Neisseria

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 of myoplasmas, particularly ureaplasmas, and suggest the infections will remain common.

Rev Argent Microbiol, 2004 Apr-Jun, 36(2), 78 - 80
Unusual phenotypic characteristic of Neisseria gonorrhoeae from male patients who have sex with men; Garcia SD et al.; We describe four isolates of Ng from men who have sex with men (MSM) patients that were able to grow in the abscense of CO2, as previously was described for N . gonorrhoeae ssp . kochii . These isolates were able to grow aerobically (without any added CO2) at 37 degrees C giving small colonies after 48 hs; two of them isolated from pharynx and urethra of one patient, were also able to grow without the blood supplement in the same conditions . In these unusual isolates the major outer-membrane proteins are of the same molecular weight than Ng . These isolates could be taken for other members of the genus if not confirmed by means of these (or other) methods.

Parasitol Today, 1991 Mar, 7(3), 29 - 33
Molecular genetics of antigenic variation; Borst P; Antigenic variation is one of the most effective strategies developed by parasites to escape immune destruction . It requires a large wardrobe of surface coats and mechanisms to exchange one coat for an unrelated one . The molecular principles of antigenic variation are now largely known in the bacterial species Borrelia and Neisseria and in the protozoa of the African trypanosome group and these three examples are discussed here by Piet Borst.

Commun Dis Intell, 2004, 28(2), 187 - 93
Annual report of the Australian Gonococcal Surveillance Programme, 2003; Enhanced immunocompetent cells in chlamydial cervicitis; Institute of Pathology, Indian Council of Medical Research, Safdarjang Hospital Campus, New Delhi, India . amittal_iop@yahoo.com

OBJECTIVE: To investigate changes in cell-mediated immunophenotypes by flow cytometry in endocervical secretions and peripheral blood in women with Chlamydia trachomatis infection . STUDY DESIGN: Fifty women attending the gynaecology outpatient department of Safdarjang Hospital, New Delhi, India, with signs and symptoms of cervicitis were enrolled . All patients underwent endocervical screening for C trachomatis (direct fluorescence antibody test {DFA}), and any coinfection with Candida (Gram stain), bacterial vaginosis (Gram stain), Neisseria gonorrhoeae (Gram stain), Trichomonas vaginalis (wet mount) and HIV (enzyme-linked immunosorbent assay) was ruled out . Flow cytometry was done to investigate changes in immunophenotypes in endocervical secretions and peripheral blood using monoclonal antibodies for surface markers (CD3, CD4, CD8, CD19, CD45 and CD83) . Data were analyzed by chi2 test, while means were compared using Student's t test . RESULTS: C trachomatis positivity was found to be 36% (n = 18) . Forty-eight patients constituted the study population since 2 patients coinfected with Candida, bacterial vaginosis and T vaginalis were excluded . A statistically significant enhancement in CD4+, CD8+ and dendritic cellular phenotypes was observed in the endocervical secretions of Chlamydia-positive patients, while B cells showed no marked difference . In the parallel study of matched peripheral blood, immunophenotypes did not show statistically significant results . CONCLUSION: Increased influx of CD4+, CD8+ and dendritic cells in the endocervix is an indication of cell-mediated immunity in response to C trachomatis infection . Local immune response in the cervical region is independent of systemic response . The mechanism by which local mucosal and systemic immune cells interact to repel or enhance susceptibility to C trachomatis infection requires further study.

Sex Transm Dis, 2004 Oct, 31(10), 628 - 30
Auxo-, sero-, and opa-typing of Neisseria gonorrhoeae strains isolated in Istanbul, Turkey; Aydin D et al.; BACKGROUND: Typing methods are essential in understanding of the transmission dynamics of Neisseria gonorrhoeae . Several typing methods were described including opa-typing . GOALS: The goal of this study was to type all of the strains we isolated in the recent past by using auxo-, sero-, and opa-typing, and to compare the discriminatory power of these methods . STUDY: Auxotyping, serotyping, and opa-typing were performed for 56 N . gonorrhoeae strains isolated from male patients with urethritis . RESULTS: A total of 9 auxotypes and 33 serovars were detected . Combining the 2 systems, a total of 45 distinct auxotype/serovar (A/S) classes were identified . The most common A/S class was NR/Bsty (5 strains) . Fifty-five distinct patterns were detected by opa-typing . Two strains that have been isolated 16 months apart gave identical patterns with opa-typing and their A/S class was also identical (NR/Bsty) . Simpson's index of diversity was found as 0.664, 0.961, 0.987, and 0.999 for auxotyping, serotyping, A/S class, and opa-typing, respectively . CONCLUSIONS: Opa-typing is a potential useful method for typing N . gonorrhoeae as a result of its high discriminatory power, rapidity, ease and relatively lower cost.

Sex Transm Dis, 2004 Oct, 31(10), 616 - 8
High-frequency of quinolone-resistant Neisseria gonorrhoeae in Austria with a common pattern of triple mutations in GyrA and ParC genes; Uthman A et al.; OBJECTIVE: Quinolones have a broad spectrum of antimicrobial activity and are widely used for the treatment of uncomplicated Neisseria gonorrhoeae infections . A dramatic increase in the number of reported N . gonorrhoeae infections as well as quinolone-resistant isolates in Vienna prompted us to investigate the pattern of mutations in these isolates . GOALS: The goal of this study was to investigate the pattern of mutations in GyrA and ParC genes in quinolone-resistant N . gonorrhoeae clinical isolates in Vienna from 1999 to 2002 . STUDY: The antibiotic susceptibility of N . gonorrhoeae clinical isolates and point mutations of the GyrA and ParC genes of 104 clinical isolates were analyzed . RESULTS: Quinolone-resistant N . gonorrhoeae isolates increased from 3.9% (3 of 77) in 1999 to 59.4% (120 of 202) in 2002 . As expected, none of the 46 N . gonorrhoeae quinolone-sensitive strains showed mutations at these positions of GyrA and ParC genes with the exception of 1 isolate, which had a single mutation at GyrA 91 . Unlike what has been previously reported for other geographic areas, 96.6% (56 of 58) of the quinolone-resistant isolates harbored common triple mutations at Gyr 91, 95, and ParC 86 . The majority of these isolates (76.8%) belong to the PPNG phenotype . CONCLUSIONS: Our data indicate that the pattern of mutations in GyrA and ParC subunits of N . gonorrhoeae in Austria differs from that reported from other geographic areas . The differences may either be the result of the difference in bacterial subtypes or various antibiotic regimens used in these regions.

J Biotechnol, 2004 Sep 30, 113(1-3), 15 - 32
Biotechnology and vaccines: application of functional genomics to Neisseria meningitidis and other bacterial pathogens; Serruto D et al.; Since its introduction, vaccinology has been very effective in preventing infectious diseases . However, in several cases, the conventional approach to identify protective antigens, based on biochemical, immunological and microbiological methods, has failed to deliver successful vaccine candidates against major bacterial pathogens . The recent development of powerful biotechnological tools applied to genome-based approaches has revolutionized vaccine development, biological research and clinical diagnostics . The availability of a genome provides an inclusive virtual catalogue of all the potential antigens from which it is possible to select the molecules that are likely to be more effective . Here, we describe the use of "reverse vaccinology", which has been successful in the identification of potential vaccines candidates against Neisseria meningitidis serogroup B and review the use of functional genomics approaches as DNA microarrays, proteomics and comparative genome analysis for the identification of virulence factors and novel vaccine candidates . In addition, we describe the potential of these powerful technologies in understanding the pathogenesis of various bacteria.

J Infect Dis, 2004 Oct 15, 190(8), 1448 - 55 Epub 2004 Sep 13.
Trichomoniasis in men and HIV infection: data from 2 outpatient clinics at Lilongwe Central Hospital, Malawi; Price MA et al.; BACKGROUND: Little is known about the epidemiologic profile of trichomoniasis in men and its relationship to human immunodeficiency virus (HIV) infection . Among men presenting for care for symptomatic sexually transmitted infections (STIs) in Malawi, trichomoniasis is not considered for first-line treatment . METHODS: We conducted a cross-sectional survey of 1187 men attending either a dermatology or STI outpatient clinic in the capital of Malawi . Men were interviewed, and the etiologies of the STIs were determined . RESULTS: At the STI clinic (n = 756 men), we identified 150 men (20%) with Trichomonas vaginalis infection, 358 men (47%) with HIV infection, and 335 men (44%) with Neisseria gonorrhoeae infection . At the dermatology clinic (n = 431 men), we identified 54 (13%), 118 (27%), and 2 (0.5%) men, respectively . At both clinics, a lower education level and reporting never having used a condom were predictive of T . vaginalis infection . Only at the dermatology clinic was older age associated with infection, and only at the STI clinic were marital, genital ulcer disease, and HIV-infection status associated with T . vaginalis infection . At the STI clinic, urethral symptoms attributable to trichomoniasis were more severe among HIV-positive men than among HIV-negative men . CONCLUSIONS: Given its high prevalence and the increased risk for HIV transmission, T . vaginalis infection should be reconsidered for inclusion in the Malawi STI-treatment regimen for men .

J Obstet Gynaecol, 2004 Aug, 24(5), 513 - 5
Prevalence of HIV, syphilis, Chlamydia trachomatis, Neisseria gonorrhoea, Trichomonas vaginalis and candidiasis among pregnant women attending an antenatal clinic in Khartoum, Sudan; Ortashi OM et al.; Antenatal screening and treatment for sexually transmitted infections (STIs) has shown some benefits in countries where the practice is adopted . To date the prevalence of STIs in the Sudan is unknown . Our aims were to establish the prevalence of STIs among pregnant women in Khartoum (Sudan), to identify any risk factors and to suggest management . One hundred and fifty-one pregnant women were recruited for the study in an antenatal clinic in Khartoum in 1999 . The prevalence of Chlamydia trachomatis, HIV, Trichomonas vaginalis, syphilis, Neisseria gonorrhoea and vulvo-vaginal candidiasis among the study population was established; 7.3% of patients suffered multiple infections . No risk factors for acquiring an STI were identified apart from genital ulcer disease . These findings suggest that a syndromic approach for treating STIs during pregnancy might not be possible . We conclude that STIs have a high prevalence among pregnant Sudanese women in Khartoum . The providers of health care in the Sudan need to revise their priority list.

J Infect Chemother, 2004 Aug, 10(4), 208 - 11
Evaluation of a rapid antigen detection test for Neisseria gonorrhoeae in urine sediment for diagnosis of gonococcal urethritis in males; Suzuki K et al.; We evaluated a rapid antigen detection method with an immunochromatographic assay for Neisseria gonorrhoeae (NOW Gonorrhea Test) by using urine samples from patients with urethritis to diagnose gonococcal infection in males . Among 58 male patients who underwent urethral swab culture, 34 cases (58.6%) were found to have N . gonorrhoeae infection . The sensitivity and specificity of the NOW Test compared with the results of standard culture were 94.1% (32/34) and 95.8% (23/24), respectively . The predictive values of positive NOW and negative NOW were 96.9% (32/33) and 92.0% (23/25), respectively . The detection limit of this assay was determined as 5 x 10(4) cfu/ml using N . gonorrhoeae suspension as an antigen . In contrast to standard cultures, gonococcal antigens in specimens were still detectable by this method up to 45 h of storage at either room temperature or 4 degrees C . Considering the rapidity and ease of this method, our results suggest that the NOW method might be a useful and reliable diagnostic screening tool for gonococcal urethritis.

J Clin Microbiol, 2004 Sep, 42(9), 4332 - 4
Multicenter validation of the cppB gene as a PCR target for detection of Neisseria gonorrhoeae; Bruisten SM et al.; The cppB gene is often used as a target for detection of Neisseria gonorrhoeae by PCR . Using a coded panel of 500 DNA samples, we determined that the cppB gene is missing in 5.8% of N . gonorrhoeae strains, and therefore we consider the cppB gene to be an unsuitable target.

Int J Antimicrob Agents, 2004 Sep, 24 Suppl 1, S15 - 22
Antimicrobial resistance of Neisseria gonorrhoeae in Japan, 1993-2002: continuous increasing of ciprofloxacin-resistant isolates; Tanaka M et al.; Susceptibility testing was conducted on 1357 isolates of Neisseria gonorrhoeae isolated from 1993 through 2002 in Japan to assess the antimicrobial resistance . Selected isolates were characterised by auxotype and analysis was done for mutations within the quinolone resistance-determining region (QRDR) in the gyrA and parC genes, which confer fluoroquinolone resistance to the organism . Isolates with ciprofloxacin resistance increased significantly from 6.6% (1993-1994) to 73.5% (2002) . The proportion of plasmid-mediated penicillin-resistant isolates (PPNG) decreased significantly from 7.9% (1993-1994) to 0.9% (2002) . The percentage of chromosomal-mediated resistance to penicillin decreased from 27.4% in 2000 to 12.0% in 2001 but increased to 28.9% in 2002 . The proportion of isolates with any type of resistance to tetracycline decreased from 24.7% in 2000 to 13.9% in 2001 and then increased to 22.3% in 2002 . The proportion of prototrophic isolates significantly decreased from 84.4% in 1992-1993 to 7.7% in 2001, while that of the proline-requiring isolates significantly increased from 4.4% in 1992-1993 and 80.8% in 1998 . The proline-requiring isolates were less susceptible to ciprofloxacin than the prototrophic or arginine-requiring isolates . Of 87 isolates resistant to ciprofloxacin, 2 (2.3%) contained five amino acid substitutions within the GyrA and ParC proteins, 76 (87.4%) contained three or four amino acid substitutions and 9 (10.3%) contained one or two amino acid substitutions.

Int J Antimicrob Agents, 2004 Sep, 24 Suppl 1, S8 - 14
Aetiology of male urethritis in patients recruited from a population with a high HIV prevalence; Sturm PD et al.; The aetiology of urethritis, the significance of potential pathogens and the relation of urethritis to HIV infection were determined in 335 men (cases) with and 100 men (controls) without urethral symptoms . Urethral swab specimens were tested for different organisms by PCR or by culture for Neisseria gonorrhoeae . The prevalence of N . gonorrhoeae and Chlamydia trachomatis was 52 and 16%, respectively . The potential pathogens: Mycoplasma genitalium, Ureaplasma urealyticum, Trichomonas vaginalis and herpes simplex virus (HSV), were present in 5, 36, 6 and 6% of the cases respectively . M . genitalium was the only potential pathogen associated with microscopic urethritis . After excluding gonococcal infections, U . urealyticum was more frequent in symptomatic patients, while the prevalence of T . vaginalis was similar among cases and controls . These results strongly suggest an a etiological role for M . genitalium in male urethritis, a possible role for U . urealyticum, but not for T . vaginalis . The control group, with 97% genital ulcer disease patients, was not suitable for the investigation of the role of HSV . The sero-prevalence of HIV was 45% . Current infections were not associated with HIV . However, a history of previous urethral discharge was associated with HIV in a multivariate analysis and supported the hypothesis that non-ulcerative sexually transmitted diseases facilitate HIV transmission.

FEMS Immunol Med Microbiol, 2004 Oct 1, 42(2), 261 - 6
Retinoic acid treated HL60 cells express CEACAM1 (CD66a) and phagocytose Neisseria gonorrhoeae; Pantelic M et al.; Neisseria gonorrhoeae (gonococci, GC) are phagocytosed by neutrophils through the interaction between opacity proteins (Opa) and the CEA (CD66) family of antigens . In order to study this interaction, we used the human myeloid leukemia HL60 cell line, which differentiates into granulocyte-like cells upon treatment with dimethylsulfoxide (DMSO) or retinoic acid (RA) . We found that RA-, but not DMSO- or untreated-HL60 cells, can phagocytose OpaI-expressing gonococci as well as Escherichia coli . The interaction of OpaI E . coli with RA-treated HL60 cells was inhibited by antibodies against CEACAM1 . Phagocytosis of OpaI E . coli was found to be a result of the expression of CEACAM1 in RA-treated HL60 cells . Our results indicate that the level of expression of CEACAM1 in HL60 cells can be regulated by treatment with RA in a differentiation-dependent manner, and that this is important for phagocytosis of OpaI-expressing gonococci or E . coli.

Clin Nephrol, 2004 Aug, 62(2), 144 - 8
Vasculitic purpura with antineutrophil cytoplasmic antibody-positive acute renal failure in a patient with Streptococcus bovis case and Neisseria subflava bacteremia and subacute endocarditis; Bauer A et al.; Subacute bacterial endocarditis is frequently associated with extracardiac manifestations and renal failure . Clinical variety of endocarditis manifestation is wide and has the potential to mimic vasculitis . Whereas Streptococcus bovis is often isolated and associated with colonic tumors, Neisseriaceae are rarely found . An association of subacute bacterial endocarditis and antineutrophil cytoplasmic antibodies has been described . We report on a 62-year-old man who was admitted to our hospital with acute oliguric renal failure and a nonpruritic purpural rush without fever . Antineutrophil cytoplasmic antibody diagnostic revealed perinuclear staining with a titre of 1 : 512 and antiproteinase-3 specificity . Immune complex-mediated glomerulonephritis without extracapillary proliferation was diagnosed in renal biopsy . Finally, blood cultures became positive for Streptococcus bovis and Neisseria flava . Echocardiography showed mobile vegetations on tricuspid valve . Under treatment with penicillin G and gentamicin, skin efflorescences and renal function recovered, but vegetations increased . A colonic tumor could be excluded, a disastrous dental status may have been a predisposal factor . When classical findings of subacute bacterial endocarditis are less clear, the presence of renal failure and antineutrophil cytoplasmic antibodies in absence of fever may lead to misdiagnosis and deleterious immunosuppressive therapy . Neisseria subflava, an upper respiratory tract commensal, may cause subacute bacterial endocarditis without typical symptoms.

Enferm Infecc Microbiol Clin, 2004 Aug-Sep, 22(7), 392 - 411
{Up to date in sexually transmitted infections: epidemiology, diagnostic approaches and treatments}; Vazquez F et al.; In the last years, there have been important advances in sexually transmitted infections such as genome sequencing of Treponema pallidum, Chlamydia trachomatis or Mycoplasma genitalium; the new taxonomic position of Calymmatobacterium granulomatis; commercial diagnostic systems based on nucleic acid amplification; the emergence of quinolone resistance in Neisseria gonorrhoeae; new therapeutic approaches in vulvovaginal candidiasis that include boric acid; the demonstration that valacyclovir reduces the risk of transmission of genital herpes or the availability of immune-response modifier in the treatment of genital warts, and that are questions in the goal of this review . Viral hepatitis and HIV were no reviewed by space reasons.

Biochemistry, 2004 Sep 14, 43(36), 11616 - 22
Kinetic characterization of the catalytic mechanism of methionine sulfoxide reductase B from Neisseria meningitidis; Olry A et al.; Methionine sulfoxide reductases catalyze the thioredoxin-dependent reduction of methionine sulfoxide back to methionine . The methionine sulfoxide reductases family is composed of two structurally unrelated classes of enzymes named MsrA and MsrB, which display opposite stereoselectivities toward the sulfoxide function . Both enzymes are monomeric and share a similar three-step chemical mechanism . First, in the reductase step, a sulfenic acid intermediate is formed with a concomitant release of 1 mol of methionine per mol of enzyme . Then, an intradisulfide bond is formed . Finally, Msrs return back to reduced forms via reduction by thioredoxin . In the present study, it is shown for the Neisseria meningitidis MsrB that (1) the reductase step is rate-determining in the process leading to formation of the disulfide bond and (2) the thioredoxin-recycling process is rate-limiting . Moreover, the data suggest that within the thioredoxin-recycling process, the rate-limiting step takes place after the two-electron chemical exchange and thus is associated with the release of oxidized thioredoxin.

Microbiology, 2004 Sep, 150(Pt 9), 3001 - 12
Involvement of genes of genome maintenance in the regulation of phase variation frequencies in Neisseria meningitidis; Martin P et al.; In Neisseria meningitidis, the reversible expression of surface antigens, i.e . phase variation, results from changes within repeated simple sequence motifs located in coding or promoter regions of the genes involved in their biosynthesis . The mutation rates of these simple sequences, which have a major influence on the generation of phenotypic diversity, can affect the fitness of the population . The aim of the present study was to investigate the involvement of genetic factors involved (mutS and dam) and not yet analysed (drg and dinB) in the regulation of phase variation frequencies of genes associated with a variety of repeat tracts . The frequency of frameshifts occurring in the polycytidine (polyC) tracts associated with siaD, spr and lgtG and in the tetranucleotide (TAAA) repeat tract associated with nadA was determined by colony immunoblotting or using the lacZ gene as a reporter . Inactivation of mutS increased the frequency of phase variation of genes presenting homopolymeric tracts of diverse length . Overexpression of dinB enhanced the instability of the homopolymeric tract associated with siaD . Investigation of the dam locus in a population of genetically distinct N . meningitidis strains revealed that 27 % of strains associated with invasive disease contained the dam gene . In all strains where a Dam function was absent, the drg gene had been inserted into the dam locus . Disruption of dam and drg in strains representative of each genotype, i.e . dam(+)/drg and dam/drg(+), did not modify phase variation frequencies . In contrast to the effects of certain genes on homopolymeric tracts, none of the genetic factors investigated affected the stability of tetranucleotide repeat tracts.

Plasmid, 2004 Sep, 52(2), 89 - 101
Molecular analysis of oriT and MobA protein in the 7.4 kb mobilizable beta-lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae; Rodriguez-Bonano NM et al.; The mobilization region of the 7.4 kb beta-lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae was characterized . The 3.2 kb HindIII-BamHI fragment of pSJ7.4 was mobilized between Escherichia coli strains by conjugative plasmid RK2 . Selected restriction enzyme-generated deletions of this fragment were subcloned in pACYC177 to obtain constructs that were suitable for analysis of the mobilization region . Mating experiments showed that a highly conserved 1.9 kb DNA region within coordinates 4096-5997 is required for mobilization of pSJ7.4 . This region contains two genes encoding the mobilization protein MobA, another protein, MobC, putatively involved in mobilization from some hosts, and an intergenic oriT . The 168 bp intergenic sequence also contains the promoters for mobA and mobC in an arrangement that suggests divergent transcription and autoregulation from oriT . The 56 kDa MobA was expressed in E . coli as a (6x)His-Tag fusion protein . Purified MobA specifically induced plasmid relaxation by nicking at the oriT . MobA is exceptional because the N-terminal region alone can mobilize pSJ7.4, albeit at a lower frequency than the full-length protein, even in the absence of MobC . The carboxyl terminal region of MobA did not share homology with other mobilization proteins, but may be involved in promoting efficient transfer of pSJ7.4.

Curr Biol, 1993 Sep 1, 3(9), 567 - 72
Ecological separation and genetic isolation of Neisseria gonorrhoeae and Neisseria meningitidis; Vazquez JA et al.; Background: Classifying bacteria into species is problematic . Most microbiologists consider species to be groups of isolates that share some arbitrary degree of relatedness of biochemical or molecular (such as DNA sequence) features and that, ideally, are clearly delineated from all other groups of isolates . The main problem in applying to bacteria a biological concept of species based on the ability or inability of their genes to recombine, is that recombination appears to be rare in bacteria in nature, as indicated by the strong linkage disequilibrium between alleles found in most bacterial populations . However, there are some naturally transformable bacteria in which assortative recombination appears to be so frequent that alleles are in, or close to, linkage equilibrium . For these recombining populations a biological concept of species might be applicable . Results: Populations of Neisseria gonorrhoeae and Neisseria meningitidis from Spain were analysed by multilocus enzyme electrophoresis . The data indicate that assortative recombination occurs frequently within populations, but not between populations . Similarly, the sequences of two house-keeping genes show no evidence of intragenic recombination between N . gonorrhoeae and N . meningitidis . Conclusions: N . gonorrhoeae and N . meningitidis represent extremely closely related 'sexual' populations that appear to be genetically isolated in nature, and thus conform to the biological concept of species . The extreme uniformity of N . gonorrhoeae house-keeping genes suggests that this species may have arisen recently as a clone of N . meningitidis that could colonize the genital tract . Ecological isolation - of populations that can colonize the genital tract from those that can colonize the nasopharynx - may have been an important component in speciation, leading to a lower frequency of recombination between species than within species.

J Microbiol Methods, 2004 Oct, 59(1), 23 - 32
Microscopic quantification of bacterial invasion by a novel antibody-independent staining method; Agerer F et al.; Microscopic discrimination between extracellular and invasive, intracellular bacteria is a valuable technique in microbiology and immunology . We describe a novel fluorescence staining protocol, called FITC-biotin-avidin (FBA) staining, which allows the differentiation between extracellular and intracellular bacteria and is independent of specific antibodies directed against the microorganisms . FBA staining of eukaryotic cells infected with Gram-negative bacteria of the genus Neisseria or the Gram-positive pathogen Staphylococcus aureus are employed to validate the novel technique . The quantitative evaluation of intracellular pathogens by the FBA staining protocol yields identical results compared to parallel samples stained with conventional, antibody-dependent methods . FBA staining eliminates the need for cell permeabilization resulting in robust and rapid detection of invasive microbes . Taken together, FBA staining provides a reliable and convenient alternative for the differential detection of intracellular and extracellular bacteria and should be a valuable technical tool for the quantitative analysis of the invasive properties of pathogenic bacteria and other microorganisms.

Clin Exp Immunol, 2004 Sep, 137(3), 552 - 8
Cytokine expression pattern in the genital tract of Chlamydia trachomatis positive infertile women - implication for T-cell responses; Reddy BS et al.; Human genital infection caused by Chlamydia trachomatis is thought to be immunologically mediated, resulting in local recruitment of lymphocyte subsets and inducing the production of cytokines . Little information is available about the role of lymphocyte recruitment and the regulation of cytokine production in the genital tract of C . trachomatis positive infertile women . We have evaluated the recruitment of lymphocyte subsets in the genital tract and production of Th1/Th2 cytokines in cervical secretions and laparoscopic specimens from the fallopian tubes of C . trachomatis positive infertile women (n = 17) and compared them with controls, viz . C . trachomatis negative infertile women (n = 20) using ELISA and flow cytometry . None of these patients were found to be infected either with Candida sps., bacterial vaginosis, Trichomonas vaginalis, Neisseria gonorrhoeae, Mycoplasma hominis or Ureaplasma urealyticum in the cervix . Flow cytometric analysis of cervical secretions in Chlamydia positive women revealed recruitment of both CD4 and CD8 lymphocytes to the genital tract was up-regulated and a variation in the production rates of different cytokines in cervical secretions and fallopian tube was observed . We found that the immune responses in cervical secretions were of Th0 type, since all the analysed cytokines, viz . IFN-gamma, TNF-alpha, IL-10 and IL-12 were up-regulated . As, both CD4 and CD8 cells contribute to the production of IFN-gamma and IL-10, these results suggest that along with CD4 cells, CD8 lymphocytes also may be important for local regulation of Th1/Th2 responses in the genital tract during C . trachomatis infection.

J Contemp Dent Pract, 2004 Aug 15, 5(3), 150 - 7
Streptococcal gingivitis: a report of case with a description of a unique gingival prosthesis; Cicek Y et al.; Acute streptococcal gingivitis is an acute inflammation of the oral mucosa . Specific bacterial infections of the gingiva may be due to neisseria gonorrhea, treponema pallidum, streptococci, and other organisms . Streptococcal infections are seen rarely . This case report describes a patient who presented with severe gingival inflammation and pain that was diagnosed as an acute streptococcal infection . Bacterial cultures were obtained from the lesion, and biopsies were obtained from the gingiva of lower incisors for histopathologic evaluation . The patient was successfully treated using conventional periodontal therapy (scaling, root planning, curettage) and antibacterial agents . The reconstructive phase for this patient consisted of the fabrication of a heat-cured acrylic gingival facade to mask the gingival recession . The treatment of acute gingivostomatitis is of importance because of the possibility of systemic secondary infections . When esthetics is important, a gingival prostheses can be considered . The differential diagnosis, etiology, and treatment of acute streptococcal gingivitis are discussed and the literature is reviewed in this report.

Microb Pathog, 2004 Aug, 37(2), 55 - 63
A Neisseria gonorrhoeae catalase mutant is more sensitive to hydrogen peroxide and paraquat, an inducer of toxic oxygen radicals; Soler-Garcia AA et al.; Catalase is hypothesized to be critical in the protection of Neisseria gonorrhoeae from H2O2 produced during aerobic respiration and by phagocytes during infection . Here we cloned the catalase (kat) gene of gonococcal strain FA1090 and constructed a genetically defined N . gonorrhoeae kat mutant to assess the role of catalase in defense against oxidative stress . The gonococcal kat gene conferred increased H2O2 resistance to a catalase-deficient Escherichia coli strain . Mutation of the kat gene in strain FA1090 via an in-frame deletion resulted in increased sensitivity to H2O2 and paraquat, an inducer of toxic oxygen radicals . Expression of catalase in trans from a shuttle vector restored catalase activity and paraquat resistance to the kat mutant, but not resistance to H2O2 . The inability to fully complement the mutant was perhaps due to a modification in the catalase, as evidenced by altered mobility of the recombinant catalase on activity gels when expressed from the shuttle vector in N . gonorrhoeae . Additionally, we showed a 262 base pair region upstream of the kat gene is required for expression in E . coli and a putative fumarate-nitrate regulator (FNR) binding site is located in this region .

J Korean Med Sci, 2004 Aug, 19(4), 495 - 500
Screening for Chlamydia and gonorrhea by strand displacement amplification in homeless adolescents attending youth shelters in Korea; Lee SJ et al.; We conducted the screening of sexually transmitted infections to define the prevalence of genital Chlamydia trachomatis and Neisseria gonorrhoeae infections and status of sexual risk behavior among homeless adolescents (10-19 yr old) in Korea . Adolescents who ran away from home and are under the care of youth shelters in ten cities in Korea served as the study population . Participants filled out a self-administered questionnaire related to sexuality . First-void urine was analyzed for chlamydial and gonococcal infection by strand displacement amplification (BDProbTec ET, BD Diagnostic Systems, MD, U.S.A.) . A total of 175 adolescents from 15 youth shelters took part in the study . Their median age was 16 yr, and 54.9% of them reported having sexual intercourse at least once . The prevalence of C . trachomatis and N . gonorrhoeae among homeless adolescents was 12.6% and 15.4%, respectively . Factors significantly associated with the infections were number of sexual partners during the past year and lifetime . This is the first community-based sexually transmitted infection (STI) screening among adolescent in Korea . Screening programs targeting sexually active adolescents are important for detection of STIs . They should be considered an alternative population-based surveillance system in order to control STIs nationally . Copyright The Korean Academy of Medical Sciences

Nucleic Acids Res . 2004 Aug 10;32(14):e114.
An in vitro strategy for the selective isolation of anomalous DNA from prokaryotic genomes; van Passel MW et al.; In sequenced genomes of prokaryotes, anomalous DNA (aDNA) can be recognized, among others, by atypical clustering of dinucleotides . We hypothesized that atypical clustering of hexameric endonuclease recognition sites in aDNA allows the specific isolation of anomalous sequences in vitro . Clustering of endonuclease recognition sites in aDNA regions of eight published prokaryotic genome sequences was demonstrated . In silico digestion of the Neisseria meningitidis MC58 genome, using four selected endonucleases, revealed that out of 27 of the small fragments predicted (<5 kb), 21 were located in known genomic islands . Of the 24 calculated fragments (>300 bp and <5 kb), 22 met our criteria for aDNA, i.e . a high dinucleotide dissimilarity and/or aberrant GC content . The four enzymes also allowed the identification of aDNA fragments from the related Z2491 strain . Similarly, the sequenced genomes of three strains of Escherichia coli assessed by in silico digestion using XbaI yielded strain-specific sets of fragments of anomalous composition . In vitro applicability of the method was demonstrated by using adaptor-linked PCR, yielding the predicted fragments from the N.meningitidis MC58 genome . In conclusion, this strategy allows the selective isolation of aDNA from prokaryotic genomes by a simple restriction digest-amplification-cloning-sequencing scheme.

Vaccine, 2004 Aug 13, 22(23-24), 3045 - 52
Very small size proteoliposomes derived from Neisseria meningitidis: an effective adjuvant for Th1 induction and dendritic cell activation; Mesa C et al.; Recent findings about pathogens and innate immune system interactions have opened new opportunities for adjuvants designs . We have elaborated a new approach, in which gangliosides are incorporated into the outer membrane complex of Neisseria meningitidis (Nm) to form very small size proteoliposomes (VSSP) . VSSP, used as monotherapy, demonstrated a unique ability to render immunogenic highly tolerated gangliosides . These results drove our attention to the immunopotentiating properties of VSSP . Here, we examined the VSSP adjuvant effect on the humoral and cellular responses, dendritic cell (DC) activation, and differentiation of Th cells . Also, the role of LPS in VSSP effect was dissected . This study reveals that VSSP is a potent adjuvant for dendritic cells activation and Th1 differentiation.

Vaccine, 2004 Aug 13, 22(23-24), 2993 - 3003
Preclinical study of influenza virus A M2 peptide conjugate vaccines in mice, ferrets, and rhesus monkeys; Fan J et al.; A universal influenza virus vaccine that does not require frequent updates and/or annual immunizations will offer significant advantages over current seasonal flu vaccines . The highly conserved influenza virus A M2 membrane protein has been previously suggested as a potential antigen target for such a vaccine . Here, we report systematic evaluation of M2 peptide conjugate vaccines (synthetic peptides of M2 extracellular domain conjugated to keyhole limpet hemocyanin (KLH) or Neisseria meningitidis outer membrane protein complex (OMPC)) in mice, ferrets, and rhesus monkeys . The conjugate vaccines were highly immunogenic in all species tested and were able to confer both protection against lethal challenge of either H1N1 or H3N1 virus in mice and reduce viral shedding in the lower respiratory tracts of mice and ferrets . The protection against lethal challenge in mice could also be achieved by passive transfer of monkey sera containing high M2 antibody titers . In addition, we showed that M2 antisera were cross reactive with M2 peptides derived from a wide range of human influenza A strains, but they failed to react with M2 peptides of the pathogenic H5N1 virus (A/Hong Kong/97) . The data presented here will permit better understanding of the potential of an M2-based vaccine approach.

Sex Transm Infect, 2004 Aug, 80(4), 280 - 5
Sexual network analysis of a gonorrhoea outbreak; De P et al.; OBJECTIVES: Sexual partnerships can be viewed as networks in order to study disease transmission . We examined the transmission of Neisseria gonorrhoeae in a localised outbreak in Alberta, Canada, using measures of network centrality to determine the association between risk of infection of network members and their position within the sexual network . We also compared risk in smaller disconnected components with a large network centred on a social venue . METHODS: During the investigation of the outbreak, epidemiological data were collected on gonorrhoea cases and their sexual contacts from STI surveillance records . In addition to traditional contact tracing information, subjects were interviewed about social venues they attended in the past year where casual sexual partnering may have occurred . Sexual networks were constructed by linking together named partners . Univariate comparisons of individual network member characteristics and algebraic measures of network centrality were completed . RESULTS: The sexual networks consisted of 182 individuals, of whom 107 were index cases with laboratory confirmed gonorrhoea and 75 partners of index cases . People who had significantly higher information centrality within each of their local networks were found to have patronised a popular motel bar in the main town in the region (p = 0.05) . When the social interaction through the bar was considered, a large network of 89 individuals was constructed that joined all eight of the largest local networks . Moreover, several networks from different communities were linked by individuals who served as bridge populations as a result of their sexual partnering . CONCLUSION: Asking clients about particular social venues emphasised the importance of location in disease transmission . Network measures of centrality, particularly information centrality, allowed the identification of key individuals through whom infection could be channelled into local networks . Such individuals would be ideal targets for increased interventions.

Can J Microbiol, 2004 Jun, 50(6), 433 - 43
Genetic analysis of Canadian isolates of C:2a:P1.2,5 and B:2a:P1.2,5 Neisseria meningitidis strains belonging to the hypervirulent clone of ET-15; Tyler S et al.; Isolates of the hypervirulent Neisseria meningitidis clone ET-15 found to express the serogroup B antigen were investigated and compared with representative members of serogroup B and C isolates . Clonal-clustering methods clearly grouped the B:ET15 isolates with C:ET15 isolates, indicating the only major difference between the two groups was in the capsule expressed . The organization of the cps operon from the B:ET15 isolates was found to be consistent with typical serogroup B isolates and differed from serogroup C isolates only in the sialyl transferase gene present . This suggests that these strains arose via recombination of the sialyl transferase gene . Specific points of recombination could not be identified, however, the majority (64%) of the B:ET15 isolates contained a copy of pseudo-IS1106 downstream of the cps operon indicating the potential for a common ancestral origin . The combination of pulsed-field gel electrophoresis (PFGE) and sequence analysis of targeted regions of the cps operon permitted the differentiation of most B:ET15 isolates indicating that they likely arose from separate genetic events and do not represent the emergence and spread of a new clone . However, two isolates that appeared identical by all methods employed were temporally and geographically related although no epidemiological evidence is available indicating a link between these strains.

Scand J Clin Lab Invest, 2004, 64(5), 479 - 96
Strategies developed by bacteria and virus for protection from the human complement system; Blom AM; The complement system is an important part of innate immunity providing immediate protection against pathogens without a need for previous exposure . Its importance is clearly shown by the fact that patients lacking complement components suffer from fulminant and recurring infections . Complement is an explosive cascade, and in order to control it there are inhibitors present on every human cell and also circulating in blood . However, many infectious agents have developed strategies to prevent clearance and destruction by complement . Some pathogens simply hijack the host's complement inhibitors, while others are able to produce their own homologues of human inhibitors . Knowledge of these mechanisms on a molecular level may aid development of vaccines and novel therapeutic strategies that would be more specific than the use of antibiotics that, apart from causing resistance problems, also affect the normal flora, the outcome of which could be devastating . In this study the structural requirements and functional consequences of interactions between the major soluble inhibitor of complement C4b-binding protein and Neisseria gonorrhoeae, Bordetella pertussis, Streptococcus pyogenes, Escherichia coli K1, Moraxella catarrhalis and Candida albicans are described . Furthermore, a novel inhibitor produced by Kaposi's sarcoma-associated herpesvirus is identified and characterized in detail: KCP . It is shown that KCP inhibits classical C3-convertase and presents activated complement factors C4b and C3b for destruction by a serine proteinase, factor I . Using molecular modelling and site-directed mutagenesis, it was possible to localize sites on the surface of KCP required for complement inhibition and it is concluded that KCP uses molecular mechanisms identical to human inhibitors.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3185 - 7
Remarkable increase in central Japan in 2001-2002 of Neisseria gonorrhoeae isolates with decreased susceptibility to penicillin, tetracycline, oral cephalosporins, and fluoroquinolones; Ito M et al.; Four hundred sixty-two clinical isolates of Neisseria gonorrhoeae recovered from 1999 through 2002 in central Japan were examined for MICs of antimicrobial agents . The majority was sensitive to ceftriaxone and spectinomycin, but a remarkable increase in isolates with decreased susceptibility to penicillin, tetracycline, oral cephalosporins, and fluoroquinolones was observed from 2001 through 2002.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3103 - 6
Emergence in Italy of a Neisseria meningitidis clone with decreased susceptibility to penicillin; Stefanelli P et al.; A rise in invasive diseases due to Neisseria meningitidis C:2b:P1.5 with decreased penicillin susceptibility occurred in Italy during the last 2 years . Real-time PCR identified the Peni phenotype, and the penA sequence revealed the mosaicism of the gene . Molecular analyses assigned the isolates to a single emergent clone of the hypervirulent A4 cluster.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 2987 - 92
Pharmacodynamic evaluation of the neutralization of endotoxin by PMX622 in mice; Lake P et al.; Polymyxin B (PMB) binds to and neutralizes endotoxin, but its systemic clinical utility is limited by neuro- and nephrotoxicity . PMX622 is a covalent conjugate of PMB and Dextran-70 designed to retain the ability of PMB to neutralize endotoxin and to retain the favorable colloidal, pharmacokinetic, and metabolic properties of Dextran-70 . PMX622 has demonstrated efficacy in a number of animal models and effectively neutralized endotoxin in phase I clinical trials . Here, we systematically evaluated the pharmacodynamic properties of PMX622 in a murine model of endotoxin-induced lethality in galactosamine-sensitized mice . PMX622 completely and dose dependently inhibited lethality in this model . A stoichiometric relationship was found between the endotoxin challenge dose and the dose of PMX622 needed for protection . PMX622 neutralized endotoxin from four different genera of gram-negative bacteria but not Neisseria meningitidis . PMX622 was significantly less toxic than PMB in the mouse, suggesting that PMX622 has a better margin of safety than PMB . The timing of PMX622 administration relative to endotoxin was crucial . PMX622 was active for several hours prior to the endotoxin challenge; however, PMX622 did not protect mice if administered >/=15 min after endotoxin challenge . This suggests that PMX622 would best be clinically used prophylactically rather than therapeutically . These studies will be crucial in designing and interpreting human clinical trials assessing PMX622 efficacy.

Biochem J, 2004 Nov 15, 384(Pt 1), 111 - 7
Distinct classes of glyoxalase I: metal specificity of the Yersinia pestis, Pseudomonas aeruginosa and Neisseria meningitidis enzymes; Sukdeo N et al.; The metalloisomerase glyoxalase I (GlxI) catalyses the conversion of methylglyoxal-glutathione hemithioacetal and related derivatives into the corresponding thioesters . In contrast with the previously characterized GlxI enzymes of Homo sapiens, Pseudomonas putida and Saccharomyces cerevisiae, we recently determined that Escherichia coli GlxI surprisingly did not display Zn2+-activation, but instead exhibited maximal activity with Ni2+ . To investigate whether non-Zn2+ activation defines a distinct, previously undocumented class of GlxI enzymes, or whether the E . coli GlxI is an exception to the previously established Zn2+-activated GlxI, we have cloned and characterized the bacterial GlxI from Yersinia pestis, Pseudomonas aeruginosa and Neisseria meningitidis . The metal-activation profiles for these additional GlxIs firmly establish the existence of a non-Zn2+-dependent grouping within the general category of GlxI enzymes . This second, established class of metal activation was formerly unidentified for this metalloenzyme . Amino acid sequence comparisons indicate a more extended peptide chain in the Zn2+-dependent forms of GlxI (H . sapiens, P . putida and S . cerevisiae), compared with the GlxI enzymes of E . coli, Y . pestis, P . aeruginosa and N . meningitidis . The longer sequence is due in part to the presence of additional regions situated fairly close to the metal ligands in the Zn2+-dependent forms of the lyase . With respect to sequence alignments, these inserts may potentially contribute to defining the metal specificity of GlxI at a structural level.

J Med Chem, 2004 Jul 29, 47(16), 3916 - 9
Phosphorylcholine-carbohydrate-protein conjugates efficiently induce hapten-specific antibodies which recognize both Streptococcus pneumoniae and Neisseria meningitidis: a potential multitarget vaccine against respiratory infections; Bay S et al.; Phosphorylcholine (ChoP) is commonly expressed at the surface of pathogens of the respiratory tract, including Streptococcus pneumoniae and Neisseria meningitidis . We designed a synthetic hapten comprising ChoP and part of its native carrier structure in S . pneumoniae, i.e . N-acetyl-D-galactosamine (GalNAc) . Protein conjugates of this hapten induced GalNAc-ChoP-specific antibodies which recognized ChoP on both S . pneumoniae and N . meningitidis . GalNAc-ChoP could therefore lead to the rational design of a novel multipurpose vaccine against respiratory infections.

Gynecol Obstet Invest, 2004, 58(4), 189 - 93 Epub 2004 Jul 14.
Increased frequency of bacterial vaginosis and Chlamydia trachomatis in pregnant women with human papillomavirus infection; da Silva CS et al.; The aim of this study was to verify the presence of bacterial vaginosis (BV), Candida sp, Chlamydia trachomatis, Neisseria gonorrhoeae, to determine the prevalence of tobacco use and measure vaginal pH (VpH) in pregnant women with (n = 26) and without (n = 26) human papillomavirus (HPV) infection, and make comparisons between these 2 groups . HPV, C . trachomatis and N . gonorrhoeae were diagnosed using hybrid capture, BV using clinical criteria, and Candida sp via cultures . A digital pH meter was used to measure VpH . The frequencies of Candida sp were 19.2 and 23.1% (p = 1), and VpH was 4.4 +/- 0.4 and 4.3 +/- 0.4 (p = 0.23), in the HPV-positive and HPV-negative groups, respectively . Compared to the group of pregnant women without HPV infection, those with HPV infection had a significantly higher prevalence of tobacco use (50 vs . 11.5%; p = 0.006), BV (53.8 vs . 15.4%; p = 0.007), and C . trachomatis (34.6 vs . 7.7%; p = 0.039) . No case of N . gonorrhoeae was diagnosed . All cases of C . trachomatis and BV had high-grade HPV infection . Copyright (c) 2004 S . Karger AG, Basel.

Proc Natl Acad Sci U S A, 2004 Jul 27, 101(30), 10961 - 6 Epub 2004 Jul 15.
A force-dependent switch reverses type IV pilus retraction; Maier B et al.; Type IV pilus dynamics is important for virulence, motility, and DNA transfer in a wide variety of prokaryotes . The type IV pilus system constitutes a very robust and powerful molecular machine that transports pilus polymers as well as DNA through the bacterial cell envelope . In Neisseria gonorrhoeae, pilus retraction is a highly irreversible process that depends on PilT, an AAA ATPase family member . However, when levels of PilT are reduced, the application of high external forces (F = 110 +/- 10 pN) induces processive pilus elongation . At forces of >50 pN, single pili elongate at a rate of v = 350 +/- 50 nm/s . For forces of <50 pN, elongation velocity depends strongly on force and relaxation causes immediate retraction . Both pilus retraction and force-induced elongation can be modeled by chemical kinetics with same step length for the rate-limiting translocation step . The model implies that a force-dependent molecular switch can induce pilus elongation by reversing the retraction mechanism.

Mol Microbiol, 2004 Aug, 53(3), 917 - 27
Analysis in vitro and in vivo of the transcriptional regulator CrgA of Neisseria meningitidis upon contact with target cells; Deghmane AE et al.; Contact between CrgA, a LysR-like regulatory protein in Neisseria meningitidis, and DNA is involved in the repression of several bacterial genes upon contact with epithelial cells . We used a defined in vitro system containing crgA promoter, purified RNA polymerase (RNAP) and purified CrgA protein to demonstrate that CrgA was directly responsible for this transcriptional repression . Interaction between the C-terminal domain of CrgA and the RNAP led to the production of short abortive transcripts, suggesting that CrgA may act by preventing RNAP from clearing the promoter . We probed the regulation by CrgA of its own production by analysing CrgA-DNA contacts during cell-bacteria interaction by assaying in vivo protection against dimethyl sulphate (DMS) methylation . Comparison of DMS footprints in vitro and in vivo suggested that CrgA repressed transcription through specific base contacts, probably in the major groove of the DNA double helix, resulting in DNA looping . Upon contact with target cells, CrgA was released from the DNA, allowing transcription of the target gene to proceed to elongation and facilitating tight control of the expression of genes regulated by CrgA .

J Bioenerg Biomembr, 2000 Dec, 32(6), 563 - 70
Functional Characterization of the Conserved "GLK" Motif in Mitochondrial Porin from Neurospora crassa; Runke G et al.; Mitochondrial porin facilitates the diffusion of small hydrophilic molecules across the mitochondrial outer membrane . Despite low sequence similarity among porins from different species, a "glycine-leucine-lysine" (GLK) motif is conserved in mitochondrial and Neisseria porins . To investigate the possible roles of these conserved residues, including their hypothesized participation in ATP binding by the protein, we replaced the lysine residue of the GLK motif of Neurospora crassa porin with glutamic acid through site-directed mutagenesis of the corresponding gene . Although the pores formed by this protein have size and gating characteristics similar to those of the wild-type protein, the channels formed by GLEporin are less anion selective than the wild-type pores . The GLEporin retains the ability to be cross linked to {alpha-(32)P}ATP, indicating that the GLK sequence is not essential for ATP binding . Furthermore, the pores formed by both GLEporin and the wild-type protein become more cation selective in the presence of ATP . Taken together, these results support structural models that place the GLK motif in a part of the ion-selective beta-barrel that is not directly involved in ATP binding.

J Biol Chem, 2004 Sep 17, 279(38), 39750 - 6 Epub 2004 Jul 14.
Structure of the Neisseria meningitidis outer membrane PilQ secretin complex at 12 A resolution; Collins RF et al.; The bacterial pathogen Neisseria meningitidis expresses long, thin, retractile fibers (called type IV pili) from its cell surface and uses these adhesive structures to mediate primary attachment to epithelial cells during host colonization and invasion . PilQ is an outer membrane protein complex that is essential for the translocation of these pili across the outer membrane . Here, we present the structure of the PilQ complex determined by cryoelectron microscopy to 12 A resolution . The dominant feature of the structure is a large central cavity, formed by four arm features that spiral upwards from a squared ring base and meet to form a prominent cap region . The cavity, running through the center of the complex, is continuous and is effectively sealed at both the top and bottom . Analysis of the complex using self-orientation and by examination of two-dimensional crystals indicates a strong C4 rotational symmetry, with a much weaker C12 rotational symmetry, consistent with PilQ possessing true C4 symmetry with C12 quasi-symmetry . We therefore suggest that the complex is a homododecamer, formed by association of 12 PilQ polypeptide chains into a tetramer of trimers . The structure of the PilQ complex, with its large and well defined central chamber, suggests that it may not function solely as a passive portal in the outer membrane, but could be actively involved in mediating pilus assembly or modification.

Proc Natl Acad Sci U S A, 2004 Jul 20, 101(29), 10798 - 803 Epub 2004 Jul 12.
Unique modifications with phosphocholine and phosphoethanolamine define alternate antigenic forms of Neisseria gonorrhoeae type IV pili; Hegge FT et al.; Several major bacterial pathogens and related commensal species colonizing the human mucosa express phosphocholine (PC) at their cell surfaces . PC appears to impact host-microbe biology by serving as a ligand for both C-reactive protein and the receptor for platelet-activating factor . Type IV pili of Neisseria gonorrhoeae (Ng) and Neisseria meningitidis, filamentous protein structures critical to the colonization of their human hosts, are known to react variably with monoclonal antibodies recognizing a PC epitope . However, the structural basis for this reactivity has remained elusive . To address this matter, we exploited the finding that the PilE pilin subunit in Ng mutants lacking the PilV protein acquired the PC epitope independent of changes in pilin primary structure . Specifically, we show by using mass spectrometry that PilE derived from the pilV background is composed of a mixture of subunits bearing O-linked forms of either phosphoethanolamine (PE) or PC at the same residue, whereas the wild-type background carries only PE at that same site . Therefore, PilV can influence pilin structure and antigenicity by modulating the incorporation of these alternative modifications . The disaccharide covalently linked to Ng pilin was also characterized because it is present on the same peptides bearing the PE and PC modifications and, contrary to previous reports, was found to be linked by means of 2,4-diacetamido-2,4,6-trideoxyhexose . Taken together, these findings provide new insights into Ng type IV pilus structure and antigenicity and resolve long-standing issues regarding the nature of both the PC epitope and the pilin glycan.

Biochemistry, 2004 Jul 20, 43(28), 9195 - 203
Specificity of the synergistic anion for iron binding by ferric binding protein from Neisseria gonorrhoeae; Bekker EG et al.; Ferric binding protein in Neisseria gonorrhoeae (nFbpA) transports iron from outer membrane receptors for host proteins across the periplasm to a permease in an alternative pathway to the use of siderophores in some pathogenic bacteria . Phosphate and nitrilotriacetate, both at pH 8, and vanadate at pH 9 are shown to be synergistic in promoting ferric binding to nFbpA, in contrast to carbonate and sulfate . Interestingly, only phosphate produces the fully closed conformation of nFbpA as defined by native electrophoresis . The role of phosphate was probed by constructing three mutants: Q58E, Q58R, and G140H . The anion and iron binding properties of the Q58E mutant are similar to the wild-type protein, implying that one phosphate oxygen is a hydrogen bond donor and may in part define the specificity of nFbpA for phosphate over sulfate . Phosphate is a weakly synergistic anion in the Q58R and G140H mutants, and these mutants do not form completely closed structures . Ferric binding was investigated by both isothermal titration and differential scanning calorimetry . The apparent affinity of nFbpA for iron in a solution of 30 mM citrate is 1 order of magnitude larger in the presence (K(app)= 1.7 x 10(5) M(-1)) of phosphate than in its absence (K(app) = 1.6 x 10(4) M(-1)) at pH 7 . Similar results were obtained at pH 8 . This increase in affinity with phosphate as well as the formation of closed structure allows nFbpA to compete for free ferric ions in solution and suggests that ferric binding to nFbpA is regulated by the synergistic phosphate anion at sites of iron uptake.

Eur J Clin Microbiol Infect Dis, 2004 Sep, 23(9), 705 - 10 Epub 2004 Jul 10.
A new confirmatory Neisseria gonorrhoeae real-time PCR assay targeting the porA pseudogene; Whiley DM et al.; The Roche Cobas Amplicor system is widely used for the detection of Neisseria gonorrhoeae but is known to cross react with some commensal Neisseria spp . Therefore, a confirmatory test is required . The most common target for confirmatory tests is the cppB gene of N . gonorrhoeae . However, the cppB gene is also present in other Neisseria spp . and is absent in some N . gonorrhoeae isolates . As a result, laboratories targeting this gene run the risk of obtaining both false-positive and false-negative results . In the study presented here, a newly developed N . gonorrhoeae LightCycler assay (NGpapLC) targeting the N . gonorrhoeae porA pseudogene was tested . The NGpapLC assay was used to test 282 clinical samples, and the results were compared to those obtained using a testing algorithm combining the Cobas Amplicor System (Roche Diagnostics, Sydney, Australia) and an in-house LightCycler assay targeting the cppB gene (cppB-LC) . In addition, the specificity of the NGpapLC assay was investigated by testing a broad panel of bacteria including isolates of several Neisseria spp . The NGpapLC assay proved to have comparable clinical sensitivity to the cppB-LC assay . In addition, testing of the bacterial panel showed the NGpapLC assay to be highly specific for N . gonorrhoeae DNA . The results of this study show the NGpapLC assay is a suitable alternative to the cppB-LC assay for confirmation of N . gonorrhoeae-positive results obtained with Cobas Amplicor .

Tissue Antigens, 2004 Aug, 64(2), 111 - 8
CD46: a complement regulator and pathogen receptor that mediates links between innate and acquired immune function; Russell S; In the last 10 years, the human cell-surface molecule, CD46, has evolved from 'just another complement regulator' to a receptor for a striking array of pathogens . CD46 not only protects cells from complement-mediated attack and facilitates infection by a large number of pathogens, but also exerts complex effects on cellular immune function . It has been proposed that CD46 links innate and adaptive immunity by affecting cellular immune function in response to complement binding, and the role of CD46 in the pathogenesis of many infectious pathogens is now the subject of intense investigation . So far, the flood of information that implicates CD46 in modifying a host response to measles, Neisseria, human herpes virus 6, and pathogens that activate complement has not yet been matched with a comprehensive understanding of the molecular mechanisms by which CD46 affects immune function . This review summarizes the evidence that points to a significant role for CD46 in a range of pathological processes and describes how CD46 might exert its effects by altering signal transduction and antigen presentation pathways.

J Clin Microbiol, 2004 Jul, 42(7), 3300 - 2
Performance of the MagNA pure LC robot for extraction of Chlamydia trachomatis and Neisseria gonorrhoeae DNA from urine and swab specimens; Dalesio N et al.; DNA from uncentrifuged urines (n = 195 and n = 202) and cervical swabs (n = 221 and n = 601) was extracted by the MagNA Pure LC robot and the Amplicor method to validate the robot's extraction for Chlamydia trachomatis and Neisseria gonorrhoeae testing by Roche PCR . The robot provided a highly sensitive and specific method of DNA extraction.

J Clin Microbiol, 2004 Jul, 42(7), 3281 - 3
Use of applied biosystems 7900HT sequence detection system and Taqman assay for detection of quinolone-resistant Neisseria gonorrhoeae; Giles J et al.; Mutations in quinolone resistance-determining regions (QRDRs) have been associated with quinolone-resistant Neisseria gonorrhoeae (QRNG) . Since diagnostic nucleic acid amplification tests for gonococci are now in frequent use, molecular detection of QRNG could facilitate surveillance in the absence of culture . Here we describe a real-time molecular assay for detecting QRDR mutations in gonococci.

J Clin Microbiol, 2004 Jul, 42(7), 3089 - 93
Comparison of the APTIMA CT and GC assays with the APTIMA combo 2 assay, the Abbott LCx assay, and direct fluorescent-antibody and culture assays for detection of Chlamydia trachomatis and Neisseria gonorrhoeae; Boyadzhyan B et al.; The Gen-Probe APTIMA Combo 2 (AC2) is a Food and Drug Administration-cleared nucleic acid amplification test (NAAT) for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae from urine and urogenital swab specimens . The Centers for Disease Control and Prevention have recommended confirmation of positive NAAT results in low-prevalence populations . APTIMA CT (ACT) and APTIMA GC (AGC) are two discrete NAATs for C . trachomatis and N . gonorrhoeae detection that are suitable for confirming AC2-positive results because they target different nucleic acid sequences . Our objective was to determine if ACT and AGC could be used as confirmatory tests for AC2 and to correlate the APTIMA assays with culture, direct fluorescent-antibody (DFA), and LCx CT and GC assays . Urine and swab specimens (1,304) were initially tested with either culture, DFA, or LCx, followed by AC2 . A subset (675) was then tested with ACT and AGC . There was absolute concordance between ACT-AGC and AC2 . LCx did not detect 1 of 14 AC2-ACT- and 1 of 6 AC2-AGC-positive urine samples, and it yielded one C . trachomatis- and one N . gonorrhoeae-positive swab result that were not detected by AC2 and ACT-AGC . Culture failed to detect 5 of 20 AC2-ACT and 3 of 4 AC2-AGC positives, and DFA missed 4 of 4 AC2-ACT positives . Thus, ACT and AGC relative sensitivity compared to that of AC2 was 100% . All APTIMA assays detected more confirmed positive results than culture, DFA, and LCx . The performance of APTIMA assays was not altered by the use of various swab types and by long-term storage of specimens . All APTIMA assays are highly sensitive and rapid . ACT and AGC can be recommended for confirmation of positive results from other NAATs, such as AC2 and LCx.

J Clin Microbiol, 2004 Jul, 42(7), 2926 - 34
Molecular typing of Neisseria gonorrhoeae isolates by pyrosequencing of highly polymorphic segments of the porB gene; Unemo M et al.; For prevention and control of gonorrhea, an objective, highly discriminating, and reproducible molecular epidemiological characterization of Neisseria gonorrhoeae is essential . In the present study, in pursuance of providing such qualities, pyrosequencing technology, a fast real-time DNA sequence analysis, was applied to six short, highly polymorphic porB gene segments, with subsequent genetic variant (genovar) determination of the bacterial isolates . The sequencing templates were obtained by real-time PCR amplification, which also included fluorescence melting curve analysis of the entire porB gene in order to determine the genogroup (porB1a or porB1b allele) prior to pyrosequencing analysis . The PSQ 96 MA system used allowed rapid (in approximately 1.5 h) determination of 96 sequences of 20 to 65 correct nucleotides each . The results were reproducible and mostly in concordance with the results of conventional Sanger dideoxy sequencing, with the exception of shorter read lengths and some uncertainty in determining the correct number of identical nucleotides in homopolymeric segments . The number of sequence variants identified in each of the six highly polymorphic segments of the porB1a and porB1b alleles (encoding surface-exposed amino acid loops of the mature PorB protein) ranged from 5 to 11 and from 8 to 39, respectively . Among porB1a isolates (n = 22) and porB1b isolates (n = 65), 22 and 64 unique genovars, respectively, were identified . All isolates were typeable . The present results provide evidence of a high discriminatory ability, practically the same as that for sequencing of the entire porB gene . In conclusion, the fast and high-throughput pyrosequencing technology can be used for molecular epidemiological characterization of N . gonorrhoeae.

Electrophoresis, 2004 Jul, 25(13), 2017 - 25
Application of capillary electrophoresis- electrospray-mass spectrometry to the separation and characterization of isomeric lipopolysaccharides of Neisseria meningitidis; Li J et al.; A capillary electrophoresis-electrospray-mass spectrometry technique for the characterization of lipopolysaccharides (LPSs) was developed, permitting the separation of trace-level O-deacylated LPS isoforms for subsequent structural characterization using tandem mass spectrometry (MS/MS) . The separation buffer and electrospray interface were optimized first using O-deacylated LPS samples from large-scale preparations . It was found that with microelectrospray or sheath-solution interface, we could separate LPS in anionic forms and detect them using either negative or positive ion mode MS . For negative ion detection mode MS, 30 mM morpholine with addition of 5% v/v methanol was employed as separation buffer . When positive ion detection mode MS was required, 10 mM ammonium acetate with addition of 5% methanol was used as separation buffer . The structural assignments obtained from MS/MS and capillary zone electrophoresis-electrospray-MS (CZE-ESMS) analyses enabled the identification of isomeric glycoforms . Application of this technique to the analysis of LPS from the galE mutants of Neisseria meningitidis strain BZ157 B5+ revealed the presence of isomeric glycoforms, in which the location of a functional group phosphoethanolamine was found to be in either inner core or lipid A-OH regions . The described technique was also applied to the analysis of LPS samples from the galE mutant of N . meningitidis strains F1576 A4+ and A4- . The occurrence of isomeric LPS glycoforms differing by the location or presence of neutral sugar residues, such as hexoses, can also be characterized using MS/MS.

J Antimicrob Chemother, 2004 Aug, 54(2), 451 - 5 Epub 2004 Jul 01.
Epidemiological characteristics and molecular basis of fluoroquinolone-resistant Neisseria gonorrhoeae strains isolated in Korea and nearby countries; Yong D et al.; OBJECTIVES: This study was performed to examine the cause of the increase in quinolone-resistant Neisseria gonorrhoeae (QRNG) observed in Korea . METHODS: The antimicrobial susceptibilities of 190 isolates of gonococci from Korea in 2000 were examined by NCCLS methods, and subsets of these isolates underwent mutation analysis of the quinolone resistance-determining regions (QRDRs) of gyrA and parC . Molecular epidemiological characterization of 25 Korean isolates and 54 isolates from overseas was performed by pulsed-field gel electrophoresis (PFGE) and the results compared . RESULTS: Most (172, 90.5%) of the 190 gonococci tested displayed reduced susceptibility to ciprofloxacin . All strains with high-level ciprofloxacin resistance (ciprofloxacin MIC >/= 4 mg/L) contained a double amino acid alteration at the 91 and 95 positions in the QRDR of GyrA and a single alteration in ParC . PFGE types of high-level QRNG in Korea were mostly different from those of other nearby countries . CONCLUSIONS: These results suggest that the observed increase in ciprofloxacin-resistant isolates is due to the mutation and spread of Korean multiclonal isolates rather than importation from overseas.

Zhonghua Liu Xing Bing Xue Za Zhi, 2004 May, 25(5), 391 - 5
{Epidemiologic surveillance for bacterial meningitis in 140 000 children under 5 years of age in Nanning district, Guangxi province}; Dong BQ et al.; OBJECTIVE: To characterize the incidence, epidemiologic features, etiologic agents and sequelae of bacterial meningitis in children under 5 years of age in Nanning, Guangxi . METHODS: A population-based surveillance was conducted to evaluate children with signs and symptoms of meningitis . All hospitals, township health centers and village clinics in the surveillance area were structured to participate in the case referral and evaluation . Cerebrospinal fluid (CSF) and blood specimens were obtained and processed using standardized microbiologic methods . RESULTS: During the 26-month surveillance period, among the children under 5 years old, a total of 1272 cases who met the screening criteria of meningitis were studied . 265 of 1272 cases were identified as clinically diagnosed meningitis, with an incidence rate of 86.36 per 100 000 population . The annual incidence rate under the 38 cases of confirmed bacterial meningitis appeared to be 12.38/100 000 . Staphylococcus species accounted for the largest proportion of laboratory-confirmed bacterial meningitis, followed by E . coli and S . pneumoniae . The highest attack rate occurred in neonates < 1 month, followed by children aged 1 - 12 months in the confirmed patients . Meningitis caused by Sp and Hi mainly occurred in children aged 1 - 12 months . All cases of meningitis due to Hi and Sp were children aged 1 - 24 months . 13.16% and 0.00% of the cases survived with complications and sequelae, and the case-fatality rate was 18.42% . 40 bacterial isolates were identified from 1193 blood cultures and 23 from 1211 cerebrospinal fluid samples, but no Neisseria meningitidis was found . CONCLUSION: Meningitis due to Hi was first confirmed in Guangxi with the incidence of 0.98 per 100 000 population . The annual incidence rate of confirmed bacterial meningitis was 12.38 per 100 000, which was considered an important public health problem in children . Staphylococci was the predominant pathogen in confirmed bacterial meningitis.

Int J STD AIDS, 2004 Jul, 15(7), 453 - 8
The management of uncomplicated adult gonococcal infection: should test of cure still be routine in patients attending genitourinary medicine clinics?
Harry C.
In this retrospective study of the outcome of treatment of 245 patients (87 females and 158 males) with a diagnosis of Neisseria gonorrhoeae infection seen between 1996 and 2002, 81% (95% confidence interval (CI) 74%-86.8%) of the males and 88.5% (95% CI 79.9%-94.3%) of the females attended for a test of cure . At initial presentation, 93% (95% CI 87.9%-96.5%) and 94.3% (95% CI 89.5%-97.4%) of males respectively, had symptoms and signs of gonococcal infection compared with 48.3% (95% CI 37.4%-59.2%) and 44.8% (95% CI 34.1%-55.9%) of females, and this difference was statistically significant (P = 0.005) . Initial diagnosis at first visit was made by Gram-stained smear in 88.6% (95% CI 82.6%-93.1%) of males and 32.2% (95% CI 22.6%-43.1%) of females, a statistically significant difference P = 0.001 . There were 12 (4.9%) cases of reinfection that rebooked to attend after failing to attend for their test of cure in two females and 10 males . There were two (0.8%) treatment failures amongst the 245 episodes in two males who still had symptoms when they returned for their test of cure . One male had a ciprofloxacin-resistant strain (CRNG) acquired locally and the other one had a betalactamase-producing CRNG/penicillinase-producing N . gonorrhoeae (PPNG) isolate acquired abroad in South America . These patients would have sought to return, as they still had signs and symptoms of gonococcal infection, and they would have been recalled following receipt of the antimicrobial susceptibility report . Post-gonococcal urethritis occurred in 36.3% (95% CI 27.8%-45.4%) of the males who attended for their test of cure, 74.8% (95% CI 67.2%-81.5%) received anti-chlamydial therapy with their standard treatment . In men who received anti-chlamydial therapy the odds ratio of having post-gonococcal urethritis was 0.42 (95% CI 0.17-1.06), P = 0.04 . Co-infection with Chlamydia trachomatis was more likely to occur amongst females (43.9%), odds ratio 3.97 (95% CI 2.07-7.67), P < 0.001 than males (16.5%) . We have now discontinued routine attendance for a test of cure and encourage our patients to telephone for their results with recall only of patients whose antimicrobial susceptibility indicate inappropriate first line therapy or who are still symptomatic.

J Leukoc Biol, 2004 Sep, 76(3), 577 - 84 Epub 2004 Jun 24.
Activation of murine macrophages by Neisseria meningitidis and IFN-gamma in vitro: distinct roles of class A scavenger and Toll-like pattern recognition receptors in selective modulation of surface phenotype; Mukhopadhyay S et al.; Innate and adaptive immune activation of macrophages (Mphi) by microorganisms and antigen-activated lymphoid cells, respectively, plays an important role in host defense and immunopathology . Antigen-presenting cells express a range of pattern recognition receptors including the class A types I and II scavenger receptors (SR-A) and Toll-like receptors (TLR) . Recognition of microbial products by SR-A and TLR controls uptake, killing, altered gene expression, and the adaptive immune response; however, the contribution of each receptor and interplay with cytokine stimuli such as interferon-gamma (IFN-gamma) are not defined . We used Neisseria meningitidis (NM), a potent activator of innate immunity, and IFN-gamma, a prototypic T helper cell type 1 proinflammatory cytokine, to compare surface antigens, secretion of mediators, and receptor functions in elicited peritoneal Mphi from wild-type and genetically modified mouse strains . We show that these stimuli regulate major histocompatibility complex type II (MHC-II) and costimulatory molecules differentially, as well as expression of the mannose receptor and of Mphi receptor with collagenous structure (MARCO), a distinct SR-A, which provides a selective marker for innate activation . In combination, NM inhibited up-regulation of MHC-II by IFN-gamma while priming enhanced release of tumor necrosis factor alpha and nitric oxide . The SR-A contributes to phagocytosis of the organisms but not to their ability to induce CD80, CD86, and MARCO or to inhibit MHC-II . Conversely, studies with lipopolysaccharide (LPS)-deficient organisms and/or TLR-4 mutant mice showed that LPS and TLR-4 are at least partially required to induce CD80, CD86, and MARCO, but LPS is not required to inhibit MHC-II . These studies provide an experimental model and identify surface markers for analysis of innate and acquired immune activation of Mphi.

Sex Transm Dis, 2004 Jul, 31(7), 443 - 7
Feasibility and short-term impact of linked education and urine screening interventions for Chlamydia and gonorrhea in male army recruits; Arcari CM et al.; OBJECTIVE: The objective of this study was to assess the feasibility of an intervention for sexually transmitted diseases (STDs) and a screening program for Chlamydia trachomatis and Neisseria gonorrhoeae infections in male Army recruits . GOALS: The goals of this study were to identify and treat chlamydia and gonorrhea infections in recruits, assess their perceptions of risk, and increase their STD knowledge and behavioral intentions . STUDY DESIGN: Volunteers (n = 3911) entering basic training (July 1999-June 2000) at Fort Jackson, South Carolina, attended an educational intervention, completed pre- and post-questionnaires, and provided a urine specimen for chlamydia and gonorrhea screening by nucleic acid amplification testing . RESULTS: Chlamydia and gonorrhea prevalences were 4.7% and 0.4%, respectively . The mean STD knowledge score, intent to use condoms, and confidence in using condoms correctly increased (P <0.001) . Participants reported increased risk perception and considered the educational program valuable (96.9%) and a learning experience (94.6%) . CONCLUSIONS: A linked educational and screening program is feasible and acceptable in male Army recruits.

Sex Transm Dis, 2004 Jul, 31(7), 437 - 42
Do people really know their sex partners? Concurrency, knowledge of partner behavior, and sexually transmitted infections within partnerships; Drumright LN et al.; BACKGROUND AND OBJECTIVE: An individual's risk of sexually transmitted infections (STIs) has been associated empirically with the individual having concurrent sexual partners (individual's concurrency) and, theoretically, with the individual's partner having concurrent partners (partner's concurrency) . GOALS: The goals of this study were to assess the relationship of STI to individual's concurrency, the partner's concurrency, and awareness of the partner's concurrency . STUDY DESIGN: We recruited 192 individuals aged 18 to 30 from sexually transmitted disease and family planning clinics in 96 partner dyads that reported first sexual contact during the previous 3 months . All individuals underwent computer interviews and testing for Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis . Individuals' self-reports of concurrency and perceptions of partners' concurrency were compared within dyads by kappa statistic; associations of STI with individual's concurrency, partner's concurrency, and knowledge of partner's concurrency were examined using chi-squared analysis, t tests, and multivariate logistic regression . RESULTS: Only 26% of individuals whose partners had other partners were aware of this (kappa = 0.17 for agreement of perceptions vs . partners' reports of concurrency) . In multivariate models, STI in individuals was independently associated with partners' concurrency (odds ratio {OR}, 3.6), lack of awareness of partner's concurrency (OR, 4.5), perceiving a partner to have concurrent partners when the partner did not (OR, 4.7), living in south San Diego, and sexual contact within 1 week of acquaintance, but not with individuals' concurrency . CONCLUSION: This study demonstrates that STI is associated with partner's concurrency and with not knowing one's partner's behavior.

BMC Microbiol . 2004 Jun 22;4(1):23.
The outer membrane protein Omp35 affects the reduction of Fe(III), nitrate, and fumarate by Shewanella oneidensis MR-1; Maier TM et al.; BACKGROUND: Shewanella oneidensis MR-1 uses several electron acceptors to support anaerobic respiration including insoluble species such as iron(III) and manganese(IV) oxides, and soluble species such as nitrate, fumarate, dimethylsulfoxide and many others . MR-1 has complex branched electron transport chains that include components in the cytoplasmic membrane, periplasm, and outer membrane (OM) . Previous studies have implicated a role for anaerobically upregulated OM electron transport components in the use of insoluble electron acceptors, and have suggested that other OM components may also contribute to insoluble electron acceptor use . In this study, the role for an anaerobically upregulated 35-kDa OM protein (Omp35) in the use of anaerobic electron acceptors was explored . RESULTS: Omp35 was purified from the OM of anaerobically grown cells, the gene encoding Omp35 was identified, and an omp35 null mutant (OMP35-1) was isolated and characterized . Although OMP35-1 grew on all electron acceptors tested, a significant lag was seen when grown on fumarate, nitrate, and Fe(III) . Complementation studies confirmed that the phenotype of OMP35-1 was due to the loss of Omp35 . Despite its requirement for wild-type rates of electron acceptor use, analysis of Omp35 protein and predicted sequence did not identify any electron transport moieties or predicted motifs . OMP35-1 had normal levels and distribution of known electron transport components including quinones, cytochromes, and fumarate reductase . Omp35 is related to putative porins from MR-1 and S . frigidimarina as well as to the PorA porin from Neisseria meningitidis . Subcellular fraction analysis confirmed that Omp35 is an OM protein . The seven-fold anaerobic upregulation of Omp35 is mediated post-transcriptionally . CONCLUSION: Omp35 is a putative porin in the OM of MR-1 that is markedly upregulated anaerobically by a post-transcriptional mechanism . Omp35 is required for normal rates of growth on Fe(III), fumarate, and nitrate, but its absence has no effect on the use of other electron acceptors . Omp35 does not contain obvious electron transport moieties, and its absence does not alter the amounts or distribution of other known electron transport components including quinones and cytochromes . The effects of Omp35 on anaerobic electron acceptor use are therefore likely indirect . The results demonstrate the ability of non-electron transport proteins to influence anaerobic respiratory phenotypes.

Chemotherapy, 2004 Jun, 50(2), 92 - 7
A comparison of the microbiological characteristics of Neisseria gonorrhoeae isolated from male and female patients with gonorrhea; Saika T et al.; BACKGROUND: A trend towards a decrease in gonorrhea in Japan was observed until the mid-1990s, but soon after that period, an increased incidence of gonorrhea was reported . METHODS: Antimicrobial susceptibility, auxotype and the type of gyrA and parC mutations were compared between 200 and 132 Neisseria gonorrhoeae isolates from male and female patients with gonorrhea, respectively . RESULTS AND CONCLUSION: The isolation frequency of ciprofloxacin-resistant isolates from male patients was higher than that from female patients (19.5 and 13.6%, respectively) . All ciprofloxacin-resistant isolates had single to triple mutations in gyrA and/or parC genes . N . gonorrhoeae isolates showed a predominance of Pro-requiring auxotypes (46.5 and 46.2% of the isolates from male and female patients, respectively) . Of the Pro-requiring isolates, the incidence of ciprofloxacin-resistant isolates was higher in the isolates from male patients than in the isolates from female patients .

Emerg Infect Dis, 2004 May, 10(5), 936 - 7
Neisseria meningitidis W135, Turkey; Doganci L et al.; We describe the first case of Neisseria meningitidis W135 meningitis in Turkey . The strain was genotypically unrelated to the clone (W)ET-37, isolated from Hajj pilgrims in 2000.

J Infect Dis, 2004 Jul 1, 190(1), 136 - 47 Epub 2004 Jun 08.
Defenses against oxidative stress in Neisseria gonorrhoeae and Neisseria meningitidis: distinctive systems for different lifestyles; Seib KL et al.; Defenses against oxidative stress are crucial for the survival of the pathogens Neisseria meningitidis and Neisseria gonorrhoeae . An Mn(II) uptake system is involved in manganese (Mn)-dependent resistance to superoxide radicals in N . gonorrhoeae . Here, we show that accumulation of Mn also confers resistance to hydrogen peroxide killing via a catalase-independent mechanism . An mntC mutant of N . meningitidis is susceptible to oxidative killing, but supplementation of growth media with Mn does not enhance the organism's resistance to oxidative killing . N . meningitidis is able to grow in the presence of millimolar levels of Mn ion, in contrast to N . gonorrhoeae, whose growth is retarded at Mn concentrations >100 micromol/L, indicating that Mn homeostasis in the 2 species is probably quite different . N . meningitidis superoxide dismutase B plays a role in protection against oxidative killing . However, a sodC mutant of N . meningitidis is no more sensitive to oxidative killing than is the wild type . A cytochrome c peroxidase (Ccp) is present in N . gonorrhoeae but not in N . meningitidis . Investigations of a ccp mutant revealed a role for Ccp in protection against hydrogen peroxide killing . These differences in oxidative defenses in the pathogenic Neisseria are most likely a result of their localization in different ecological niches.

Mol Ther, 2004 Jun, 9(6), 775 - 85
Novel mouse model for carcinoembryonic antigen-based therapy; Chan CH et al.; Many novel cancer therapies, including immunotherapy and gene therapy, are specifically targeted to tumor-associated molecules, among which carcinoembryonic antigen (CEA) represents a popular example . Discrepancies between preclinical experimental data in animal models and clinical outcome in terms of therapeutic response and toxicity, however, often arise . Preclinical testing can be compromised by the lack of CEA and other closely related human CEA family members in rodents, which lack analogous genes for most human CEA family members . Here, we report the construction of a transgenic mouse with a 187-kb human bacterial artificial chromosome (CEABAC) that contains part of the human CEA family gene cluster including complete human CEA (CEACAM5), CEACAM3, CEACAM6, and CEACAM7 genes . The spatiotemporal expression pattern of these genes in the CEABAC mice was found to be remarkably similar to that of humans . This novel mouse will ensure better assessment than previously utilized models for the preclinical testing of CEA-targeted therapies and perhaps allow the testing of CEACAM6, which is overexpressed in many solid tumors and leukemias, as a therapeutic target . Moreover, expression of CEA family genes in gastrointestinal, breast, hematopoietic, urogenital, and respiratory systems could facilitate other clinical applications, such as the development of therapeutic agents against Neisseria gonorrhoeae infections, which use CEA family members as major receptors.

Proc Natl Acad Sci U S A, 2004 Jun 22, 101(25), 9417 - 22 Epub 2004 Jun 10.
Identification of an outer membrane protein required for the transport of lipopolysaccharide to the bacterial cell surface; Bos MP et al.; Lipopolysaccharide (LPS), also known as endotoxin due to its severe pathophysiological effects in infected subjects, is an essential component of the outer membrane (OM) of most Gram-negative bacteria . LPS is synthesized in the bacterial inner membrane, a process that is now well understood . In contrast, the mechanism of its transport to the outer leaflet of the OM has remained enigmatic . We demonstrate here that the OM protein, known as increased membrane permeability (Imp) or organic solvent tolerance protein, is involved in this process . An Imp-deficient mutant of Neisseria meningitidis was viable and produced severely reduced amounts of LPS . The limited amount of LPS that was still produced was not accessible to LPS-modifying enzymes expressed in the OM or added to the extracellular medium . We conclude therefore that Imp mediates the transport of LPS to the cell surface . The role of Imp in LPS biogenesis and its high conservation among Gram-negative bacteria make it an excellent target for the development of novel antibacterial compounds.

Cell Microbiol, 2004 Jul, 6(7), 663 - 70
Epithelial cell responses induced upon adherence of pathogenic Neisseria; Plant L et al.; Neisseria meningitidis and Neisseria gonorrhoeae colonize human mucosal surfaces and cause sepsis/meningitis and gonorrhoea respectively . The first step in the infection process is pilus-mediated adhesion of the bacteria to epithelial cells, followed by host cell invasion . Adhesion of pathogenic Neisseria elicits multiple responses in host cells, including cellular signalling events, cytokine production and modulation of the eukaryotic cell surface . We used microarrays to assess the respective involvement of 375 human cytokine and adhesion related genes during adhesion of piliated and non-piliated N . gonorrhoeae, and piliated encapsulated N . meningitidis to the epithelial cell line ME-180 . We identified 29 differentially regulated genes not previously reported to respond to neisserial infections, many of which encode membrane proteins . Selected genes were further analysed by semiquantitative RT-PCR, and protein expression was examined by flow cytometry . We found that N . gonorrhoeae elicited a different inflammatory response than N . meningitidis and we also demonstrated that early adhesion events are responsible for the induction of specific genes . Our data create a new platform for elucidating the interaction between pathogenic Neisseria and target cells.

Cell Microbiol, 2004 Jul, 6(7), 625 - 37
Human dendritic cell activation by Neisseria meningitidis: phagocytosis depends on expression of lipooligosaccharide (LOS) by the bacteria and is required for optimal cytokine production; Uronen-Hansson H et al.; Group B Neisseria meningitidis is a human pathogen, for which a universally effective vaccine is still not available . Immune responses to bacteria are initiated by dendritic cells (DC), which internalize and process bacterial antigens for presentation to T cells . We show here that optimal IL-12 and TNF-alpha production by human monocyte derived DC in response to killed serogroup B N . meningitidis depends on physical contact and internalization of the bacteria by DC . The majority of DC producing cytokines had internalized N . meningitidis while inhibition of bacterial internalization markedly impaired IL-12 and TNF-alpha, but not IL-6 production . Internalization of N . meningitidis was shown to depend on lipooligosaccharide (LOS) expressed by the bacteria with poor internalization of LOS deficient bacteria compared to wild-type bacteria . Restoration of LOS biosynthesis in a LOS regulatory strain also restored both internalization and cytokine production and was enhanced in the presence of LPS binding protein (LBP) . These results suggest that DC phagocytosis depends on expression of LOS within the bacteria and that optimal cytokine production, particularly IL-12, requires internalization of the bacteria . These findings have important implications for designing vaccines that will induce protective immune responses to group B N . meningitidis.

J Community Health, 2004 Aug, 29(4), 265 - 9
Prevalence of chlamydial and gonorrheal infections among females in a juvenile detention facility, Honolulu, Hawaii; Katz AR et al.; Limited studies have shown high rates of gonorrheal and chlamydial infections among incarcerated adolescents, however, this population has not been routinely targeted for sexually transmitted disease (STD) screening . This study was done to ascertain the feasibility of screening females for STDs at a juvenile detention facility in Honolulu, Hawaii, and to determine the prevalence of chlamydial and gonorrheal infections in this population . Between January 2000 and December 2001, pelvic examination-based STD screening was offered to all female detainees, on selected dates . Specimens from multiple anatomical sites were cultured for Neisseria gonorrhoeae . Endocervical swabs were tested for Chlamydia trachomatis using a DNA probe (GenProbe) . One-hundred one of 204 (50%) eligible females were screened . Fourteen of 101 (13.9%) females were screen positive for chlamydial infection, while six of 101 (5.9%) were culture positive for infection with N . gonorrhoeae . Three females were co-infected with both organisms . The high STD rates justify the institution of a routine screening program for this high risk and previously underserved group.

Microbiology, 2004 Jun, 150(Pt 6), 1713 - 22
DNA methyltransferases from Neisseria meningitidis and Neisseria gonorrhoeae FA1090 associated with mismatch nicking endonucleases; Kwiatek A et al.; The genes encoding the DNA methyltransferases M.NmeDI and M.NmeAI from Neisseria meningitidis associated with the genes encoding putative Vsr endonucleases were overexpressed in Escherichia coli . The enzymes were purified to apparent homogeneity on Ni-NTA agarose columns, yielding proteins of 49+/-1 kDa and 39.6+/-1 kDa, respectively, under denaturing conditions . M.NmeDI recognizes the degenerate sequence 5'-RCCGGB-3' . It methylates the first 5' cytosine residue on both strands within the core sequence CCGG . The enzyme shows higher affinity with the hemimethylated degenerate sequence than with the unmethylated degenerate sequence . Comparison of the amino acid sequence of the target-recognizing domain of M.NmeDI with the closest neighbours recognizing the sequence 5'-RCCGGY-3' showed the presence of the homologous domain and an additional domain that may be responsible for recognizing the degenerate sequence . M.NmeAI recognizes the sequence 5'-CCGG-3' and methylates the second 5' cytosine residue on both DNA strands . In Neisseria gonorrhoeae strain FA1090 the homologues of these ORFs are truncated due to a variety of mutations.

Sex Transm Infect, 2004 Jun, 80(3), 230 - 5
Low prevalence of cervical infections in women with vaginal discharge in west Africa: implications for syndromic management; Pepin J et al.; OBJECTIVES: To measure prevalence and risk factors for cervical infections among a large sample of women consulting for vaginal discharge in west Africa and to evaluate its syndromic management through a two visit algorithm . METHODS: In 11 health centres in Benin, Burkina Faso, Ghana, Guinee, and Mali 726 women who presented with a vaginal discharge without abdominal pain and who denied being a sex worker (SW) were enrolled . Cervical samples were tested for the detection of Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) with polymerase chain reaction (PCR) assays . All participants were treated with single dose (2 g) metronidazole and clotrimazole cream for 3 days . They were randomised to be told either to come back on day 7 only if there was no improvement in the discharge (group A), or to come back on day 7 regardless of response to treatment (group B) . RESULTS: Overall, the prevalence of NG and CT was only 1.9% (14/726) and 3.2% (23/726) respectively . Risk factors previously recommended by the WHO were not associated with the presence of cervical infection, with the exception of the number of sex partners in the past 3 months . When taken together, these risk factors had a positive predictive value of only 6.4% to identify cervical infections . Prevalence of cervical infection was not higher in women who came back on day 7, regardless of the strategy used . Prevalence of NG/CT was lower in Ghana and Benin (5/280, 1.8%), where comprehensive interventions for SW have been ongoing for years, than in the three other countries (27/446, 6.1%, p = 0.01) . CONCLUSIONS: NG and CT infections are uncommon in west African women who consult for vaginal discharge and who are not SW . Syndromic management of vaginal discharge should focus on the proper management of vaginitis . The control of gonococcal and chlamydial infection should be redesigned around interventions focusing on sex workers.

Sex Transm Dis, 2004 Jun, 31(6), 353 - 9
High occurrence of simultaneous mutations in target enzymes and MtrRCDE efflux system in quinolone-resistant Neisseria gonorrhoeae; Dewi BE et al.; BACKGROUND: Emergence of multidrug-resistant Neisseria gonorrhoeae resulting from new genetic mutations is a serious threat to controlling gonorrhea . GOAL: To determine 1) antimicrobial susceptibilities and the corresponding genetic mutations and 2) the role of MtrRCDE efflux system in gonococcal resistance to fluoroquinolones . STUDY DESIGN: Antimicrobial susceptibility testing and sequence analysis of gyrA, parC, and mtrR loci of 131 N . gonorrhoeae isolates from Japan . RESULTS: The proportion of N . gonorrhoeae strains resistant and intermediate-resistant to antimicrobials was 25.2% and 48.9% for ciprofloxacin, 25.2% and 30.5% for ofloxacin, 12.2% and 53.4% for penicillin; and 17.6% and 51.1% for tetracycline, respectively . Strains were categorized into 22 mutation profiles, with GyrA-S91F/ParC-D86N/MtrR-G45D being the most predominant profile . The frequency of mutation in gyrA, parC, mtrR, and the mtrR promoter was 71%, 47.3%, 77.1%, and 23.7%, respectively . Seventy-one percent of strains carried mutations in both gyrA and mtrR . CONCLUSION: This study reports simultaneous mutations in fluoroquinolone target enzymes and the MtrRCDE efflux system as a fluoroquinolone-resistant mechanism in N . gonorrhoeae.

AIDS, 2004 May 21, 18(8), 1195 - 9
Long-term effects of condom promotion programmes for vaginal and oral sex on sexually transmitted infections among sex workers in Singapore; Wong ML et al.; OBJECTIVES: To evaluate the long-term impact of condom promotion programmes for vaginal and oral sex among female brothel-based sex workers in Singapore . DESIGN: A pre-test/post-test comparison group followed by a time series design was used to compare trends in condom use for vaginal sex and cervical gonorrhoea incidence from 1990 to 2002 across cross-sectional samples of sex workers before and after programme implementation in 1995 . The subsequent condom promotion programme for oral sex was evaluated using the interrupted time series with a retrospective pre-test to post-test matched control group design . METHODS: Sex workers completed a questionnaire before and 6 months after participation in educational sessions . Cervical and pharyngeal swabs were taken monthly for cultures for Neisseria gonorrhoeae . RESULTS: Consistent condom use for vaginal sex increased significantly from < 45.0% before 1995 (pre-intervention period) to 95.1% in 2002, with a corresponding decline in cervical gonorrhoea incidence from > 30 to 2/1000 person-months . Adjustment for temporal changes in sociodemographic characteristics did not materially alter the trends . Consistent oral condom use increased significantly from < 50% before 1996 to 97.2% in 2002, with a corresponding decline in pharyngeal gonorrhoea from > 12 to 4.7/1000 person-months . CONCLUSION: The interventions produced sustained high levels of condom use for vaginal and oral sex with corresponding declines in cervical and pharyngeal gonorrhoea incidence.

Methods Mol Biol, 2004, 276, 305 - 24
Integrated system for rapid proteomics analyses using microfluidic devices coupled to nanoelectrospray mass spectrometry; Li J et al.; This chapter presents an integrated and modular microsystem providing rapid analyses of low femtomole of in-gel digests for proteomics applications . Enhancement of sample throughput is facilitated using an autosampler, a microfabricated device comprising a large (2.4-microL total volume) separation channel together with a low-dead-volume interface to nES mass spectrometry . Sample preconcentration is achieved by packing C18 reverse phase or immobilized metal affinity chromatography (IMAC) beads into the large channel of this microfluidic device to adsorb peptides or enrich the sample in phosphopeptides prior to capillary electrophoresis separation and MS detection . This integrated microfluidic systems enables a sample throughput of up to 12 samples/h with a detection limit of approx 5 nM (25 fmol inj.) . Replicate injections of peptide standards indicated that reproducibility of migration time was typically 1.2-1.8%, whereas relative standard deviation (RSD) values of 9.2-11.8% were obtained on peak heights . The application of this device is demonstrated for 2D gel spots obtained from protein extracts of human astrocyte cells and for excised bands of membrane proteins from Neisseria meningitidis . A stepped acetonitrile gradient can be incorporated with the present microfluidic system to enhance selectivity during sample analysis.

Shanghai Kou Qiang Yi Xue, 1996 Sep, 5(3), 143 - 5
{A preliminary study on the relationship between Streptococcus and Neisseria,Veillonella in an artificial mouth model}; Zhu M et al.; The viable cell count of monobacterial plaque of Streptococcus and mixed bacterial plaque of streptococcus and Neisseria Veillonella,also the demineralization caused by the plaques were observed.The assumption that Neisseria and Veillonella might reduce demineralization of enamel caused by streptococcus was confirmed.The relationship between streptococcus and Neisseria,Veillonella were preliminarily found out.

Shanghai Kou Qiang Yi Xue, 1992 Dec, 1(2), 103 - 5
{Isolation of Neisseria from human dental plaque and study of characteristics of acid degradation}; Zhu M et al.; The Neisseria was isolated and identified from human dental plaque.The incidence of Neisseria was 100%.The N.Sicca was the predominant group.The results of sugar metabolism of Neisseria indicated that they were not only able to produce acid while sugar was offered sufficiently,but also degrade lactate evenacetate to supply energy after sugar was consumed.

Microbes Infect, 2004 Jun, 6(7), 693 - 701
Pathogenic consequences of Neisseria gonorrhoeae pilin glycan variation; Ghosh SK et al.; A Neisseria gonorrhoeae (gonococcus, GC) pilin glycosylation gene, pgtA, can either possess or lack phase-variation ability . Many GC, particularly the disseminated strains, carry a phase-variable pgtA . However, other GC, predominantly the uncomplicated gonorrhea isolates, carry a pgtA lacking phase-variability . These and other results suggest GC pilin glycan's pathogenic involvement.

Clin Infect Dis, 2004 Jun 1, 38(11), 1635 - 7 Epub 2004 May 12.
Clinical features and outcome of pediatric Neisseria meningitidis serogroup W135 infection: a report of 5 cases; Faye A et al.; We describe 5 pediatric cases of Neisseria meningitidis serogroup W135 infection . Infectious and/or reactive extrameningeal involvement was frequent . One patient had a persistent postmeningococcal inflammatory syndrome . Four of 5 isolates belonged to the clonal complex 37 . The important risk of extrameningeal complications must be borne in mind when treating children with N . meningitidis W135 infection.

J Antibiot (Tokyo), 2004 Mar, 57(3), 210 - 7
Antibiotics GE23077, novel inhibitors of bacterial RNA polymerase . I . Taxonomy, isolation and characterization; Ciciliato I et al.; GE 23077 factors A1, A2, B1 and B2 are novel antibiotics isolated from fermentation broths of an Actinomadura sp . strain . GE23077 antibiotics are cyclic peptides, which inhibit Escherichia coli RNA polymerase at nM concentrations . Both rifampicin-sensitive and rifampicin-resistant polymerases are inhibited, whereas E . coli DNA polymerase and wheat germ RNA polymerase are substantially not affected . In spite of the potent activity on the enzyme, the antibiotics generally show poor activity against whole cell bacteria . The spectrum of activity is restricted to Moraxella catarrhalis, including clinical isolates, with partial activity against Neisseria gonorrhoeae and Mycobacterium smegmatis.

Zhonghua Nan Ke Xue, 2004 Apr, 10(4), 269 - 71, 274
{Fusion expression of Neisseria gonorrhoeae outmembrane protein with a mucosal adjuvant}; Pan J et al.; OBJECTIVE: To express a fusion protein of Neisseria gonorrhoeae with a mucosal adjuvant . METHODS: The gene coding Loop VI-VIII(PL678) of porin, an out-membrane protein of Neisseria gonorrhoeae, was obtained by PCR . It was inserted into a plasmid fused with subunit B of heat labile enterotoxin . The recombinant was transformed in E . coli . The expression of fusion protein was analysed by ELISA, SDS-PAGE and Western-blot . RESULT: Fusion protein with LTB was successfully expressed, and displayed both the ability of binding GM1 and the reactogenicity with polyclonal antibodies against Neisseria gonorrhoeae . CONCLUSION: The expression of fusion protein laid a foundation for the study of the intramolecular vaccine against Neisseria gonorrhoeae.

Res Microbiol, 2004 Apr, 155(3), 129 - 35
Omp85, an evolutionarily conserved bacterial protein involved in outer-membrane-protein assembly; Voulhoux R et al.; The insertion of proteins into membranes generally requires the assistance of membrane proteins . A protein, designated Omp85 in Neisseria meningitidis, was shown to be required for the assembly of bacterial outer-membrane proteins . The protein is essential for the viability of the bacteria and is ubiquitous among Gram-negative bacteria . Omp85 depletion results in the accumulation of aggregates of unfolded outer-membrane proteins, and we argue that Omp85 is directly involved in outer-membrane-protein assembly . Omp85 shows sequence similarity with Toc75 of the chloroplast protein-import machinery, suggesting a common evolutionary origin.

J Infect Dis, 2004 Jun 1, 189(11), 2085 - 93 Epub 2004 Apr 30.
Quinolone resistance-determining region mutations and por type of Neisseria gonorrhoeae isolates: resistance surveillance and typing by molecular methodologies; Giles JA et al.; Quinolone resistance is increasing rapidly in Neisseria gonorrhoeae and is a significant public health problem that requires ongoing surveillance . To examine the feasibility of molecular surveillance of quinolone resistance, and to further characterize an outbreak of resistant N . gonorrhoeae in Israel, the quinolone resistance-determining region (QRDR) sequences and the por types of 80 N . gonorrhoeae isolates were determined using molecular techniques . QRDRs of gyrA and parC were amplified by polymerase chain reaction and were sequenced directly . The por type was determined by checkerboard hybridizations performed using oligonucleotide probes to regions encoding 5 variable loops of the porin protein . All 42 ciprofloxacin-resistant (CipR) isolates had mutations in QRDRs of both gyrA and parC, and identical mutations were found in 93% of these isolates . One intermediately resistant isolate had 1 mutation in gyrA, and susceptible isolates showed no mutations . Forty isolates had 1 of 2 por types that differed only by an in-frame deletion in variable region 5; all but 1 of these isolates were CipR . QRDR sequencing and por type determination showed that the outbreak of CipR N . gonorrhoeae in Israel was clonal . QRDR mutations were consistent with those previously characterized; this indicates that DNA probes can be developed for rapid detection and surveillance of quinolone-resistant N . gonorrhoeae in settings in which nonculture diagnostic methods are used.

Appl Microbiol Biotechnol, 2004 Nov, 65(6), 671 - 7 Epub 2004 May 12.
Design of mineral medium for growth of Actinomadura sp . ATCC 39727, producer of the glycopeptide A40926: effects of calcium ions and nitrogen sources; Technikova-Dobrova Z et al.; Actinomadura sp . ATCC 39727 produces the glycopeptide antibiotic A40926, structurally similar to teicoplanin, with significant activity against Neisseria gonorrhoeae and precursor of the semi-synthetic antibiotic dalbavancin . In this study the production of A40926 by Actinomadura under a variety of growth conditions was investigated . The use of chemically defined mineral media allowed us to analyze the influence of carbon and nitrogen sources, phosphate, ammonium and calcium on the growth and the antibiotic productivity of Actinomadura . We confirm recent data {Gunnarsson et al . (2003) J Ind Microbiol Biotechnol 30:150-156} that low initial concentrations of phosphate and ammonium are beneficial for growth and A40926 production, and we provide new evidence that the production of A40926 is depressed by calcium, but promoted when L-glutamine or L-asparagine are used as nitrogen sources instead of ammonium salts.

Microbiology, 2004 May, 150(Pt 5), 1427 - 37
The signal peptide sequence of a lytic transglycosylase of Neisseria meningitidis is involved in regulation of gene expression; Serruto D et al.; The 60 nucleotides encoding the signal peptide of the Neisseria meningitidis membrane-bound lytic transglycosylase (MltA) homologue GNA33 were found to exert a negative regulatory effect on expression of GNA33 from either a T7- or a P(lac)-driven system in Escherichia coli . Down-regulation was observed to occur at the transcriptional/post-transcriptional level and could possibly be ascribed to the formation of a stem-loop secondary structure within the signal peptide sequence . Slowing down the transcription rate through inhibition/titration of the RNA polymerase resulted in a considerable increase in mRNA accumulation, suggesting that a better coupling of translation to transcription would impede the formation of the putative secondary structure . Screening of synonymous mutations in the signal peptide sequence that showed high-level expression of an in-frame fusion to a reporter resulted in the isolation of several deletion mutants lacking most of the sequence participating in the putative secondary structure . Interestingly, the increase in the steady-state mRNA level observed in deletion mutants was higher, reaching a 300-fold increment, than that found in substitution mutants . Our results support the hypothesis that the rate of transcription controls the formation of a secondary structure in the region of the GNA33 transcript corresponding to the signal peptide sequence and this, when formed, negatively regulates expression.

Zhonghua Liu Xing Bing Xue Za Zhi, 2004 Feb, 25(2), 146 - 9
{Studies on the infection status of seven species Mycoplasma, three species of Chlamydia, Neisseria gonorrhoeae and Garderella vaginalis in 76 patients with sexual transmitted diseases}; Sun R et al.; OBJECTIVE: To study the infectious status of seven species of Mycoplasma, three species of Chlamydia, Neisseria gonorrhoeae and Garderella vaginalis in the 76 male sexual transmitted disease (STD) patients in Yangzhou city . METHODS: Twelve species of pathogens including Ureaplasma urealyticum (Uu), Mycoplasma hominis (Mh), Mycoplasma pneumoniae (Mpn), Mycoplasma genitalium (Mg), Mycoplasma fermentans (Mf), Mycoplasma penetrans (Mpe), Mycoplasma prium (Mpi), Chlamydia trachomatis (Ct), Chlamydia pneumoniae (Cpn), Chlamydia psittaci (Cps), Neisseria gonorrhoeae (Ng) and Garderella vaginalis (GV) were detected by nested polymerase chain reaction including PPNG . RESULTS: The positive rates of Uu, Mh, Mpn, Mg, Mf, Mpe, Ct, Ng were 64.5%, 27.6%, 26.3%, 18.4%, 2.6%, 2.6%, 31.6%, 36.8%, in which Penicillinase-producing neisseria gonorrhoeae (PPNG) accounted for 14.3%, GV 15.8% . No Mpi, Cpn or Cps were found . There was more significant therapeutic effects on the detectable rate of Mycoplasma nucleic acid between positive gonococcus and negative gonococcus in male STDs patients (chi(2) = 3.848, P < 0.05) . CONCLUSION: The infection rates of Mycoplasma, Chlamydia, Ng and GV were high among male STD patients in Yangzhou city . In clinical practice, more attention should be paid on correct diagnosis and treatment for patients, with Gonococcus, Chlamydia, Mycoplasma and GV.

Mol Microbiol, 2004 May, 52(4), 1081 - 90
Fur functions as an activator and as a repressor of putative virulence genes in Neisseria meningitidis; Delany I et al.; Fur is a well-known iron-responsive repressor of gene transcription, which is used by many bacteria to respond to the low-iron environment that pathogens encounter during infection . Four promoters of Neisseria meningitidis predicted to have Fur-binding boxes were selected to study the molecular interactions between Fur and the promoter regions of genes expected to play a central role in survival and pathogenesis . We demonstrate that Fur acts not only as a repressor, but also as an activator of gene expression both in vivo and in vitro . We report that Fur binds to operators located upstream of three promoters that are positively regulated in vivo by Fur and iron, whereas Fur binds to an operator overlapping the classically iron-repressed tbp promoter . Deletion of the upstream operator in the norB promoter abolished activation of transcription in vivo in response to iron and in vitro in response to Fur . The role of such a dual mechanism of Fur regulation during infection is discussed.

Mol Microbiol, 2004 May, 52(4), 1069 - 80
Structural tolerance of bacterial autotransporters for folded passenger protein domains; Veiga E et al.; In this report we investigate the capacity of bacterial autotransporters (AT) to translocate folded protein domains across the outer membrane (OM) . Polypeptides belonging to the AT family contain a C-terminal domain that supports the secretion of the N-domain (the passenger) across the OM of Gram-negative bacteria . Despite some controversial data, it has been widely accepted that N-passenger domains of AT must be unfolded and devoid of disulphide bonds for efficient translocation . To address whether or not AT are able to translocate folded protein domains across the OM, we employed several types of recombinant antibodies as heterologous N-passengers of the transporter C-domain of IgA protease (C-IgAP) of Neisseria gonorroheae . The N-domains used were single chain Fv fragments (scFv) and variable mono-domains derived from camel antibodies (V(HH)) selected on the basis of their distinct and defined folding properties (i.e . enhanced solubility, stability and presence or not of disulphide bonds) . Expression of these hybrids in Escherichia coli shows that stable scFv and V(HH) domains are efficiently (>99%) translocated towards the bacterial surface regardless of the presence or not of disulphide bonds on their structure . Antigen-binding assays demonstrate that surface-exposed scFv and V(HH) domains are correctly folded and thus able to bind their cognate antigens . Expression of scFv- or V(HH)-C-IgAP hybrids in E . coli dsbA or fkpA mutant cells reveals that these periplasmic protein chaperones fold these N-domains before their translocation across the OM . Furthermore, large N-passengers composed of strings of V(HH) domains were secreted in a folded state by AT with no loss of efficacy (>99%) despite having multiple disulphide bonds . Thus AT can efficiently translocate toward the cell surface folded N-passengers composed of one, two or three immunoglobulin (Ig) domains, each with a folded diameter between approximately 2 nm and having disulphide bonds . This tolerance for folded protein domains of approximately 2 nm fits with the diameter of the central hydrophilic channel proposed for the ring-like oligomeric complex assembled by C-IgAP in the OM.

J Bacteriol, 2004 May, 186(10), 3266 - 9
Characterization of a nucleotide-binding domain associated with neisserial iron transport; Lau GH et al.; The fbpABC operon in Neisseria gonorrhoeae encodes an ATP-binding cassette transporter required for iron uptake from the host ferric binding proteins . The gene for the nucleotide-binding domain (fbpC) expressed in Escherichia coli has intrinsic ATPase activity (0.5 mmol/min/mg) uncoupled from the iron transport process . The FbpC E164D mutant is found to have a 10-fold reduction in specific activity . FbpC is covalently modified by 8-azido-{gamma32P}ATP, indicating that FbpC is a functional ATPase that likely combines with FbpB to form a ferric iron transporter.

MMWR Morb Mortal Wkly Rep, 2004 Apr 30, 53(16), 335 - 8
Increases in fluoroquinolone-resistant Neisseria gonorrhoeae among men who have sex with men--United States, 2003, and revised recommendations for gonorrhea treatment, 2004; Centers for Disease Control and Prevention (CDC); In the United States, an estimated 700,000-800,000 persons are infected with Neisseria gonorrhoeae each year . Since 1993, CDC has recommended use of fluoroquinolones (i.e., ciprofloxacin, floxacin, or levofloxacin) for gonorrhea treatment . Fluoroquinolone therapy is used frequently because it is an inexpensive, oral, and single-dose therapy . However, because of increased prevalence of fluoroquinolone-resistant N . gonorrhoeae (QRNG) in Asia, the Pacific Islands (including Hawaii), and California, fluoroquinolones are no longer recommended for treating gonorrhea acquired in those locations . This report describes increases in QRNG among men who have sex with men (MSM) in Massachusetts, New York City, and 30 sites surveyed by the Gonococcal Isolate Surveillance Project (GISP) during 2003 . CDC recommends that clinicians no longer use fluoroquinolones as a first-line treatment for gonorrhea in MSM.

Biochemistry, 2004 May 11, 43(18), 5239 - 45
Structural basis for novel delta-regioselective heme oxygenation in the opportunistic pathogen Pseudomonas aeruginosa; Friedman J et al.; The Gram-negative bacterium Pseudomonas aeruginosa contains a heme oxygenase (pa-HO) that primarily oxygenates the delta-meso heme carbon {Caignan, G . A., Deshmukh, R., Wilks, A., Zeng, Y., Huang, H . W., Moenne-Loccoz, P., Bunce, R . A., Eastman, M . A., and Rivera, M . (2002) J . Am . Chem . Soc . 124, 14879-14892} . This differs from other previously characterized heme oxygenases, which display regioselectivity for the alpha-meso heme carbon . Here we report the crystal structure of pa-HO at 1.60 A resolution and compare it to the 1.50 A structure of nm-HO from Neisseria meningitidis {Schuller, D . J., Zhu, W., Stojiljkovic, I., Wilks, A., and Poulos, T . L . (2001) Biochemistry 40, 11552-11558} . The crystal structure of pa-HO maintains the same overall fold as other bacterial and mammalian heme oxygenases, including a conserved network of hydrogen-bonded solvent molecules important for dioxygen activation . The novel delta-regioselectivity of heme oxygenation observed by pa-HO is due to the heme being rotated by approximately 100 degrees, which places the delta-meso heme carbon in the same position as the alpha-meso heme carbon in other heme oxygenases . The main interaction in pa-HO that stabilizes the unique heme orientation is a salt bridge between Lys132 and the heme 7-propionate, as well as hydrophobic contacts involving Leu29, Val33, and Phe189 with the heme methyl and vinyl groups.

J Infect Dis, 2004 May 1, 189(9), 1547 - 55 Epub 2004 Apr 16.
Refinement of a therapeutic Shiga toxin-binding probiotic for human trials; Pinyon RA et al.; We have previously constructed a recombinant bacterium expressing a modified lipopolysaccharide (LPS) mimicking the Shiga toxin receptor, which binds toxin with high avidity . This involved cloning Neisseria galactosyl transferase genes (lgtC and lgtE) in pK184 in a derivative of Escherichia coli R1 (CWG308) . Such constructs have considerable potential for prevention of disease caused by Shiga toxin-producing E . coli (STEC) . However, neither the E . coli host strain nor the expression plasmid is suitable for human use, because the former is derived from a clinical isolate and the latter contains a kanamycin-resistance gene . We have constructed, as a prelude to human trials, a nonpathogenic E . coli K-12 C600 derivative with deletions in waaO and waaB, such that it has the same LPS core structure as CWG308 . We also deleted the thyA gene from this strain, rendering it thymine dependent . The kanamycin-resistance gene was also deleted from pK184 and was replaced with Salmonella typhimurium thyA . Neisseria lgtCE was then cloned into this plasmid and transformed into C600 Delta waaOB Delta thyA . The plasmid was stably maintained, and the construct produced a modified LPS and neutralized Stx1 and Stx2c . Moreover, mice challenged with an otherwise fatal dose of STEC were completely protected by oral administration of the novel construct.

FEBS Lett, 2004 Apr 30, 564(3), 294 - 300
The plug domain of a neisserial TonB-dependent transporter retains structural integrity in the absence of its transmembrane beta-barrel; Oke M et al.; Transferrin binding protein A (TbpA) is a TonB-dependent outer membrane protein expressed by pathogenic bacteria for iron acquisition from human transferrin . The N-terminal 160 residues (plug domain) of TbpA were overexpressed in both the periplasm and cytoplasm of Escherichia coli . We found this domain to be soluble and monodisperse in solution, exhibiting secondary structure elements found in plug domains of structurally characterized TonB-dependent transporters . Although the TbpA plug domain is apparently correctly folded, we were not able to observe an interaction with human transferrin by isothermal titration calorimetry or nitrocellulose binding assays . These experiments suggest that the plug domain may fold independently of the beta-barrel, but extracellular loops of the beta-barrel are required for ligand binding.

N Z Med J . 2004 Apr 02;117(1191):U817.
Antimicrobial susceptibility among Neisseria gonorrhoeae in New Zealand in 2002; Heffernan H et al.; AIM: To estimate the prevalence of antimicrobial resistance among Neisseria gonorrhoeae, and to determine whether the increase in ciprofloxacin resistance observed in Auckland in 2001 had occurred in other parts of the country . METHODS: The antimicrobial susceptibility of N . gonorrhoeae (isolated in New Zealand over a 4-month period between April and August 2002) was tested at either LabPlus, Auckland District Health Board, or at ESR, using the same agar dilution method . RESULTS: The prevalence of resistance to the antimicrobials tested was: ceftriaxone, 0%; ciprofloxacin, 6.8%; penicillin, 9.0%; spectinomycin, 0%; and tetracycline, 27.8% . There were few statistically significant geographical differences in resistance within New Zealand . Gonococcal infections acquired in Asia were more likely to be ciprofloxacin and penicillin resistant than infections acquired in New Zealand . CONCLUSIONS: Ciprofloxacin resistance among N . gonorrhoeae in New Zealand has reached a level where this antibiotic is no longer the most appropriate first-line treatment . In fact, ceftriaxone should now be considered the most reliable option for the treatment and control of gonorrhoea in New Zealand, particularly in the Northland/Auckland region.

Ann Dermatol Venereol, 2004 Mar, 131(3), 251 - 4
{Prevalence of asymptomatic sexually transmitted infections among high-risk patients attending a free anonymous HIV-screening center}; Toskine I et al.; INTRODUCTION: The aim of this study was to assess the prevalence of sexually transmitted infections among patients attending an anonymous HIV Screening Center . PATIENTS AND METHODS: This prospective study was performed in the HIV Screening Center of University hospital in Reims (France) from May 1997 to December 1997 . The inclusion criteria were the asymptomatic clinical presentation and the presence of risk factors for sexually transmitted infections referring to WHO criteria . The methods included clinical examination after application of acetic acid and urethral and endocervical swabs to identify:Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, Gardnerella vaginalis, Trichomonas vaginalis in specific culture . Treponema pallidum and HIV-1 infection were both detected by Enzym Linked Immuno Sorbent Assay (ELISA) . RESULTS: One hundred and one patients (62 men and 39 women) were included in the study . Their mean age was 27 +/- 4 Years . Risk factors for sexually transmitted infections were: multiple sexual partners 81 p . 100; homo or bisexuality 16 p . 100; intravenous drug use 3 p . 100 . The sexually transmitted infections were: HIV-1 infection 1 p . 100;Ureaplasma urealyticum 25 p . 100; genital warts 5 p . 100;Chlamydia trachomatis 3 p . 100; Gardnerella vaginalis 3 p . 100; Mycoplasma hominis 2 p . 100; Treponema pallidum 0 p . 100; Neisseria gonorrhoeae 0 p . 100; Trichomonas vaginalis 0 p . 100 . The prevalence of sexually transmitted infections was significantly higher among women (p<0.05) . DISCUSSION: Classical sexually transmitted infections and HIV infection were rarely detected in this study; but prevalence of other sexually transmitted infections (genital warts, Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum) was high . Ureaplasma urealyticum is considered as a possible pathogenic agent in pregnant women (preterm delivery, decrease of birth weight, chorioamniotitis) . These results suggest that other than sexually transmitted infections in high risk patients attending a HIV Screening Center other sexually transmitted infections should also be systematically screened for.

Sex Transm Dis, 2004 May, 31(5), 305 - 10
Antimicrobial resistance patterns (1999-2002) and characterization of ciprofloxacin-resistant Neisseria gonorrhoeae in Korea; Yoo J et al.; BACKGROUND: Antimicrobial susceptibilities of Neisseria gonorrhoeae were monitored during 4 years . In Korea, ciprofloxacin-resistant N . gonorrhoeae has dramatically increased after recommendation as a therapeutic drug . GOAL: The goal of this study was to determine the resistance patterns and characterize Korean ciprofloxacin-resistant N . gonorrhoeae . STUDY DESIGN: Antimicrobial susceptibilities were performed . PFGE profile and DNA sequencing of gyrA and parC genes were used to characterize the ciprofloxacin-resistant isolates in Korea . RESULTS: Tetracycline, ofloxacin, ciprofloxacin-resistant isolates were increased and among them, the proportion of isolates resistant to ciprofloxacin increased remarkably from 1% in 1999 to 48.8% in 2002 . Fifteen different types by PFGE profile were identified . Major alteration type was M12 (67%), which have amino acid substitution in gyrA (S-91-->F, D-95-->G) and parC (S-87-->A) . CONCLUSION: We could conclude that resistance for ciprofloxacin was remarkably increased during 4 years . Ciprofloxacin-resistant N . gonorrhoeae was supposed by the spread of several strains that had a small number of origins.

Sex Transm Dis, 2004 May, 31(5), 273 - 7
The effect of urine testing in evaluations of the sensitivity of the Gen-Probe Aptima Combo 2 assay on endocervical swabs for Chlamydia trachomatis and neisseria gonorrhoeae: the infected patient standard reduces sensitivity of single site evaluation; Moncada J et al.; BACKGROUND: The Gen-Probe APTIMA Combo 2 (AC2) assay is a second-generation transcription-mediated amplification assay for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) . GOAL: The goal of this study was to evaluate AC2 performance of endocervical (cx) swabs for the detection of CT and NG using either a specimen or an infected patient standard . STUDY DESIGN: In a multicenter clinical study, we compared AC2 with Abbott's ligase chain reaction (LCR) and Roche's polymerase chain reaction (PCR; Amplicor or COBAS) for CT, and we compared AC2 with Abbott's LCR and culture for NG . A total of 1569 females were enrolled in the study; we collected cx and first-catch urine (FCU) specimens . RESULTS: CT prevalence was 13.3% for cx specimens and 13.7% for FCU specimens . NG prevalence was 8.7% and 7.9% for cx and FCU specimens, respectively . When based only on cx specimens, AC2, LCR, and PCR sensitivities for CT were 99.4%, 95.6%, and 95.6%, respectively . However, cx sensitivity for CT was reduced to 92.1%, 86.6%, and 87.1% for each respective assay when based on both cx and FCU specimen results (infected patient standard) . NG sensitivities for AC2, LCR, and culture based solely on cx specimen results were 99.2%, 96.1%, and 85.9%, respectively . Based on infected patient standard, the sensitivities of each respective assay were 98.5%, 93.9%, and 84.0% . CONCLUSIONS: The infected patient standard reduces the sensitivity of the endocervical evaluation because some infected patients are positive only with FCU . The reduction in sensitivity is greater when testing for CT . Specificities improved slightly, because some unique cx positives, initially classified as false-positive were confirmed by a positive FCU result . Sensitivity of AC2 was higher than LCR, PCR, and culture . Specificity was slightly lower, but discrepant analysis (using alternate TMA targets) of apparent AC2 false-positives showed that 75% to 80% were true-positives.

Infect Immun, 2004 May, 72(5), 2742 - 52
Engulfment of Neisseria gonorrhoeae: revealing distinct processes of bacterial entry by individual carcinoembryonic antigen-related cellular adhesion molecule family receptors; McCaw SE et al.; Individual Neisseria gonorrhoeae colony opacity-associated (Opa) protein variants can bind up to four different carcinoembryonic antigen-related cellular adhesion molecule (CEACAM) receptors . Most human cells encountered by gonococci express a combination of CEACAM receptors, thereby complicating the elucidation of intracellular signaling pathways triggered by individual receptors . Here, we compare the process of bacterial engulfment by a panel of stably transfected HeLa epithelial cell lines expressing each CEACAM receptor in isolation . CEACAM1 and CEACAM3 each contain proteinaceous transmembrane and cytoplasmic domains; however, the processes of neisserial uptake mediated by these receptors differ with respect to their susceptibilities to both tyrosine kinase inhibitors and the actin microfilament-disrupting agent cytochalasin D . Neisserial uptake mediated by glycosylphosphatidylinositol (GPI)-anchored CEACAM5 and CEACAM6 was not significantly affected by any of a broad spectrum of inhibitors tested . However, cleavage of the GPI anchor by phosphatidylinositol-specific phospholipase C reduced bacterial uptake by HeLa cells expressing CEACAM5, consistent with a single zipper-like mechanism of uptake mediated by this receptor . Regardless of the CEACAM receptor expressed, internalized gonococci were effectively killed by a microtubule-dependent process that required acidification of the bacterium-containing phagosome . Given the phase-variable nature of neisserial Opa proteins, these results indicate that the mechanism of bacterial engulfment and the cellular response to gonococcal infection depend on both the receptor specificities of the neisserial Opa protein variants expressed and the spectrum of CEACAM receptors present on target cells, each of which determines the combination of receptors ultimately engaged.

Infect Immun, 2004 May, 72(5), 2528 - 37
Local and systemic antibody responses in mice immunized intranasally with native and detergent-extracted outer membrane vesicles from Neisseria meningitidis; Guthrie T et al.; The mouse humoral immune response toward native or detergent-extracted outer membrane vesicles (NOMVs and DOMVs, respectively) from Neisseria meningitidis was determined after intranasal immunization . Both preparations elicited high frequencies of NOMV-specific antibody-forming cells (AFCs) locally in the nasal associated lymphoid tissue (NALT) after three or four weekly doses . The diffuse NALT (D-NALT) contained ca . 10-fold more NOMV-specific AFCs than those observed in the mediastinal lymph node, spleen, and bone marrow . AFCs observed in the D-NALT were primarily immunoglobulin A positive (IgA(+)) and were maintained for at least 1 month . In contrast, the organized NALT (O-NALT) contained low numbers of AFCs, and the response was relatively short-lived . In other lymphoid tissues, AFCs producing various IgG subclasses and IgM were present with IgG2b-producing AFCs being dominant or codominant with IgA or IgG2a . In serum and in all of the tissues examined, with the exception of the NALT, NOMVs clearly induced a stronger antibody response and a broader range of antibody isotypes than DOMVs . The development of NOMV-specific AFCs in spleen and bone marrow after intranasal immunization was slow compared to intravenous immunization but, once established, the intranasally elicited responses increased steadily for at least 75 days . NOMV-specific antibodies induced via several routes of immunization had high bactericidal activities in serum . Our results indicated that intranasally administered OMVs induced strong local and systemic antibody responses in mice that were relatively long-lived.

Infect Immun, 2004 May, 72(5), 2507 - 12
Intranasal immunization with multivalent group A streptococcal vaccines protects mice against intranasal challenge infections; Hall MA et al.; We have previously shown that a hexavalent group A streptococcal M protein-based vaccine evoked bactericidal antibodies after intramuscular injection . In the present study, we show that the hexavalent vaccine formulated with several different mucosal adjuvants and delivered intranasally induced serum and salivary antibodies that protected mice from intranasal challenge infections with virulent group A streptococci . The hexavalent vaccine was formulated with liposomes with or without monophosphorylated lipid A (MPL), cholera toxin B subunit with or without holotoxin, or proteosomes from Neisseria meningitidis outer membrane proteins complexed with lipopolysaccharide from Shigella flexneri . Intranasal immunization with the hexavalent vaccine mixed with these adjuvants resulted in significant levels of antibodies in serum 2 weeks after the final dose . Mean serum antibody titers were equivalent in all groups of mice except those that were immunized with hexavalent protein plus liposomes without MPL, which were significantly lower . Salivary antibodies were also detected in mice that received the vaccine formulated with the four strongest adjuvants . T-cell proliferative assays and cytokine assays using lymphocytes from cervical lymph nodes and spleens from mice immunized with the hexavalent vaccine formulated with proteosomes indicated the presence of hexavalent protein-specific T cells and a Th1-weighted mixed Th1-Th2 cytokine profile . Intranasal immunization with adjuvanted formulations of the hexavalent vaccine resulted in significant levels of protection (80 to 100%) following intranasal challenge infections with type 24 group A streptococci . Our results indicate that intranasal delivery of adjuvanted multivalent M protein vaccines induces protective antibody responses and may provide an alternative to parenteral vaccine formulations.

Expert Opin Pharmacother, 2004 Apr, 5(4), 829 - 54
The use of fluoroquinolones in gonorrhoea: the increasing problem of resistance; Dan M; The recent re-emergence of gonorrhoea in developed countries has been accompanied by the rise and spread of gonococcal resistance to the fluoroquinolones . In the 1980s fluoroquinolones were considered an important addition to the arsenal of agents used to treat gonorrhoea . They proved to be excellent drugs for this indication, including infections caused by penicillinase-producing and tetracycline-resistant strains of Neisseria gonorrhoeae . However, as gonococci have a well-recognised potential to develop resistance to antibiotics, the first reports of reduced susceptibility to fluoroquinolones appeared a few years after their introduction . Gonococcal resistance to fluoroquinolones is now well-established in the Far East, from where it has spread to Australia, Hawaii, California and Europe . In Africa and Latin America, gonococci continue to be susceptible to fluoroquinolones.

Proc Natl Acad Sci U S A, 2004 May 18, 101(20), 7530 - 5 Epub 2004 Apr 19.
An aminoacyl-tRNA synthetase-like protein encoded by the Escherichia coli yadB gene glutamylates specifically tRNAAsp; Dubois DY et al.; The product of the Escherichia coli yadB gene is homologous to the N-terminal part of bacterial glutamyl-tRNA synthetases (GluRSs), including the Rossmann fold with the acceptor-binding domain and the stem-contact fold . This GluRS-like protein, which lacks the anticodon-binding domain, does not use tRNA(Glu) as substrate in vitro nor in vivo, but aminoacylates tRNA(Asp) with glutamate . The yadB gene is expressed in wild-type E . coli as an operon with the dksA gene, which encodes a protein involved in the general stress response by means of its action at the translational level . The fate of the glutamylated tRNA(Asp) is not known, but its incapacity to bind elongation factor Tu suggests that it is not involved in ribosomal protein synthesis . Genes homologous to yadB are present only in bacteria, mostly in Proteobacteria . Sequence alignments and phylogenetic analyses show that the YadB proteins form a distinct monophyletic group related to the bacterial and organellar GluRSs (alpha-type GlxRSs superfamily) with ubiquitous function as suggested by the similar functional properties of the YadB homologue from Neisseria meningitidis.

FEMS Immunol Med Microbiol, 2004 May 1, 41(1), 43 - 50
Analysis of the role of pglI in pilin glycosylation of Neisseria meningitidis; Warren MJ et al.; Pilin is the major subunit of the essential virulence factor pili and is glycosylated at Ser63 . In this study we investigated the gene pglI to determine whether it is involved in the biosynthesis of the pilin-linked glycan of Neisseria meningitidis strain C311#3 . A N . meningitidis C311#3pglI mutant resulted in a change of apparent molecular weight in SDS-PAGE and altered binding of antisera, consistent with a role in the biosynthesis of the pilin-linked glycan . These data, in conjunction with homology with well-characterised acyltransferases suggests a specific role for pglI in the biosynthesis of the basal 2,4-diacetamido-2,4,6-trideoxyhexose residue of the pilin-linked glycan.

J Adolesc Health, 2004 May, 34(5), 376 - 83
A descriptive analysis of STD prevalence among urban pregnant African-American teens: data from a pilot study; Diclemente RJ et al.; OBJECTIVE: To assess the prevalence of sexually transmitted diseases (STDs) among a sample of African-American adolescent females at the time of their first prenatal visit and to assess key characteristics of those testing positive for sexually transmitted diseases . The study also determined differences in these characteristics between adolescents who were and those who were not diagnosed with an STD . METHODS: One-hundred-and-seventy pregnant African-American adolescents (aged 14-20 years; mean = 17.5 years) receiving their first prenatal visit were recruited at a prenatal clinic located in a large urban hospital . Biological assessment included nucleic acid amplification testing for gonococcal, chlamydial, and trichomonal infections . Rapid plasma reagin testing assessed infection with syphilis . A self-administered survey and in-depth face-to-face interview were used to collect detailed information assessing adolescents' sociodemographic characteristics, psychosocial indices, and their recent sexual risk behaviors . Data were analyzed using Student's t-tests and contingency table analyses, respectively, for continuous and categorical variables . RESULTS: Overall, 23.5% tested positive for one of the four STDs . Thirteen percent were infected with Chlamydia trachomatis, 1.2% with Neisseria gonorrhoeae, 8.9% with Trichomonas vaginalis, and 1.2% with Treponema pallidum . More than one-half reported recent (past 6 months) treatment for an STD, 30% of these tested positive for at least one of the four STDs assessed . Adolescents testing positive for STDs held favorable attitudes toward condom use, but levels of sexual risk were generally high . There were no sociodemographic, psychosocial, and sexual-risk differences between those testing positive and negative . CONCLUSION: Findings support STD screening efforts targeting pregnant adolescents . Providing clinic-based counseling and prevention education programs to pregnant adolescents regardless of apparent risk factors may also be warranted.

Curr Opin Infect Dis, 2004 Feb, 17(1), 49 - 52
Tubal factor infertility, with special regard to chlamydial salpingitis; Mardh PA; PURPOSE OF REVIEW: This article will highlight recent research into tubal factor infertility as one of the main causes of involuntary childlessness in women . There will be a focus on chlamydial infections . RECENT FINDINGS: The most common cause of tubal factor infertility is occlusion of the fallopian tubes due to an infection by a sexually transmitted agent, by Chlamydia trachomatis or Neisseria gonorrhoeae.The prevalence of diagnosed cases of tubal factor infertility (TFI) can be correlated to the epidemiological situation regarding these agents that was prevailing several years ago . This is partly due to the trend seen in many Western countries that women often postpone to try to get pregnant . Therefore, there is often a time lag between the acute primary pelvic inflammatory disease (PID) and when women first consult because of fertility problems . Sub-clinical salpingitis is today regarded as even more common than symptomatic PID . Persistent tubal infections by C . trachomatis are also a common feature, even despite courses of antibiotic therapy . The current focus on TFI has been on the immunopathology of tubal chlamydial infections, for which differences in host factors, such as genetic polymorphism in cytokine response and human leukocyte antigen type, may play a role in the outcome of pelvic inflammatory disease . Hysterosonography is a more convenient mode for diagnosing tubal occlusion than hysterosalpingography . The use of new species-specific antibody tests for C . trachomatis has decreased previous specificity problems found when used to detect tubal occlusion in work-up of women consulting because of infertility . SUMMARY: Infection by C . trachomatis is a major cause of TFI . Many cases of chlamydial salpingitis have a more or less subclinical course . The tubal infection may become chronic in spite of antibiotic therapy . Immunological processes may continue after microbiological cure, which stresses the importance of screening for C . trachomatis in order to detect and treat carriers to hinder spread to still uninfected women.

J Bacteriol, 2004 May, 186(9), 2841 - 55
Conserved glycines in the C terminus of MinC proteins are implicated in their functionality as cell division inhibitors; Ramirez-Arcos S et al.; Alignment of 36 MinC sequences revealed four completely conserved C-terminal glycines . As MinC inhibits cytokinesis in Neisseria gonorrhoeae and Escherichia coli, the functional importance of these glycines in N . gonorrhoeae MinC (MinC(Ng)) and E . coli MinC (MinC(Ec)) was investigated through amino acid substitution by using site-directed mutagenesis . Each mutant was evaluated for its ability to arrest cell division and to interact with itself and MinD . In contrast to overexpression of wild-type MinC, overexpression of mutant proteins in E . coli did not induce filamentation, indicating that they lost functionality . Yeast two-hybrid studies showed that MinC(Ec) interacts with itself and MinD(Ec); however, no interactions involving MinC(Ng) were detected . Therefore, a recombinant MinC protein, with the N terminus of MinC(Ec) and the C terminus of MinC(Ng), was designed to test for a MinC(Ng)-MinD(Ng) interaction . Each MinC mutant interacted with either MinC or MinD but not both, indicating the specificity of glycine residues for particular protein-protein interactions . Each glycine was mapped on the C-terminal surfaces (A, B, and C) of the solved Thermotoga maritima MinC structure . We found that MinC(Ec) G161, residing in close proximity to the A surface, is involved in homodimerization, which is essential for MinC function . Glycines corresponding to MinC(Ec) G135, G154, and G171, located within or adjacent to the B-C surface junction, are critical for MinC-MinD interactions . Circular dichroism revealed no gross structural perturbations of the mutant proteins, although the contribution of glycines to protein flexibility and stability cannot be discounted . Using molecular modeling, we propose that exposed conserved MinC glycines interact with exposed residues of the alpha-7 helix of MinD.

Sex Transm Dis, 2004 Mar, 31(3), 180 - 4
Mutations in the gyrA and parC genes and in vitro activities of fluoroquinolones in 91 clinical isolates of Neisseria gonorrhoeae in Japan; Shigemura K et al.; BACKGROUND AND OBJECTIVES: Fluoroquinolone resistance in Neisseria gonorrhoeae has been associated with alternations in the quinolone-resistance determining regions in the gyrA and parC genes . GOAL: The goal of this study was to investigate the correlation between fluoroquinolone minimum inhibitory concentrations (MICs) and mutations in the gyrA and parC genes of 91 N . gonorrhoeae clinical isolates from Japan . STUDY DESIGN: The MICs of fluoroquinolones ciprofloxacin, levofloxacin, and gatifloxacin for 91 clinical isolates from male gonococcal urethritis in Hyogo or Osaka, Japan, were measured, and the gyrA and parC genes of these isolates were sequenced . RESULTS: Among 91 isolates tested, over 70% isolates were resistant to ciprofloxacin . We found that 4 mutations (Ser-91-Phe, Ser-91-Ile, Asp-95-Gly in gyrA, and Ser-88-Pro in parC) had significant correlation to MICs of fluoroquinolone (ciprofloxacin, levofloxacin, and gatifloxacin) . CONCLUSION: Some mutations in QRDR had a significant relationship to the fluoroquinolone resistance of N . gonorrhoeae clinical isolates from Japan.

J Biol Chem, 2004 Jul 2, 279(27), 28339 - 44 Epub 2004 Apr 09.
Intermediate trapping on a mutant retaining alpha-galactosyltransferase identifies an unexpected aspartate residue; Lairson LL et al.; Lipopolysaccharyl-alpha-1,4-galactosyltransferase C (LgtC), a glycosyltransferase family 8 alpha-1,4-galactosyltransferase from Neisseria meningitidis, catalyzes the transfer of galactose from UDP galactose to terminal lactose-containing acceptor sugars with net retention of anomeric configuration . To investigate the potential role of discrete nucleophilic catalysis suggested by the double displacement mechanism generally proposed for retaining glycosyltransferases, the side chain amide of Gln-189, which is suitably positioned to act as the catalytic nucleophile of LgtC, was substituted with the more nucleophilic carboxylate-containing side chain of glutamate in the hope of accumulating a glycosyl-enzyme intermediate . The resulting mutant was subjected to kinetic, mass spectrometric, and x-ray crystallographic analysis . Although the K(m) for UDP-galactose is not significantly altered, the k(cat) was reduced to 3% that of the wild type enzyme . Electrospray mass spectrometric analysis revealed that a steady state population of the Q189E variant contains a covalently bound galactosyl moiety . Liquid chromatographic/mass spectrometric analysis of fragmented proteolytic digests identified the site of labeling not as Glu-189 but, surprisingly, as the sequentially adjacent Asp-190 . However, the side chain carboxylate of Asp-190 is located 8.9 A away from the donor substrate in the available crystal structure . Kinetic analysis of a D190N mutant at this position revealed a k(cat) value 3000-fold lower than that of the wild type enzyme . A 2.6-A crystal structure of the Q189E mutant with bound uridine 5'-diphospho-2-deoxy-2-fluoro-alpha-d-galactopyranose revealed no significant perturbation of the mode of donor sugar binding nor of active site configuration . This is the first trapping of an intermediate in the active site of a retaining glycosyltransferase and, although not conclusive, implicates Asp-190 as an alternative candidate catalytic nucleophile, thereby rekindling a longstanding mechanistic debate.

Int J STD AIDS, 2004 Apr, 15(4), 243 - 8
Gonorrhoea in young heterosexuals attending urban STD clinics in Britain: a cross-sectional survey; Jungmann E et al.; Young people in the UK bear the brunt of sexually transmitted infections, in particular of gonorrhoea . We aimed to assess whether young people with gonorrhoea (under 21 years) attending sexual health clinics differed from older individuals with gonorrhoea in their behavioural and clinical characteristics and management outcomes . The results of this cross-sectional study suggest that young people were more likely to be female (66.2% vs 34.1%), have concurrent infection with Chlamydia trachomatis (55.4% vs 30.2%) and a history of recent gonococcal infection (81.3% vs 35.5%) if they ever had gonorrhoea . Young women were more likely to experience treatment delay and not to attend for follow-up than older women . Resistance to ciprofloxacin was high in both age groups but the prevalence of penicillinase-producing Neisseria gonorrhoeae was higher in older patients (11.5% vs 1.3%) . Different management protocols for young and older patients with gonorrhoea may need to be considered.

Mol Immunol, 2004 Apr, 40(18), 1333 - 46
Complement inhibitor C4b-binding protein-friend or foe in the innate immune system?
Blom AM, Villoutreix BO, Dahlback B.
The complement system constitutes an important component of the defence against foreign organisms, functioning both in innate and adaptive immune systems . It is potentially harmful also to the own organism and is therefore tightly regulated by a number of membrane-bound and soluble factors . C4b-binding protein (C4BP) is a potent circulating soluble inhibitor of the classical and lectin pathways of complement . In recent years, the relationships between the structure of C4BP and its functions have been elucidated using a combination of computer-based molecular analysis and recombinant DNA technologies . Moreover, two novel functions have recently been ascribed to C4BP . One is the ability of C4BP to localize complement regulatory activity to the surface of apoptotic cells via its interaction with the membrane-binding vitamin K-dependent protein S . The other is the ability of C4BP to act as a survival factor for B cells due to an interaction with CD40 . The complement regulatory activity of C4BP is not only beneficial because it is also explored by pathogens such as Neisseria gonorrhoeae, Bordetella pertussis, Streptococcus pyogenes, Escherichia coli K1, and Candida albicans, that bind C4BP to their surfaces . This contributes to the serum resistance and the pathogenicity of these bacteria . In this review, the structural requirements and functional importance of the interactions between C4BP and its various ligands are discussed.

Rhinology, 2004 Mar, 42(1), 23 - 9
An open label, randomized comparative study of levofloxacin and amoxicillin/clavulanic acid in the treatment of purulent sinusitis in adult Thai patients; Jareoncharsri P et al.; The objective of the study was to compare the clinical efficacy and bacteriological response of levofloxacin and amoxicillin/clavulanic acid (co-amoxiclav) in the treatment of purulent maxillary sinusitis . Sixty patients randomly received either levofloxacin 300 mg orally once daily (LEV group) or co-amoxiclav 625 mg three times a day (COA group) for 14 days . Thirty four patients were in the LEV group and 26 patients were in the COA group . The mean total symptom score was significantly decreased after treatment and was comparable between both groups . Radiological improvement was 61.8% in the LEV group (41.2% resolution, 20.6% improvement) and 61.5% in the COA group (26.9% resolution, 34.6% improvement) . Pretreatment maxillary antral aspiration cultures were positive in 28 patients (82.4%) in the LEV group and 20 patients (76.9%) in the COA group . Bacteriological eradication was 78.5% in the LEV group and 70.0% in the COA group, which was not significantly different . In the LEV group, the eradication rate for major pathogens of acute sinusitis was 100% for H . influenzae (both betalactamase +ve and -ve), 100% for S . pneumoniae and S . aureus, 100% for Neisseria species, and 66.7% for P . aeruginosa . The eradication rate in the COA group was 75% for H . influenzae (both betalactamase +ve and -ve), 100% for S . pnumoniae and S . aureus, 50% for Neisseria species, and 0% for P . aeruginosa . There were no significant changes in vital sign measurements or hemato-biochemical parameters at the end of treatment as compared to baseline values, in both groups . Adverse events were found in 8.8% of patient in the LEV group and in 7.7% of patients in the COA group . Adverse events included nausea, abdominal pain, and diarrhea . All the adverse events in both groups were mild and resolved spontaneously . This study demonstrated that levofloxacin 300 mg orally once daily was as effective and safe as amoxicillin/clavulanic acid 625 mg three times a day in the treatment of maxillary sinusitis, either acute or acute exacerbation . Both drugs showed bacteriological efficacy that was not significantly different . The once daily dosage regimen is more applicable, convenience and has better compliance.

Zhonghua Liu Xing Bing Xue Za Zhi, 2004 Jan, 25(1), 58 - 9
{A molecular-epidemiologic study on TEM-1 genes of Neisseria gonorrhoeae in Wuxi area}; Mi ZH et al.; OBJECTIVE: In order to study the epidemiology of TEM-1 genes of Neisseria gonorrhoeae (Ng) in Wuxi area . METHODS: In the light from foreign literature that positive Ng strains of beta-lactamase contain plasmid sequences of TEM-1 genes, heminested PCR for TME-1 genes of Ng detection was self-designed . Ng of 195 strains were detected, that were isolated in Wuxi area from Jan to the Oct, 2002 . RESULTS: There were 138 TEM-1 genes positive in 195 isolated strains with a positive rate of 70.8% . There was one case of heminested PCR product of TEM-1 genes which showed the homogeneity was 99% in Wuxi area, when comparing with pFA7 sequences of positive Ng plasmid of beta-lactase from register GenBank . CONCLUSION: Data showed that the Ng strains with TEM-1 genes were the prevalent ones in Wuxi area.

Cell Microbiol, 2004 May, 6(5), 473 - 84
Expression of epithelial cell iron-related genes upon infection by Neisseria meningitidis; Bonnah RA et al.; Infection by the obligate human pathogens Neisseria meningitidis (MC) and Neisseria gonorrhoeae (GC) reduces the expression of host epithelial cell transferrin receptor 1 (TfR-1) (Bonnah et al., 2000, Cellular Microbiology 2: 207-218) . In addition, the rate and pattern of TfR-1 cycling is altered, leading to diminished uptake of Tf-iron by infected host cells . As Tf-iron is important for maintaining iron homeostasis in the eukaryotic cell, these findings raised the possibility that Neisseria infection might affect further pathways of epithelial cell iron metabolism . We used a specialized cDNA microarray platform, the 'IronChip', to investigate the expression of genes involved in iron transport, storage and regulation . We show that mRNA expression of several host genes involved in iron homeostasis is altered . Surprisingly, the general mRNA expression profile of infected cells closely resembled that of uninfected cells grown in an iron-limited environment . An important exception to this profile is TfR-1, the mRNA level of which is strongly reduced . Low TfR-1 expression may be explained in part by decreased activity of the iron-regulatory proteins (IRPs) in MC-infected cells, which may result in the destabilization of TfR-1 mRNA . Intriguingly, low IRP activity contrasts with the decrease in H-ferritin protein levels in infected cells . This finding suggests that low IRP activity may be responsible in part for the decrease in TfR-1 mRNA levels . A discussion of these novel findings in relation to MC infection and virulence is provided.

Sex Transm Infect, 2004 Apr, 80(2), 105 - 7
Susceptibilities of Neisseria gonorrhoeae to fluoroquinolones and other antimicrobial agents in Hyogo and Osaka, Japan; Shigemura K et al.; OBJECTIVES: Decreasing susceptibility of Neisseria gonorrhoeae to fluoroquinolones has been reported in several countries . Knowledge of local N gonorrhoeae susceptibilities to various antimicrobials is important for establishing a rational treatment strategy in each region . METHODS: Isolates of N gonorrhoeae from male urethritis patients attending four urological clinics in Hyogo and Osaka prefectures in Japan were collected during 2002 . The MICs for nine antimicrobials: penicillin G, tetracycline, cefixime, ceftriaxone, levofloxacin, gatifloxacin, ciprofloxacin, moxifloxacin, and spectinomycin were determined for each isolate . All isolates were also tested for beta lactamase producing profiles . RESULTS: Among the 87 isolates obtained, only one isolate was revealed to produce beta lactamase . MIC90 values for ciprofloxacin, levofloxacin, gatifloxacin, and moxifloxacin were over 8 microg/ml, over 8 microg/ml, 4 microg/ml, and 2 microg/ml, respectively . The proportion of isolates resistant to fluoroquinolones was over 60% (ciprofloxacin, 70.1%; levofloxacin, 65.5%; gatifloxacin, 70.1%) . Chromosomally mediated penicillin and tetracycline resistance was identified in 12.6% and 33.3% of the isolates . MIC90 values for cefixime and ceftriaxone and were 0.5 microg/ml and 0.0063 microg/ml . All isolates were sensitive to ceftriaxone and 90.8% of them were sensitive to cefixime . MIC90 for spectinomycin was 32 microg/ml and all isolates were sensitive to it . Fluoroquinolone resistance correlated significantly with MICs for penicillin G but not tetracycline . CONCLUSION: Ceftriaxone and spectinomycin demonstrated lower MICs and so are recommended for N gonorrhoeae . Susceptibilities of N gonorrhoeae should be monitored periodically by region.

Schweiz Rundsch Med Prax, 2004 Mar 3, 93(10), 372 - 6
{Unexpected complication in an elderly lady suffering from rheumatoid arthritis}; Hattenschwiler A et al.; A 66-year old female suffering from rheumatoid arthritis was treated with methotrexate and intra-articular steroid injections . She had gone through pulmonary tuberculosis at the age of 2 years, also, surgery had been performed 2 years ago because of perforated sigmoid diverticulitis . The patient now presented with episodes of abdominal pain and diarrhea as well as occasional night sweats . Laboratory investigation (normal BSR, CRP and white blood cell counts) did not indicate the presence of an inflammatory process, such as reoccurrence of diverticulitis . However, leukocyturia was repetitively found in this patient with the conventional urine culture yielding no significant bacterial growth . Further urine investigation did not indicate infection with Chlamydia trachomatis or Neisseria gonorrhoeae . Ziehl Neelson stains of morning urinary samples did not show acid-fast rods, however, Mycobacterium tuberculosis was finally isolated by culture . Thus, urogenital tuberculosis was finally diagnosed in this patient . Infection, hematogenic dissemination, and spontaneous remission of pulmonary tuberculosis had occurred more than 60 years ago . After a long latent period, reactivation of tuberculosis happened during drug-induced immunosuppression . The patient was successfully treated with an anti-tuberculosis triple-drug therapy during 2 months followed by a double-drug therapy during 4 months.

Ann Ig, 2003 Nov-Dec, 15(6), 845 - 50
{Carriers of Neisseria meningitidis among teen-agers in Apulia (Italy)}; Montagna MT et al.; An investigation about N . meningitidis carriers was carried out in Apulia (Italy) among teen-agers . 85 healthy carriers were identified (1.4%): the 81.2% of isolates resulted polyagglutinable, 3.5% autoagglutinable and 1.2% were not classifiable with antisera . The other strains belonged to serogroup B (9.4%), W135 (2.3%), Y (1.2%) and 29E (1.2%) . Some Authors report that non-groupable strains are usually isolated from healthy carriers pharynx . The most recent molecular methods permit to subdivide these bacteria in serotype and serosubtype by membrane protein antigens, too . So, today the only identification as serogroup is not able to study N . meningitidis epidemiology.

Mem Inst Oswaldo Cruz, 2003 Dec, 98(8), 1089 - 91 Epub 2004 Mar 09.
Evaluation of four methods for detecting the beta-lactamase activity in Neisseria gonorrhoeae isolated in Cuba; Llanes R et al.; Four methods (chromogenic, acidimetric, inhibition, and iodometric) for demonstration of the beta-lactamase production by 70 isolates of Neisseria gonorrhoeae, were evaluated in Cuba . There was 100% correlation between all beta-lactamase methods and the standardized penicillin dilution susceptibility test for penicillinase-non-producing N . gonorrhoeae . For penicillinase-producing N . gonorrhoeae strains, there was a perfect correlation between the chromogenic method and penicillin susceptibility testing, but one and two strains failed to give a positive result for beta-lactamase with the inhibition/acidimetric and the iodometric methods, respectively . There was a high concordance between the chromogenic method, considered as gold standard and the rest of penicillinase tests evaluated: Kappa Index (KI) = 0.98 for inhibition/acidimetric methods and KI = 0.97 for the iodometric method . The four methods evaluated were accurate, reproducible, easily readable, economical, and ease to use for screening primary isolates of N . gonorrhoeae in Cuba . We recommended the use of the inhibition method, when testing the penicillinase activity in gonococcal isolates in provincial and municipal reference laboratories.

J Biol Chem, 2004 Jun 11, 279(24), 24944 - 56 Epub 2004 Mar 24.
A novel "clip-and-link" activity of repeat in toxin (RTX) proteins from gram-negative pathogens . Covalent protein cross-linking by an Asp-Lys isopeptide bond upon calcium-dependent processing at an Asp-Pro bond; Osicka R et al.; Clinical isolates of Neisseria meningitidis produce a repeat in toxin (RTX) protein, FrpC, of unknown biological activity . Here we show that physiological concentrations of calcium ions induce a novel type of autocatalytic cleavage of the peptide bond between residues Asp(414) and Pro(415) of FrpC that is insensitive to inhibitors of serine, cysteine, aspartate, and metalloproteases . Moreover, as a result of processing, the newly generated amino-terminal fragment of FrpC can be covalently linked to another protein molecule by a novel type of Asp-Lys isopeptide bond that forms between the carboxyl group of its carboxyl-terminal Asp(414) residue and the epsilon-amino group of an internal lysine of another FrpC molecule . Point substitutions of negatively charged residues possibly involved in calcium binding (D499K, D510A, D521K, and E532A) dramatically reduced the self-processing activity of FrpC . The segment necessary and sufficient for FrpC processing was localized by deletion mutagenesis within residues 400-657, and sequences homologous to this segment were identified in several other RTX proteins . The same type of calcium-dependent processing and cross-linking activity was observed also for the purified ApxIVA protein of Actinobacillus pleuropneumoniae . These results define a protein cleavage and cross-linking module of a new class of RTX proteins of Gram-negative pathogens of man, animals, and plants . In the calcium-rich environments colonized by these bacteria this novel activity is likely to be of biological importance.

Glycobiology, 2004 Jun, 14(6), 537 - 46 Epub 2004 Mar 24.
Characterization of the alpha-1,2-N-acetylglucosaminyltransferase of Neisseria gonorrhoeae, a key control point in lipooligosaccharide biosynthesis; Wakarchuk W et al.; The biosynthesis of the lipooligosaccharide (LOS) in Neisseria meningitidis has a control point that regulates the extension of the alpha-chain on heptose (I) of the LOS . The gene that encodes the protein responsible for this control had been identified elsewhere, but the enzyme encoded by the gene was not characterized . We have now shown that this same control mechanism operates in the related species, Neisseria gonorrhoeae, using a gene knockout and subsequent characterization of the LOS species produced . We also cloned and expressed the enzyme from both of these pathogens . Using a synthetic acceptor substrate, we have shown unequivocally that the enzyme is an alpha-1,2-N-acetylglucosaminyltransferase . Experiments with both the core oligosaccharide and the synthetic acceptors suggests that the addition of the alpha-1,2-N-acetylglucosamine moiety on the heptose (II) residue precedes the addition of the ethanolamine phosphate at the O3 position on this heptose (II), and that in the absence of the alpha-1,2-N-acetylglucosamine moiety leads to the addition of an extra ethanolamine phosphate on the heptose (II) residue . Our data do not support the hypothesis that ethanolamine phosphate at O3 of heptose (II) is added and is then required for the addition of the N-acetylglucosamine at O2 by the LgtK enzyme . This enzyme represents a control point in the biosynthesis of the LOS of this pathogen and is a potential target for therapeutic intervention.

J Natl Med Assoc, 2004 Mar, 96(3), 367 - 71
Prevalence of HIV among women with vaginal discharge in a gynecological clinic; Anorlu R et al.; OBJECTIVE: Vaginal discharge is a common complaint among women with sexually transmitted diseases (STDs) in Nigeria . Given the association between STDs and HIV and the alarming rise in the prevalence of HIV in Nigeria, we performed this study to determine the prevalence of HIV in patients with vaginal discharge . STUDY DESIGN: Between April 2000 and August 2000, women who attended the Lagos University Teaching Hospital and complained of vaginal discharge were adequately counseled for HIV serotest . High vaginal/endocervical swabs and venous blood were taken for microbiological studies and HIV serotest, respectively . RESULTS: Out of the 230 women counseled for HIV serotest 140 (60.9%) consented . The mean age of the 140 was 31.1+/-6.9 years (range 18-50 years) . In 31 (22.1%), Candida albicans was isolated, in 26 Gardnerella vaginalis (18.8%), in 17 Staphylococcal spp . (12.1%); and E . coli, Trichomonas vaginalis, and Neisseria gonococcus in 7.9%, 4.3%, and 1.4%, respectively . Four (2.9%) of the 140 patients were HIV seropositive, and all had Candida albicans isolated . The prevalence of HIV among patients with Candida albicans was 12.9% . Association between HIV seropositivity and Candida albicans infection is significant (X2=14.48; p=0.002) . CONCLUSION: Candida albicans is a common cause of vaginal discharge among HIV-seropositive women in Lagos.

Infect Immun, 2004 Apr, 72(4), 2088 - 100
Vaccine potential of the Neisseria meningitidis 2086 lipoprotein; Fletcher LD et al.; A novel antigen that induces cross-reactive bactericidal antibodies against a number of Neisseria meningitidis strains is described . This antigen, a approximately 28-kDa lipoprotein called LP2086, was first observed within a complex mixture of soluble outer membrane proteins (sOMPs) following a series of fractionation, protein purification, and proteomics steps . Approximately 95 different neisserial isolates tested positive by Western blotting and PCR screening methods for the presence of the protein and the gene encoding LP2086 . The strains tested included isolates of N . meningitidis serogroups A, B, C, W135, and Y, Neisseria gonorrhoeae, and Neisseria lactamica . To better understand the microheterogeneity of this protein, the 2086 genes from 63 neisserial isolates were sequenced . Two different subfamilies of LP2086 were identified based on deduced amino acid sequence homology . A high degree of amino acid sequence similarity exists within each 2086 subfamily . The highest degree of genetic diversity was seen between the two subfamilies which share approximately 60 to 75% homology at the nucleic acid level . Flow cytometry (fluorescence-activated cell sorting) analyses and electron microscopy indicated that the LP2086 is localized on the outer surface of N . meningitidis . Antiserum produced against a single protein variant was capable of eliciting bactericidal activity against strains expressing different serosubtype antigens . Combining one recombinant lipidated 2086 (rLP2086) variant from each subfamily with two rPorA variants elicited bactericidal activity against all strains tested . The rLP2086 family of antigens are candidates worthy of further vaccine development.

Sex Transm Dis, 2004 Apr, 31(4), 209 - 14
Gonorrhoea surveillance system in France: 1986-2000; Herida M et al.; OBJECTIVE: The objectives of this study were to monitor epidemiologic trends in Neisseria gonorrhoeae (NG) infections and to assess antimicrobial resistance patterns of NG . METHODS: RENAGO, a voluntary-based laboratory surveillance system, including private and public laboratories, was set up in France in 1986 . Every NG strain isolated by the participating laboratories was sent to the National Reference Center, Institut Alfred Fournier, Paris, where susceptibility to 6 antibiotics was tested . These laboratories reported monthly numbers of positive NG isolates and associated epidemiologic data . Because the number of laboratories varied every year, the average number of NG isolates per laboratory per year was computed to monitor annual trends in gonorrhea incidence . RESULTS: The average number of NG isolated per laboratory per year decreased steadily from 10.6 in 1986 to 0.6 in 1997 but then yearly increased and reached 1.9 in 2000 . Increasing gonorrhea was observed mostly in men and in the Paris area . The proportion of rectal strains increased significantly from 0.9% in 1986 to 9.2% in 2000 . From 1998 to 2000, 5.3% of the isolates had reduced susceptibility or were resistant to ciprofloxacin, 7.3% were PPNG strains and 27.4% were resistant to tetracycline, 18.5% had a reduced susceptibility to thiamphenicol, and 1 strain was resistant to the latter . Neither strain was resistant to ceftriaxone or to spectinomycin . CONCLUSION: From 1998 to 2000, gonorrhea markedly increased in France and seemed to affect mainly gay men in Paris, which suggests relapses to risky sexual behaviors and highlights the need for strengthening prevention . During the same period, NG ciprofloxacin resistance has sharply increased and should be carefully monitored because of the wide use of ciprofloxacin single-dose treatment.

Immunology, 2004 Feb, 111(2), 173 - 8
Toll-like receptor 2 (TLR2) and TLR4 are present inside human dendritic cells, associated with microtubules and the Golgi apparatus but are not detectable on the cell surface: integrity of microtubules is required for interleukin-12 production in response to internalized bacteria; Uronen-Hansson H et al.; The activation of dendritic cells (DCs) by microbes is mediated by pattern recognition receptors including the Toll-like receptors (TLR) . Bacterial lipopolysaccharide acts via TLR4 whereas peptidoglycan and lipoprotein responses are mediated by TLR2 . It is generally accepted that TLR binding to microbes occurs at the cell surface but this has not been directly demonstrated for human DCs . We show here that TLR2 and TLR4 are expressed inside DCs in an abundant tubulovesicular pattern with a focus of intense staining adjacent to the nucleus . In contrast, there was no detectable expression on the cell surface . TLR2 and TLR4 were readily found both intracellularly and on the surface of monocytes . They were shown to be closely associated with the Golgi complex and colocalized with alpha-tubulin, displaying a high focal concentration at the microtubule organizing centre . Alignment of TLR2 and TLR4 with microtubules was observed, suggesting that microtubules serve as transport tracks for TLR vesicles . Depolymerization of the microtubule network disrupted the intracellular expression of TLR2 and TLR4 and profoundly inhibited interleukin-12 (IL-12) production in response to Neisseria meningitidis but did not prevent phagocytosis . These data are consistent with the bacterial signalling through TLR2 and TLR4 required for IL-12 production occurring inside DCs after phagocytosis.

J Med Chem, 2004 Mar 25, 47(7), 1775 - 82
Antineoplastic agents . 499 . Synthesis of hystatin 2 and related 1H-benzo{de}{1,6}-naphthyridinium salts from aaptamine; Pettit GR et al.; The marine sponge constituent aaptamine (1) has been converted to the cancer cell growth inhibitor and antibiotic designated hystatin 2 (8a) . Herein, we also report results of an initial SAR evaluation of new benzyl derivatives of aaptamine (1) . Single benzylation was found to occur at nitrogen N-4 and led to the formation of the 4-benzylaaptamine derivatives 7a-c, whereas double benzylation gave the quaternary 1H-benzo{de}{1,6}-naphthyridinium salts 8a-c . The anticancer and antimicrobial properties of these aaptamine derivatives are described . The quaternary ammonium salts 8a (hystatin 2) and 8b exhibited significant inhibitory activity against the murine P388 lymphocytic leukemia and a minipanel of human cancer cell lines . Salts 8a and 8b also had broad spectrum antimicrobial activities and were most potent against Mycobacterium tuberculosis, Neisseria gonorrhoeae, and Micrococcus luteus . Naphthyridinium chloride 8a was selected for further development, and results of an initial cell cycle analysis and a cDNA microarray study showed effects consistent with inhibition of the S-phase of cell growth.

J Comput Chem, 2004 May, 25(7), 956 - 67
Geometric algorithms for the conformational analysis of long protein loops; Cortes J et al.; The efficient filtering of unfeasible conformations would considerably benefit the exploration of the conformational space when searching for minimum energy structures or during molecular simulation . The most important conditions for filtering are the maintenance of molecular chain integrity and the avoidance of steric clashes . These conditions can be seen as geometric constraints on a molecular model . In this article, we discuss how techniques issued from recent research in robotics can be applied to this filtering . Two complementary techniques are presented: one for conformational sampling and another for computing conformational changes satisfying such geometric constraints . The main interest of the proposed techniques is their application to the structural analysis of long protein loops . First experimental results demonstrate the efficacy of the approach for studying the mobility of loop 7 in amylosucrase from Neisseria polysaccharea . The supposed motions of this 17-residue loop would play an important role in the activity of this enzyme .

Curr Infect Dis Rep, 2004 Apr, 6(2), 129 - 134
Gonorrhea Update; Bash MC; This article provides a brief summary of recent US epide-miology, antimicrobial resistance, and treatment of Neisseria gonorrhoeae infections . Selected research regarding infections caused by N . gonorrhoeae is described, with particular emphasis on the advances made by new molecular methods.

Biochemistry, 2004 Mar 23, 43(11), 3104 - 10
Crystal structure of the covalent intermediate of amylosucrase from Neisseria polysaccharea; Jensen MH et al.; The alpha-retaining amylosucrase from the glycoside hydrolase family 13 performs a transfer reaction of a glucosyl moiety from sucrose to an acceptor molecule . Amylosucrase has previously been shown to be able to use alpha-D-glucopyranosyl fluoride as a substrate, which suggested that it could also be used for trapping the reaction intermediate for crystallographic studies . In this paper, the crystal structure of the acid/base catalyst mutant, E328Q, with a covalently bound glucopyranosyl moiety is presented . Sucrose cocrystallized crystals were soaked with alpha-D-glucopyranosyl fluoride, which resulted in the trapping of a covalent intermediate in the active site of the enzyme . The structure is refined to a resolution of 2.2 A and showed that binding of the covalent intermediate resulted in a backbone movement of 1 A around the location of the nucleophile, Asp286 . This structure reveals the first covalent intermediate of an alpha-retaining glycoside hydrolase where the glucosyl moiety is identical to the expected biologically relevant entity . Comparison to other enzymes with anticipated glucosylic covalent intermediates suggests that this structure is a representative model for such intermediates . Analysis of the active site shows how oligosaccharide binding disrupts the putative nucleophilic water binding site found in the hydrolases of the GH family 13 . This reveals important parts of the structural background for the shift in function from hydrolase to transglycosidase seen in amylosucrase.

Genetics, 2004 Jan, 166(1), 25 - 32
Strong positive selection and recombination drive the antigenic variation of the PilE protein of the human pathogen Neisseria meningitidis; Andrews TD et al.; The PilE protein is the major component of the Neisseria meningitidis pilus, which is encoded by the pilE/pilS locus that includes an expressed gene and eight homologous silent fragments . The silent gene fragments have been shown to recombine through gene conversion with the expressed gene and thereby provide a means by which novel antigenic variants of the PilE protein can be generated . We have analyzed the evolutionary rate of the pilE gene using the nucleotide sequence of two complete pilE/pilS loci . The very high rate of evolution displayed by the PilE protein appears driven by both recombination and positive selection . Within the semivariable region of the pilE and pilS genes, recombination appears to occur within multiple small sequence blocks that lie between conserved sequence elements . Within the hypervariable region, positive selection was identified from comparison of the silent and expressed genes . The unusual gene conversion mechanism that operates at the pilE/pilS locus is a strategy employed by N . meningitidis to enhance mutation of certain regions of the PilE protein . The silent copies of the gene effectively allow "parallelized" evolution of pilE, thus enabling the encoded protein to rapidly explore a large area of sequence space in an effort to find novel antigenic variants.

J Clin Virol, 2004 Apr, 29(4), 230 - 40
Optimization of PCR based detection of human papillomavirus DNA from urine specimens; Brinkman JA et al.; BACKGROUND: Human papillomavirus (HPV) causes cervical cancer . Current screening requires a yearly pelvic exam and Pap smear . However, these procedures are impractical for screening all women at risk for disease . Urine sampling has been successfully utilized to screen for Chlamydia trachomatis (CT) and Neisseria gonorrhoreae (NG) infections and has been considered for HPV DNA detection by several investigators . However, no study to date has been performed to specifically optimize HPV detection in urine . OBJECTIVES: To compare handling and extraction techniques in order to optimize the HPV specific PCR system in urine specimens . STUDY DESIGN: Examination of 10 characteristics that may contribute to PCR inhibition in urine was performed utilizing 10SG mulitstixs . Five different DNA extraction methods were compared in spiked specimens and in 10 clinical specimens . After the optimal extraction technique was identified, concentration of the sample with and without prior dilution was compared to the original protocol . Lastly, specimen handling was compared between immediate processing, refrigerating overnight, or freezing overnight . RESULTS AND CONCLUSIONS: the presence of protein in urine enhanced amplification while nitrites decreased amplification . Of the extraction methods tested, the QIAamp DNA Mini Kit demonstrated the best amplification from urine samples spiked with HPV DNA and clinical specimens . The addition of a dilution step and a concentration step before applying the Qiagen protocol further increased amplification of beta-globin (from 50 to 63%) and the HPV L1 gene (from 13 to 33%) . Lastly, refrigerating the specimens at 4 degrees C overnight appears to produce better amplification (62% beta-globin and 17% HPV positive) than either immediate processing (46% beta-globin and 13% HPV+) or freezing the specimen for 24h prior to processing (46% beta-globin and 10% HPV+) . In these studies, amplification was low despite optimization . Additional improvements are required prior to clinical application of a urine-based HPV DNA detection system.

Bull Soc Pathol Exot, 2003 Jan, 96(5), 372 - 5
{Fighting principles against sexually transmitted infections in tropical areas}; Ly F et al.; Owing to their frequency and their own complications, sexually transmitted infections (STI) constitute a very classical health problem, especially in developing areas . The interest for those diseases has been on the rise since the beginning of the AIDS pandemic . Conventional sexually transmitted diseases (such as syphilis, chancroid, infections due to Neisseria gonorrhoeae and to Chlamydia trachomatis) are potent cofactors of HIV transmission . It has been established that the treatment of patients with STI was able to lower the HIV incidence at the community level . Today the fight against STI relies mainly on preventive measures (promotion of lower risk sexual behaviours, blood transfusion safety), and on medical management of patients with STI . In order to treat rapidly cheaply and efficiently a great number of patients, standardised syndromic approaches, using generic drugs, have been elaborated for the most common STI . Other important complementary measures are based on the treatment of sexual partners and on the promotion of condom use . In addition to prevention of their specific complications, the fight against STI has become a major component of health policies against AIDS in developing countries.

EMBO J, 2004 Mar 24, 23(6), 1257 - 66 Epub 2004 Mar 11.
Structure of the translocator domain of a bacterial autotransporter; Oomen CJ et al.; Autotransporters are virulence-related proteins of Gram-negative bacteria that are secreted via an outer-membrane-based C-terminal extension, the translocator domain . This domain supposedly is sufficient for the transport of the N-terminal passenger domain across the outer membrane . We present here the crystal structure of the in vitro-folded translocator domain of the autotransporter NalP from Neisseria meningitidis, which reveals a 12-stranded beta-barrel with a hydrophilic pore of 10 x 12.5 A that is filled by an N-terminal alpha-helix . The domain has pore activity in vivo and in vitro . Our data are consistent with the model of passenger-domain transport through the hydrophilic channel within the beta-barrel, and inconsistent with a model for transport through a central channel formed by an oligomer of translocator domains . However, the dimensions of the pore imply translocation of the secreted domain in an unfolded form . An alternative model, possibly covering the transport of folded domains, is that passenger-domain transport involves the Omp85 complex, the machinery required for membrane insertion of outer-membrane proteins, on which autotransporters are dependent.

Carbohydr Res, 2004 Feb 25, 339(3), 693 - 8
Enzymatic supported synthesis of lacto-N-neotetraose using dendrimeric polyethylene glycol; Renaudie L et al.; The lacto-N-neotetraose tetrasaccharide was synthesized on a new dendrimeric support, based on polyethylene glycol . Starting from 1-thio-beta-D-lactose, the trisaccharide (2-acetamido-2-deoxy-beta-D-glucopyranosyl)-(1-->3)-O-beta-D-galactopyranosyl-(1-->4)-1-thio-beta-D-glucopyranose was obtained using Neisseria meningitidis beta-(1-->3)-N-acetylglucosaminyltransferase according to a soluble synthesis approach, bound on the support and galactosylated using the milk beta-(1-->4)-galactosyl transferase to give after cleavage the tetrasaccharide lacto-N-neotetraose.

Carbohydr Res, 2004 Apr 2, 339(5), 1007 - 14
Molecular modeling insights into the catalytic mechanism of the retaining galactosyltransferase LgtC; Tvaroska I; The bacterial enzyme lipopolysaccharyl alpha-galactosyltransferase C (EC 2.4.1.x, LgtC) is involved in the synthesis of lipooligosaccharides displayed on the cell surfaces of Neisseria meningitidis . LgtC catalyzes the transfer of a galactosyl residue from UDP-Gal to the terminal galactose residue of glycoconjugates with an overall retention of stereochemistry at the anomeric center . Several hypothetical catalytic mechanisms of the LgtC enzyme were examined herein using DFT quantum chemical methods up to the B3LYP/6-311++G**//B3LYP/6-31G* level . The computational model used to follow the reaction is based on the crystallographic structure of LgtC in complex with both the nucleotide-galactose donor and the oligosaccharide-acceptor analogues . The 136 atoms included in this model represent fragments of residues critical for the substrate binding and catalysis . From our calculations, the preferred pathway is predicted to be a one step mechanism with the nucleophilic attack of the acceptor oxygen onto the anomeric carbon and the proton transfer to a phosphate oxygen occurring simultaneously . This mechanism has an A(N)D(N)A(H)D(H) character, with the unique transition state structure in which the attacking galactose group is more closely bound to the anomeric carbon than to the UDP leaving group and where the hydrogen bond between the nucleophile and the leaving group oxygens facilitates the attack of the acceptor O4(') from the same side of the transferred galactose.

Carbohydr Res, 2004 Apr 2, 339(5), 995 - 1006
Molecular dynamics simulations of glycosyltransferase LgtC; Snajdrova L et al.; Molecular dynamics simulations have been performed on fully solvated alpha-(1-->4)-galactosyltransferase LgtC from Neisseria meningitidis with and without the donor substrate UDP-Gal and in the presence of the manganese ion . The analysis of the trajectories revealed a limited movement in the loop X (residues 75-80) and a larger conformational change in the loop Y (residues 246-251) in the simulation, when UDP-Gal was not present . In this case, the loops X and Y open by almost 10A, exposing the active site to the solvent . The 'hinge region' responsible for the opening is composed of residues 246-247 . We have also analyzed the behavior of the manganese ion in the simulations . The coordination number is 6 when UDP-Gal is present and it increases to 7 when it is absent . In the latter case, three water molecules become coordinated to the ion . In both cases, the coordination is very stable implying that the manganese ion is tightly bound in the active site of the enzyme even if UDP-Gal is not present . Further analysis of the structural water molecules location confirmed that the mobility of water molecules in the active site and the accessibility of this site for solvent are higher in the absence of the substrate.

Int J STD AIDS, 2004 Feb, 15(2), 116 - 9
Prevalence of sexually transmitted diseases and human immunodeficiency virus among women attending prenatal services in Apia, Samoa; Sullivan EA et al.; There is no routine prenatal screening for sexually transmitted diseases (STDs) and human immunodeficiency virus (HIV) in pregnancy in Samoa . Testing for chlamydial infection is not available . To gather information on pregnant women, a prevalence survey was conducted in Apia, Samoa, utilizing two prenatal hospital clinics . Pregnant (n=427) women were tested for Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis using polymerase chain reaction (PCR), and for syphilis (n=441) by rapid plasmid reagin (RPR) and HIV (n=441) by enzyme-linked immunosorbent assay (ELISA) . Results were: chlamydia 30.9% (132); trichomoniasis 20.8%; gonorrhoea 3.3%; syphilis 0.5%; and HIV 0% . Overall 42.7% had at least 1 STD . Young women aged <25 years were three times more likely to have a STD than older women (odds ratio=3.0, 95% confidence intervals 2.0, 4.5) . The lack of inexpensive, reliable field diagnostics remain a barrier to sustainable STD control programmes for pregnant women living in developing countries.

Int J STD AIDS, 2004 Feb, 15(2), 94 - 8
Vaginal tampons as specimen collection device for the molecular diagnosis of non-ulcerative sexually transmitted infections in antenatal clinic attendees; Sturm PD et al.; Self-inserted vaginal tampons for the molecular diagnosis of non-ulcerative STIs were evaluated . Cervical and vaginal swabs, tampons and urines were collected from 185 first-time antenatal clinic attendees . Cultures and nucleic acid amplification assays (NAA) were performed . The sensitivity of PCR on tampons for Trichomonas vaginalis was with 94% (CI 85-98%) significantly higher (P<0.001) than culture (50%, CI 38-62%) or urine (53%, CI 41-65%) . Neisseria gonorrhoeae culture had a sensitivity of 64% (CI 36-86%), strand displacement assay (SDA) had a sensitivity of 79% (CI 49-94%) using tampon specimens, 57% (CI 30-81%) using endocervical swabs and 43% (CI 19-70%) using urines . There was no difference in sensitivity of SDA for Chlamydia trachomatis using tampon specimens, urine or endocervical swabs . The specificity approached 100% for all assays on all specimens . NAA on tampons for the detection of T . vaginalis, N . gonorrhoeae and C . trachomatis identified more infections than assays on swabs or urines . This reached statistical significance for T . vaginalis only.

Protein Expr Purif, 2004 Apr, 34(2), 176 - 82
Cysteine mediated multimerization of a recombinant dengue E fragment fused to the P64k protein following immobilized metal ion affinity chromatography; Lopez C et al.; A fragment of the Envelope protein of Dengue 2 virus encoding the amino acid (aa) 286-426 was fused to the P64k protein from Neisseria meningitidis . The hybrid gene (PD5) was expressed in Escherichia coli . The influence of using immobilized metal ion affinity chromatography (IMAC) in the purification process of PD5 was examined . After the elution, PD5 protein formed aggregates of high molecular weight depending on disulfide-bonds . The study of different conditions affecting the redox potential in the system revealed the influence of the copper ions on the multimerization of the protein, whereas metals with minor redox potential-for instance, zinc or nickel ions-did not cause this effect . It was also demonstrated that cysteines involved in this process belonged to the P64k protein . Finally, a PD5 purification process was established reaching a 85% of purity using the Zn(2+) as metal ion in the IMAC . This is a very useful finding due to the wide use of P64k as a carrier protein and the advantages of the IMAC as a chromatographic process.

Microb Drug Resist, 2003 Winter, 9(4), 353 - 60
Molecular epidemiology of Neisseria gonorrheae isolates with plasmid-mediated tetracycline resistance in Canada: temporal and geographical trends (1986-1997); Greco V et al.; Plasmid-mediated resistance to tetracycline in Neisseria gonorrhoeae (TRNG) isolates is caused by the acquisition of a 25.2-MDa conjugative, tetM-containing plasmid (TetM plasmid) . The presence of the TetM plasmid is the leading cause of gonococcal resistance to tetracycline in most countries . Between 1986 and 1997, 6,306 TRNG isolates were isolated in different Canadian provincial laboratories and subsequently submitted to the national laboratory for further strain characterization . Because nonculture-based identification of N . gonorrhoeae was more widely used after 1995, this snapshot of the molecular epidemiology of TRNG in Canada, which is only possible if bacteria are cultured, represents a comprehensive data baseline that may no longer be achievable . Temporal trends indicate that TRNG isolations peaked in 1994 (18.9% of isolates tested) . Antimicrobial susceptibilities (MIC) to tetracycline and penicillin were determined for 4,064 TRNG isolated between 1986 and 1994 . The MICs of TRNG isolates ranged from 16 microg/ml to 32 microg/ml of tetracycline, although one isolate had an MIC of 8 microg/ml and the MICs of four isolates were 2 microg/ml . Penicillinase-producing TRNG (i.e., PP/TRNG) comprised 34.1% of all TRNG (n = 1,386) and 52 TRNG isolates exhibited chromosomal resistance to penicillin . Most of the PP/TRNG (94.1%) carried Africa type (3.2 MDa) beta-lactamase-producing plasmids; only 76 (5.5%) PP/TRNG carried Asia type (4.4 MDa) penicillinase-producing plasmids and three isolates carried Toronto type (3.05 MDa) plasmids . TRNG isolates were also retrospectively typed by auxotype (A), serovar (S), and plasmid (P) content analysis . Eleven auxotype/serovar (A/S) groups comprised the majority (93%) of 4,064 typed TRNG isolates with A/S classes NR/IB-2, NR/IB-3, and NR/IB-1 accounting for 75.1% of the strains characterized . Classification of 670 TRNG for tetM type demonstrated that the Dutch (n = 531) type TetM plasmids predominated over the American (n = 139) type TetM plasmids.

J Antimicrob Chemother, 2004 Apr, 53(4), 653 - 6 Epub 2004 Mar 03.
Fluoroquinolone-resistant Neisseria gonorrhoeae isolates from Russia: molecular mechanisms implicated; Vereshchagin VA et al.; OBJECTIVES: During a longitudinal study of the prevalence of antimicrobial resistance in Neisseria gonorrhoeae, a number of high-level fluoroquinolone-resistant isolates were obtained from the sexually transmitted diseases clinic in the Moscow region in 2002 . The aim of the present study was to determine the molecular mechanisms of resistance and to assess the clonal relationship of these strains METHODS: For the 32 clinical strains of N . gonorrhoeae studied, the MIC values were determined for four fluoroquinolones . The gyrA, parC, por and mtrR genes were studied for the presence of mutations associated with fluoroquinolone resistance . RESULTS: We detected strains of N . gonorrhoeae showing high-level resistance to fluoroquinolones (21 strains, with MICs 1-32 mg/L) . Mutations in gyrA and parC known to cause fluoroquinolone resistance were detected in a majority of strains . There were four strains (among 21) without known changes in gyrA and parC . However, amino acid changes in the Por protein and mutations in the promoter or encoding region of the mtrR gene were detected in three of them . One strain had no alteration in gyrA, parC, por or mtrR . CONCLUSIONS: The present study documents the first case of fluoroquinolone-resistant N . gonorrhoeae in Russia.

J Nat Prod, 2004 Feb, 67(2), 214 - 20
Antineoplastic agents . 522 . Hernandia peltata (Malaysia) and Hernandia nymphaeifolia (Republic of Maldives); Pettit GR et al.; Bioassay (P388 lymphocytic leukemia cell line and human tumor cell lines)-guided separation of the extracts prepared from the tropical and coastal trees Hernandia peltata (Malaysia) and Hernandianymphaeifolia (Republic of Maldives) led to the isolation of a new lignan designated as hernanol (1) and 12 previously known lignans: (-)-deoxypodophyllotoxin (2), deoxypicropodophyllin (3), (+)-epiaschantin (4), (+)-epieudesmin (5), praderin (6), 5'-methoxyyatein (7), podorhizol (8), deoxypodorhizone (9), bursehernin (10), kusunokinol (11), clusin (12), and (-)-maculatin (13) . The oxidative cyclization (with VOF(3)) of lignans 8, 9, and 10 resulted in a new and unusual benzopyran (14), isostegane (15), and a new dibenzocyclooctadiene lactone (16), respectively . The structure and relative stereochemistry of hernanol (1) and lignans 3, 7, 8, 9, 10, 11, and 12 were determined by 1D and 2DNMR and HRMS analyses . The structures and absolute stereochemistry of structures 2, 4, 5, 6, 13, 14, 15, and 16 were unequivocally determined by single-crystal X-ray diffraction analyses . Evaluation against the murine P388 lymphocytic leukemia cell line and human tumor cell lines showed podophyllotoxin derivatives 2 and 3 to be strong cancer cell line growth inhibitors and substances 4, 5, 8, and 15 to have marginal cancer cell line inhibitory activities . Seven of the lignans and one of the synthetic modifications (14) inhibited growth of the pathogenic bacterium Neisseria gonorrhoeae.

Clin Infect Dis, 2004 Mar 1, 38(5), 649 - 54 Epub 2004 Feb 18.
The epidemiology of fluoroquinolone-resistant Neisseria gonorrhoeae in Hawaii, 2001; Newman LM et al.; Increases in the number of infections with fluoroquinolone-resistant Neisseria gonorrhoeae in Asia and the United States threaten the efficacy of fluoroquinolones as inexpensive, single-dose, orally administered treatments for gonorrhea . This report describes the findings of a field investigation of an increase in the number of infections with ciprofloxacin-resistant N . gonorrhoeae (CipRGC) in Hawaii in 2001 . We conducted a case review of 53 patients with CipRGC, who constituted 20% of the 267 patients with cultures positive for N . gonorrhoeae during this period . Nearly one-half of patients with CipRGC were seen by clinicians in private practice, one-third were seen by clinicians at a sexually transmitted diseases (STD) clinic, and only 2% were seen by clinicians in the military . Among the 117 patients with culture-confirmed gonorrhea who attended the public STD clinic, we found a prevalence of infection with CipRGC of 17% . The demographic and clinical characteristics of patients with CipRGC were similar to those of patients with gonorrhea that was not resistant to ciprofloxacin, suggesting that fluoroquinolone-resistant gonorrhea has become endemic in Hawaii.

Jpn J Infect Dis, 2004 Feb, 57(1), 17 - 20
Prevalence and treatment of Chlamydia trachomatis, Ureaplasma urealyticum, and Mycoplasma hominis in patients with non-gonococcal urethritis; Kilic D et al.; The aim of present study was to evaluate the occurrence of Chlamydia trachomatis, Mycoplasma hominis, and Ureaplasma urealyticum in non-gonococcal urethritis (NGU) and to determine the bacterial resistance to six antibiotics in order to determine the most suitable treatment strategy . A total of 50 patients were enrolled into the study . Urethral samples were taken with a dacron swab placed into urethra 2 - 3 cm in males, and vaginal samples were taken from the endocervical region in women . The patient samples that did not grow Neisseria gonorrhoeae were accepted as NGU . Direct immunofluorescence technique was used for the investigation of C . trachomatis . Mycoplasma IST was used for the isolation of M . hominis and U . urealyticum . U . urealyticum was isolated from 24 patients . Thirteen of them had only U . urealyticum, and the rest had mixed pathogen organisms (7 U . urealyticum + M . hominis; 3 U . urealyticum + C . trachomatis, and 1 U . urealyticum + M . hominis + C . trachomatis) . C . trachomatis was detected in 12 patients . While 8 patients had C . trachomatis only, the rest had a mixture of the pathogen organisms listed above . Partner examinations could be performed for only 22 patients' partners . In the evaluation of antibiotic susceptibility, higher resistance was obtained against ofloxacin in U . urealyticum, and against erythromycin with M . hominis . Our results indicated that doxycycline or ofloxacin should be the first choice when empirical treatment is necessary.

Am J Obstet Gynecol, 2004 Feb, 190(2), 305 - 13
The antimicrobial treatment of subacute endometritis: a proof of concept study; Eckert LO et al.; OBJECTIVE: Our purpose was to evaluate the antimicrobial therapy effect on clinical and laboratory findings among women at risk for endometritis . STUDY DESIGN: A prospective antimicrobial treatment trial of 153 women was performed to characterize subacute endometritis and to determine the treatment effect on endometritis resolution . RESULTS: After antimicrobial treatment, significant reductions occurred in abnormal bleeding (60% vs 29%), mucopurulent cervicitis (20% vs 6%), uterine tenderness (20% vs 6%), and histologic endometritis (38% vs 4%), all P<.001 . In women with prior pelvic inflammatory disease (PID), endometritis was present in 43% with and 28% without current Chlamydia trachomatis or Neisseria gonorrhoeae . In women without prior PID, endometritis was present in 23% with and 12% without current C trachomatis or N gonorrhoeae (P=.002 for trend) . CONCLUSIONS: In women without a clinical diagnosis of PID, antimicrobial therapy decreased abnormal clinical findings and histologic endometritis . Prior PID is additive with current cervical infection as a risk for endometritis.

Infect Immun, 2004 Mar, 72(3), 1333 - 40
Identification of novel virulence-associated genes via genome analysis of hypothetical genes; Garbom S et al.; The sequencing of bacterial genomes has opened new perspectives for identification of targets for treatment of infectious diseases . We have identified a set of novel virulence-associated genes (vag genes) by comparing the genome sequences of six human pathogens that are known to cause persistent or chronic infections in humans: Yersinia pestis, Neisseria gonorrhoeae, Helicobacter pylori, Borrelia burgdorferi, Streptococcus pneumoniae, and Treponema pallidum . This comparison was limited to genes annotated as hypothetical in the T . pallidum genome project . Seventeen genes with unknown functions were found to be conserved among these pathogens . Insertional inactivation of 14 of these genes generated nine mutants that were attenuated for virulence in a mouse infection model . Out of these nine genes, five were found to be specifically associated with virulence in mice as demonstrated by infection with Yersinia pseudotuberculosis in-frame deletion mutants . In addition, these five vag genes were essential only in vivo, since all the mutants were able to grow in vitro . These genes are broadly conserved among bacteria . Therefore, we propose that the corresponding vag gene products may constitute novel targets for antimicrobial therapy and that some vag mutants could serve as carrier strains for live vaccines.

Salud Publica Mex, 2003, 45 Supp 5, S681 - 9
{Prevalence of anti-Chlamydia trachomatis and anti-Neisseria gonorrhoeae antibodies in Mexican populations}; Cravioto Mdel C et al.; OBJECTIVE: To estimate the prevalence of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infection in groups of individuals at different risks of sexually transmitted infections (STI) . MATERIAL AND METHODS: Between January 1992 and December 1993, a cross-sectional multicentric study was carried out at the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran (National Institute of Medical Sciences and Nutrition "Salvador Zubiran") in Mexico City . The study population consisted of 945 reproductive age subjects (585 females and 360 males) . Low and high risk groups were classified according to their risk for STI . High risk groups included infertile women with tubal damage, women with a history of ectopic pregnancy or abortion, infertile men, HIV/AIDS patients, homo- or bisexual men, and female commercial sex workers . Low risk groups included primigravidae, fertile men, and infertile women with no tubal damage . Serum anti-NG and anti-CT IgG and IgA were determined, in duplicate by immune-enzymatic assay, using as antigens NG pili and the LI fraction of CT . Descriptive analysis is presented as percentages . RESULTS: NG prevalence in females was 13.7% by IgG and 14.3% by IgA . CT prevalence was 11.4% by IgG and 4.4% by IgA . In males, NG prevalences were 3.3% and 13.3% by IgG and IgA, respectively; CT prevalences were 7.2% and 5.5%, respectively . In commercial sex workers, NG prevalences were 31.2% by IgG and 28.4% by IgA, and CT 25.0% and 5.7% by IgG and IgA, respectively . In women with infertility due to tubal damage the prevalences of NG were 5.6% and 9.8%, respectively, and those of CT were 8.4% and 1.4%, respectively . In 110 young primigravid NG prevalences were 4.5% and 10.0%, respectively, and CT 3.6% and 9.1% . CONCLUSIONS: These data confirm the high prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis in female commercial sex workers and homo- or bisexual men, but not in other high-risk groups like infertile women or women with a history of ectopic pregnancy or abortion . The English version of this paper is available at:http://www.insp.mx/salud/index.html.

Pol Merkuriusz Lek, 2003 Nov, 15(89), 459 - 62
{Epidemiology and therapy of infections caused by Neisseria meningitidis}; Skoczynska A et al.; In spite of the progress in antibiotic therapy and immunoprophylaxis Neisseria meningitidis remains one of the major causes of meningitis and sepsis burdened by high mortality . In this paper characteristics of N . meningitidis and in particular its epidemiology, virulence factors, antibiotic therapy and resistance mechanisms, immuno- and chemoprophylaxis, are discussed . Data from the National Reference Centre for Bacterial Meningitis are also presented . These data indicate that this pathogen is the most common laboratory confirmed bacterial etiologic agent of community-acquired meningitis in Poland.

J Clin Microbiol, 2004 Feb, 42(2), 923 - 4
Culture-negative pericarditis caused by Neisseria meningitidis serogroup C; Moumile K et al.; We describe a case of primary purulent culture-negative pericarditis caused by Neisseria meningitidis serogroup C occurring in an 8-month-old previously healthy boy, which was detected in pericardial fluid by broad-spectrum PCR amplification.

J Clin Microbiol, 2004 Feb, 42(2), 618 - 21
Epidemiology and etiology of sexually transmitted infection among hotel-based sex workers in Dhaka, Bangladesh; Nessa K et al.; The prevalence of sexually transmitted infections (STIs) and reproductive tract infections (RTIs) among hotel-based sex workers (HBSWs) in Dhaka, Bangladesh, was studied . A total of 400 HBSWs were enrolled in the study during April to July 2002 . Endocervical swabs, high vaginal swabs, and blood samples from 400 HBSWs were examined for Neisseria gonorrhoeae (by culture), Chlamydia trachomatis (by PCR), Trichomonas vaginalis (by microscopy), antibody to Treponema pallidum (by both rapid plasma reagin and Treponema pallidum hemagglutination tests), and antibody to herpes simplex virus type 2 (HSV-2) (by enzyme-linked immunosorbent assay) . Sociodemographic information as well as gynecological and obstetric information was collected . Among the HBSWs, 228 women (57%) were symptomatic and 172 (43%) were asymptomatic, 35.8% were positive for N . gonorrhoeae, 43.5% were positive for C . trachomatis, and 4.3% were positive for T . vaginalis . A total of 8.5% had syphilis, 34.5% were positive for HSV-2, and 86.8% were positive for at least one RTI or STI . There was no significant difference between the prevalences of STIs among the symptomatic and asymptomatic HBSWs . These data suggested a high prevalence of STIs, particularly gonorrhea and chlamydia, among HBSWs in Dhaka.

J Clin Microbiol, 2004 Feb, 42(2), 591 - 4
Comparison of mismatch amplification mutation assay with DNA sequencing for characterization of fluoroquinolone resistance in Neisseria gonorrhoeae; Sultan Z et al.; A mismatch amplification mutation assay (MAMA) was developed for identification of point mutations in quinolone resistance-determining region (QRDR) of gyrA at codons 91 and 95 . MAMA PCR was used to detect mutations at codons 91 and 95 of gyrA in 117 Neisseria gonorrhoeae isolates (with ciprofloxacin MICs of 0.004 to >32 microg/ml) from Bangladesh during 1997 to 2001 . The QRDR regions of the gyrA genes from 31 randomly selected isolates were sequenced, and the results were compared with those of MAMA PCR . Using mismatch PCR, a mutation at Ser91 could be detected in all 27 (resistant and intermediate) isolates, and an Asp95-to-Gly95 mutation could be detected in all 15 isolates, as detected by sequencing . MAMA PCR offers a simple, inexpensive, rapid, and easier alternative for detection of point mutations in fluoroquinolone resistance in N . gonorrhoeae.

J Pak Med Assoc, 2003 Dec, 53(12), 580 - 3
Spectrum of complications and mortality of bacterial meningitis: an experience from a developing country; Rabbani MA et al.; OBJECTIVE: The aim of this study was to obtain data on predisposing factors, causative organisms and their associated mortality and complications related to acute bacterial meningitis . METHODS: The chart review of all patients in whom acute bacterial meningitis was diagnosed at The Aga Khan University Hospital from January 1995 through December 2001 . RESULTS: One hundred ninety-four patients were included in study . There were 146 males and 48 females . The mean age of our study population was 41 +/- 12.3 years . One hundred and ninety (97.9%) patients had community-acquired meningitis; only 4 (2.0%) patients developed meningitis nosocomially . The two most common predisposing factors were diabetes mellitus (13.9%) and otitis media (7.7%) among all 194 patients . A significant proportion of patients with complications had diabetes mellitus (24.6%, p < 0.001) . CSF and blood cultures were positive in 53 (27.3%) and 42 (21.6%) patients respectively; there was no statistical difference found . The most common organisms isolated were Streptococcus pneumoniae in 35 (36.8%) patients followed by Neisseria meningitides in 30 (31.5%) patients . Approximately 68% of positive cultures yielded S . pneumoniae and N . meningitides (p < 0.0001) . The overall mortality rate was 22.1% . The mortality rate for Streptococcus pneumoniae was 17.1% . The highest mortality was observed in patients with Pseudomonal meningitis where all four patients expired followed by mortality rate of 85.7% in Escherichia coli afflicted patients (p < 0.001) . Complications occurred in 73 (37.6%) patients with persistent complications in 31 (42.4%) patients . Complications resolved in 34 (46.5%) patients . The most common complications were seizures (12.8%) and cranial nerve palsies (11.3%) . Seizures were more likely to occur in older patients (p < 0.05) whereas hydrocephalus was more common in younger patients (p < 0.05) . CONCLUSION: Bacterial Meningitis remains a serious disease associated with substantial morbidity and mortality . Most cases are community acquired with S . Pneumoniae being the most common pathogen . Old age, diabetes mellitus, a positive culture, seizures as a complication and late stage in the disease are the important predictors of a poor outcome.

Mol Microbiol, 2004 Feb, 51(4), 1027 - 37
Structure of the OmpA-like domain of RmpM from Neisseria meningitidis; Grizot S et al.; RmpM is a putative peptidoglycan binding protein from Neisseria meningitidis that has been shown to interact with integral outer membrane proteins such as porins and TonB-dependent transporters . Here we report the 1.9 A crystal structure of the C-terminal domain of RmpM . The 150-residue domain adopts a betaalphabetaalphabetabeta fold, as first identified in Bacillus subtilis chorismate mutase . The C-terminal RmpM domain is homologous to the periplasmic, C-terminal domain of Escherichia coli OmpA; these domains are thought to be responsible for non-covalent interactions with peptidoglycan . From the structure of the OmpA-like domain of RmpM, we suggest a putative peptidoglycan binding site and identify residues that may be essential for binding . Both the crystal structure and solution experiments indicate that RmpM may exist as a dimer . This would promote more efficient peptidoglycan binding, by allowing RmpM to interact simultaneously with two glycan chains through its C-terminal, OmpA-like binding domain, while its (structurally uncharacterized) N-terminal domain could stabilize oligomers of porins and TonB-dependent transporters in the outer membrane.

Rev Fac Cien Med Univ Nac Cordoba, 2003, 60(2), 77 - 81
{Pelvic inflammatory disease associated with Neisseria meningitis bacteremia}; Leoni AF et al.; The Pelvic Inflammatory disease (PID) caused by Neisseria meningitidis (Nm), as an extraneurological manifestation considered as a case study with few precedents . It is so far known that the occasional transposition of ecological niches occurs . There are several bibliographical reports that show an increase in the colonization of the genitourinary tract due to Nm . These may or may not cause disease in individuals during colonization . In adults, the symptoms and any complications which may arise are similar to those caused by Neisseria gonorrhoeae (Ng) . It is highly necessary to correctly identify the agents of Neisseria gender which are obtained from the genital tract . This permits an accurate diagnosis to be made and results in a better understanding of the epidemiology . Morbimortality arising from Nm infection is worth studying to produce a better knowledge of the pathogenia of the infection caused by this microorganism.

J Bacteriol, 2004 Feb, 186(4), 1038 - 49
The lgtABCDE gene cluster, involved in lipooligosaccharide biosynthesis in Neisseria gonorrhoeae, contains multiple promoter sequences; Braun DC et al.; Biosynthesis of the variable core domain of lipooligosaccharide (LOS) in Neisseria gonorrhoeae is mediated by glycosyl transferases encoded by lgtABCDE . Changes within homopolymeric runs within lgtA, lgtC, and lgtD affect the expression state of these genes, with the nature of the LOS expressed determined by the functionality of these genes . However, the mechanism for modulating the amount of multiple LOS chemotypes expressed in a single cell is not understood . Using mutants containing polar disruptions within the lgtABCDE locus, we determined that the expression of this locus is mediated by multiple promoters and that disruption of transcription from these promoters alters the relative levels of simultaneously expressed LOS chemotypes . Expression of the lgtABCDE locus was quantified by using xylE transcriptional fusions, and the data indicate that this locus is transcribed in trace amounts and that subtle changes in transcription result in phenotypic changes . By using rapid amplification of 5' cDNA ends, transcriptional start sites and promoter sequences were identified within lgtABCDE . Most of these promoters possessed 50 to 67% homology with the consensus gearbox promoter sequence of Escherichia coli.

Clin Microbiol Infect, 2004 Feb, 10(2), 137 - 42
Carriage of Neisseria meningitidis by Greek children: risk factors and strain characteristics; Pavlopoulou ID et al.; Oropharyngeal swabs were cultured from 554 children aged 2-19 years attending nurseries, primary schools and secondary schools in the central Athens area . A questionnaire was completed to identify risk factors for carriage . Susceptibility to antimicrobial agents was determined by Etest . The genetic relatedness of the strains was examined by pulsed-field gel electrophoresis (PFGE), and isolate serogrouping was performed by slide agglutination . Twenty-two (4%) children were carriers of Neisseria meningitidis; seven isolates belonged to serogroup C, and five to serogroup B . One isolate was resistant to co-trimoxazole, and five showed intermediate resistance to penicillin . DNA analysis of 16 isolates revealed six distinct PFGE patterns . Clusters with indistinguishable PFGE patterns were noted in the same school . More than one serogroup was included in the same clonal group . On multivariate logistic regression analysis, only age > 12 years remained independently associated with the carrier state (odds ratio, 7.96; 95% CI, 2.24-28.33; p < 0.001) . Overall, the N . meningitidis carriage rate among Greek schoolchildren increased with age, and the predominant serogroups in the Athens region were groups C and B . These findings may have important implications for future immunisation strategies with conjugate vaccines.

Sex Transm Infect, 2004 Feb, 80(1), 68 - 71
Evaluation of real time polymerase chain reaction assays for confirmation of Neisseria gonorrhoeae in clinical samples tested positive in the Roche Cobas Amplicor assay; Tabrizi SN et al.; OBJECTIVE: Development of a rapid, sensitive, and accurate assay for confirmation of Neisseria gonorrhoeae in clinical samples . METHOD: Two real time polymerase chain reaction (PCR) assays, developed on the LightCycler for amplification of the N gonorrhoeae cppB gene, were utilised for confirmation of this bacterial pathogen in samples positive by the Roche Cobas Amplicor assay . Performance characteristics of the two assays were compared with other commercial nucleic acid amplification assays, including the Abbott LCx and Roche 16S rRNA tests . RESULTS: All related Neisseria as well as other bacterial species tested negative by both cppB gene based assays, whereas 120 N gonorrhoeae clinical isolates from various geographical regions gave in positive results . Both assays had a sensitivity of one copy per reaction . 122 clinical samples positive and another 50 samples negative for N gonorrhoeae by Roche Cobas Amplicor were selected from a specimen pool of more than 3000 women tested previously . Overall, 73 of 122 (59.8%) samples were confirmed as positive . The two real time assays had sensitivities of 99% and 100% and specificities of 98% and 100%, respectively . The 16S and LCx assays produced similar results to the real time assays, indicating a similar sensitivity to and specificity of both real time assays . CONCLUSION: The data from this study highlight the need to confirm N gonorrhoeae positive Cobas Amplicor PCR results as an important part of the testing algorithm of all diagnostic laboratories utilising this assay.

Sex Transm Infect, 2004 Feb, 80(1), 35 - 42
The sexual health of pupils in years 4 to 6 of primary schools in rural Tanzania; Todd J et al.; BACKGROUND/OBJECTIVES: There is an urgent need for effective interventions to improve the sexual and reproductive health of adolescents . Reliable data on the sexual health of adolescents are needed to guide the development of such interventions . The aim was to describe the sexual health of pupils in years 4 to 6 of 121 rural primary schools in north western Tanzania, before the implementation of an innovative sexual health intervention in 58 of the schools . METHODS: A cross sectional survey of primary school pupils in rural Tanzania was carried out . The study population comprised pupils registered in years 4 to 6 of 121 primary schools in 20 rural communities in 1998 . Basic demographic information was collected from all pupils seen . Those born before 1 January 1985 (aged approximately 14 years and over) were invited to participate in the survey, and asked about their knowledge and attitudes towards sexual health issues, and their sexual experience . A urine specimen was requested and tested for HIV, Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and, for females, pregnancy . RESULTS: 9283 pupils born before 1 January 1985 were enrolled and provided demographic information and a urine sample . Male pupils were significantly older than females (mean age 15.5 years v 14.8 years, p<0.001), but all other demographic characteristics were similar between the sexes . 14 (0.2%) of the enrolled pupils (four male and 10 female) were HIV positive, 83 (0.9%) were positive for CT, and 12 (0.1%) for NG . 32 female pupils (0.8%) were positive by pregnancy test . Sexual experience was reported by one fifth of primary school girls, and by almost half of boys . Only 45/114 (39%) girls with biological markers of sexual activity reported having had sex . CONCLUSIONS: HIV, CT, NG, and pregnancy were present though at relatively low levels among pupils in years 4 to 6 of primary school . A high proportion of pupils with a biological marker of sexual activity denied ever having had sex . Alternative ways of collecting sensitive data about the sexual behaviour of school pupils should be explored.

Lett Appl Microbiol, 2004, 38(2), 87 - 92
Chemical composition and microbicidal activity of extracts from Brazilian and Bulgarian propolis; Salomao K et al.; AIMS: The chemical composition of ethanol extracts from a Brazilian (Et-Bra) and a Bulgarian (Et-Blg) propolis, and their activity against the protozoan Trypanosoma cruzi, several fungi and bacteria species were determined . METHODS AND RESULTS: The chemical composition was determined by high temperature high resolution gas chromatography coupled to mass spectrometry . Microbiological activity was assayed in vitro against T . cruzi, Candida albicans, Sporothrix schenckii, Paracoccidioides brasiliensis, Neisseria meningitidis, Streptococcus pneumoniae and Staphylococcus aureus . CONCLUSIONS: Et-Bra and Et-Blg, although with totally distinct compositions, were active against T . cruzi and the three species of fungi . Et-Blg was more effective than Et-Bra against bacteria, particularly N . meningitidis and Strep . pneumoniae . SIGNIFICANCE AND IMPACT OF THE STUDY: Although with different classes of components, both propolis extracts showed microbicidal activity . For the bactericidal activity it was possible to establish a positive correlation with the high content of flavonoids of the Bulgarian extract.

J Infect Dis, 2004 Feb 1, 189(3), 410 - 9 Epub 2004 Jan 20.
Inhibition of Neisseria gonorrhoeae genital tract infection by leading-candidate topical microbicides in a mouse model; Spencer SE et al.; The development of effective vaginal microbicides is paramount in the fight against the spread of sexually transmitted infections . Preclinical testing of candidate microbicides for the prevention of gonorrhea has been seriously hindered by the lack of an animal model . We assessed the efficacy of 7 promising formulated agents--CarraGuard, Ushercell, {poly}sodium 4-styrene sulfonate (T-PSS), PRO 2000, ACIDFORM, cellulose acetate phthalate (CAP), and BufferGel--by use of a mouse model of Neisseria gonorrhoeae genital tract infection . Mice received test agent, relevant placebo, or no treatment, followed by intravaginal N . gonorrhoeae challenge . N . gonorrhoeae colonization was tested by vaginal culture . CarraGuard, Ushercell, and T-PSS demonstrated significant protection, compared with control agents and no treatment . PRO 2000, ACIDFORM, and CAP showed significant protection, compared with no treatment but not compared with respective control agents . Mice that received BufferGel were provided significant protection, compared with untreated control mice; no placebo was tested . The findings of the present study suggest that topical agents may effectively reduce N . gonorrhoeae infection and that further evaluation is warranted.

J Infect Dis, 2004 Feb 1, 189(3), 402 - 9 Epub 2004 Jan 20.
Neisseria meningitidis undergoes PilC phase variation and PilE sequence variation during invasive disease; Rytkonen A et al.; Neisseria meningitidis colonizes the upper respiratory tract (URT), enters the blood stream, and reaches the cerebrospinal fluid (CSF) . In the present study, we show that bacteria isolated from the URT adhere better to human epithelial cells, compared with bacteria from blood or CSF, which suggests that important changes of virulence-associated proteins take place during bacterial dissemination . Phase variation in the pilus adhesin PilC and sequence variation in the pilus subunit PilE occurred among strains from 1 patient . Changes were not found in the invasion-associated opacity proteins or in lipooligosaccharides . PilC was frequently expressed in serogroup B strains and in URT strains but was often switched off in other serogroups and in CSF strains . Strains lacking PilC showed impaired adhesion to epithelial cells . These data argue that N . meningitidis undergoes PilC phase variation and PilE sequence variation during invasive disease.

Sex Transm Dis, 2004 Feb, 31(2), 113 - 6
Rescreening for gonorrhea and chlamydial infection through the mail: a randomized trial; Sparks R et al.; BACKGROUND: Rescreening patients after treatment of Chlamydia trachomatis or Neisseria gonorrhoeae infection has had high yield but low rates of participation . GOAL: The goal of this study was to determine if rescreening for gonorrhea and chlamydial infection in a largely urban sexually transmitted disease population would be more successful if individuals were given the option of submitting a specimen for testing through the mail . STUDY DESIGN: We conducted a randomized clinical trial involving 122 patients of whom 62 were assigned to clinic rescreening and 60 were given the option of either mailing a specimen for testing or going to a clinic for rescreening . RESULTS: Twenty-seven patients (45%) given the option of either rescreening in the clinic or through the mail and 20 (32%) assigned to clinic rescreening were rescreened within 28 days of enrollment in the study (odds ratio, 1.7; 95% confidence interval, 0.8-3.8) . Of the 60 patients randomized to the clinic rescreening or mailing option, 11 of 18 (61%) who opted to mail in a specimen and 16 of 42 (38%) who chose clinic rescreening were rescreened within 28 days of enrollment (P = 0.10) . CONCLUSIONS: Although not statistically significant, this study indicates that mailed rescreening could be a successful method to increase rescreening rates.

Sex Transm Dis, 2004 Feb, 31(2), 73 - 8
The emergence of Neisseria gonorrhoeae with decreased susceptibility to Azithromycin in Kansas City, Missouri, 1999 to 2000; McLean CA et al.; BACKGROUND AND OBJECTIVES: We describe the first cluster of persons with Neisseria gonorrhoeae with decreased susceptibility to azithromycin (AziDS; minimum inhibitory concentration >/=1.0 microg/mL) in the United States.GOAL The goal of this study was to identify risk factors for AziDS N . gonorrhoeae and to describe isolate microbiology . STUDY DESIGN: Persons with AziDS N . gonorrhoeae (cases) were identified in Kansas City, Missouri, through the Gonococcal Isolate Surveillance Project (GISP) in 1999 and expanded surveillance, January 2000 to June 2001 . A case-control study using 1999 GISP participants was conducted; control subjects had azithromycin-susceptible N . gonorrhoeae . RESULTS: Thirty-three persons with AziDS N . gonorrhoeae were identified . Case patients were older than control patients (median age, 33 years vs . 23 years; P <0.001) . Fifty percent of cases and 13% of control subjects had a history of sex with a female commercial sex worker (odds ratio, 7.0; 95% confidence interval, 1.3-36.0); 50% of cases and 4% of control subjects met sex partners on street A (P <0.01) . AziDS N . gonorrhoeae isolates were phenotypically and genotypically similar and contained an mtrR gene mutation . CONCLUSIONS: With few treatment options remaining, surveillance for antimicrobial-resistant N . gonorrhoeae is increasingly important, especially among persons at high risk.

Vaccine, 2004 Jan 26, 22(5-6), 660 - 9
DNA immunization of mice with a plasmid encoding Neisseria gonorrhea PorB protein by intramuscular injection and epidermal particle bombardment; Zhu W et al.; Immunogenicity of a DNA vaccine encoding PorB from Neisseria gonorrhoeae strain FA1090 was analyzed in BALB/C mice immunized by intramuscular needle injection or epidermal gene gun bombardment . Both delivery routes generated measurable specific antibodies although the gene gun response was slower . Antibody isotypes were indicative of Th2 activation following gene gun immunization and of Th1 activation following intramuscular injection . In both immunization protocols, boosting with either renatured recombinant (rr) PorB protein or PorB expressed from Venezuelan equine encephalitis virus replicon particles (VRPs) significantly increased anti-PorB antibody levels . Boosting with rrPorB protein had little effect on antibody isotypes, while boosting with VRPs expressing PorB-enhanced a Th1 type response . Whole cell binding experiments showed that a portion of the antibodies recognized the surface of the homologous N . gonorrhoeae strain . Serum from groups with high antibody levels showed some opsonization of the homologous strain using human neutrophils . These results showed the potential of DNA vaccination for the purpose of priming an antibody response against PorB of N . gonorrhoeae . When combined with a protein or VRP boost, DNA priming resulted in high-titer and long-lasting responses . Based on different prime-boost protocols, we could polarize immune responses to predominantly Th1 or Th2, which should enable future studies of the types of immune responses that are protective in mouse models of gonorrhea.

J Endotoxin Res, 2003, 9(6), 349 - 60
Effects of bacterial cell wall components (PAMPs) on the expression of monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP-1alpha) and the chemokine receptor CCR2 by purified human blood monocytes; Moller AS et al.; Regulation of chemokine production and the expression of chemokine receptors play an important role during inflammation and infectious diseases . The present study was designed to study the effects of five different bacterial cell wall components (PAMPs) on the production of MCP-1 and MIP-1alpha and the expression of CCR2 by highly purified human blood monocytes . All five PAMPs induced high expression of mRNA and protein synthesis of both chemokines . Generally, MCP-1 mRNA and protein levels were higher than MIP-1alpha levels . Expression of MCP-1 and MIP-1alpha differed both at the mRNA and at the protein levels, MIP-1alpha always showing a more rapid initial increase, attaining lower protein levels than MCP-1 . Antibodies against CD14 significantly inhibited the inducing effects of all the PAMPs used . Antibody against TLR2 inhibited the chemokine production induced by LTA and AraLAM by more than 36% (P < 0.05) while chemokine production induced by Escherichia coli-LPS, purified E . coli-LPS and Neisseria meningitidis-LPS was inhibited by more than 60% by antibody against TLR4 (P < 0.05) . The inducing effects of all five PAMPs could be inhibited by rIL-4, rIL-10 and rIL-13 . rIL-4 was the most effective . Generally, IC(50) of these anti-inflammatory cytokines were lower for the MIP-1alpha than for the MCP-1 production . The cell surface expression of CCR2 was significantly down-regulated by all five PAMPs in addition to a decrease in cytosolic free calcium and binding of rMCP-1 . We conclude that MCP-1 and MIP-1alpha as well as the MCP-1 receptor CCR2 will be substantially regulated upon monocyte contact with various cell wall components (PAMPs) from Gram-negative and Gram-positive bacteria as well as from mycobacteria.

Biochemistry, 2004 Jan 27, 43(3), 710 - 7
Mechanistic and kinetic study of the ATP-dependent DNA ligase of Neisseria meningitidis; Magnet S et al.; The gene from Neisseria meningitidis serogroup A, encoding a putative, secreted ATP-dependent DNA ligase was cloned and overexpressed, and the soluble protein was purified . Mass spectrometry indicated that the homogeneous protein was adenylated as isolated, and sedimentation velocity experiments suggested that the enzyme exists as a monomer in solution . The 31.5 kDa protein can catalyze the ATP-dependent ligation of a singly nicked DNA duplex but not blunt-end joining . The first step of the overall reaction, the ATP-dependent formation of an adenylated ligase, was studied by measuring the formation of the covalent intermediate and isotope exchange between {alpha-32P} ATP and PPi . Mg2+ was absolutely required for this reaction and was the best divalent cation to promote catalysis . Electrophoretic gel mobility shift assays revealed that the enzyme bound both unnicked and singly nicked double stranded DNA with equivalent affinity (Kd approximately 50 nM) but cannot bind single stranded DNA . Preadenylated DNA was synthesized by transferring the AMP group from the enzyme to the 5'-phosphate of a 3'-dideoxy nicked DNA . The rate of phosphodiester bond formation at the preadenylated nick was also Mg(2+)-dependent . Kinetic data showed that the overall rate of ligation, which occurred at 0.008 s(-1), is the result of three chemical steps with similar rate constants (approximately 0.025 s(-1)) . The Km values for ATP and DNA substrates, in the overall ligation reaction, were 0.4 microM and 30 nM, respectively.

J Bacteriol, 2004 Feb, 186(3), 850 - 7
Transferrin-binding protein B of Neisseria meningitidis: sequence-based identification of the transferrin-Binding site confirmed by site-directed mutagenesis; Renauld-Mongenie G et al.; A sequence-based prediction method was employed to identify three ligand-binding domains in transferrin-binding protein B (TbpB) of Neisseria meningitidis strain B16B6 . Site-directed mutagenesis of residues located in these domains has led to the identification of two domains, amino acids 53 to 57 and 240 to 245, which are involved in binding to human transferrin (htf) . These two domains are conserved in an alignment of different TbpB sequences from N . meningitidis and Neisseria gonorrhoeae, indicating a general functional role of the domains . Western blot analysis and BIAcore and isothermal titration calorimetry experiments demonstrated that site-directed mutations in both binding domains led to a decrease or abolition of htf binding . Analysis of mutated proteins by circular dichroism did not provide any evidence for structural alterations due to the amino acid replacements . The TbpB mutant R243N was devoid of any htf-binding activity, and antibodies elicited by the mutant showed strong bactericidal activity against the homologous strain, as well as against several heterologous tbpB isotype I strains.

J Bacteriol, 2004 Feb, 186(3), 730 - 9
A mutant form of the Neisseria gonorrhoeae pilus secretin protein PilQ allows increased entry of heme and antimicrobial compounds; Chen CJ et al.; A spontaneous point mutation in pilQ (pilQ1) resulted in phenotypic suppression of a hemoglobin (Hb) receptor mutant (hpuAB mutant), allowing gonococci to grow on Hb as the sole source of iron . PilQ, formerly designated OMP-MC, is a member of the secretin family of proteins located in the outer membrane and is required for pilus biogenesis . The pilQ1 mutant also showed decreased piliation and transformation efficiency . Insertional inactivation of pilQ1 resulted in the loss of the Hb utilization phenotype and decreased entry of free heme . Despite the ability of the pilQ1 mutant to use Hb for iron acquisition and porphyrin, there was no demonstrable binding of Hb to the cell surface . The pilQ1 mutant was more sensitive to the toxic effect of free heme in growth medium and hypersensitive to the detergent Triton X-100 and multiple antibiotics . Double mutation in pilQ1 and tonB had no effect on these phenotypes, but a double pilQ1 pilT mutant showed a reduction in Hb-dependent growth and decreased sensitivity to heme and various antimicrobial agents . Insertional inactivation of wild-type pilQ also resulted in reduced entry of heme, Triton X-100, and some antibiotics . These results show that PilQ forms a channel that allows entry of heme and certain antimicrobial compounds and that a gain-of function point mutation in pilQ results in TonB-independent, PilT-dependent increase of entry.

Saudi Med J, 2003 Dec, 24(12), 1300 - 7
Disease burden and case management of bacterial meningitis among children under 5 years of age in Saudi Arabia; Al-Mazrou YY et al.; OBJECTIVE: This study was designed to explore the case management and disease burden of bacterial meningitis among children below the age of 5 years in the Kingdom of Saudi Arabia . METHODS: A prospective descriptive study was conducted during June 1999 through to May 2001 in 8 hospitals from 5 cities in different areas of the country . Those were, King Fahad Specialist Hospital and Maternity & Children Hospital, Buraidah, Belgorashi General Hospital and King Fahad Specialist Hospital, Al-Baha, Maternity & Children Hospital and Ohud Hospital, Al-Madina, Al-Qatif General Hospital in the Eastern Region and Prince Abdulrahman Bin Ahmed Alsudairy Central Hospital, Sakaka . The study population was 171,818 children under the age of 5 years . RESULTS: During the study period 208 cases of bacterial meningitis were identified, 141 (67.8%) with a definite causative organism: Hemophilus influenzae type b, Neisseria meningitidis, Streptococcus pneumoniae and other bacterial species . The remaining 67 cases (32%) were labeled as aseptic meningitis . A considerable proportion of cases was found to have received an antibiotic prior to presentation . While symptoms such as fever or poor feeding were common among cases, meningeal signs were less prominent . A lumbar puncture was carried out on all cases to reach diagnosis by gram stain, latex agglutination test and cerebrospinal fluid (CSF) and blood cultures following standard procedures . The immediate burden of meningitis cases was found to be the lengthy stay of patients in the hospital wards and intensive care units . Some of the main causative agents were resistant to the conventional antimicrobial therapy, but susceptible to newer antibiotics . CONCLUSION: The study being based on a population based surveillance gave a better overview on causative organisms of meningitis emphasizing that Gram stain, serology of CSF and culture (of CSF and blood) should be used . A high index of suspicion is needed to diagnose meningitis in children . Lumbar tap should be encouraged and supported in terms of training and more authorization to apply in diagnostic tests of such conditions . Audiometric measurement is a crucial need in the assessment of meningitis cases and in the process of their follow up . This type of service is clearly missing in our system . Influential steps are to be planned to avail this service.

Med J Aust, 2004 Jan 5, 180(1), 40 - 1
An unusual cause of an epidural abscess; van Hal SJ et al.; A previously well 30-year-old man presented with severe progressive back pain, joint pain and fever . Magnetic resonance imaging confirmed an epidural abscess . A sexual history revealed both risk factors for and previous symptoms of a sexually acquired infection . Neisseria gonorrhoeae was isolated from a rectal swab and from a wrist aspirate, consistent with disseminated gonococcal infection . The epidural abscess resolved clinically and radiologically after treatment for N . gonorrhoeae with ceftriaxone.

J Pharm Sci, 2004 Feb, 93(2), 273 - 82
Anionic microparticles are a potent delivery system for recombinant antigens from Neisseria meningitidis serotype B; Singh M et al.; The adsorption behavior of model proteins onto anionic poly(lactide-co-glycolide) (PLG) microparticles was evaluated . PLG microparticles were prepared by a w/o/w solvent evaporation process in the presence of the anionic surfactant dioctyl sodium sulfosuccinate (DSS) . The effect of surfactant concentration and adsorption conditions on the adsorption efficiency and release rates in vitro was also studied . Subsequently, the microparticle formulation was tested to evaluate the efficacy of anionic microparticles as delivery systems for recombinant antigens from Neisseria meningitides type B (Men B), with and without CpG adjuvant . Protein (antigen) binding to anionic PLG microparticles was influenced by both electrostatic interaction and by other mechanisms, including hydrophobic attraction . The Men B antigens adsorbed efficiently onto anionic PLG microparticles and, following immunization in mice, induced potent enzyme-linked immunosorbent assay (ELISA) and serum bactericidal activity in comparison to alum-adsorbed formulations . These Men B antigens represent an attractive approach for vaccine development .

Sex Transm Dis, 2004 Jan, 31(1), 38 - 51
Testing for chlamydial and gonorrheal infections outside of clinic settings: a summary of the literature; Ford CA et al.; BACKGROUND: Nucleic acid amplification tests (NAATs) provide new technology that makes it feasible to initiate testing for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) infection outside of clinic settings . METHODS: We summarized the English-language literature describing chlamydial or gonorrheal testing with self-collection of urine or vaginal specimens outside of clinic settings in developed countries published between January 1995 and August 2002 . RESULTS: Testing for CT or GC infection has been initiated in school, community, and home settings . Purposes include screening of asymptomatic populations, improving quality of clinic-based health care, and research . Challenges include defining and reaching target populations, overcoming logistic issues, developing communication and counseling strategies, and determining whether alternative testing strategies are effectively reducing infection rates . CONCLUSIONS: The use of NAATs to detect CT and GC infection outside of clinic settings will undoubtedly continue . Future research should focus on how to best use this technology to reduce rates of infection.

Biophys J, 2004 Jan, 86(1 Pt 1), 3 - 9
Water accessibility, aggregation, and motional features of polysaccharide-protein conjugate vaccines; Berti F et al.; A relaxometric investigation of a nontoxic mutant of diphtheria toxin and of its conjugates with capsular polysaccharides of different groups of Neisseria meningitidis was performed . The insertion of polysaccharides chains alters dramatically the hydrodynamic properties of the protein . The model-free analysis of the (1)H nuclear magnetic relaxation dispersion profiles of their water solutions shows: i), a reduced protein hydration with respect to the carrier protein alone; ii), a much larger flexibility of the conjugates with respect to a compact macromolecule of the same molecular weight; and iii), a strong tendency to aggregate . The above findings are largely independent on the nature of the polysaccharide and thus provide a fairly general picture of the dynamic properties of glycoconjugate proteins.

J Infect Chemother, 2003 Dec, 9(4), 310 - 3
Frequency of alterations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV in 19 clinical isolates of Neisseria gonorrhoeae in Tokyo in 2002; Abe H et al.; The frequency of alterations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV in 19 clinical isolates of Neisseria gonorrhoeae obtained in Tokyo in 2002 was studied . The frequencies of GyrA and ParC mutations in these 19 isolates were 100% (19 of 19) and 84.2% (16 of 19), respectively, and these results were 1.48-fold (100%/67.6%) and 3.58-fold (84.2%/23.5%) higher, respectively, than the frequencies reported in 1998 in 68 isolates obtained in Fukuoka during the period from 1992 to 1996 . Isolates with increasing numbers of mutations were more resistant not only to levofloxacin but also to other antibiotics . The 50% and 90% minimum inhibitory concentrations (MICs) to levofloxacin during the period from 1995 to 1996 were 0.063 and 1 micro g/ml, and they increased to 4 and 8 micro g/ml, respectively, in the present study . All 19 cases of gonoccocal urethritis in the present study were cured with a single intramuscular injection of 2 g spectinomycin.

Eur J Biochem, 2004 Jan, 271(1), 23 - 32
Overexpression and enzymatic characterization of Neisseria gonorrhoeae penicillin-binding protein 4; Stefanova ME et al.; The penicillin-binding proteins (PBPs) are ubiquitous bacterial enzymes involved in cell wall biosynthesis, and are the targets of the beta-lactam antibiotics . The low molecular mass Neisseria gonorrhoeae PBP 4 (NG PBP 4) is the fourth PBP revealed in the gonococcal genome . NG PBP 4 was cloned, overexpressed, purified, and characterized for beta-lactam binding, DD-carboxypeptidase activity, acyl-donor substrate specificity, transpeptidase activity, inhibition by a number of active site directed reagents, and pH profile . NG PBP 4 was efficiently acylated by penicillin (30,000 m-1.s-1) . Against a set of five alpha- and epsilon-substituted l-Lys-D-Ala-D-Ala substrates, NG PBP 4 exhibited wide variation in specificity with a preference for N epsilon-acylated substrates, suggesting a possible preference for crosslinked pentapeptide substrates in the cell wall . Substrates with an N epsilon-Cbz group demonstrated pronounced substrate inhibition . NG PBP 4 showed 30-fold higher activity against the depsipeptide Lac-ester substrate than against the analogous peptide substrate, an indication that k2 (acylation) is rate determining for carboxypeptidase activity . No transpeptidase activity was apparent in a model transpeptidase reaction . Among a number of active site-directed agents, N-chlorosuccinimide, elastinal, iodoacetamide, iodoacetic acid, and phenylglyoxal gave substantial inhibition, and methyl boronic acid gave modest inhibition . The pH profile for activity against Ac2-l-Lys-D-Ala-d-Ala (kcat/Km) was bell-shaped, with pKa values at 6.9 and 10.1 . Comparison of the enzymatic properties of NG PBP 4 with other DD-carboxypeptidases highlights both similarities and differences within these enzymes, and suggests the possibility of common mechanistic roles for the two highly conserved active site lysines in Class A and C low molecular mass PBPs.

Rinsho Byori, 2003 Nov, 51(11), 1061 - 7
{Evaluation of the simultaneous detection system for Chlamydia trachomatis/Neisseria gonorrhoeae DNA by the isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN)}; Shimada M et al.; The isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) is a new isothermal DNA amplification method composed of exo Bca DNA polymerase, RNaseH and DNA-RNA chimeric primers . We developed the simultaneous detection system for Chlamydia trachomatis/Neisseria gonorrhoeae DNA, combined with luminescence detection by a probe hybridization . In the performance tests, this system was able to detect 10 to 100 copies of C . trachomatis/N . gonorrhoeae DNA for only 3.5 hours, and was highly specific to C . trachomatis/N . gonorrhoeae without any cross-reaction to C . pneumoniae, N . lactamica, N . sicca or N . meningitidis . When we tested 60 clinical samples of urine and cervical swabs, the interpretive results were completely consistent with those obtained by Roche PCR system . Of 13 positive samples by the ICAN and PCR systems for C . trachomatis, four were negative by EIA method(IDEIA Chlamydia) . These results indicate that the ICAN system is an efficient and sensitive system to simultaneously detect C . trachomatis/N . gonorrhoeae DNA.

Int J STD AIDS, 2003 Dec, 14(12), 830 - 4
'You can't tell by looking': pilot study of a community-based intervention to detect asymptomatic sexually transmitted infections; Low N et al.; Innovative and non-stigmatizing interventions are required to reduce ethnic inequalities in rates of sexually transmitted infections among young people . We therefore designed an intervention, 'You can't tell by looking,' which combined health promotion with testing for gonorrhoea and chlamydia using nucleic acid amplification technology and treatment and partner notification delivered in the non-clinical settings . One hundred and eighty-one participants were seen in 13 sessions in local further education colleges . Forty-three percent of participants were from Black Caribbean or Black Other ethnic groups and 39% were Black African: 125 of 181 participants were sexually active and 109 of these (87%) provided a urine specimen . 10/109 (9.2%, 95% confidence interval 4.5-16.2%) samples were confirmed positive for Chlamydia trachomatis and two were also positive for Neisseria gonorrhoeae . Only 7% of those tested found it embarrassing . The intervention was both feasible and acceptable to young people . It could be tested in a wider variety of non-clinical settings and evaluated in a cluster randomized trial.

Afr J Reprod Health, 2003 Aug, 7(2), 100 - 5
Antibiogram and plasmid profiles of Neisseria gonorrhoeae isolates from Cameroon: useful tools for epidemiological survey; Ndip RN et al.; A prospective laboratory-based investigative study was carried out on clinical isolates of N . gonorrhoea to determine their antibiotic susceptibility patterns and plasmid profile using standard microbiological and molecular techniques . All the 32 isolates studied showed total resistance to penicillin, spectinomycin and amoxyclinE . On the other hand, susceptibilities of 100%, 98.6% and 98.6% were noted for ciprofloxacin, ofloxacin and norfloxacin respectively . Thirty (93.8%) of the 32 isolates were found to harbour plasmids of molecular weights ranging from 9.2 to 25.2 Mdal . Three distinct groups of N . gonorrhoea isolates were identified based on the molecular weights of the plasmids, namely, group one (9.2 Mdal), group two (12.6 Mdal) and group three (25.2 Mdal) . These results suggest that different strains of N . gonorrhoea may be circulating in Fako Division of Cameroon, a finding that is of clinical and epidemiological significance.

Obstet Gynecol, 2003 Nov, 102(5 Pt 1), 962 - 9
Histologic endometritis in asymptomatic human immunodeficiency virus-infected women: characterization and effect of antimicrobial therapy; Eckert LO et al.; OBJECTIVE: To estimate the prevalence, risk factors, clinical symptoms and signs, and response to antimicrobial therapy of histologic endometritis in human immunodeficiency virus (HIV)-infected women without clinical salpingitis . METHODS: This was a cross-sectional study of 42 HIV-infected women enrolled from a single clinic . Subjects underwent standardized history, examination, and laboratory determinations, including endometrial biopsy . Women with suspected pelvic inflammatory disease were excluded . All women were given antibiotics and repeat evaluation in 5-7 weeks . Histologic endometritis was defined by at least one stromal plasma cell per 120x field and five or more surface polymorphonuclear leukocytes per 400x field . Chi-square and Fisher exact tests were used as appropriate . RESULTS: Histologic endometritis was present among 16 (38%) of 42 evaluable HIV-infected women, none of whom had Chlamydia trachomatis or Neisseria gonorrhoeae . Douching three or more times per month, history of ectopic pregnancy, and two or more prior urinary tract infections were associated with endometritis, as was elevated erythrocyte sedimentation rate (P < or = .05) . Physical examination findings and mean CD4+ lymphocyte count were similar among those with and without endometritis . In the nine HIV-infected women with a repeat biopsy, endometritis decreased from four (44%) to two (22%) after treatment (P = .30) . CONCLUSION: The prevalence of histologic endometritis in HIV-infected women was high despite few examination findings and no demonstrated pathogens . Endometritis in HIV-infected women might be related to pathogens not evaluated, to prior infection, or to reduced immunity from HIV.

Sex Transm Infect, 2003 Dec, 79(6), 484 - 6
Evaluation of ligase chain reaction for the non-cultural detection of rectal and pharyngeal gonorrhoea in men who have sex with men; Young H et al.; OBJECTIVES: To compare a nucleic acid amplification test (ligase chain reaction) with culture for detecting rectal and pharyngeal gonorrhoea in men who have sex with men (MSM) . METHODS: Duplicate rectal and throat swabs from MSM attending a genitourinary medicine clinic were collected for culture on modified New York City medium and detection of gonococcal nucleic acid by the Abbott ligase chain reaction (LCR) utilising probes based on opa 1 gene sequences . LCR positive culture negative specimens were tested by a second LCR utilising probes based on pilin gene sequences . Patients with rectal and/or pharyngeal cultures yielding Gram negative diplococci confirmed as Neisseria gonorrhoeae by biochemical and immunological methods were diagnosed with rectal and/or pharyngeal gonorrhoea . The criteria for diagnosing rectal and pharyngeal infection by LCR included a positive opa LCR with a positive culture from the same site or, in the case of a negative culture, a positive opa LCR and a positive pilin LCR . RESULTS: Duplicate rectal samples were obtained from 227 MSM . The results of LCR and culture were concordant in 219 samples (96.5%) . The prevalence of rectal gonorrhoea by LCR and culture was 7.0% (16/227) and 4.0% (9/227), respectively . Duplicate throat samples were obtained from 251 MSM . The results of LCR and culture were concordant in 230 (91.6%) cases . The prevalence of pharyngeal gonorrhoea by LCR and culture was 12.7% (32/251) and 6.0% (15/251), respectively . The specificity of LCR was 99.5% (210/211) for rectal and 98.2% (215/219) for pharyngeal specimens . CONCLUSIONS: The high prevalence and asymptomatic nature of pharyngeal and rectal gonococcal infection suggests that routine screening for infection at these sites by a nucleic acid amplification test method such as LCR should be considered as part of the overall strategy to control gonorrhoea in MSM.

Sex Transm Infect, 2003 Dec, 79(6), 469 - 73
High postpartum rates of sexually transmitted infections among teens: pregnancy as a window of opportunity for prevention; Ickovics JR et al.; OBJECTIVES: To identify incidence and predictors of Chlamydia trachomatis and Neisseria gonorrhoeae among postpartum adolescents . These estimates are compared to similar estimates among a cohort of non-pregnant, sexually active teens . METHODS: 203 pregnant and 208 non-pregnant adolescents aged 14-19 years were recruited from 10 community based health clinics in Connecticut, United States . Structured interviews and sexually transmitted infection (STI) testing using ligase chain reaction (LCR) were conducted at a baseline visit (during the third trimester for the pregnant adolescents), and at 6 and 12 month follow up visits (3 and 9 months post partum, for those pregnant at baseline) . RESULTS: Among pregnant teens, new infections of C trachomatis and N gonorrhoeae increased from 7.1% at the 6 month follow up interview to 14.3% at the 12 month follow up interview; among non-pregnant teens, new infections remained relatively stable over the 6 and 12 month follow up interviews (9.0% to 8.3%) (group by time interaction, p = 0.005) . C trachomatis and N gonorrhoeae prevalence was 1.9 times higher (95% CI: 0.97 to 3.89, p = 0.06) among teens in the late postpartum follow up compared to the non-pregnant teens, controlling for baseline STIs . Predictors of postpartum STIs included having a new partner and number of partners per year of sexual activity . CONCLUSIONS: Postpartum adolescents are vulnerable to STIs . Routine prenatal and postpartum care provide unique opportunities to promote condom use and other risk reduction interventions among adolescents . If sustained post partum, long term reproductive health can be promoted.

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