(11), 6408 - 17 Gonococcal porin IB activates NF-kappaB in human urethral epithelium and increases the expression of host antiapoptotic factors; Binnicker MJ et al.; Infection of human urethral epithelial cells (UECs) with Neisseria gonorrhoeae increases the transcription of several host antiapoptotic genes, including bfl-1, cox-2, and c-IAP-2 . In order to identify the bacterial factor(s) responsible for eliciting these changes, the transcriptional status of apoptotic machinery was monitored in UECs challenged with certain gonococcal membrane components . Initially, we observed that infection of UECs with gentamicin-killed gonococci increased the expression of the antiapoptotic Bcl-2 family member, bfl-1 . This observation indicated that viable, replicating bacteria are not required for induction of antiapoptotic gene expression . Confirming this observation, treatment of UECs with purified gonococcal membrane increased the expression of bfl-1, cox-2, and c-IAP-2 . This finding suggested that a factor or multiple factors present in the outer membrane (OM) are responsible for altering UEC antiapoptotic gene expression . Interestingly, treatment of UECs with gonococcal porin IB (PorB IB), a major constituent of the OM, significantly increased the transcription of bfl-1, cox-2, and c-IAP-2 . The upregulation of these genes by PorB IB was determined to be dependent on NF-kappaB activation, as inhibiting NF-kappaB blocked induced expression of these genes . This work demonstrates the altered expression of host apoptotic factors in response to gonococcal PorB IB and supports a model whereby UEC cell death may be modulated as a potential mechanism of bacterial survival and proliferation. Clin Infect Dis, 2004 Nov 1, 39(9), 1387 - 90 Epub 2004 Oct 11. Positive predictive value of Gen-Probe APTIMA Combo 2 testing for Neisseria gonorrhoeae in a population of women with low prevalence of N . gonorrhoeae infection; Golden MR et al.; We collected specimens from women who tested positive for Neisseria gonorrhoeae using the Gen-Probe APTIMA Combo 2 (AC2), and we performed confirmatory tests using a nucleic acid amplification test (NAAT) that targets alternate gonococcal nucleic acid sequences . Among 59,664 specimens, 280 (0.47%) had positive results using AC2; 265 of these specimens were tested using the confirmatory NAAT, of which 258 yielded positive results (positive predictive value, 97.4%; 95% confidence interval, 95.1%-98.8%) . Routine confirmatory testing of specimens with positive AC2 gonorrhea test results is not indicated. Int J Med Microbiol, 2004 Sep, 294(2-3), 67 - 73 Population structure of pathogenic bacteria revisited; Achtman M; This minireview summarizes the historical development of bacterial population genetic concepts since the early 1980s . Initially multilocus enzyme electrophoresis was used to determine population structures but this technique is poorly portable between laboratories and was replaced in 1998 by multilocus sequence typing . Diverse population structures exist in different bacterial species . Two distinctive structures are described in greater detail . "Young" organisms, such as Yersinia pestis, have evolved or undergone a severe bottleneck in recent millennia and have not yet accumulated much sequence diversity . "genoclouds" in subgroup III Neisseria meningitidis arise because of the accumulation of diversity due to herd immunity, which is then purified during subsequent epidemic spread. Acta Paediatr Taiwan, 2004 May-Jun, 45(3), 158 - 62 Acute flaccid paralysis due to myelopathy in childhood bacterial meningitis; Tu YF et al.; Spinal cord damage is a rare neurological complication of bacterial meningitis . From 1988 to 2001, three of 186 pediatric patients with acute bacterial meningitis presented with acute flaccid paralysis due to myelopathy . The disease-onset ages and causative pathogens of the three patients were 2 days (group B streptococcus), 3 months (Streptococcus pneumoniae), and 13 years (Neisseria meningitidis), respectively . Spinal MR imaging was normal in the 13-year-old patient, who was left with mild residual motor weakness . Severe necrotizing myelitis and syringomyelia over the thoracic spinal cord were found in the other two younger patients . Both had severe paralysis at follow-up . All three patients required mechanical ventilation or vasopressor agents during antibiotic treatment for bacterial meningitis. J Clin Periodontol, 2004 Nov, 31(11), 996 - 1002 Subgingival microbiota of chronic periodontitis subjects from different geographic locations; Haffajee AD et al.; BACKGROUND: Most clinical studies assume that the subgingival microbiota is similar from one geographic location to another . The purpose of the present investigation was to examine the composition of the subgingival microbiota in chronic periodontitis subjects from four countries . METHOD: Subjects with chronic periodontitis (N, Sweden=101; USA=115; Brazil=58; Chile=26) were recruited . Subjects were measured at baseline for plaque, gingivitis, bleeding on probing (BOP), suppuration, pocket depth (PD) and attachment level (AL) at six sites per tooth . Subgingival plaque samples taken from the mesial aspect of each tooth at baseline were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples=6036) . % DNA probe counts comprised by each species was determined for each site and averaged across sites in each subject . Significance of differences in proportions of each species among countries was determined using ancova adjusting for age, mean pocket depth, gender and smoking status . p-Values were adjusted for multiple comparisons . RESULTS: On average, all species were detected in samples from subjects in the four countries . Thirteen species differed significantly in adjusted mean proportions among countries even after adjusting for multiple comparisons . Porphyromonas gingivalis, one species that differed in proportions among countries, comprised adjusted means of 7.5, 11.9, 1.6 and 6.6% of the microbiota in subjects from Brazil, Chile, Sweden and USA (p<0.001), while mean proportions of Treponema denticola were 6.7, 4.2, 0.8 and 2.3, respectively (p<0.001) . In contrast, a key periodontal pathogen, Tannerella forsythensis, exhibited mean proportions ranging from 6.2-8.5% and did not differ significantly among countries . Besides these species, prominent species in Brazil were Actinomyces naeslundii genospecies 1 and 2 (8.4%, 7.2%) and Prevotella intermedia (6.5%); in Chile, Prevotella melaninogenica (6.4%) and Neisseria mucosa (5.3%); in Sweden A . naeslundii genospecies 2 (8.4%), Capnocytophaga gingivalis (7.1%) and Peptostreptococcus micros (5.0%); in USA A . naeslundii genospecies 2 (7.5%), P . intermedia (6.8%) and C . gingivalis (6.1%) . CONCLUSIONS: The microbial profiles of subgingival plaque samples from chronic periodontitis subjects in four countries showed surprisingly marked differences . These differences persisted after adjusting for age, mean pocket depth, gender and smoking status . (c) Blackwell Munksgaard, 2004 J Bacteriol, 2004 Nov, 186(21), 7175 - 85 The N terminus of MinD contains determinants which affect its dynamic localization and enzymatic activity; Szeto J et al.; MinD is involved in regulating the proper placement of the cytokinetic machinery in some bacteria, including Neisseria gonorrhoeae and Escherichia coli . Stimulation of the ATPase activity of MinD by MinE has been proposed to induce dynamic, pole-to-pole oscillations of MinD in E . coli . Here, we investigated the effects of deleting or mutating conserved residues within the N terminus of N . gonorrhoeae MinD (MinD(Ng)) on protein dynamism, localization, and interactions with MinD(Ng) and with MinE(Ng) . Deletions or mutations were generated in the first five residues of MinD(Ng), and mutant proteins were evaluated by several functional assays . Truncation or mutation of N-terminal residues disrupted MinD(Ng) interactions with itself and with MinE . Although the majority of green fluorescent protein (GFP)-MinD(Ng) mutants could still oscillate from pole to pole in E . coli, the GFP-MinD(Ng) oscillation cycles were significantly faster and were accompanied by increased cytoplasmic localization . Interestingly, in vitro ATPase assays indicated that MinD(Ng) proteins lacking the first three residues or with an I5E substitution possessed higher MinE(Ng)-independent ATPase activities than the wild-type protein . These results indicate that determinants found within the extreme N terminus of MinD(Ng) are implicated in regulating the enzymatic activity and dynamic localization of the protein. Clin Microbiol Rev, 2004 Oct, 17(4), 965 - 81, table of contents The molecular mechanisms used by Neisseria gonorrhoeae to initiate infection differ between men and women; Edwards JL et al.; The molecular mechanisms used by the gonococcus to initiate infection exhibit gender specificity . The clinical presentations of disease are also strikingly different upon comparison of gonococcal urethritis to gonococcal cervicitis . An intimate association occurs between the gonococcus and the urethral epithelium and is mediated by the asialoglycoprotein receptor . Gonococcal interaction with the urethral epithelia cell triggers cytokine release, which promotes neutrophil influx and an inflammatory response . Similarly, gonococcal infection of the upper female genital tract also results in inflammation . Gonococci invade the nonciliated epithelia, and the ciliated cells are subjected to the cytotoxic effects of tumor necrosis factor alpha induced by gonococcal peptidoglycan and lipooligosaccharide . In contrast, gonococcal infection of the lower female genital tract is typically asymptomatic . This is in part the result of the ability of the gonococcus to subvert the alternative pathway of complement present in the lower female genital tract . Gonococcal engagement of complement receptor 3 on the cervical epithelia results in membrane ruffling and does not promote inflammation . A model of gonococcal pathogenesis is presented in the context of the male and female human urogenital tracts. J Microbiol Methods, 2004 Dec, 59(3), 415 - 21 Rapid detection of gyrA and parC mutations in fluoroquinolone-resistant Neisseria gonorrhoeae by denaturing high-performance liquid chromatography; Shigemura K et al.; The detection of DNA sequence variation is fundamental to the identification of the genomic basis of phenotypic variability . Denaturing high-performance liquid chromatography (DHPLC) is a novel technique that is used to detect mutations in human DNA . This is the first report that this technique is used as a tool to detect mutations in genes encoding fluoroquinolone resistance in Neisseria gonorrhoeae . Eighty-one strains of N . gonorrhoeae were used in this study . Genomic DNA from each strain was subjected to PCR amplification of 225 bp in gyrA and 166 bp in parC spanning the fluoroquinolone-resistance determining regions (QRDRs) . After we performed DNA sequencing of these amplicons and identification of mutations in the QRDRs, DHPLC was undertaken to investigate whether its results correlate the distinctive chromatogram with their DNA mutations pattern . The profilings detected by DHPLC completely corresponded to the results of the DNA sequencing in mutation patters in gyrA and parC genes . They resulted in the following amino acid substitutions: Ser-91Phe, Asp-95Gly, and Asp-95Asn in gyrA; and Gly-85Asp, Asp-86Asn, Ser-87Arg, and Ser-88Pro in parC, respectively . These mutations existed alone or as combinations, and we identified five mutations patterns in gyrA and six in parC including wild-type . These mutations and their patterns could be rapidly and reproducibly identified from the PCR products using DHPLC, producing specific peak patterns that correlate with genotypes . This novel detection system facilitates the detection of resistance alleles, providing a rapid (5 min per sample), economic (96 sample per run), and reliable technique for characterizing fluoroquinolone resistance in N . gonorrhoeae. Sex Transm Dis, 2004 Sep, 31(9), 542 - 6 Prevalence and risk behaviors for chlamydial infection in a population-based study of female adolescents in Brazil; Miranda AE et al.; BACKGROUND: Adolescents are vulnerable to sexually transmitted infections (STIs) and unplanned pregnancy . Prevention measures and assistance are of significant public health importance in this population . OBJECTIVE: The objective of this study was to identify demographic, behavioral, and clinical factors for STIs and to determine the prevalence of Chlamydia trachomatis infection (CT) among female adolescents in Vitoria, Brazil . METHODS: We performed a cross-sectional study among female adolescents (15-19 years) served by the Health Family Program . Participants were screened for CT and Neisseria gonorrhoeae (GC) using ligase chain reaction applied to urine and answered a face-to-face questionnaire to assess demographic, behavioral, and clinical factors . All participants and their parents signed the informed consent . RESULTS: Four hundred sixty-four young women were sampled . The prevalence of CT was 8.9% (95% confidence interval {CI}, 6.5-11.9%) overall . Among sexually active women, CT and gonorrhea prevalence were 12.2 (95% CI, 9.4-17.0%) and 1.9% (95% CI, 1.1-2.7%), respectively . Previously diagnosed STI was reported by 12.8% . Women who reported regular condom use and having condoms at home were significantly less likely to have CT, and having never purchased condoms was significantly associated with a positive CT result . CONCLUSION: A high prevalence of CT was found in this population, and behavioral risk was high despite readily available STI prevention information . Women who reported positive condom use behaviors were less likely to have CT . These results demonstrate the need for ongoing STI prevention activities, including STI screening and continued successful risk reduction activities such as condom use to further decrease CT and other STI among adolescents. J Biochem Mol Biol, 2004 Sep 30, 37(5), 597 - 602 A mutagenic study of beta-1,4-galactosyltransferases from Neisseria meningitidis; Park JE et al.; N-terminal His-tagged recombinant beta-1,4-galactosyltransferase from Neisseria meningitidis was expressed and purified to homogeneity by column chromatography using Ni-NTA resin . Mutations were introduced to investigate the roles of, Ser68, His69, Glu88, Asp90, and Tyr156, which are components of a highly conserved region in recombinant beta-1,4 galactosyltransferase . Also, the functions of three other cysteine residues, Cys65, Cys139, and Cys205, were investigated using site-directed mutagenesis to determine the location of the disulfide bond and the role of the sulfhydryl groups . Purified mutant galactosyltransferases, His69Phe, Glu88Gln and Asp90Asn completely shut down wild-type galactosyltransferase activity (1-3 %) . Also, Ser68Ala showed much lower activity than wild-type galactosyltransferase (19 %) . However, only the substitution of Tyr156Phe resulted in a slight reduction in galactosyltransferase activity (90 %) . The enzyme was found to remain active when the cysteine residues at positions 139 and 205 were replaced separately with serine . However, enzyme reactivity was found to be markedly reduced when Cys65 was replaced with serine (27 %) . These results indicate that conserved amino acids such as Cys65, Ser68, His69, Glu88, and Asp90 may be involved in the binding of substrates or in the catalysis of the galactosyltransferase reaction. Int J STD AIDS, 2004 Oct, 15(10), 707 - 8 Pseudomonas orchitis in puberty; Rajagopal AS; Acute epididymo-orchitis is a common cause of 'acute scrotum' in adolescence and young adults, and the common causative pathogens are Chlamydia trachomatis and Neisseria gonorrhoeae . This is a rare case of acute epididymo-orchitis due to Pseudomonas aeruginosa in a pubertal boy with a history of 'ano-receptive' intercourse . On Medline search there are no reports of pseudomonas orchitis in this age group. Electrophoresis, 2004 Oct, 25(18-19), 3132 - 8 Critical aspects of analysis of Micrococcus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary electrophoresis; Hoerr V et al.; Within the frame of our study we investigated Microccocus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary zone electrophoresis (CZE) . They form chains and clusters on a different scale, which can be reflected in the electropherograms . A low buffer concentration of Tris-borate and Na2EDTA containing a polymeric matrix of 0.0125% poly(ethylene) oxide (PEO) was used . Key factors were the standardization and optimization of CE conditions, buffer solution, and pretreatment of bacterial samples, which are not transferable to different bacterial strains, in general . The different compositions of the cell wall of on the one hand Gram-positive (M . luteus) and Gram-negative (N . cinerea) cocci and on the other hand Gram-negative, rod-shaped bacteria (P . fluorescens), are probably responsible for the different pretreatment conditions . J Clin Microbiol, 2004 Oct, 42(10), 4803 - 4 Survival of Neisseria gonorrhoeae isolates of different auxotypes in six commercial transport systems; Graver MA et al.; We determined the rates of survival of six clinical and two control strains of Neisseria gonorrhoeae in six commercial transport systems, using NCCLS standard M40-A methodology . Differences in strain recovery and system performance were marked . A strain less robust than the recommended ATCC 43069 would provide a more exacting standard. J Clin Microbiol, 2004 Oct, 42(10), 4636 - 40 High prevalence of genital mycoplasmas among sexually active young adults with urethritis or cervicitis symptoms in La Crosse, Wisconsin; Schlicht MJ et al.; Sexually active young adults in the small college town of La Crosse, Wisconsin, were evaluated for conventional sexually transmitted pathogens and tested for infections with mycoplasmas . The prevalence in 65 symptomatic men or women and 137 healthy volunteers (67 men and 70 women) was compared . Urine specimens from both cohorts were tested by ligase chain reaction for Chlamydia trachomatis or Neisseria gonorrhoeae . In addition, the urethral or cervical swabs from the symptomatic subjects were tested by PCR for Mycoplasma genitalium and cultured for Mycoplasma hominis and the ureaplasmas . The results confirmed a relatively low prevalence of gonorrhea among symptomatic men (12%) and chlamydia among symptomatic men (15%) and normal women (3%) . In contrast, infections with mycoplasmas, especially the ureaplasmas (57%), were common and the organisms were the only potential sexually transmitted pathogen detected in 40 (62%) symptomatic subjects . Because of the high prevalence, we also evaluated urethral swabs from an additional 25 normal female volunteers and recovered ureaplasmas from 4 (16%) subjects . Additionally, the participants rarely used protection during sexual intercourse and some symptomatic subjects apparently acquired their infections despite using condoms regularly . The findings demonstrate a strong association between abnormal urogenital findings and detection