Neisseria

Sex Transm Dis, 2002 Jan, 29(1), 25 - 31
Pulsed-field gel electrophoresis as an epidemiologic tool for Neisseria gonorrhoeae: identification of clusters within serovars; Unemo M et al.; BACKGROUND: The increasing incidence of gonorrhea in Sweden in 1998 was due to mostly domestic cases . Among these, two core groups were identified: homosexual men with serovar IB-2 and young heterosexuals with serovar IB-3 . GOALS: To explore the genetic homogeneity/heterogeneity within the predominant serovars, IB-2 and IB-3, of Neisseria gonorrhoeae in Sweden by pulsed-field gel electrophoresis (PFGE) and to compare these results to epidemiologic information, as well as examine the genetic diversity within and between the 25 other represented serovars of N gonorrhoeae . STUDY DESIGN: By PFGE, 237 N gonorrhoeae isolates were examined, and the results were compared with epidemiologic data for the IB-2 and IB-3 isolates . RESULTS: In 79% of the domestic IB-2 cases involving homosexuals and 66% of the domestic IB-3 cases involving young heterosexuals, the isolates were genetically indistinguishable by PFGE . A high genetic diversity was identified within and between the 27 included serovars . CONCLUSIONS: Examination by means of PFGE indicated that one N gonorrhoeae clone each of the serovars IB-2 and IB-3 created the majority of the two core groups of domestic cases.

J Clin Microbiol, 2002 Jan, 40(1), 281 - 3
Evaluation of a strand displacement amplification assay (BD ProbeTec-SDA) for detection of Neisseria gonorrhoeae in urine specimens; Akduman D et al.; The performance of a strand displacement amplification assay (the BDProbeTec-SDA assay) in detecting Neisseria gonorrhoeae in urine specimens was evaluated . When performed under stringent quality control conditions, the BDProbeTec-SDA assay is a sensitive, specific, and efficient method for the screening of large numbers of noninvasively obtained specimens . Because the predictive value of an assay is a function of the prevalence of the disease, culture confirmation is needed for samples with positive results from populations in which the prevalence of a disease is low or in situations in which false-positive results may have important medical, psychosocial, or medicolegal consequences.

Zhonghua Fu Chan Ke Za Zhi, 2001 Sep, 36(9), 539 - 41
{Pathogens of transmitted disease in the pathogenesis of acute pelvic inflammatory disease}; Zhou B et al.; OBJECTIVE: To study the pathogenesis of pathogens of sexually transmitted disease (STD) in acute pelvic inflammatory disease (PID) . METHODS: Neisseria gonorrhoeae, chlamydia trchomatis, ureaplasma urealyticum were cutured respectively in 130 patients with acute PID . RESULTS: The pathogens of STD had been found in 55 patients . The detective rate of Neisseria gonorrhoeae was 6.9% (9/130), about half of them were mixed infection; the detective rate of chlamydia trachomatis was 4.6% (6/130), all of them were mixed infection; the positive percentage of ureaplasma urealyticum was 37.7% (49/130), one third of them were mixed infection . CONCLUSION: The pathogens of STD might cause acute PID or only play a synergetic role in it.

West Indian Med J, 2001 Sep, 50(3), 198 - 202
Single dose (direct observed) azithromycin therapy for Neisseria gonorrhoeae and Chlamydia trachomatis in STD clinic attenders with genital discharge in Trinidad and Tobago; Swanston WH et al.; The purpose of this study was to determine the prevalence and to assess the efficacy of a single one gram oral dose of azithromycin under direct observed therapy of genital discharge due to Neisseria gonorrhoeae and Chlamydia trachomatis infections in STD clinic attenders in Trinidad and Tobago . All patients with genital discharge and their contacts were given one gram oral dose of azithromycin under direct supervision after collection of urethral and cervical swabs for N gonorrhoeae culture and smear and for C trachomatis antigen detection by ELISA . Clinical and microbiological evaluation was done on those who returned after 7-10 days for follow-up . Of the 735 patients who were enrolled in the study, 319 (43.4%) had N gonorrhoeae and 100 (13.6%) had C trachomatis . Only 151 (36%) of the 419 patients with a pathogenic isolate returned for clinical and microbiological assessment . The remaining 268 (64%) of the 419 patients were lost to follow-up . One hundred and forty-three patients (94.7%) had total abatement of signs and symptoms after taking azithromycin . One patient (0.65%), who had both N gonorrhoeae and C trachomatis, improved clinically with the drug . Seven patients (six with N gonorrhoeae and one with C trachomatis) failed to respond clinically to azithromycin . Microbiological eradication was achieved in 115 (100%) patients who had single infection with N gonorrhoeae and in 23 patients (96%) with C trachomatis infection . Of 12 patients with combined infections, N gonorrhoeae and C trachomatis were eradicated in 10 and 12 patients, respectively, after initial treatment . In two patients with combined infection, N gonorrhoeae continued to be isolated after treatment with azithromycin . A single one gram oral dose of azithromycin under direct supervision is useful in the treatment of uncomplicated genital infection with N gonorrhoeae and C trachomatis in STD clinic attenders in Trinidad.

Res Microbiol, 2001 Nov, 152(9), 781 - 91
Expression of Neisseria gonorrhoeae cell division genes ftsZ, ftsE and minD is influenced by environmental conditions; Ramirez-Arcos S et al.; The activity of the promoter regions of the cell division genes ftsZ, ftsE, minC, minD and minE from Neisseria gonorrhoeae (Ng) was studied under different environmental conditions using lacZ translational fusions . The promoters of the minNg genes have not been previously determined and we identified promoter regions upstream of each gene (minCp, minDp and minEp) . We determined that minDp had the strongest activity . Expression of the promoter regions of ftSZ(Ng) and ftsE(Ng), which we had previously identified, as well as minD(Ng), were then studied under conditions reflecting the environment of the genitourinary tract . These conditions included anaerobiosis, presence of isoleucine or urea (3 mM and 400 mM, respectively) and acidity of pH 6 . Both beta-galactosidase expression and northern blot analysis indicated that all three genes were upregulated under anaerobiosis . The addition of isoleucine as well as media at pH 6 did not have any significant effects on the promoter activity of these genes while the presence of urea significantly decreased ftsZ(Ng) promoter activity . The expression of the minD(Ng) promoter region was analyzed during different growth phases and shown to follow the growth behavior of the culture . By contrast, the ftSZ(Ng) promoter activity continued to rise after the onset of the stationary phase . When gonococcal ftsZ promoter 1, (Pz1) was altered by site-directed mutagenesis, a significant decrease in the expression of ftsZ(Ng) was observed under both aerobic and anaerobic conditions . These data infer that gonococci regulate their cell division in response to different environments.

Carbohydr Res, 2002 Jan 7, 337(1), 11 - 20
Synthesis of alpha-lactosyl-(1-->3)-L-glycero-alpha-D-manno-heptopyranoside, a partial oligosaccharide structure expressed within the lipooligosaccharide produced by Neisseria gonorrhoeae strain 15253; Ishii K et al.; The glycosyl donor, hepta-O-benzyl-beta-lactosyl trichloroacetimidate (4) was prepared by treating hepta-O-benzyl-lactose with trichloroacetonitrile in the presence of potassium carbonate . The acceptor, methyl 2-O-benzyl-4,6-O-benzylidene-7,8-dideoxy-alpha-D-manno-oct-7-enopyranoside (8) was synthesized by hydrolysis of a 3,4-butane diacetal of methyl L-glycero-alpha-D-manno-oct-enopyranoside and subsequent benzylidenation . Glycosidation of the donor 4 with the acceptor 8 in 1,4-dioxane using Me(3)SiOTf as a promoter for 1 h at room temperature gave methyl (2,3,4,6-tetra-O-benzyl-beta-D-galactopyranosyl)-(1-->4)-(2,3,6-tri-O-benzyl-alpha-D-glucopyranosyl)-(1-->3)-2-O-benzyl-4,6-O-benzylidene-7,8-dideoxy-alpha-D-manno-oct-7-enopyranoside (9) as a major product (59%) . The oct-enopyranoside moiety of the trisaccharide 9 was converted to a heptopyranoside (80%) by oxidative cleavage with OsO(4)-NaIO(4) and subsequent reduction . Hydrogenolysis of the resulting trisaccharide and subsequent acetylation gave the peracetate of alpha-lactosyl-(1-->3)-Hep . Deacetylation of the peracetate afforded the title trisaccharide.

Obstet Gynecol, 2001 Dec, 98(6), 1117 - 23
Chlamydial infection and unplanned pregnancy in women with ready access to health care; Thomas AG et al.; OBJECTIVE: To apply urine-based ligase chain reaction for Chlamydia trachomatis (C . trachomatis) and Neisseria gonorrhoeae (N . gonorrhoeae) detection and standard urine-based pregnancy testing for Navy-enlisted women, and to compare the prevalence and epidemiologic correlates of these adverse reproductive outcomes . METHODS: Participants were surveyed and urine was collected for pregnancy testing using standard laboratory methods and detection of C . trachomatis and N . gonorrhoeae infection by ligase chain reaction . Self-administered surveys facilitated collection of demographics, sexual behavior, including contraceptive use, sexual partners, sexually transmitted disease, and pregnancy history . RESULTS: Among 299 of 314 participants, the prevalence of chlamydial infection was 4.7% and of pregnancy was 9.7%, with 48.3% of the pregnancies unplanned . Chlamydia trachomatis infection was univariately associated with having a new sex partner within the last 6 months, more sexual partners, single marital status, condom use, drinking until passing out or vomiting in the past 30 days (alcohol misuse), and current pregnancy . Unplanned pregnancy was univariately associated with young age, single marital status, inconsistent condom use, having a new sex partner within the last 6 months, and more recent sexual partners . Among the pregnant women, four (13.8%) were infected with C . trachomatis . CONCLUSION: The high rates of chlamydial infection and unplanned pregnancy found in this population of employed young women with ready access to health care and health education underscore the challenge of enhancing reproductive health via compliance with effective contraceptive and sexually transmitted disease prevention methods . This is a challenge that remains unmet.

Proc Natl Acad Sci U S A, 2001 Dec 18, 98(26), 15276 - 81
Neisseria gonorrhoeae PilV, a type IV pilus-associated protein essential to human epithelial cell adherence; Winther-Larsen HC et al.; Type IV pili (Tfp) of Neisseria gonorrhoeae, the Gram-negative etiologic agent of gonorrhea, facilitate colonization of the human host . Tfp are assumed to play a key role in the initial adherence to human epithelial cells by virtue of the associated adhesin protein PilC . To examine the structural and functional basis for adherence in more detail, we identified potential genes encoding polypeptides sharing structural similarities to PilE (the Tfp subunit) within the N . gonorrhoeae genome sequence database . We show here that a fiber subunit-like protein, termed PilV, is essential to organelle-associated adherence but dispensable for Tfp biogenesis and other pilus-related phenotypes, including autoagglutination, competence for natural transformation, and twitching motility . The adherence defect in pilV mutants cannot be attributed to reduced levels of piliation, defects in fiber anchoring to the bacterial cell surface, or to unstable pilus expression related to organelle retraction . PilV is expressed at low levels relative to PilE and copurifies with Tfp fibers in a PilC-dependent fashion . Purified Tfp from pilV mutants contain PilC adhesin at reduced levels . Taken together, these data support a model in which PilV functions in adherence by promoting the functional display of PilC in the context of the pilus fiber.

J Biol Chem, 2002 Mar 1, 277(9), 7438 - 46 Epub 2001 Dec 19.
Nuclear factor-kappa B directs carcinoembryonic antigen-related cellular adhesion molecule 1 receptor expression in Neisseria gonorrhoeae-infected epithelial cells; Muenzner P et al.; The human-specific pathogen Neisseria gonorrhoeae expresses opacity-associated (Opa) protein adhesins that bind to various members of the carcinoembryonic antigen-related cellular adhesion molecule (CEACAM) family . In this study, we have analyzed the mechanism underlying N . gonorrhoeae-induced CEACAM up-regulation in epithelial cells . Epithelial cells represent the first barrier for the microbial pathogen . We therefore characterized CEACAM expression in primary human ovarian surface epithelial (HOSE) cells and found that CEACAM1-3 (L, S) and CEACAM1-4 (L, S) splice variants mediate an increased Opa(52)-dependent gonoccocal binding to HOSE cells . Up-regulation of these CEACAM molecules in HOSE cells is a direct process that takes place within 2 h postinfection and depends on close contact between microbial pathogen and HOSE cells . N . gonorrhoeae-triggered CEACAM1 up-regulation involves activation of the transcription factor nuclear factor kappaB (NF-kappaB), which translocates as a p50/p65 heterodimer into the nucleus, and an NF-kappaB-specific inhibitory peptide inhibited CEACAM1-receptor up-regulation in N . gonorrhoeae-infected HOSE cells . Bacterial lipopolysaccharides did not induce NF-kappaB and CEACAM up-regulation, which corresponds to our findings that HOSE cells do not express toll-like receptor 4 . The ability of N . gonorrhoeae to up-regulate its epithelial receptor CEACAM1 through NF-kappaB suggests an important mechanism allowing efficient bacterial colonization during the initial infection process.

J Bacteriol, 2002 Jan, 184(2), 420 - 6
Point mutations in HpuB enable gonococcal HpuA deletion mutants to grow on hemoglobin; Chen CJ et al.; Neisseria gonorrhoeae ordinarily requires both HpuA and HpuB to use hemoglobin (Hb) as a source of iron for growth . Deletion of HpuA resulted in reduced Hb binding and failure of growth on Hb . We identified rare Hb-utilizing colonies (Hb(+)) from an hpuA deletion mutant of FA1090, which fell into two phenotypic classes . One class of the Hb(+) revertants required expression of both TonB and HpuB for growth on Hb, while the other class required neither TonB nor HpuB . All TonB/HpuB-dependent mutants had single amino acid alterations in HpuB, which occurred in clusters, particularly near the C terminus . The point mutations in HpuB did not restore normal Hb binding . Human serum albumin inhibited Hb-dependent growth of HpuB point mutants lacking HpuA but did not inhibit growth when expression of HpuA was restored . Thus, HpuB point mutants internalized heme in the absence of HpuA despite reduced binding of Hb . HpuA facilitated Hb binding and was important in allowing use of heme from Hb for growth.

Infect Immun, 2002 Jan, 70(1), 407 - 11
Influence of the length of the lipooligosaccharide alpha chain on its sialylation in Neisseria meningitidis; Tsai CM et al.; The sialylation of lipooligosaccharide (LOS) in Neisseria meningitidis plays a role in the resistance of the organism to killing by normal human serum . The length of the alpha chain extending out from the heptose I {Hep (I)} moiety of LOS influenced sialylation of N . meningitidis LOS in vitro and in vivo . The alpha chain required a terminal Gal and a trisaccharide or longer oligosaccharide to serve as an acceptor for sialylation . The disaccharide lactose (Galbeta1-4Glc) in the alpha chain of immunotype L8 LOS could not function as an acceptor for the sialyltransferase, probably due to steric hindrance imposed by the neighboring Hep (II) with phosphorylethanolamine and another group attached.

Infect Immun, 2002 Jan, 70(1), 335 - 44
Neisserial immunoglobulin A1 protease induces specific T-cell responses in humans; Tsirpouchtsidis A et al.; We have previously shown that immunoglobulin A1 (IgA1) protease, an exoenzyme of pathogenic neisseriae, can trigger the release of proinflammatory cytokines from human monocytic subpopulations . Here, we demonstrate a dose-dependent T-cell response to recombinant gonococcal IgA1 protease (strain MS11) in healthy human blood donors . This response was delayed in comparison to the immune response against tetanus toxoid . Stimulation with IgA1 protease led to the activation of CD4(+) and CD8(+) T cells, as well as CD19(+) B cells and CD56(+) NK cells, indicated by de novo expression of CD69 . Only CD4(+) T cells proliferated and stained positive for intracellular gamma interferon (IFN-gamma) . Both proliferation and IFN-gamma production were dependent on antigen presentation via major histocompatibility complex class II . Peripheral blood mononuclear cells stimulated with IgA1 protease produce IFN-gamma and tumor necrosis factor alpha but no, or very low amounts of, interleukin-10 (IL-10) or IL-4, indicating a Th1-based proinflammatory immune response . These findings support the significance of IgA1 protease as a virulence determinant of bacterial meningitis and its function as a dominant proinflammatory T-cell antigen.

J Infect Dis, 2001 Dec 15, 184(12), 1538 - 47 Epub 2001 Dec 03.
Analysis of the ontogeny of the murine humoral response to Neisseria meningitidis B capsular polysaccharide reveals levels of complexity relevant to vaccine development; Colino J et al.; Although purified capsular polysaccharide of Neisseria meningitidis group B (CpsB) is not immunogenic at any age, CpsB on the bacterial surface elicits antibody responses late in ontogeny . Therefore, a detailed analysis of the ontogeny of the murine anti-CpsB response to N . meningitidis could determine key parameters regarding the poor immunogenicity of CpsB . The effects of bacterial dose, hyperimmunization, age, and sex on the induction of primary and secondary anti-CpsB immunoglobulin isotype profiles were studied . It was demonstrated that the timing and repetition of immunization and of the bacterial dose have a marked differential effect on the primary induction of anti-CpsB immunoglobulin isotypes and on the ability to induce anti-CpsB antibody responses after subsequent rechallenge . It is noteworthy that the ontogeny of the response is related to the appearance of natural anti-CpsB antibodies, but this is not associated with the presence of CpsB cross-reactive antigens in the microflora.

Clin Infect Dis, 2002 Jan 15, 34(2), 173 - 6 Epub 2001 Dec 07.
Increased sensitivity of DNA amplification testing for the detection of pharyngeal gonorrhea in men who have sex with men; Page-Shafer K et al.; We compared ligase chain reaction (LCR) assay with standard culture for the detection of pharyngeal Neisseria gonorrhoeae infection in men who have sex with men (MSM) presenting at a sexually transmitted diseases clinic in San Francisco . Pharyngeal specimens were obtained from 200 MSM who reported performing fellatio during the previous 2 weeks . Confirmatory testing of discrepant specimens was conducted using N . gonorrhoeae pilin proteins . Prevalence of pharyngeal N . gonorrhoeae was 6% by culture or 11% by LCR . The sensitivity and specificity of LCR were 94.7% and 97.8%, respectively, compared with values of 47.4% and 100% for culture . Prevalence of pharyngeal N . gonorrhoeae infection, as determined by DNA amplification testing, was higher than that suggested by traditional culture . Results support the use of DNA amplification testing in the oropharynx . The high prevalence of pharyngeal N . gonorrhoeae infection among MSM suggests that routine screening should be considered in efforts to reduce the burden of gonorrhea in this population.

J Antimicrob Chemother, 2001 Dec, 48(6), 853 - 9
Evolution in the trends of antimicrobial resistance in Neisseria gonorrhoeae isolated in Durban over a 5 year period: impact of the introduction of syndromic management; Moodley P et al.; Antimicrobial susceptibility testing was performed on isolates of Neisseria gonorrhoeae obtained from patients attending the City Health STD clinic in Durban, KwaZuluNatal, using the following drugs: penicillin, tetracycline, ciprofloxacin, ofloxacin, ceftriaxone, spectinomycin, erythromycin and azithromycin . These isolates were collected over a 6 year period from 1995 to 2000 . Four hundred and fifteen strains were tested: 61 in 1995, 198 in 1997, 98 in 1998/99 and 58 in 1999/2000 . A shift to the right is observed in the susceptibilities of N . gonorrhoeae to the currently recommended drugs in the syndromic management guidelines viz . penicillin, tetracycline, ceftriaxone, ciprofloxacin, spectinomycin and erythromycin . The prevalence of penicillinase-producing N . gonorrhoeae is currently c . 30%, whereas that of plasmid-mediated tetracycline-resistant N . gonorrhoeae is c . 50% . There is a definite association between the MICs of strains falling within the penicillin and tetracycline chromosomally resistant group, and strains exhibiting a decreased susceptibility to ciprofloxacin and ceftriaxone . The MICs of azithromycin showed a similar distribution when compared with erythromycin for 1999/2000 isolates . We postulate that the presence of efflux pumps might play a role in the increasing MICs that we observe among structurally unrelated groups of drugs . Furthermore, widespread use of these antimicrobials in the community may offer a selective advantage to the development of resistance . The implications of this are far reaching and the local susceptibility trends of N . gonorrhoeae need to be monitored constantly to direct therapy.

Presse Med, 2001 Nov 3, 30(32), 1589 - 91
{Purpura fulminans and coxsackie B2 co-infection}; Fourier C et al.; INTRODUCTION: During septic shock in children, myocardial deficiency usually lasts 3 to 6 days . When prolonged, an other etiology should be looked for . OBSERVATION: A 29 month-old child presented with purpura fulminans, probably due to Neisseria meningitidis . Positivity of the serodiagnostic for coxsackie B2, performed because of persisting myocardial deficiency, evoked diagnosis of a coxsackie B2 co-infection . DISCUSSION: Abnormally prolonged myocardial deficiency during purpura fulminans must lead to the search for a congenital, metabolic or infectious cardiomyopathy.

Pediatrics . 2001 Dec;108(6):E103.
Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae infection in pediatric private practice; Best D et al.; BACKGROUND: Universal screening of sexually active adolescents for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) has been recommended in settings in which prevalence is 2% or greater . Although believed to be above 2%, the prevalence of CT and GC infection in private practice settings has not been clearly established and may affect screening practices . OBJECTIVES: To determine CT and GC infection prevalence in 2 pediatric private practices . DESIGN: Cross-sectional study . SETTING: Two pediatric private practices in suburban North Carolina . PATIENTS: Convenience sample of patients aged 15 to 24 years who were seen from August 1998 through June 1999 . MAIN OUTCOME MEASURES: Prevalence of CT and GC infection . RESULTS: Of 1114 eligible patients, 803 (72%) completed questionnaires and provided urine specimens tested for CT and GC infection using ligase chain reaction assays . Mean age was 17.1 years (standard deviation: 1.8) . Most participants were female (63%), white (87%), and from highly educated families (64% of their mothers graduated from college) . Sexual activity was reported by 41% . Prevalence of CT infection in all participants was 0.9% (confidence interval {CI}: 0.4%-1.8%); in sexually active participants, 2.1% (CI: 0.9%-4.3%); in sexually active females, 2.7% (CI: 1.0%-5.7%); and in sexually active males, 0.9% (CI: 0.0%-5.1%) . One case of GC infection was found . CONCLUSIONS: The prevalence of CT and GC infection in this private practice population was much lower than reported in other settings . Screening recommendations may need to be reassessed if other low prevalence populations are found.

Vet Microbiol, 2002 Jan 3, 84(1-2), 53 - 67
A putative iron-regulated TonB-dependent receptor of Mannheimia (Pasteurella) haemolytica A1: possible mechanism for phase variation; Graham MR et al.; A recombinant plasmid that codes for a novel iron receptor protein (Irp) of Mannheimia (Pasteurella) haemolytica A1 was isolated by the partial complementation of an Escherichia coli fur mutant . The deduced amino acid sequence of Irp exhibited characteristics typical of TonB-dependent receptors . These include: a TonB-box at the N-terminal; a 50 amino acid region homologous to the "plug" domain of the E . coli FhuA and FepA receptors; and a C-terminal TonB-dependent signature which likely functions as an outer membrane anchoring domain . Previously uncharacterized Irp homologues were detected by BLAST analysis of available databases and incomplete microbial genomes . When the irp homologues from Neisseria gonorrhoeae and N . meningitidis were cloned by PCR and expressed in E . coli, novel proteins of the predicted size (84kDa) were detected in cell lysates, demonstrating that these are functional genes . The M . haemolytica A1 irp gene undergoes phase variation at a nucleotide region which contain the sequence AAAAAAATTAAAA (7A-2T-4A) flanked by a short inverted repeat . Site-specific mutagenesis of the 7A-2T-4A sequence as well as replacement of the inverted repeats resulted in a stable construct that expressed the Irp protein without phase variation . The expression of irp in M . haemolytica A1 was regulated by iron concentrations and most likely a Fur homologue, consistent with the proposed function of Irp in iron metabolism . The irp genes may represent contingency loci that play a role in iron acquisition during infection.

Sex Transm Dis, 2001 Dec, 28(12), 714 - 24
Risk-based versus alternative algorithms for antibiotic prophylaxis among women seeking early suction abortion: a cost-effectiveness simulation; Friedman HB et al.; BACKGROUND: Particularly in resource-poor settings, simple, inexpensive, and cost-effective algorithms are needed to direct antibiotic prophylaxis to prevent sequelae of infections with Chlamydia trachomatis, Neisseria gonorrhoeae, and bacterial vaginosis-associated organisms among women undergoing abortion . GOAL: To assess the prevalence of and risk factors for infections among women seeking abortion in Bali, Indonesia, and to use these data in designing a cost-effective risk-based prophylaxis algorithm . STUDY DESIGN: A cross-sectional analysis and data-based simulation of risk-based and alternative prophylaxis algorithms were performed . RESULTS: The risk-based algorithm would have provided prophylaxis to 70% (95% CI, 53-83%) of women with cervical infection, 64% (95% CI, 54-74%) of those with bacterial vaginosis, and 57% (95% CI, 42-72%) of those with trichomoniasis . For cervical infection, the algorithm was more cost effective than all others evaluated . The cost-effectiveness was acceptable for bacterial vaginosis . CONCLUSIONS: Risk-based algorithms may be cost effective in identifying women likely to benefit from preabortion prophylaxis . Prospective evaluation is needed to validate these findings.

J Clin Microbiol, 2001 Dec, 39(12), 4575 - 6
Improved rate of isolation of Neisseria meningitidis by direct plating of pharyngeal swabs; Cunningham R et al.; Culturing of pharyngeal swabs for Neisseria meningitidis is an important clinical and epidemiological tool . Routine methods include direct plating onto solid medium or later plating in the laboratory . A comparison of these methods used with 490 high school students found a significantly higher carriage rate with direct plating (11.8 versus 6.1%; P < 0.001).

J Clin Microbiol, 2001 Dec, 39(12), 4227 - 32
Laribacter hongkongensis gen . nov., sp . nov., a novel gram-negative bacterium isolated from a cirrhotic patient with bacteremia and empyema; Yuen KY et al.; A bacterium was isolated from the blood and empyema of a cirrhotic patient . The cells were facultatively anaerobic, nonsporulating, gram-negative, seagull shaped or spiral rods . The bacterium grows on sheep blood agar as nonhemolytic, gray colonies 1 mm in diameter after 24 h of incubation at 37 degrees C in ambient air . Growth also occurs on MacConkey agar and at 25 and 42 degrees C but not at 4, 44, and 50 degrees C . The bacterium can grow in 1 or 2% but not 3, 4, or 5% NaCl . No enhancement of growth is observed with 5% CO(2) . The organism is aflagellated and nonmotile at both 25 and 37 degrees C . It is oxidase, catalase, urease, and arginine dihydrolase positive, and it reduces nitrate . It does not ferment, oxidize, or assimilate any sugar tested . 16S rRNA gene sequencing showed that there are 91 base differences (6.2%), 112 base differences (7.7%), and 116 base differences (8.2%) between the bacterium and Microvirgula aerodenitrificans, Vogesella indigofera, and Chromobacterium species, respectively . The G+C content (mean and standard deviation) is 68.0% +/- 2.43%, and the genomic size is about 3 Mb . Based on phylogenetic affiliation, the bacterium belongs to the Neisseriaceae family of the beta-subclass of Proteobacteria . For these reasons, a new genus and species, Laribacter hongkongensis gen . nov., sp . nov., is proposed, for which HKU1 is the type strain . Further studies should be performed to ascertain the potential of this bacterium to become an emerging pathogen.

Curr Infect Dis Rep, 2001 Dec, 3(6), 540 - 545
Nongonococcal and Nonchlamydial Cervicitis; Nyirjesy P; Cervicitis encompasses a variety of gynecologic conditions . Although inflammation on a Papanicolaou smear may be indicative of true cervical inflammation, establishing a diagnosis of mucopurulent cervicitis (MCP) is more clinically useful . MCP seems to be analogous to nongonococcal urethritis in men . Many cases of MCP are related to Chlamydia trachomatis infection, but the majority of cases have other potential causes . Other possible pathogens include Neisseria gonorrhoeae, Trichomonas vaginalis, Ureaplasma urealyticum, Mycoplasma genitalium, herpes simplex virus, and cytomegalovirus . Many cases may be related to cervical ectopy . MCP is a marker for endometritis, salpingitis, and adverse pregnancy outcomes . Given the current shortcomings in our understanding of this disease, an empiric approach to evaluation and treatment is suggested.

Mol Microbiol, 2001 Nov, 42(3), 659 - 72
Receptor-mediated endocytosis of Neisseria gonorrhoeae into primary human urethral epithelial cells: the role of the asialoglycoprotein receptor; Harvey HA et al.; Urethral epithelial cells are invaded by Neisseria gonorrhoeae during gonococcal infection in men . To understand further the mechanisms of gonococcal entry into host cells, we used the primary human urethral epithelial cells (PHUECs) tissue culture system recently developed by our laboratory . These studies showed that human asialoglycoprotein receptor (ASGP-R) and the terminal lactosamine of lacto-N-neotetraose-expressing gonococcal lipooligosaccharide (LOS) play an important role in invasion of PHUECs . Microscopy studies showed that ASGP-R traffics to the cell surface after gonococcal challenge . Co-localization of ASGP-R with gonococci was observed . As ASGP-R-mediated endocytosis is clathrin dependent, clathrin localization in PHUECs was examined after infection . Infected PHUECs showed increased clathrin recruitment and co-localization of clathrin and gonococci . Preincubating PHUECs in 0.3 M sucrose or monodansylcadaverine (MDC), which both inhibit clathrin-coated pit formation, resulted in decreased invasion . N . gonorrhoeae strain 1291 produces a single LOS glycoform that terminates with Gal(beta1-4)GlcNac(beta1-3)Gal(beta1-4)Glc (lacto-N-neotetraose) . Invasion assays showed that strain 1291 invades significantly more than four isogenic mutants expressing truncated LOS . Sialylation of strain 1291 LOS inhibited invasion significantly . Preincubation of PHUECs in asialofetuin (ASF), an ASGP-R ligand, significantly reduced invasion . A dose-response reduction in invasion was observed in PHUECs preincubated with increasing concentrations of NaOH-deacylated 1291 LOS . These studies indicated that an interaction between lacto-N-neotetraose-terminal LOS and ASGP-R allows gonococcal entry into PHUECs.

Am J Obstet Gynecol, 2001 Nov, 185(5), 1209 - 17
Randomized trial of presumptive sexually transmitted disease therapy during pregnancy in Rakai, Uganda; Gray RH et al.; OBJECTIVE: The purpose of this study was to assess presumptive sexually transmitted disease treatment on pregnancy outcome and HIV transmission . STUDY DESIGN: In a randomized trial in Rakai District, Uganda, 2070 pregnant women received presumptive sexually transmitted disease treatment 1 time during pregnancy at varying gestations, and 1963 control mothers received iron/folate and referral for syphilis . Maternal-infant sexually transmitted disease/HIV and infant outcomes were assessed . Intent-to-treat analyses estimated adjusted rate ratios and 95% confidence intervals . RESULTS: Sexually transmitted diseases were reduced: Trichomonas vaginalis (rate ratio, 0.28; 95% CI, 0.18%-0.49%), bacterial vaginosis (rate ratio, 0.78; 95% CI, 0.69-0.87), Neisseria gonorrhoeae /Chlamydia trachomatis (rate ratio, 0.43; 95% CI, 0.27-0.68), and infant ophthalmia (rate ratio, 0.37; 95% CI, 0.20-0.70) . There were reduced rates of neonatal death (rate ratio, 0.83; 95% CI, 0.71-0.97), low birth weight (rate ratio, 0.68; 95% CI, 0.53-0.86), and preterm delivery (rate ratio, 0.77; 95% CI, 0.56-1.05); but there were no effects on maternal HIV acquisition or perinatal HIV transmission . CONCLUSION: Reductions of maternal sexually transmitted disease improved pregnancy outcome but not maternal HIV acquisition or perinatal HIV transmission.

Sex Transm Infect, 2001 Dec, 77(6), 423 - 6
Comparison of urine, first and second endourethral swabs for PCR based detection of genital Chlamydia trachomatis infection in male patients; Sugunendran H et al.; OBJECTIVES: To compare endourethral swabs and urine as diagnostic specimens for the detection of genital Chlamydia trachomatis infection using the polymerase chain reaction (PCR), in male patients attending a genitourinary clinic and to assess whether the first endourethral swab used solely for diagnosing gonococcal infection could be used for C trachomatis detection as well . METHODS: Two endourethral swabs were taken from 80 male patients, in whom the likelihood of genital C trachomatis infection was high . The first swab was used for microscopy and culture for Neisseria gonorrhoeae, before being used for C trachomatis detection . First voided urine specimens were collected from 61 of these patients . All three specimens were processed for C trachomatis DNA detection using the Roche Cobas Amplicor PCR . A diagnosis of genital C trachomatis infection was made if any one of the specimens tested reproducibly positive . Samples from 13 patients showing discrepant PCR results between swabs and/or urine were retested by ligase chain reaction (LCR) . RESULTS: Chlamydia trachomatis DNA was detected in 35 (43.8%) of the 80 patients . In 17 of the 35 patients (48.6%), all the genital specimens were positive . However, in 18 (51.4%) patients, one or more of the genital specimens had negative PCR results . Among the 18 patients with discrepant results, urine was found to be a more sensitive diagnostic specimen than the second urethral swab picking up 13 out of 16 positives (81.3%) as opposed to five out of 18 (27.8%) . There was no significant difference between the two swabs . Retesting by LCR, of the samples from 13 of the 18 patients with discrepant PCR results confirmed them all as true positives, although as with PCR, not all specimens in the set were concordantly positive . LCR detected all the 13 positives in urine, while there was no difference in the detection rate between the first and the second urethral swabs . CONCLUSIONS: Urine appeared to be a better diagnostic specimen than the conventional second endourethral swab for C trachomatis detection by PCR in this cohort of male patients . There was no difference between the first swab, intended primarily for N gonorrhoeae testing and the second swab intended for C trachomatis detection . This raises questions over the need for the conventional second swab for detecting C trachomatis.

Acta Cardiol, 2001 Oct, 56(5), 327 - 9
Acute ST-segment elevation in Neisseria meningitis; Gach O et al.; Meningitis due to Neisseria meningitidis occurred in a young man . On admission, he was in septic shock and the electrocardiogram revealed convex upwards ST-segment elevation in inferior and lateral leads . Rapid and complete normalisation of the ECG was observed and the patient had a favourable evolution with intensive therapy.The mechanisms of the ECG abnormalities in this disease are unclear . In this patient, ST-segment elevation was probably related to severe transmural ischaemia or prolonged coronary artery spasm as suggested by increase and decrease of cardiac enzymes and transient echocardiographic wall motion abnormalities without pericarditis . However, myocarditis could not be completely ruled out.

Kansenshogaku Zasshi, 2001 Oct, 75(10), 894 - 7
{A case of bacterial peritonitis caused by Neisseria gonorrhoeae}; Akahane T et al.; On January 17, 2001, a 39-year old female with sudden abdominal pain was admitted to her neighboring outpatient clinic and diagnosed as suspicious of infectious enteritis . However, on the next day (January 18, 2001) she was soon transferred to Toyoshina Red Cross Hospital with the chief complaint of severe abdominal pain, high fever, and of conspicuous leucocytosis . Laboratory data on her admission demonstrated apparent signs of inflammation and she was soon undergone an emergency operation . Neisseria gonorrhoeae was recovered from her ascetic fluid, otherwise Chlamydia EIA was negative . The antibiotic chemotherapy of minocycline (200 mg/day) was continued for the first 9 days and sulbactam/cefoperazone (2 g/day) had been administered for the first 5 days . Her symptoms were discontinued on her 10th hospital day, and she was discharged on the 14th hospital day . There have been few reported cases in Japan of bacterial peritonitis due to N . gonorrhoeae . This is, to the best of our knowledge, the first reported case of N . gonorrhoeae peritonitis in Japan.

Int J Antimicrob Agents, 2001 Nov, 18(5), 437 - 40
Antimicrobial activity of BMS 284756 (T-3811) against Neisseria gonorrhoeae tested by three methods; Deshpande L et al.; The potency of BMS 284756, a novel des-F(6)-quinolone, was tested against 137 clinical isolates of Neisseria gonorrhoeae including 50 strains observed to be resistant to ciprofloxacin and other newer quinolones . The gonococci were tested using NCCLS methods (agar dilution, disk diffusion) and Etest . BMS 284756 potency versus N . gonorrhoeae was generally two- to four-fold greater than ciprofloxacin . Penicillin resistance in the absence of ciprofloxacin resistance did not affect BMS 284756 activity . However, elevated ciprofloxacin MICs were associated with higher BMS 284756 MIC results as follows (BMS 284756 MIC(50)/MIC range in mg/l): ciprofloxacin-susceptible strains (0.016 or 0.03/0.004-0.06), ciprofloxacin-intermediate strains (0.06 or 0.12/0.008-0.25) and ciprofloxacin-resistant strains (0.12 or 0.5/0.12-1) . Etest MICs were routinely lower than those produced by the reference agar dilution method, but the correlation coefficient remained acceptable (r = 0.87) . Similarly acceptable correlation was achieved with 5 microg disk zone diameters (r = 0.78), where all zones were > or = 28 mm (MIC < or = 1 mg/l) . In conclusion, BMS 284756 was very active against N . gonorrhoeae (MIC(50) 0.03 mg/l overall) including ciprofloxacin-resistant strains and could be considered as a single-dose therapeutic option for gonorrhoea.

Antimicrob Agents Chemother, 2001 Dec, 45(12), 3603 - 6
Outbreak of cefozopran (penicillin, oral cephems, and aztreonam)-resistant Neisseria gonorrhoeae in Japan; Muratani T et al.; We have previously reported that the Neisseria gonorrhoeae isolates from clinical failure cases treated with cefdinir and aztreonam, beta-lactams exhibited high MICs . These resistant isolates were clearly separated from the isolates exhibiting a low level of resistance to beta-lactams as shown by the MIC distribution of cefozopran . Restriction fragment length polymorphism DNA typing revealed that the outbreak of cefozopran-resistant isolates in Kitakyushu, Japan, occurred as a result of clonal spread.

Niger J Med, 2001 Apr-Jun, 10(2), 72 - 4
Antimicrobial susceptibility pattern of Neisseria gonorrhoeae isolated at the Jos University Teaching Hospital, Jos, Central Nigeria; Banwat EB et al.; Widespread, inadequate and inappropriate treatment of gonococcal infections are facilitating antibiotic resistance in Jos Nigeria . We present an antimicrobial susceptibility pattern of Neisseria gonorrhoeae strains in this study . Thirty-one strains of Neisseria gonorrhoeae were isolated by inoculating swabs onto prewarmed heated blood agar and Thayer Martins medium and incubated overnight at 35 to 37 degrees C . The isolates were then screened for antimicrobial susceptibility by the disk diffusion method . About 93.6% were susceptible to azithromycin, 90.3% to ciprofloxacin, 87.1% to ceftriaxone, 80.7% and 64.3% to erythromycin and ofloxacin respectively . Less than half were each susceptible to spectinomycin (38.7%) and gentamicin (32.8%), while 12.9% and 4.6% were susceptible to tetracycline and penicillin respectively . These findings suggest that gentamicin, tetracycline and penicillin may not be recommended in this environment for treating gonococcal infections . Azithromycin, ciprofloxacin and ceftriaxone are the suggested alternatives . Prompt diagnosis, treatment and indeed continued surveillance are hereby recommended for an effective management of gonococcal infections.

Contraception, 2001 Sep, 64(3), 169 - 72
High rate of bacterial vaginosis among women with intrauterine devices in Manado, Indonesia; Joesoef MR et al.; Recent research reported that bacterial vaginosis (BV) might enhance the acquisition and transmission of HIV . BV is also associated with an increased risk of pelvic inflammatory disease, a disease also associated with intrauterine device (IUD) insertion . To measure the magnitude of this problem, we conducted a prevalence survey of BV and sexually transmitted diseases (STDs; defined as current infections with Neisseria gonorrhoeae, Chlamydia trachomatis, and/or Trichomonas vaginalis) among all patients attending a family planning clinic in Manado from May to July 1999 . BV was diagnosed by Gram stain using Nugent's criteria and vaginal trichomoniasis by wet mount or culture . Cervical infections with C . trachomatis and N . gonorrhoeae were diagnosed by DNA probe . Of 357 patients, 116 (32.5%) had BV, 83 (23.3%) had trichomoniasis, 9 (2.5%) had chlamydia, and 8 (2.2%) had gonorrhea . The prevalence of STD was similar among users of all types of contraception . However, BV was more common among IUD users (47.2%) than among non-IUD users (29.9%) . This association persisted after controlling for age, education, ever had douching, and any STD (odds ratio 2.0, 95% CI 1.1-3.8) . BV was also associated with STD (41.3% in women with STD vs . 29.4% in women without) . This association remained significant after adjusting for age, education, ever had douching, and IUD use (odds ratio 1.7, 95% CI 1.1-2.9) . Because we found that BV was associated with IUDs and that other studies reported that both BV and IUDs were associated with pelvic inflammatory disease, a Gram stain evaluation of BV may be considered prior to IUD insertion

Mol Microbiol, 2001 Oct, 42(2), 293 - 307
Structural alterations in a type IV pilus subunit protein result in concurrent defects in multicellular behaviour and adherence to host tissue; Park HS et al.; The ability of bacteria to establish complex communities on surfaces is believed to require both bacterial-substratum and bacterial-bacterial interactions, and type IV pili appear to play a critical but incompletely defined role in both these processes . Using the human pathogen Neisseria gonorrhoeae, spontaneous mutants defective in bacterial self-aggregative behaviour but quantitatively unaltered in pilus fibre expression were isolated by a unique selective scheme . The mutants, carrying single amino acid substitutions within the conserved amino-terminal domain of the pilus fibre subunit, were reduced in the ability to adhere to a human epithelial cell line . Co-expression of the altered alleles in the context of a wild-type pilE gene confirmed that they were dominant negative with respect to aggregation and human cell adherence . Strains expressing two copies of the altered alleles produced twice as much purifiable pili but retained the aggregative and adherence defects . Finally, the defects in aggregative behaviour and adherence of each of the mutants were suppressed by a loss-of-function mutation in the twitching motility gene pilT . The correlations between self-aggregation and the net capacity of the microbial population to adhere efficiently demonstrates the potential significance of bacterial cell-cell interactions to colonization.

Acta Trop, 2001 Dec 21, 80(3), 261 - 4
Neisseria gonorrhoeae and Chlamydia trachomatis infections in patients attending STD and family planning clinics in Bissau, Guinea-Bissau; Gomes JP et al.; Accurate clinical and laboratory data about sexually transmitted diseases (STD) prevalence in Guinea-Bissau are not available . These data are important, since HIV2 is prevalent in this country, rates of HIV1 are increasing and STDs facilitate HIV transmission . Since DNA amplification methods have demonstrated to accurately diagnose chlamydial infections and gonorrhoea, the Amplicor CT/NG PCR Assay with Internal Control of Amplification (Roche Diagnostic System, Branchburg, NJ, USA) was used to estimate the prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis genital infections in STDs and Family Planning Clinic attenders in Bissau, from March to July 1997 . Two hundred and two cervical swabs and 31 urethral swabs were examined . Two women were excluded from this study because their cervical swabs contained inhibitory substances . N . gonorrhoeae was identified in 34/200 (17%) women and in 12/31 (38.7%) men . C . trachomatis was detected in 8/200 (4%) women there were no positive C . trachomatis results among the 31 men with urethritis . One woman presented a mixed infection with both organisms . The prevalence difference between men and women was not statistically significant (P=0.6) for C . trachomatis infection, but it was significant for N . gonorrhoeae infection (P=0.01) . The prevalence rates of these infections found in this study, support the need for an urgent strategy to control STD in the region.

J Med Microbiol, 2001 Nov, 50(11), 991 - 5
A comparison of multilocus sequence typing and fluorescent fragment-length polymorphism analysis genotyping of clone complex and other strains of Neisseria meningitidis; Hookey JV et al.; Five National Collection of Type Culture (NCTC) strains and 14 isolates of Neisseria meningitidis, representing 13 outbreak isolates from within the UK, were examined by multilocus sequence typing (MLST) for seven house-keeping genes . The results were compared with those of fluorescent amplified fragment-length polymorphism (FAFLP) analysis . Phylogenetic inferences were made from 3284-nucleotide lengths of sequence for the 19 isolates, by distance and parsimony methods . Two clusters of isolates were delineated . The larger, comprising eight isolates--S1, S3, Ironville, P9, ET-37 (M99-241951), P7, P10 and P60--shared 100-99.2% similarity and varied in only 40 nucleotides (approximately 1.22% variation) from the consensus sequence alignment . This cluster could be equated to the ET-37 complex because it had allelic signatures identical to MLST sequence types 11 and 50 . These eight isolates were also assigned to one group by FAFLP . The reference ET-5 complex isolate 'ET-5 (NG144/82)' and an isolate (X9) from an outbreak in the north of England were also grouped together by MLST . They shared 99.2% similarity and differed within the aroE and fumC genes by 4 and 17 nucleotides, respectively . Their MLST sequence types were 32 and 661 and, therefore, these two isolates could be equated to the ET-5 complex . They also grouped together by FAFLP . A comparison of the resources required to apply MLST to the 19 isolates examined with those needed to characterise them by FAFLP indicated that FAFLP (a fragment-based genotyping method) is more cost-effective than the partial sequencing approach, MLST.

Bioorg Khim, 2000 Nov, 26(11), 817 - 24
{Inhibition of binding of activated compliment component C4b with its target}; Kozlov LV et al.; The inhibition of covalent binding of the nascent C4b fragment of the human complement component to its natural target, immunoglobulin G, was studied . To this end, an immunoenzyme system was developed . In this ELISA method, the complement was activated on the sorbed IgG molecules and the resulting nascent C4b fragment acylated IgG or interacted with a competitive inhibitor added to the system . The inhibition constants for binding of the nascent C4b to its target were determined for immunoglobulins G1, G2, G3, G4, M, and A1, as well as for ferritin, yeast mannan, capsid polysaccharides of the Neisseria meningitidis A, B, and C serotypes, diphtheria anatoxin, epinephrine, and salicylic acid . On the basis of the experimental data, the immunoglobulin role at the activation stage of the complement regulation cascade, the relationship between the antigen immunogenicity and its ability to interact with C4b, and the direct effect of a number of therapeutic agents on the complement system were discussed . Lectins of various specificities were shown to inhibit the enzymic activation of C4 by the first complement component and the subsequent C4b sorption to its target, which allowed us to suggest that some oligosaccharide fragments of the C1s and C4 molecules are spatially close to the C1s active site and to the thioester bond of C4.

Sex Transm Dis, 2001 Oct, 28(10), 555 - 64
Experimental gonococcal urethritis and reinfection with homologous gonococci in male volunteers; Schmidt KA et al.; BACKGROUND: Reinfection, a common occurrence with gonorrhea, may result from a lack of protective immune response, or from the tremendous gonococcal strain variation . GOAL: A two-phase study in human volunteers tested whether experimental infection with Neisseria gonorrhoeae MS11mkC would protect against reinfection with the same organisms . STUDY DESIGN: In phase 1, an intraurethral inoculum of 57,000 piliated, transparent (opacity protein-negative {Opa-}) MS11mkC N gonorrhoeae infected 14 of 15 (93%) volunteers . The volunteers were encouraged to delay treatment for at least 5 days . In phase 2, which began 2 weeks after treatment for the initial infection, volunteers were inoculated with 7,100 piliated, Opa- MS11mkC . RESULTS: The phase 2 challenge infected 6 of 14 (43%) previously infected volunteers and 5 of 10 (50%) naive control subjects . Phase 1 volunteers who resisted reinfection were significantly more likely to have had a fourfold or greater increase in lipooligosaccharide immunoglobulin G during phase 1 than those who did not resist reinfection (P = 0.026) . CONCLUSIONS: Although infection did not provide protection from reinfection under the conditions used, the results suggest that immunity to reinfection is more complex than anticipated by the experimental design.

Diagn Microbiol Infect Dis, 2001 Sep-Oct, 41(1-2), 83 - 8
Neisseria meningitidis with decreased susceptibility to penicillin: report from the SENTRY antimicrobial surveillance program, North America, 1998-99; Richter SS et al.; The purpose of this study was to estimate the prevalence of Neisseria meningitidis with decreased susceptibility to penicillin (MIC, >0.06 microg/mL) in North America (NA) . Antimicrobial susceptibility testing by Etest (AB BIODISK, Solna, Sweden) was performed on 53 invasive clinical isolates obtained from 11 SENTRY Antimicrobial Surveillance Program participants in NA (9 states, 2 provinces) during 1998-99 . All strains were markedly susceptible to ciprofloxacin (MIC(90), 0.008 microg/mL) and cefotaxime (MIC(90), < or = 0.002 microg/mL) . Only 54.7% were susceptible to trimethoprim-sulfamethoxazole (TMP/SMX) at < or = 0.5/9.5 microg/mL . One strain was resistant to rifampin (MIC, > 32 microg/mL) and 16 isolates (30.2%) were relatively resistant to penicillin with MICs ranging from 0.094 to 0.25 microg/mL . No beta-lactamase production was detected . The serogroup distribution was 40% Y, 28% B, 24% C, 2% W-135, and 6% of strains were nongroupable . The prevalence of N . meningitidis with decreased susceptibility to penicillin in NA appears higher than previous reports.

J Am Chem Soc, 2001 Nov 7, 123(44), 10909 - 18
Microbial glycosyltransferases for carbohydrate synthesis: alpha-2,3-sialyltransferase from Neisseria gonorrheae; Izumi M et al.; The alpha-2,3-sialyltransferase from Neisseria gonorrheae was overproduced in E . coli for exploitation of its substrate specificity and synthetic utility . Several potential acceptor substrates were synthesized in this study, including mono- and oligosaccharides, glycolipids, and glycopeptides and their sulfate derivatives . Some CMP-sialic acid derivatives with modification at the C-5 position were also prepared for evaluation as donor substrates . It was found that the enzyme exhibits a broader acceptor substrate specificity when compared to other sialyltransferases, though the donor specificity is quite limited . Application of the enzyme to the preparative synthesis of representative sialyl glycoconjugates has been demonstrated . On the basis of this work and the work of others, this enzyme is the most versatile and synthetically useful among all sialyltransferases known to date, especially for the synthesis of sulfate-containing glycoconjugates.

AIDS, 2001 Aug, 15 Suppl 4, S15 - 30
Ecological and individual level analysis of risk factors for HIV infection in four urban populations in sub-Saharan Africa with different levels of HIV infection; Auvert B et al.; OBJECTIVE: To identify factors that could explain differences in rate of spread of HIV between different regions in sub-Saharan Africa . DESIGN: Cross-sectional study . METHODS: The study took place in two cities with a relatively low HIV prevalence (Cotonou, Benin and Yaounde, Cameroon), and two cities with a high HIV prevalence (Kisumu, Kenya and Ndola, Zambia) . In each of these cities, a representative sample was taken of about 1000 men and 1000 women aged 15-49 years . Consenting men and women were interviewed about their socio-demographic background and sexual behaviour; and were tested for HIV, herpes simplex virus type 2 (HSV-2), syphilis, Chlamydia trachomatis and Neisseria gonorrhoea infection, and (women only) Trichomonas vaginalis . Analysis of risk factors for HIV infection was carried out for each city and each sex separately . Adjusted odds ratios (aOR) were obtained by multivariate logistic regression . RESULTS: The prevalence of HIV infection in sexually active men was 3.9% in Cotonou, 4.4% in Yaounde, 21.1% in Kisumu, and 25.4% in Ndola . For women, the corresponding figures were 4.0, 8.4, 31.6 and 35.1% . High-risk sexual behaviour was not more common in the high HIV prevalence cities than in the low HIV prevalence cities, but HSV-2 infection and lack of circumcision were consistently more prevalent in the high HIV prevalence cities than in the low HIV prevalence cities . In multivariate analysis, the association between HIV infection and sexual behavioural factors was variable across the four cities . Syphilis was associated with HIV infection in Ndola in men {aOR = 2.7, 95% confidence interval (CI) = 1.5-4.91 and in women (aOR = 1.7, 95% CI = 1.1-2.6) . HSV-2 infection was strongly associated with HIV infection in all four cities and in both sexes (aOR ranging between 4.4 and 8.0) . Circumcision had a strong protective effect against the acquisition of HIV by men in Kisumu (aOR = 0.25, 95% CI = 0.12-0.52) . In Ndola, no association was found between circumcision and HIV infection but sample sizes were too small to fully adjust for confounding . CONCLUSION: The strong association between HIV and HSV-2 and male circumcision, and the distribution of the risk factors, led us to conclude that differences in efficiency of HIV transmission as mediated by biological factors outweigh differences in sexual behaviour in explaining the variation in rate of spread of HIV between the four cities.

Proteomics, 2001 Aug, 1(8), 975 - 86
Integrated system for high-throughput protein identification using a microfabricated device coupled to capillary electrophoresis/nanoelectrospray mass spectrometry; Li J et al.; An integrated microsystem providing rapid analyses of trace-level tryptic digests for proteomics application is presented . This modular microsystem includes an autosampler and a microfabricated device comprising a sample introduction port and an array of separation channels together with a low dead-volume facilitating the interface to nanoelectrospray mass spectrometry . Sequential injection and separation of peptide standards and tryptic digests was achieved with a throughput of up to 30 samples per hour with less than 3% sample carryover . Replicate injections of peptide mixtures indicated that reproducibility of migration time was typically better than 2.3% relative standard deviation (RSD) whereas RSD values of 3.7-11.8% were observed on peak height . Mass spectral detection of submicromolar protein digests (< 7 femtomoles/injection) was achieved using a quadrupole/time of flight instrument in less than 2 min/per sample with peak widths of 1.8-7.0 s . The analytical potential of this integrated device for the identification of gel isolated proteins from Neisseria meningitidis immunotype L3 has been demonstrated using both peptide mass-fingerprint database searching and on-line tandem mass spectrometry.

Can J Microbiol, 2001 Sep, 47(9), 871 - 6
Acid stress upregulated outer membrane proteins in clinical isolates of Neisseria gonorrhoeae, but not most commensal Neisseria; Pettit RK et al.; Human immune serum recognition of outer membrane components from commensal and pathogenic Neisseria cultured under neutral and acidic conditions was investigated . Acid stress caused no detectable alterations in lipooligosaccharide migration and (or) staining, in outer membrane protein profiles, or in immune serum recognition of outer membrane components from Neisseria mucosa or Neisseria sicca . There was also no difference in the lipoologosaccharide electrophoretic pattern of acid- and neutral-grown Neisseria lactamica, but there were differences in outer membrane protein expression . The outer membrane protein alterations induced by acid stress in N . lactamica were not the same as those seen in isolates from patients with uncomplicated gonococcal infection, pelvic inflammatory disease, and disseminated gonococcal infection . Many differences were detected in the immune serum recognition of outer membrane components from acid- and neutral-cultured N . lactamica and from the clinical isolates of Neisseria gonorrhoeae, and these should be considered in vaccine design.

J Clin Microbiol, 2001 Nov, 39(11), 3819 - 22
TaqMan-based detection of Trichomonas vaginalis DNA from female genital specimens; Jordan JA et al.; A double-labeled fluorescent probe was designed and evaluated for detecting Trichomonas vaginalis DNA in a 5' nuclease (TaqMan) assay . The T . vaginalis-specific probe contains a 5'-fluorescein (5'-FAM) and a 3'-rhodamine (TAMRA) derivative . Female genital secretions were collected on Amplicor (Roche Molecular, Indianapolis, Ind.) swabs and by a transport system used for Chlamydia trachomatis and/or Neisseria gonorrhoeae DNA detection by PCR . Five hundred fifty-two female genital specimens, of which 248 (45%) were vaginal specimens and 304 (55%) were introital, were tested for both T . vaginalis DNA and viable microorganisms using the 5' nuclease assay and broth culture, respectively . Of these, 304 of 552 (55%) were also evaluated by direct microscopic examination for the characteristic motile organism . After resolving discrepancies, the comparisons produced an analytical sensitivity and specificity for the TaqMan-based PCR assay of 97.8 and 97.4%, respectively . As a result, DeltaRQ values (differences in fluorescence due to probe hybridization and resulting 5'-FAM cleavage from the specific PCR product) of > or =2.0 and < or =1.5 were established for T . vaginalis-positive and -negative cutoffs, respectively . DeltaRQ values between 1.5 and 2.0 were considered indeterminate . Overall findings revealed a high level of agreement between PCR and culture for detecting T . vaginalis . Potential benefits of the 5' nuclease assay include a greater sensitivity compared to direct microscopic examination and the ease of testing large numbers of clinical specimens in a significantly shorter turnaround time compared to culture.

Int J Med Microbiol, 2001 Sep, 291(4), 307 - 14
Signal transduction pathways induced by virulence factors of Neisseria gonorrhoeae; Popp A et al.; The obligate human pathogen Neisseria gonorrhoeae infects a variety of human tissues . In recent years, several host cell receptors for the major bacterial adhesins have been identified . While the knowledge of the molecular mechanism of colonisation has helped to understand special aspects of the infection, like the explicit tropism of gonococci for human tissues, the long-term consequences of engaging these receptors are still unknown . A variety of signalling pathways initiated by the activated receptors and by bacterial proteins transferred to the infected cell have been defined which include lipid second messenger, protein kinases, proteases and GTPases . These pathways control important steps of the infection, such as tight adhesion and invasion, the induction of cytokine release, and apoptosis . The detailed knowledge of bacteria-induced signalling pathways could allow the design of new therapeutic approaches which might be advantageous over the classical antibiotics therapy.

Euro Surveill, 2001 May, 6(5), 86 - 90
The laboratory surveillance system of Chlamydia trachomatis and Neisseria gonorrhoeae infections in Denmark; Hoffmann S; This article presents results of surveillance of C . trachomatis and N . gonorrhoeae infection in Denmark . In 1999 and 2000, there were 291 and 309 male cases of laboratory-confirmed gonorrhoeae, whereas the number of female cases was 43 for both years . The penicillin resistance was steady, with a prevalence of 40% in 1999, whereas the prevalence of fluoroquinolone resistance increased from 0% in 1990 to 27% in 1999 . The number of laboratory-confirmed C . trachomatis infections remains around 13 000 cases per year . Men aged 20-29 years and women aged 15-24 are the most at risk.

Euro Surveill, 2001 May, 6(5), 69 - 70
Europe-wide surveillance for sexually transmitted infections: a timely and appropriate intervention; Fenton K et al.; Sexually transmitted infections (STIs), including HIV, pose a significant threat to the health and wellbeing of Europeans and require concerted and sustained intervention . Over the past decade, the number of reported cases of STIs, including antimicrobial resistant Neisseria gonorrhoeae, has increased appreciably, and outbreaks of syphilis have recently been reported in many Member states of the European Union (EU) . Young people, homosexual men, and those with poor access to health care (including preventive and educational services) seem to be particularly affected, with increased travel and migration, high risk sexual behaviours, and a deterioration of public health responses to STI control (for example, partner notification) being key driving factors . Many of these demographic and behavioural trends are similar across EU states, as is the desire to develop and implement effective STI prevention and control programmes.

Mol Microbiol, 2001 Oct, 42(1), 167 - 81
The Neisseria gonorrhoeae lpxLII gene encodes for a late-functioning lauroyl acyl transferase, and a null mutation within the gene has a significant effect on the induction of acute inflammatory responses; Ellis CD et al.; LPS is a fundamental constituent of the outer membrane of all Gram-negative bacteria, and the lipid A domain plays a central role in the induction of inflammatory responses . We identified genes of the Neisseria gonorrhoeae lipid A biosynthetic pathway by searching the complete gonococcal genome sequence with sequences of known enzymes from other species . The lpxLII gene was disrupted by an insertion-deletion in an attenuated aroA mutant of the gonococcal strain MS11 . Lipopolysaccharide (LPS) and lipid A analysis demonstrated that the lpxLII mutant had synthesized an altered LPS molecule lacking a single lauric fatty acid residue in the GlcN II of the lipid A backbone . LPS of the lpxLII mutant had a markedly reduced ability to induce the proinflammatory cytokines tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6 and IL-8 from human macrophages and IL-8 from polymorphonuclear cells . This study demonstrates that the lpxLII gene in gonococci encodes for a late-functioning lauroyl acyl transferase that adds a lauric acid at position 2' in the lipid A backbone . The presence of lauric acid at such a position appears to be crucial for the induction of full inflammatory responses by N . gonorrhoeae LPS.

Sex Transm Dis, 2001 Nov, 28(11), 630 - 2
Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae in Turkey among men With urethritis; Agacfidan A et al.; BACKGROUND: Chlamydia trachomatis and Neisseria gonorrhoeae are known to cause urethritis . However, only a small number of studies in Eastern European countries have investigated the causes of urethritis . GOALS: To determine the prevalence of C trachomatis and N gonorrhoeae among men with symptomatic urethritis in Istanbul, Turkey, and to determine whether contact with a commercial sex worker increased the likelihood of chlamydial infections . STUDY DESIGN: Men with a diagnosis of urethritis at the Istanbul Faculty of Medicine were screened for C trachomatis and N gonorrhoeae by Abbott's ligase chain reaction (LCR) using either urethral swabs or first-void urine . N gonorrhoeae cultures were done on a subset of these patients . RESULTS: The study enrolled 813 men . All of the men denied condom use during their previous sexual exposures . The overall prevalence of C trachomatis, as determined by LCR, was 15.7% . Only 192 patients were screened for both organisms . N gonorrhoeae prevalence was 9.4% . There was no difference in the chlamydia prevalence between men who had contact with commercial sex workers (CSWs) and men who had no such contact (15.3% versus 17.2%) . However, clients of foreign CSWs were more likely to have chlamydia than clients of registered Turkish CSWs . CONCLUSIONS: C trachomatis and N gonorrhoeae are commonly found in Turkish men with urethritis . The findings did not show more chlamydial infection among men who had contact with CSWs than among men who had no such contact . The failure to use condoms among these men must be addressed.

Sex Transm Dis, 2001 Nov, 28(11), 624 - 9
A case-control study of beliefs and behaviors associated with sexually transmitted disease occurrence in Estonia; Wilson TE et al.; BACKGROUND: Epidemiologic data document rapidly increasing sexually transmitted disease (STD) rates throughout Eastern Europe . GOAL: This case-control study was designed to delineate factors contributing to the STD epidemic in Estonia . STUDY DESIGN: For this study, 189 study participants and 112 control subjects completed a behavioral questionnaire and underwent testing for Neisseria gonorrhoeae, Chlamydia trachomatis, and Treponema pallidum . RESULTS: The prevalence of STDs among the control subjects was 32% . Although the participants believed that condoms prevent STDs, only 17% reported consistent use . Methods believed to prevent transmission included washing the genitals (65%), urinating (26%), douching (35%), and using oral contraceptives (19%) . An interaction between sex and travel outside Estonia (odds ratio, 0.1; 95% CI, 0-0.7) reflects the fact that males with STDs were more likely to report travel (46% of participants and 45.5% of control subjects with STD) than were those without STD (16.1% of controls without STD) . CONCLUSIONS: STD rates are related to high-risk sexual behavior among males traveling outside of Estonia . Intervention is needed to promote understanding of disease transmission dynamics in this area, and to decrease sexual risk behavior, particularly in the context of travel.

Ned Tijdschr Geneeskd, 2001 Sep 29, 145(39), 1899 - 900
{Increased Neisseria gonorrhoeae quinolone resistance in Amsterdam}; Peerbooms PG et al.; In addition to a rise in the number of cases of gonorrhoea, the susceptibility of Neisseria gonorrhoeae to antibiotics is also a cause for concern . After a period of high resistance rates to penicillin and tetracycline between 1985 and 1995, resistance rates have dropped considerably in recent years, probably due to changes in treatment regimens . However, recently we have seen an increasing number of quinolone-resistant N . gonorrhoeae isolates in Amsterdam, the Netherlands, a development that has previously been reported in other parts of the world . Some form of national resistance monitoring for gonococci is therefore urgently required to allow timely detection of changes in N . gonorrhoeae resistance.

Kansenshogaku Zasshi, 2001 Sep, 75(9), 819 - 21
{Gonococcal abscess in the prepuce: a case report}; Yamada Y et al.; We reported a case of gonococcal abscess in the prepuce not accompanied with urethritis . As initial therapy fluoroquinolone was ineffective, the abscess spread to the subcutaneous tissue around the penile root . After intravenous administration of cefodizime, the clinical symptoms improved, and Neisseria gonorrhoeae was eradicated.

Transplantation, 2001 Oct 15, 72(7), 1241 - 4
Postmenopausal tubo-ovarian abscess due to Pseudomonas aeruginosa in a renal transplant patient: a case report and review of the literature; El Khoury J et al.; BACKGROUND: Pseudomonas aeruginosa is an uncommon cause of infection in the female genital tract . We report a case of postmenopausal tubo-ovarian abscess (TOA) due to P . aeruginosa in a renal transplant recipient . The presentation included mild abdominal symptoms with rapid progression of peritonitis and surgical abscess drainage . This is the first such case in an organ transplant recipient described in the English literature . METHODS AND RESULTS: Published reports of 1040 cases of TOA were reviewed . The most common features were a history of sexually transmitted disease or pelvic inflammatory disease, and symptoms including abdominal pain and fever . Escherichia coli, Bacteroides spp., and Klebsiella pneumoniae were the most frequently encountered pathogens . Neisseria gonorrhoeae and Chlamydia trachomatis, which are frequently isolated from cervical cultures, are uncommonly isolated from tubo-ovarian abscesses . Forty percent of patients were treated with antibiotics alone, 18.8% with abdominal surgery, and 32% with surgery and antimicrobial therapy . CONCLUSION: This report illustrates the muted presentation and atypical microbiology of gynecologic infection in an organ transplant recipient.

J Infect Dis, 2001 Nov 1, 184(9), 1216 - 9 Epub 2001 Sep 27.
Features of Chlamydia trachomatis and Neisseria gonorrhoeae infection in male Army recruits; Cecil JA et al.; Non-health care-seeking male United States Army recruits were tested for Chlamydia trachomatis (n=2245) and Neisseria gonorrhoeae (n=884), using a urine ligase chain reaction test to determine prevalence and potential risk factors for infection . The prevalence of chlamydial infection was 5.3% . Black race, a new sex partner, a history of trichomonas, and the presence of symptoms were associated with chlamydial infection . The prevalence of N . gonorrhoeae infection was 0.6% . Only a reported history of or positive test for C . trachomatis was associated with gonorrheal infection . Of those testing positive for chlamydia, 14% reported symptoms versus 40% of those with gonorrhea . Younger age was not a predictor of either infection, as has been shown for women . A substantial number of male army recruits are infected with C . trachomatis, but few are infected with N . gonorrhoeae . Screening on the basis of symptoms alone would miss the majority of both infections.

J Bacteriol, 2001 Nov, 183(21), 6394 - 403
Homologues of neisserial heme oxygenase in gram-negative bacteria: degradation of heme by the product of the pigA gene of Pseudomonas aeruginosa; Ratliff M et al.; The oxidative cleavage of heme to release iron is a mechanism by which some bacterial pathogens can utilize heme as an iron source . The pigA gene of Pseudomonas aeruginosa is shown to encode a heme oxygenase protein, which was identified in the genome sequence by its significant homology (37%) with HemO of Neisseria meningitidis . When the gene encoding the neisserial heme oxygenase, hemO, was replaced with pigA, we demonstrated that pigA could functionally replace hemO and allow for heme utilization by neisseriae . Furthermore, when pigA was disrupted by cassette mutagenesis in P . aeruginosa, heme utilization was defective in iron-poor media supplemented with heme . This defect could be restored both by the addition of exogenous FeSO4, indicating that the mutant did not have a defect in iron metabolism, and by in trans complementation with pigA from a plasmid with an inducible promoter . The PigA protein was purified by ion-exchange chromotography . The UV-visible spectrum of PigA reconstituted with heme showed characteristics previously reported for other bacterial and mammalian heme oxygenases . The heme-PigA complex could be converted to ferric biliverdin in the presence of ascorbate, demonstrating the need for an exogenous reductant . Acidification and high-performance liquid chromatography analysis of the ascorbate reduction products identified a major product of biliverdin IX-beta . This differs from the previously characterized heme oxygenases in which biliverdin IX-alpha is the typical product . We conclude that PigA is a heme oxygenase and may represent a class of these enzymes with novel regiospecificity.

J Bacteriol, 2001 Nov, 183(21), 6253 - 64
Gonococcal MinD affects cell division in Neisseria gonorrhoeae and Escherichia coli and exhibits a novel self-interaction; Szeto J et al.; The Min proteins are involved in determining cell division sites in bacteria and have been studied extensively in rod-shaped bacteria . We have recently shown that the gram-negative coccus Neisseria gonorrhoeae contains a min operon, and the present study investigates the role of minD from this operon . A gonococcal minD insertional mutant, CJSD1, was constructed and exhibited both grossly abnormal cell division and morphology as well as altered cell viability . Western blot analysis verified the absence of MinD from N . gonorrhoeae (MinD(Ng)) in this mutant . Hence, MinD(Ng) is required for maintaining proper cell division and growth in N . gonorrhoeae . Immunoblotting of soluble and insoluble gonococcal cell fractions revealed that MinD(Ng) is both cytosolic and associated with the insoluble membrane fraction . The joint overexpression of MinC(Ng) and MinD(Ng) from a shuttle vector resulted in a significant enlargement of gonococcal cells, while cells transformed with plasmids encoding either MinC(Ng) or MinD(Ng) alone did not display noticeable morphological changes . These studies suggest that MinD(Ng) is involved in inhibiting gonococcal cell division, likely in conjunction with MinC(Ng) . The alignment of MinD sequences from various bacteria showed that the proteins are highly conserved and share several regions of identity, including a conserved ATP-binding cassette . The overexpression of MinD(Ng) in wild-type Escherichia coli led to cell filamentation, while overexpression in an E . coli minD mutant restored a wild-type morphology to the majority of cells; therefore, gonococcal MinD is functional across species . Yeast two-hybrid studies and gel-filtration and sedimentation equilibrium analyses of purified His-tagged MinD(Ng) revealed a novel MinD(Ng) self-interaction . We have also shown by yeast two-hybrid analysis that MinD from E . coli interacts with itself and with MinD(Ng) . These results indicate that MinD(Ng) is required for maintaining proper cell division and growth in N . gonorrhoeae and suggests that the self-interaction of MinD may be important for cell division site selection across species.

J Leukoc Biol, 2001 Oct, 70(4), 543 - 50
Identification and comparison of residues critical for cell-adhesion activities of two neutrophil CD66 antigens, CEACAM6 and CEACAM8; Kuroki M et al.; CEACAM6 (CD66c) and CEACAM8 (CD66b) are cell-adhesion proteins on neutrophils that belong to the human carcinoembryonic antigen (CEA) family . CEACAM6 reveals homophilic adhesion and heterophilic adhesion to other CEACAM family antigens including CEACAM8, CEACAM1, and CEA, whereas CEACAM8 exhibits only heterophilic adhesion to CEACAM6 . Here, we investigated and compared structural requirements for the homophilic adhesion of CEACAM6 and heterophilic adhesion between CEACAM6 and CEACAM8 at the amino acid level by using CHO transfectants expressing their mutant and chimeric proteins . The NH(2)-terminal domain (N-domain) of CEACAM6 expressed on a CHO cell was suggested to bind the N-domain of CEACAM6 or CEACAM8 on the opposing cell . By homologue-scanning mutagenesis, we found that the locations of the sequences critical for the adhesion of CEACAM6 to itself and to CEACAM8 are overlapped and that they are highly similar but not identical to the locations of the residues previously shown to be essential for the binding of CEACAM antigens to Opa proteins of pathogenic NEISSERIAE: Our findings imply that subtle differences in the N-domain sequences determine the specificity of the CEACAM antigens on neutrophils for interaction with the same or different CEACAM antigens and the bacterial proteins.

FEMS Microbiol Lett, 2001 Sep 25, 203(2), 173 - 7
Genetic analysis of conservation and variation of lipooligosaccharide expression in two L8-immunotype strains of Neisseria meningitidis; Zhu P et al.; Neisseria meningitidis strains A1 and M978 both express the lipooligosaccharide (LOS) L8 immunotype {Gu et al., J . Clin . Microbiol . 30 (1992) 2047-2053} . Under different growth conditions, strain A1 did not change its LOS profile whereas strain M978 produced variable LOS profiles on SDS-PAGE . To understand the genetic basis of LOS conservation and variation, their lgt locus encoding glycosyltransferases responsible for the biosynthesis of the alpha-chain of LOS was analyzed . Strain A1 possessed only two genes, lgtA and lgtH, at the lgt locus . The lgtA gene was inactivated due to a frameshift mutation; thus strain A1 expressed only L8 LOS . In contrast, strain M978 contained five genes lgtZ, lgtC, lgtA, lgtB and lgtE at this locus, thus it had a potential to express L1, L3,7 in addition to the L8 LOS . The data showed that strain A1 is a better reference strain for the L8 immunotype because of the stability of L8 LOS expression resulting from its unique lgt locus . In addition, these two strains had two new genetic organizations, lgtAH and lgtZCABE, compared to the reported gene organization at the lgt locus in N . meningitidis.

J Cell Biol, 2001 Oct 1, 155(1), 133 - 43
Activation of ErbB2 receptor tyrosine kinase supports invasion of endothelial cells by Neisseria meningitidis; Hoffmann I et al.; ErbB2 is a receptor tyrosine kinase belonging to the family of epidermal growth factor (EGF) receptors which is generally involved in cell differentiation, proliferation, and tumor growth, and activated by heterodimerization with the other members of the family . We show here that type IV pilus-mediated adhesion of Neisseria meningitidis onto endothelial cells induces tyrosyl phosphorylation and massive recruitment of ErbB2 underneath the bacterial colonies . However, neither the phosphorylation status nor the cellular localization of the EGF receptors, ErbB3 or ErbB4, were affected in infected cells . ErbB2 phosphorylation induced by N . meningitidis provides docking sites for the kinase src and leads to its subsequent activation . Specific inhibition of either ErbB2 and/or src activity reduces bacterial internalization into endothelial cells without affecting bacteria-induced actin cytoskeleton reorganization or ErbB2 recruitment . Moreover, inhibition of both actin polymerization and the ErbB2/src pathway totally prevents bacterial entry . Altogether, our results provide new insight into ErbB2 function by bringing evidence of a bacteria-induced ErbB2 clustering leading to src kinase phosphorylation and activation . This pathway, in cooperation with the bacteria-induced reorganization of the actin cytoskeleton, is required for the efficient internalization of N . meningitidis into endothelial cells, an essential process enabling this pathogen to cross host cell barriers.

J Antimicrob Chemother, 2001 Oct, 48(4), 545 - 8
Frequency of selection of fluoroquinolone-resistant mutants of Neisseria gonorrhoeae exposed to gemifloxacin and four other quinolones; Ruiz J et al.; We studied the frequency of mutation of clinical isolates of Neisseria gonorrhoeae (two nalidixic acid susceptible and two nalidixic acid resistant), and the stability of the mutants obtained, in the presence of three different concentrations of five fluoroquinolones . The frequency of mutation was low for all the quinolones . Only one N . gonorrhoeae mutant, obtained with trovafloxacin at 4 x MIC presented a stable increase in the MIC of this quinolone, not attributable to novel mutation(s), both in the gyrA and parC genes, although not showing any increase in the MIC of the other quinolones tested . In summary, gemifloxacin was the only quinolone tested for which resistant mutants were not obtained.

Cell Microbiol, 2001 Oct, 3(10), 681 - 91
CEACAM is not necessary for Neisseria gonorrhoeae to adhere to and invade female genital epithelial cells; Swanson KV et al.; Neisseria gonorrhoeae has a repertoire of up to 11 opacity-associated (Opa) proteins that are adhesins . Most Opa proteins adhere to CEACAM antigens and when CEACAM molecules are present on the surface of transfected epithelial cells their binding by Opa is thought to induce invasion of these cells by gonococci . In this study, we investigated whether several malignant epithelial cell lines, normal cervical and fallopian tube epithelial cell cultures, as well as normal fallopian tube tissue express several of the CEACAM molecules, and whether gonococci use these molecules for adherence and invasion of these female genital epithelial cells . A primary cervical cell culture and metastatic cervical cell line ME180 both expressed CEACAM as shown by whole cell ELISA and flow cytometry, and increased the surface expression of total CEACAM during incubation with Opa+ gonococci . Opa+ gonococci both adhered to and invaded these cells; CEACAM-specific monoclonal antibody (MAb) partially abolished this interaction . Two primary fallopian epithelial tube cell cultures, a primary cervical cell culture and two malignant cell lines, HEC-1-B and HeLa, did not express CEACAM nor was CEACAM mRNA present . No evidence of either intracellular or secreted extracellular CEACAM was found with HEC-1-B and HeLa cells . Opa+ gonococci both adhered to and invaded CEACAM non-expressing cells; however, Opa+ gonococcal association with these non-expressing cell lines could not be inhibited with CEACAM-specific MAb . These data show that CEACAM is not always expressed on female genital epithelial cells and is not essential for gonococcal adherence and invasion . However, when CEACAM is expressed, Opa+ gonococci exploit it for the adherence to and invasion of these cells.

J Microbiol Methods, 2001 Nov, 47(2), 135 - 49
Novel vaccine strategies to T-independent antigens; Lesinski GB et al.; T cell independent antigens do not require T cell help to induce an immune response, and are characterized by a lack of immunologic memory . These antigens can be divided into two classes, TI-1 or TI-2 . TI-1 antigens, such as bacterial lipopolysaccharide, are potent B-cell mitogens, capable of non-specific, polyclonal activation of B cells . In contrast, TI-2 antigens can only activate mature B cells and consist of highly repetitive structures, such as capsular polysaccharides (CPS) from bacteria . Many vaccines currently in use consist of purified capsular polysaccharides from pathogenic bacteria such as Streptococcus pneumoniae and Neisseria meningitidis . These vaccines are efficacious in immune-competent adults, however, due to their TI-2 nature, are not effective in children <2 years of age . Converting polysaccharides into T cell dependent (TD) antigens, allows children, <2, to produce an effective immune response . This review focuses on various strategies used to convert the immune response to polysaccharide antigens from TI-2 to a TD response . Conjugate vaccines, anti-idiotypic antibodies, phage display library technology and DNA vaccines are discussed.

Hautarzt, 2001 Sep, 52(9), 820 - 3
{Recurrent chancriform mucous membrane ulcer in plasmacytoma with secondary IgA deficiency . Pyoderma chancriforme of the tongue}; Hegemann B et al.; A 66 year old patient presented with a nine month history of recurrent oral ulcerations involving the tongue . We diagnosed chancriform pyoderma and a previously not identified multiple myeloma with secondary immunoglobulin deficiency . Clinically and histologically we excluded a necrotizing ulcerative stomatitis as found in individuals with cellular immunodeficiency as in late-stage HIV-infection . On culture only Neisseria catarrhalis was found . Chancriform pyoderma is often associated with local bacterial infections, especially Staphylococcus aureus . The most common sites are the genitalia and periorbital region; involvement of the oral mucosa is uncommon . To the best of our knowledge, this is the fourth reported case with tongue lesions . The multiple myeloma-associated immunoglobulin deficiency might have facilitated the oral manifestations of chancriform pyoderma.

J Comput Biol, 2001, 8(4), 401 - 27
Detection of recombination in DNA multiple alignments with hidden Markov models; Husmeier D et al.; Conventional phylogenetic tree estimation methods assume that all sites in a DNA multiple alignment have the same evolutionary history . This assumption is violated in data sets from certain bacteria and viruses due to recombination, a process that leads to the creation of mosaic sequences from different strains and, if undetected, causes systematic errors in phylogenetic tree estimation . In the current work, a hidden Markov model (HMM) is employed to detect recombination events in multiple alignments of DNA sequences . The emission probabilities in a given state are determined by the branching order (topology) and the branch lengths of the respective phylogenetic tree, while the transition probabilities depend on the global recombination probability . The present study improves on an earlier heuristic parameter optimization scheme and shows how the branch lengths and the recombination probability can be optimized in a maximum likelihood sense by applying the expectation maximization (EM) algorithm . The novel algorithm is tested on a synthetic benchmark problem and is found to clearly outperform the earlier heuristic approach . The paper concludes with an application of this scheme to a DNA sequence alignment of the argF gene from four Neisseria strains, where a likely recombination event is clearly detected.

Clin Infect Dis, 2001 Nov 1, 33(9), 1455 - 61 Epub 2001 Sep 24.
Evaluation of use of a single intravaginal swab to detect multiple sexually transmitted infections in active-duty military women; Rompalo AM et al.; The accuracy and suitability of use of a single intravaginal swab (SIS) for polymerase chain reaction detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, and human papillomavirus infection was assessed in a cross-sectional study of 841 active-duty military women . The SIS, compared with standard diagnostic tests, allowed detection of more gonorrhea, more chlamydial infection, and more trichomoniasis . Sensitivity and specificity of SIS detection compared with adjudicated true-positive diagnoses were 95.8% and 97.8%, respectively, for gonorrhea, 94.6% and 99.3% for chlamydial infection, and 92.2% and 98.2% for trichomonal infection . Results with SISs were comparable to those with cervical swabs tested for human papillomavirus . Assay of clinician-collected and self-collected SISs yielded prevalences similar to those of standard diagnostic tests for all sexually transmitted infections . Therefore, the use of SISs is acceptable for the simultaneous diagnosis of multiple sexually transmitted infections and has potential for use as a self-administered diagnostic tool with widespread applicability among women.

Am J Obstet Gynecol, 2001 Sep, 185(3), 629 - 32
A randomized trial that compared oral cefixime and intramuscular ceftriaxone for the treatment of gonorrhea in pregnancy; Ramus RM et al.; OBJECTIVE: The purpose of this study was to evaluate prospectively the Centers for Disease Control recommendations for the treatment of gonococcal infection in pregnancy . STUDY DESIGN: One hundred sixty-one women who were referred with probable endocervical gonorrhea underwent pretreatment endocervical, anal, and oral cultures for Neisseria gonorrhoeae . The women were randomly assigned to receive ceftriaxone 125 mg intramuscularly or cefixime 400 mg orally . Treatment was open and in a 1:1 distribution . There were 95 evaluable patients . The tests of cure cultures were performed 4 to 10 days after treatment . RESULTS: Eighty-six women (91%) had endocervical infection; 39 women (41%) had anal infection, and 11 women (12%) had pharyngeal infection . Fifty of 95 women (53%) had concomitant endocervical chlamydial infection . The overall efficacy was 91 of 95 subjects (95.8%; 95% CI, 89.6%-98.8%) . Ceftriaxone was effective in 41 of 43 cases (95%; 95% CI, 84.2%-99.4%), and cefixime was effective in 50 of 52 cases (96%; 95% CI, 86.8%-99.5%) . No significant difference was noted in the overall efficacy or by site of infection . Three of the 4 women who experienced treatment failures admitted to unprotected intercourse before their test of cure culture . CONCLUSION: Both intramuscular ceftriaxone 125 mg and oral cefixime 400 mg appear to be effective for the treatment of gonococcal infection in pregnancy.

Biochemistry, 2001 Sep 25, 40(38), 11552 - 8
Crystal structure of heme oxygenase from the gram-negative pathogen Neisseria meningitidis and a comparison with mammalian heme oxygenase-1; Schuller DJ et al.; We report the crystal structure of heme oxygenase from the pathogenic bacterium Neisseria meningitidis at 1.5 A and compare and contrast it with known structures of heme oxygenase-1 from mammalian sources . Both the bacterial and mammalian enzymes share the same overall fold, with a histidine contributing a ligand to the proximal side of the heme iron and a kinked alpha-helix defining the distal pocket . The distal helix differs noticeably in both sequence and conformation, and the distal pocket of the Neisseria enzyme is substantially smaller than in the mammalian enzyme . Key glycine residues provide the flexibility for the helical kink, allow close contact of the helix backbone with the heme, and may interact directly with heme ligands.

Eur J Biochem, 2001 Sep, 268(18), 4908 - 17
Solution structure of the lipoyl domain of the chimeric dihydrolipoyl dehydrogenase P64K from Neisseria meningitidis; Tozawa K et al.; The antigenic P64K protein from the pathogenic bacterium Neisseria meningitidis is found in the outer membrane of the cell, and consists of two parts: an 81-residue N-terminal region and a 482-residue C-terminal region . The amino-acid sequence of the N-terminal region is homologous with the lipoyl domains of the dihydrolipoyl acyltransferase (E2) components, and that of the C-terminal region with the dihydrolipoyl dehydrogenase (E3) components, of 2-oxo acid dehydrogenase multienzyme complexes . The two parts are separated by a long linker region, similar to the linker regions in the E2 chains of 2-oxo acid dehydrogenase complexes, and it is likely this region is conformationally flexible . A subgene encoding the P64K lipoyl domain was created and over-expressed in Escherichia coli . The product was capable of post-translational modification by the lipoate protein ligase but not aberrant modification by the biotin protein ligase of E . coli . The solution structure of the apo-domain was determined by means of heteronuclear NMR spectroscopy and found to be a flattened beta barrel composed of two four-stranded antiparallel beta sheets . The lysine residue that becomes lipoylated is in an exposed beta turn that, from a {1H}-15N heteronuclear Overhauser effect experiment, appears to enjoy substantial local motion . This structure of a lipoyl domain derived from a dihydrolipoyl dehydrogenase resembles that of lipoyl domains normally found as part of the dihydrolipoyl acyltransferase component of 2-oxo acid dehydrogenase complexes and will assist in furthering the understanding of its function in a multienzyme complex and in the membrane-bound P64K protein itself.

Kansenshogaku Zasshi, 2001 Aug, 75(8), 692 - 5
{A case of sepsis by Neisseria meningitidis beginning with pneumonia during a trip abroad}; Shiraishi M et al.; A 62-year-male presented a high fever and a dry cough during a trip to Australia . He was admitted to a hospital as soon as be returned to Japan . The next day after returning to Japan, he was transferred to our hospital with septic shock and loss of consciousness . Neisseria meningitidis was cultured from his blood . N . meningitidis is rare in Japan . However its seems common, in some foreign countries . With these findings, it can be postulated that N . meningitidis might be one of the etiological agents of the imported infectious disease.

Mol Microbiol, 2001 Sep, 41(5), 1199 - 210
Analysis of type I restriction modification systems in the Neisseriaceae: genetic organization and properties of the gene products; Piekarowicz A et al.; The hsd locus (host specificity of DNA) was identified in the Neisseria gonorrhoeae genome . The DNA fragment encoding this locus produced an active restriction and modification (R/M) system when cloned into Escherichia coli . This R/M system was designated NgoAV . The cloned genomic fragment (7800 bp) has the potential to encode seven open reading frames (ORFs) . Several of these ORFs had significant homology with other proteins found in the databases: ORF1, the hsdM, a methylase subunit (HsdM); ORF2, a homologue of dinD; ORF3, a homologue of hsdS; ORF4, a homologue of hsdS; and ORF5, an endonuclease subunit hsdR . The endonuclease and methylase subunits possessed strongest protein sequence homology to the EcoR124II R/M system, indicating that NgoAV belongs to the type IC R/M family . Deletion analysis showed that only ORF3 imparted the sequence specificity of the RM.NgoAV system, which recognizes an interrupted palindrome sequence (GCAN(8-)TGC) . The genetic structure of ORF3 (208 amino acids) is almost identical to the structure of the 5' truncated hsdS genes of EcoDXXI or EcoR124II R/M systems obtained by in vitro manipulation . Genomic sequence analysis allowed us to identify hsd loci with a very high homology to RM.NgoAV in two strains of Neisseria meningitidis . However, significant differences in the organization and structure of the hsdS genes in both these systems suggests that, if functional, they would possess recognition sites that differ from the gonococcus and from themselves.

Infect Immun, 2001 Oct, 69(10), 6419 - 26
Soluble pilin of Neisseria gonorrhoeae interacts with human target cells and tissue; Rytkonen A et al.; Pili of Neisseria gonorrhoeae are phase-variable surface structures that mediate adherence to host target cells . Each pilus is composed of thousands of major pilus subunits, pilins, pilus-associated protein PilC, and possibly other components . Piliated and nonpiliated gonococcal clones may secrete a soluble smaller pilin (S-pilin) that is cleaved after amino acid 39 of the mature pilin protein . Here, purified S-pilin was found to migrate as a 61- to 64-kDa double band on nondenaturing gels, suggesting the formation of tetrameric S-pilin proteins with two isomeric forms . In situ studies of binding to formalin-fixed tissue sections demonstrated the binding of S-pilin to human tissue but not to tissue from mouse or rat organs, showing the presence of a human-specific receptor-binding domain within the pilin polypeptide . Pretreatment of the target tissues with proteinase K decreased gonococcal binding dramatically, whereas pretreatment with neuraminidase and meta-periodate, which cleave carbon-carbon linkages between vicinal hydroxyl groups in carbohydrates, did not affect gonococcal binding . In overlay assays, purified S-pilin bound to a band with a migration pattern and size similar to those of CD46, a cellular pilus receptor . Further, binding of N . gonorrhoeae to target cells and tissues could be blocked by both CD46 antibodies and purified S-pilin . These data argue that S-pilin interacts with a protein domain(s) of the CD46 receptor on human cells.

Infect Immun, 2001 Oct, 69(10), 6336 - 47
Gonococcal genes encoding transferrin-binding proteins A and B are arranged in a bicistronic operon but are subject to differential expression; Ronpirin C et al.; Neisseria gonorrhoeae is capable of utilizing host iron-binding proteins, such as transferrin, lactoferrin, and hemoglobin, as the sole source of iron . The receptor involved in transferrin iron acquisition is composed of two distinct transferrin-binding proteins, TbpA and TbpB . The genes that encode these proteins are linked on the chromosome in the order tbpB-tbpA but are separated by an inverted repeat of unknown function . In this study, we sought to understand the transcriptional organization and regulation of the tbp genes, using a combination of lacZ transcriptional fusion analysis and reverse transcriptase PCR (RT-PCR) . First, we demonstrated that tbpB and tbpA are cotranscribed and coregulated from the common upstream promoter that precedes tbpB . Using beta-galactosidase activity as a surrogate for tbp-specific transcription, we found that tbpB-specific transcripts were more prevalent than tbpA-specific transcripts after 2 h of growth under iron stress conditions . We confirmed the results obtained by fusion analysis by using RT-PCR applied to native RNA isolated from wild-type gonococci . Three different varieties of RT-PCR were employed: relative, competitive, and real time quantitative . The results of all analyses indicated that tbpB-specific transcripts were approximately twofold more prevalent than tbpA-specific transcripts at steady state . In iron-stressed cultures, the ratio of tbpB- to tbpA-specific message was approximately 2; however, in iron-replete cultures, this ratio dropped to 1 . Using these techniques, we also quantitated the effects of iron, external pH, and presence of ligand on tbp mRNA levels.

Cell Microbiol, 2001 Sep, 3(9), 611 - 22
The role of complement receptor 3 (CR3) in Neisseria gonorrhoeae infection of human cervical epithelia; Edwards JL et al.; Neisseria gonorrhoeae is an important sexually transmitted pathogen and a major cofactor in HIV-1 infection . This organism uses different mechanisms to infect male and female genital tract epithelia . Receptor-mediated endocytosis of N . gonorrhoeae is the principle mechanism of entry into male urethral epithelial cells . Infection in men leads to a pronounced inflammatory response . In contrast, N . gonorrhoeae infection in women induces ruffling of the cervical epithelia, allowing a macropinocytic mechanism of entry . Infection in women is frequently asymptomatic, suggesting suppression of the inflammatory response . N . gonorrhoeae-induced membrane ruffling and inflammation suppression are consistent with the ability of this bacterium to enter cervical epithelial cells, in vitro and in vivo, by interaction with complement receptor 3 (CR3), a receptor that does not trigger an inflammatory response . This receptor is present on cervical epithelial cells but not on male urogenital tract epithelia . N . gonorrhoeae engagement of CR3 initiates a unique mechanism of bacterial-induced membrane ruffling and internalization . These studies explain why the pathology of N . gonorrhoeae infection differs between males and females . Additionally, the observation that this receptor is present on cervical epithelia may provide insight into the pathogenesis of other sexually transmitted pathogens.

J Infect Dis, 2001 Oct 1, 184(7), 922 - 5 Epub 2001 Aug 29.
Cleavage of the hormone human chorionic gonadotropin, by the Type 1 IgA1 protease of Neisseria gonorrhoeae, and its implications; Senior BW et al.; The hormone human chorionic gonadotropin (hCG) serves to maintain the fetus during early pregnancy and regulate the onset of labor in late pregnancy . hCG also prevents Neisseria gonorrhoeae from developing invasive characteristics . Part of the beta subunit of hCG has an amino acid sequence similar to that of the hinge of human IgA1, which is the site of action of IgA1 proteases . This study examined the sensitivity of hCG to gonococcal IgA1 proteases, by means of autoradiography, immunoblotting, and RIA . hCG was cleaved in the beta subunit by the type 1 but not the type 2 IgA1 proteases of N . gonorrhoeae . hCG cleavage by gonococcal IgA1 proteases in vivo may increase the invasiveness of the pathogen and destroy its natural biologic activity, with major consequences for the fetus and the pregnancy.

Annu Rev Microbiol, 2001, 55, 673 - 707
Antigenic variation at the infected red cell surface in malaria; Kyes S et al.; Many pathogens that either rely on an insect vector to complete their life cycle (e.g., Trypanosoma spp . and Borrelia spp.) or exist in a unique ecological niche where transmission from host to host is sporadic (e.g., Neisseria spp.) have evolved strategies to maintain infection of their mammalian hosts for long periods of time in order to ensure their survival . Because they have to survive in the face of a fully functional immune system, a common feature of many of these organisms is their development of sophisticated strategies for immune evasion . For the above organisms and for malaria parasites of the genus Plasmodium, a common theme is the ability to undergo clonal antigenic variation . In all cases, surface molecules that are important targets of the humoral immune response are encoded in the genome as multicopy, nonallelic gene families . Antigenic variation is accomplished by the successive expression of members of these gene families that show little or no immunological cross-reactivity . In the case of malaria parasites, however, some of the molecules that undergo antigenic variation are also major virulence factors, adding an additional level of complication to the host-parasite interaction . In this review, we cover the history of antigenic variation in malaria and then summarize the more recent data with particular emphasis on Plasmodium falciparum, the etiological agent of the most severe form of human malaria.

Annu Rev Microbiol, 2001, 55, 561 - 90
Recombination and the population structures of bacterial pathogens; Feil EJ et al.; The population structures of bacterial species are complex and often controversial . To a large extent, this is due to uncertainty about the frequency and impact of recombination in bacteria . The existence of clones within bacterial populations, and of linkage disequilibrium between alleles at different loci, is often cited as evidence for low rates of recombination . However, clones and linkage disequilibrium are almost inevitable in species that divide by binary fission and can be present in populations where recombination is frequent . In recent years, it has become possible to directly compare rates of recombination in different species . These studies indicate that in many bacterial species, including Neisseria meningitidis, Streptococcus pneumoniae, and Staphylococcus aureus, evolutionary change at neutral (housekeeping) loci is more likely to occur by recombination than mutation and can result in the elimination of any deep-rooted phylogenetic signal . In such species, the long-term evolution of the population is dominated by recombination, but this does not occur at a sufficiently high frequency to prevent the emergence of adaptive clones, although these are relatively short-lived and rapidly diversify.

Acta Obstet Gynecol Scand, 2001 Aug, 80(8), 757 - 61
Risk for anogenital cancer and other cancer among women hospitalized with gonorrhea; Johansen C et al.; BACKGROUND: We investigated the relationship between infections with Neisseria gonorrhoeae and anogenital and other cancers . METHODS: Nationwide and population based register linkage study utilizing prospectively notified information . The observed numbers of cancers among the women were compared with those expected on the basis of national incidence rates . RESULTS: In a cohort of 4440 women hospitalized for gonorrhea we observed a total of 227 cases of cervical intraepithelial neoplasia grade III (CIN III), with 103 expected (standardized incidence ratio (SIR), 2.2; 95% confidence interval (CI), 1.9-2.5) . No significantly increased risk for other anogenital cancers or cancer at other sites was seen . CONCLUSIONS: These results support the view that the observed association between gonorrheal infection and subsequent cervical preneoplasia is due mainly to surveillance bias . However, our results also indicate that women hospitalized with a N . gonorrhoeae infection will benefit from the compliance with the regular Pap smear screening programs.

J Adolesc Health, 2001 Sep, 29(3 Suppl), 87 - 92
Correlation between urine and cervical specimens for the detection of cervical Chlamydia trachomatis and Neisseria gonorrhoeae using ligase chain reaction in a cohort of HIV infected and uninfected adolescents; Peralta L et al.; PURPOSE: (a) To examine the concordance between ligase chain reaction (LCR) results from urine and cervical samples for Chlamydia trachomatis and Neisseria gonorrhoeae in HIV infected and uninfected adolescent women, and (b) to examine factors that may influence the concordance of LCR results in this population . METHODS: Baseline specimens from 269 of 334 female subjects enrolled in a longitudinal study of HIV infection in adolescents were analyzed for C . trachomatis and N . gonorrhoeae using ligase chain reaction (LCR) assays in a central laboratory . Concordance was measured using kappa coefficient with permutation analyses to calculate the difference between HIV status groups . Discordant LCR results were examined for the co-infection with the other microorganism, bacterial vaginosis, or Trichomonas vaginalis . RESULTS: The prevalence of C . trachomatis detected by LCR in the HIV infected and uninfected groups was 19.3% and 12.2%, respectively (p = .16); the prevalence of N . gonorrhoeae was 7.0% and 2.4%, respectively (p = .16) . Urine LCR assay sensitivity to detect cervical C . trachomatis infection was 86% (95% CI: 68%-96%) in the HIV infected group and 100% (95% CI: 69%-100%) in the HIV uninfected group . Urine LCR assay sensitivity to detect cervical N . gonorrhoeae infection was 92% (95% CI: 62%-100%) in the HIV infected group . There were only 2 N . gonorrhoeae infections in the HIV uninfected group, and both were urine LCR positive . Differences in sensitivity between HIV infected and HIV uninfected subjects were not statistically significant . Coinfection with N . gonorrhoeae, bacterial vaginosis or Trichomonas vaginalis was not associated with the concordance of urine and cervical LCR results . CONCLUSION: The relatively high sensitivity of urine LCR testing overall suggests that urine screening may be reasonable for sexually active adolescent females with or without HIV infection in situations in which urine screening may be more acceptable than pelvic examinations.

Aust N Z J Public Health, 2001 Aug, 25(4), 368 - 70
C . trachomatis and N . gonorrhoeae surveillance in New Zealand: comparison of laboratory and clinic data; McNicholas AM et al.; OBJECTIVE: To determine the extent to which clinic-based sexually transmitted infection (STI) surveillance underrepresents the number of laboratory-confirmed cases of Chlamydia trachomatis and Neisseria gonorrhoeae in the Waikato and Bay of Plenty regions of New Zealand; and to estimate incidence rates for these two infections . METHODS: Data on C . trachomatis and N . gonorrhoeae were collected from diagnostic laboratories in the study regions for the year 2000, and compared with routine clinic-based STI surveillance data . RESULTS: Most laboratory-confirmed C . trachomatis (65.5%) and N . gonorrhoeae (55.7%) infections were diagnosed by healthcare providers outside the clinic-based STI surveillance system . The estimated incidence rate for C . trachomatis was 501 per 100,000, and 50 per 100,000 for N . gonorrhoeae . CONCLUSIONS AND IMPLICATIONS: Laboratory surveillance of C . trachomatis and N . gonorrhoeae provides a more complete picture of disease burden . Given the high infection rates reported, developing a national strategy for the management of STIs should be a public health priority in New Zealand.

Microbiol Mol Biol Rev, 2001 Sep, 65(3), 353 - 70, table of contents
Sodium ion cycle in bacterial pathogens: evidence from cross-genome comparisons; Hase CC et al.; Analysis of the bacterial genome sequences shows that many human and animal pathogens encode primary membrane Na+ pumps, Na+-transporting dicarboxylate decarboxylases or Na+ translocating NADH:ubiquinone oxidoreductase, and a number of Na+ -dependent permeases . This indicates that these bacteria can utilize Na+ as a coupling ion instead of or in addition to the H+ cycle . This capability to use a Na+ cycle might be an important virulence factor for such pathogens as Vibrio cholerae, Neisseria meningitidis, Salmonella enterica serovar Typhi, and Yersinia pestis . In Treponema pallidum, Chlamydia trachomatis, and Chlamydia pneumoniae, the Na+ gradient may well be the only energy source for secondary transport . A survey of preliminary genome sequences of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Treponema denticola indicates that these oral pathogens also rely on the Na+ cycle for at least part of their energy metabolism . The possible roles of the Na+ cycling in the energy metabolism and pathogenicity of these organisms are reviewed . The recent discovery of an effective natural antibiotic, korormicin, targeted against the Na+ -translocating NADH:ubiquinone oxidoreductase, suggests a potential use of Na+ pumps as drug targets and/or vaccine candidates . The antimicrobial potential of other inhibitors of the Na+ cycle, such as monensin, Li+ and Ag+ ions, and amiloride derivatives, is discussed.

Minerva Ginecol, 2000 Dec, 52(12 Suppl 1), 19 - 24
{Sexually transmitted diseases in adolescence}; De Seta F et al.; BACKGROUND: Sexually transmitted diseases (STD) have their most incidence in adolescence . We conducted an observational study on the symptomatic patients of our STD's office to evaluate epidemiological distribution, behavioral patterns and diffusion of genital infections in teen-agers in respect of the adult population . METHODS: From april 1996 to april 1999 we studied two groups of patients: 54 teen-agers (13-19 years) and 917 women in fertile age (20-40 years) . All the patients underwent to a clinical and microbiological examination of vaginal secretions (Gram slide, wet mount, cultures for bacteria, yeasts, eventually Chlamydia trachomatis, Mycoplasma spp, Neisseria gonorrhoeae, Herpes simplex virus--HSV--) and to an anamnestic and socio-epidemiological questionnaire . RESULTS: The more frequent genital infections among adolescents was vulvovaginal candidiasis (35% vs . 23.9% of adult people) . HSV was found in 1.8% of teen-agers (vs . 0.6%) and when requested, Chlamydia trachomatis and mycoplasmas was found respectively in 16.6% (vs . 1.1%) and 50% (vs . 28%) in adolescents . We observed a high percentage of pregnancy (22.2%) of voluntary pregnancy interruption (41.6%) and a specific sexual behavior among teenagers . CONCLUSIONS: Our data show a particular behavioral risk pattern in the adolescence population and indicate the necessity of a serious program of prevention and sexual education that allow young people to live with more consciousness and safety their sexuality.

J Clin Microbiol, 2001 Sep, 39(9), 3092 - 8
Enhancing the specificity of the COBAS AMPLICOR CT/NG test for Neisseria gonorrhoeae by retesting specimens with equivocal results; Van Der Pol B et al.; The COBAS AMPLICOR CT/NG test for Neisseria gonorrhoeae cross-reacts with certain strains of nonpathogenic Neisseria species . In some strains, the target sequence is identical to that of N . gonorrhoeae, whereas other strains have a small number of mismatches within the regions recognized by the primers or probe used in the COBAS AMPLICOR NG test . These cross-reactive strains are occasionally present in urogenital specimens, causing false-positive results in the COBAS AMPLICOR NG test . Analysis of the data generated in a large multicenter clinical trial showed that 2.9% of the specimens gave signals between A(660)s of 0.2 and 3.5 but that one-half of these equivocal specimens did not contain N . gonorrhoeae . Most of these equivocal specimens were correctly classified as true positive or true negative by retesting in duplicate and defining a PCR-positive result as two of three results with an A(660) of > or =2.0 . If specimens had been classified as positive or negative based on a single test result using a cutoff of an A(660) of 0.2, specificity would have ranged from 96.2 to 98.9% depending on specimen type, sex, and presence of symptoms . By employing the equivocal zone-retesting algorithm, specificity increased to 98.6 to 99.9% with little effect (0.1 to 4.9% decrease) on sensitivity in most specimen types, enabling the test to achieve a positive predictive value of at least 90% in populations with a prevalence of 4% or higher . In lower-prevalence populations, the test could be used to screen for presumptive infections that would have to be confirmed by an independent test.

Arch Pediatr Adolesc Med, 2001 Aug, 155(8), 947 - 53
Subsequent sexually transmitted infection in urban adolescents and young adults; Orr DP et al.; OBJECTIVE: To compare the rates of subsequent infection with Chlamydia trachomatis, Neisseria gonorrhoeae, or Trichomonas vaginalis in a group of high-risk adolescents and young adults . METHODS: At the time of treatment, 444 unmarried teenagers and young adults aged 13 to 25 years were enrolled from an urban sexually transmitted disease clinic and 3 community-based primary care clinics . Subjects were infected with C trachomatis, N gonorrhoeae, or T vaginalis, were diagnosed as having nongonococcal urethritis (in men), or were uninfected sexual contacts with one of these infections . Subjects returned at 1, 3, 5, and 7 months . RESULTS: The rate of subsequent infection was substantial . Forty percent of men and 53% of women who were uninfected contacts at enrollment were estimated to be infected within 7 months; 60% of men and 73% of women infected at enrollment were estimated to be reinfected . Among women, subjects who were infected at enrollment had a shorter time to subsequent infection (median, 140 days) compared with uninfected contacts (median, 209 days) (P =.04) . Among men, findings were similar, but the difference in median time to subsequent infection was not significant (P =.08) . Baseline characteristics that predicted shorter time to reinfection were female sex and infection at enrollment . When sexual behaviors in the 2 months preceding each subsequent data collection visit were included in the model, only being female and reporting at least one new interval sexual partner were significant predictors of subsequent sexually transmitted infections . CONCLUSIONS: These data support recent research that has found high rates of subsequent infection among high-risk adolescents and young adults . Contacts of a sexually transmitted infection appear to be at equally high risk for subsequent infection as those with a personal history of infection . Our data suggest that more frequent than annual screening for N gonorrhoeae, C trachomatis, and T vaginalis would be appropriate in at-risk adolescent and young adult populations, including individuals who are uninfected sexual contacts to a sexually transmitted infection.

Kansenshogaku Zasshi, 2001 Jul, 75(7), 551 - 6
{Drug susceptibility of clinically isolated strains of Neisseria gonorrhoeae}; Murakami H et al.; We investigated the drug susceptibilities of Neisseria gonorrhoeae by using 98 strains of clinical isolates at Toho University Omori Hospital from 1994 to 1998 . Minimum Inhibitory Concentrations (MICs) of 15 antimicrobial agents were determined with agar dilution methods according to the guidelines of NCCLS . Among these isolates, only 4 strains (4.1%) were found to be penicillinase producing N . gonorrhoeae . Ceftriaxone showed the most potent activity of which MICs of all strains were 0.06 microgram/ml or less . Macroride antimicrobial agents and minocycline also showed strong activities of which MICs of most of the strains were 0.06 microgram/ml or less . With the criteria of NCCLS, 10 strains (10.2%) were found to be resistant to ciprofloxacin and these 10 strains also showed cross resistance to other fluoroquiolones we tested . Our results also revealed that the number of resistant strains against fluoroquiolones abruptly increased from 1996 and indicate the needs of further surveillance of antimicrobial resistance in clinical isolates of N . gonorrhoeae.

J Endotoxin Res, 2000, 6(5), 411 - 5
Differential roles of TLR2 and TLR4 in the host response to Gram-negative bacteria: lessons from a lipopolysaccharide-deficient mutant of Neisseria meningitidis; Ingalls RR et al.; The inflammatory response to bacterial infections plays an important role in the detection and elimination of invading micro-organisms . Various components of the bacterial cell wall are capable of activating this pro-inflammatory response . In the case of Gram-negative bacteria, lipopolysaccharide (LPS) is the dominant trigger, although other bacterial factors are also capable of activating this systemic inflammatory response . Recently, Toll-like receptors (TLRs) have been implicated in host responses to bacterial pathogens . Specifically, TLR4 mediates LPS responses while TLR2 plays a broader role in the recognition of a variety of bacteria and bacterial antigens . The experiments in this study were designed to examine the role of Gram-negative cell wall components, other than LPS, and their cellular receptors in the host response to infection using an LPS-deficient mutant of Neisseria meningitidis . Although less potent than the parental strain, we found the LPS-deficient mutant to be a capable inducer of the inflammatory response in a variety of cell types . Moreover, cellular activation by this mutant required expression of CD14 and TLR2.

Sex Transm Dis, 2001 Sep, 28(9), 539 - 45
Antimicrobial susceptibilities of Neisseria gonorrhoeae in Kigali, Rwanda, and trends of resistance between 1986 and 2000; Van Dyck E et al.; BACKGROUND: Plasmid-mediated and chromosomal-mediated resistance of Neisseria gonorrhoeae to penicillin, tetracycline, thiamphenicol, and trimethoprim-sulfamethoxazole has spread dramatically in Africa . Monitoring of antimicrobial susceptibility is a key element in the control of sexually transmitted diseases . GOAL: To document antimicrobial susceptibilities of gonococci isolated during the past 15 years in Kigali, Rwanda . STUDY DESIGN: Minimal inhibitory concentrations of recently collected gonococcal isolates of eight antimicrobials were determined . The results were compared with data collected for isolates obtained since 1986 . RESULTS: In 1986, 35% of the gonococcal isolates were penicillinase-producing N gonorrhoeae . Tetracycline-resistant N gonorrhoeae appeared in 1989 . The prevalence of penicillinase-producing N gonorrhoeae and tetracycline-resistant N gonorrhoeae increased significantly to 70.5% and 89.2%, respectively . Chromosomal resistance to penicillin, tetracycline, and thiamphenicol increased temporarily, then decreased significantly . Chromosomal resistance to trimethoprim-sulfamethoxazole appeared in 1988 and increased to 21.6% . All the isolates were susceptible to ceftriaxone, ciprofloxacin, spectinomycin, and kanamycin . CONCLUSIONS: This study illustrated the rapidly increasing frequencies of penicillinase-producing N gonorrhoeae and tetracycline-resistant N gonorrhoeae . Chromosomal resistance to thiamphenicol and trimethoprim-sulfamethoxazole excludes these drugs as alternative treatment . Programs for antimicrobial susceptibility surveillance of N gonorrhoeae should urgently be established in Africa.

Sex Transm Dis, 2001 Sep, 28(9), 521 - 6
Reduced susceptibility to azithromycin and high percentages of penicillin and tetracycline resistance in Neisseria gonorrhoeae isolates from Manaus, Brazil, 1998; Dillon JA et al.; BACKGROUND: The identification of Neisseria gonorrhoeae isolates resistant to antimicrobial agents currently recommended for the treatment of gonococcal infections continues to escalate globally . Thus, in some areas, resistance to fluoroquinolone drugs is commonplace; several reports document resistance to third-generation cephalosporins, and the sporadic isolation of spectinomycin-resistant isolates continues unabated . Gonococcal resistance to azithromycin, an antibiotic used for the primary treatment of gonococcal infections in some Latin American countries, also has been described . Because the prevalence of resistant isolates is insufficiently documented in many areas of Latin America, the efficacy of locally recommended therapies for gonococcal infections is often unknown . GOAL: To determine the antimicrobial susceptibility and strain types of N gonorrhoeae isolates collected in Manaus, Brazil . These data will establish antimicrobial susceptibility baseline data for the region as a reference point for future surveillance . STUDY DESIGN: Consecutive N gonorrhoeae isolates from urethral and endocervical specimens were collected and examined for identity, antimicrobial susceptibility, and strain type (plasmid content, tetM type, auxotype, and serovar) . RESULTS: Most of the isolates (65/81; 85.2%) were resistant to tetracycline, penicillin, or both, with the majority (n = 62) carrying plasmid-mediated resistance to tetracycline (tetracycline-resistant N gonorrhoeae {TRNG}) . All of the TRNG contained the Dutch-type tetM plasmid, and 18 were A/S class NR/IA-02 . Penicillinase-producing N gonorrhoeae comprised 8.2% (7/81) of the isolates . Of these seven isolates, four also were TRNG, and two carried chromosomal resistance to tetracycline . The isolates were susceptible to ciprofloxacin, spectinomycin, and ceftriaxone . However, 23 isolates were characterized by reduced susceptibility to azithromycin (MIC, 0.25-0.5 microg/ml), and one isolate had reduced susceptibility to ciprofloxacin (MIC, 0.25 microg/ml) . CONCLUSIONS: This study supports the continued use of third-generation cephalosporins, spectinomycin, and fluoroquinolone drugs for the primary treatment of gonococcal infections in Manaus . The occurrence of isolates with reduced susceptibility to azithromycin and ciprofloxacin underscores the importance of ongoing antimicrobial susceptibility monitoring to support decisions regarding appropriate drugs for the treatment of gonococcal infections.

Sex Transm Dis, 2001 Sep, 28(9), 508 - 14
Antimicrobial susceptibility of Neisseria gonorrhoeae isolates from three Caribbean countries: Trinidad, Guyana, and St . Vincent; Dillon JA et al.; BACKGROUND: The percentage of Neisseria gonorrhoeae isolates resistant to antimicrobial agents commonly used for treatment is unknown in many Caribbean countries . GOAL: To determine the antimicrobial susceptibility of N gonorrhoeae isolates from Trinidad (144 isolates), Guyana (70 isolates), and St . Vincent (68 isolates) so baseline data can be established for further studies, and to assist in establishing effective treatment guidelines . STUDY DESIGN: Consecutive urethral and endocervical specimens from several clinics were collected and identified as N gonorrhoeae . Isolates of N gonorrhoeae were tested for their susceptibility to penicillin, tetracycline, ceftriaxone, ciprofloxacin, spectinomycin, and azithromycin . The presumptive identification of penicillinase-producing N gonorrhoeae and/or tetracycline-resistant N gonorrhoeae isolates based on MIC was confirmed by plasmid and tetM content analysis . RESULTS: High percentages of penicillin and/or tetracycline resistance were observed in N gonorrhoeae isolates from Guyana (92.9%), St . Vincent (44.1%), and Trinidad (42.4%) . Isolates from all three countries were susceptible to ceftriaxone, ciprofloxacin, and spectinomycin . One penicillinase-producing N gonorrhoeae/tetracycline-resistant N gonorrhoeae from Guyana had an MIC of 0.5 microg/l to ciprofloxacin . This and nine other isolates from Guyana also were resistant to azithromycin (defined as MIC > or = 2.0 microg/ml) as well as penicillin and tetracycline . A reduced susceptibility to azithromycin was displayed by 16% of the isolates from St . Vincent and 72% of the isolates from Guyana (MIC, 0.25-1.0 microg/ml) . Most penicillinase-producing N gonorrhoeae isolates carried Africa-type plasmids (61/90), with 28 of 90 having Toronto-type plasmids and a single isolate carrying an Asia-type plasmid . The tetM determinant in tetracycline-resistant N gonorrhoeae isolates was predominantly of the Dutch type (68/91) . CONCLUSIONS: The high prevalence of N gonorrhoeae isolates from 3 of 21 English- and Dutch-speaking Caricom countries in the Caribbean with either plasmid-mediated or chromosomal resistance to penicillin and tetracycline supports international observations that these drugs should not be used to treat gonococcal infections . The detection of isolates with reduced susceptibility to drugs such as azithromycin, which currently are recommended for treatment in the region, attest to the importance of the continued monitoring of gonococcal antimicrobial susceptibility for the maintenance of effective treatment guidelines.

Int J Antimicrob Agents, 2001 Aug, 18(2), 113 - 9
Mechanisms other than penicillin-binding protein-2 alterations may contribute to moderate penicillin resistance in Neisseria meningitidis; Orus P et al.; Penicillin resistance in Neisseria spp is thought to be generated by the interspecies transfer of genetic material from naturally penicillin-resistant, commensal species . We examined a series of successive transformants with increasing levels of penicillin resistance, obtained by co-cultivation of Neisseria meningitidis derivatives with Neisseria polysaccharea . Our results suggest that, in addition to the well-known decrease in penicillin affinity of penicillin-binding protein-2 (PBP-2), decreased expression of the class 3 porin as well as decreased affinity of PBP-1 may contribute to higher level resistance of N . meningitidis to penicillin G and other beta-lactam compounds.

Infect Dis Obstet Gynecol, 2001, 9(3), 143 - 6
Prevalence of sexually transmitted infections (Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and human papillomavirus) in female attendees of a sexually transmitted diseases clinic in Ulaanbaatar, Mongolia; Garland SM et al.; BACKGROUND: Epidemiological data suggest that the prevalence of syphilis, gonorrhea and trichomoniasis has increased in both urban and rural areas of Mongolia . These data are primarily substantiated by notifications of cases of clinically apparent disease in both rural and urban areas, plus laboratory diagnoses from the AIDS/STD Reference Center, Ulaanbaatar . In the past 5 years, however, there has been a marked decline in the total number of patients being screened for sexually transmitted infections (STIs) . An assessment of true prevalence of STIs in a female population attending an urban sexually transmitted diseases (STD) clinic was therefore commenced . METHODS: Consecutive women attending an STD clinic in Ulaanbaatar had genital samples collected by the insertion and immediate removal of a tampon, which was then tested for the presence of Neisseria gonorrhoeae, Chlamydia trachomatis, human papillomavirus (HPV) and Trichomonas vaginalis, using polymerase chain reaction (PCR) amplification . RESULTS: A total of 110 women were studied (mean age 26.7 years) . Overall, 58 (53%) patients had one or more pathogens identified; 43 (39%) had a single pathogen, while 15 (14%) had mixed pathogens . C . trachomatis was found in 15 (14%), N . gonorrhoeae in 12 (11%), T . vaginalis in nine (8%) and HPV in 39 (36%) . Among the 39 HPV-positive patients, oncogenicgenotypes (16, 18, 31, 33, 35, 39, 45, 51, 52) were found in 17(44%) patients . CONCLUSIONS: Sexually transmitted infections as defined by PCR were common, and found in 53% of female attendees of an urban STD clinic in Mongolia . As infections with conventional STIs increase the risk of human immunodeficiency virus (HIV) transmission, it is imperative that strategies be introduced to reduce the prevalence of STIs . Furthermore, detection of oncogenic HPV was common, indicating that it is vital that a strategy to reduce cervical cancer such as a pre-cancer cervical cytology screening program also be introduced.

Antimicrob Agents Chemother, 2001 Sep, 45(9), 2651 - 4
Multidrug-resistant strains of Neisseria gonorrhoeae in Greece; Mavroidi A et al.; Eighty-seven out of 575 gonococci isolated in Greece from 1991 to 1998 belonged to serovar Bropyst and exhibited resistance to penicillin, tetracycline, erythromycin, and chloramphenicol . Conventional and molecular typing showed three clusters, A, B, and C, that were associated with networks of high- frequency transmitters (cluster A with homosexuals and clusters B and C with refugees from Eastern Europe) . Study of one isolate revealed mutations in the penA, mtrR, and porB genes that may explain the multidrug-resistant phenotype.

Infect Immun, 2001 Sep, 69(9), 5840 - 8
Distinct proinflammatory host responses to Neisseria gonorrhoeae infection in immortalized human cervical and vaginal epithelial cells; Fichorova RN et al.; In this study we utilized immortalized morphologically and functionally distinct epithelial cell lines from normal human endocervix, ectocervix, and vagina to characterize gonococcal epithelial interactions pertinent to the lower female genital tract . Piliated, but not nonpiliated, N . gonorrhoeae strain F62 variants actively invaded these epithelial cell lines, as demonstrated by an antibiotic protection assay and confocal microscopy . Invasion of these cells by green fluorescent protein-expressing gonococci was characterized by colocalization of gonococci with F actin, which were initially detected 30 min postinfection . In all three cell lines, upregulation of interleukin 8 (IL-8) and IL-6, intercellular adhesion molecule 1 (CD54), and the nonspecific cross-reacting antigen (CD66c) were detected 4 h after infection with piliated and nonpiliated gonococci . Furthermore, stimulation of all three cell lines with gonococcal whole-cell lysates resulted in a similar upregulation of IL-6 and IL-8, confirming that bacterial uptake is not essential for this response . Increased levels of IL-1 were first detected 8 h after infection with gonococci, suggesting that the earlier IL-8 and IL-6 responses were not mediated through the IL-1 signaling pathway . The IL-1 response was limited to cultures infected with piliated gonococci and was more vigorous in the endocervical epithelial cells . The ability of gonococci to stimulate distinct proinflammatory host responses in these morphologically and functionally different compartments of the lower female genital tract may contribute directly to the inflammatory signs and symptoms characteristic of disease caused by N . gonorrhoeae.

Microbiology, 2001 Aug, 147(Pt 8), 2333 - 40
Inverse relationship between pilus-mediated gonococcal adherence and surface expression of the pilus receptor, CD46; Tobiason DM et al.; Pilus-mediated adherence to mucosal epithelial cells is a critical step for Neisseria gonorrhoeae to establish an infection in the human host . CD46, the defined receptor for the gonococcal pilus, is a complement-regulatory protein that is expressed on all human nucleated cells . It was observed that a piliated, Opa(-) variant of gonococcal strain FA1090 adhered with different efficiencies to the human epithelial cell lines tested (Chang, ME180, HEC-1B and PC-3) . Surprisingly, these differences in adherence levels did not correlate with levels of CD46 expressed by these cell lines . In fact, there was an inverse relationship between total surface-exposed CD46 and gonococcal adherence . Four major isoforms of CD46 are produced due to alternative RNA splicing of a surface-exposed region and the cytoplasmic tail . The relative isoform surface expression of each cell line was determined, and each was found to express different ratios of the four CD46 isoforms . No correlation could be derived between CD46 isoform surface expression and pilus-mediated gonococcal adherence, indicating that CD46 does not act as a classic receptor for gonococcal pili.

Microbiology, 2001 Aug, 147(Pt 8), 2321 - 32
Comparative whole-genome analyses reveal over 100 putative phase-variable genes in the pathogenic Neisseria spp; Snyder LA et al.; Previously, a complete genome analysis of Neisseria meningitidis strain MC58 revealed the largest repertoire of putative phase-variable genes described in any species to date . Initial comparisons with two incomplete Neisseria spp . genome sequences available at that time revealed differences in the repeats associated with these genes in the form of polymorphisms, the absence of the potentially unstable elements in some alleles, and in the repertoire of the genes that were present . Analyses of the complete genomes of N . meningitidis strain Z2491 and Neisseria gonorrhoeae strain FA1090 have been performed and are combined with a comprehensive comparative analysis between the three available complete genome sequences . This has increased the sensitivity of these searches and provided additional contextual information that facilitates the interpretation of the functional consequences of repeat instability . This analysis identified: (i) 68 phase-variable gene candidates in N . meningitidis strain Z2491, rather than the 27 previously reported; (ii) 83 candidates in N . gonorrhoeae strain FA1090; and (iii) 82 candidates in N . meningitidis strain MC58, including an additional 19 identified through cross-comparisons with the other two strains . In addition to the 18 members of the opa gene family, a repertoire of 119 putative phase-variable genes is described, indicating a huge potential for diversification mediated by this mechanism of gene switching in these species that is central to their interactions with the host and environmental transitions . Eighty-two of these are either known (14) or strong (68) candidates for phase variation, which together with the opa genes make a total of 100 identified genes . The repertoires of the genes identified in this analysis diverge from the different species groupings, indicating horizontal exchange that significantly affects the species and strain complements of these genes.

Am J Obstet Gynecol, 2001 Jul, 185(1), 163 - 5
Midtrimester vaginal Mycoplasma genitalium in women with subsequent spontaneous preterm birth; Lu GC et al.; OBJECTIVE: We sought to determine the midtrimester prevalence of Mycoplasma genitalium in women who had subsequent spontaneous preterm birth . STUDY DESIGN: In a prospective study of lower genital tract infections, we identified 127 women who subsequently had spontaneous preterm birth . Vaginal samples were obtained between 21 and 25 weeks' gestation for pH, for bacterial vaginosis Gram stain, and cultures that yielded Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis . M genitalium was identified by using validated polymerase chain reaction (PCR) primers, and the results were compared to pregnancy outcomes . RESULTS: Of 124 women with spontaneous preterm births, only five (3.9%) had PCR assays positive forM genitalium . The mean +/- SD delivery gestational age was similar for women with a positive PCR (34.6 +/- 2.2 weeks) and a negative PCR (34.0 +/- 2.7 weeks) (P =.62) . None of the women with positive PCR results tested positive for any other sexually transmitted disease, whereas 36 (30%) women with negative PCR results tested positive . CONCLUSIONS: The occurrence of M genitalium in the vagina at midtrimester is infrequent in women with subsequent spontaneous preterm birth.

J Clin Microbiol, 2001 Aug, 39(8), 2958 - 60
Efficacy of a swab transport system in maintaining viability of Neisseria gonorrhoeae and Streptococcus pneumoniae; Farhat SE et al.; The efficacy of swab transport systems in maintaining viability of Neisseria gonorrhoeae and Streptococcus pneumoniae is crucial both for establishing definitive diagnosis and for monitoring emerging resistance . We tested the efficacy of a newly modified Amies charcoal swab transport system, the StarSwab SP131X (Starplex Scientific, Inc., Etobicoke, Ontario, Canada), by using a combined total of 31 clinical and American Type Culture Collection stock reference strains of N . gonorrhoeae and S . pneumoniae in 46 suspensions of concentrations ranging from 10(5) to 10(8) CFU/ml . Triplicate swabs per strain held at room temperature for 0, 24, and 48 h were plated without prior vortexing, and their growths were graded . All 31 strains were viable at 0 and 24 h . Gonococcal viability at 48 h varied considerably, even among strains with comparable inoculum sizes, suggesting that viability might be strain dependent and confirming the different structural and growth profiles of gonococcal strains . S . pneumoniae strains showed consistent viability, with all strains recovered at all holding periods . This study demonstrates that the StarSwab SP131X is capable of maintaining the viability of N . gonorrhoeae and S . pneumoniae for at least 24 and 48 h, respectively, and reinforces the need for adequate sampling and for timely processing of specimens to maintain optimum performance.

J Clin Microbiol, 2001 Aug, 39(8), 2946 - 50
Detection of cervical infections in colposcopy clinic patients; Lanham S et al.; The purpose of this study was to determine if Neisseria gonorrhoeae; Chlamydia trachomatis; herpes simplex virus; cytomegalovirus; Epstein-Barr virus; human herpesviruses 6, 7, and 8; or adeno-associated virus influenced the production of cervical intraepithelial neoplasia . Two hundred thirty-one cervical smear samples were tested for the presence of the organisms by PCR . In addition, human papillomavirus types in the samples were determined by PCR and classified into cancer risk types of high, moderate, and low . There was no link with cervical intraepithelial neoplasia status and detection of herpes simplex virus, cytomegalovirus, Epstein-Barr virus, human herpesviruses 6 and 8, gonorrhea, or chlamydia . However, high-grade cervical intraepithelial neoplasia was found more frequently with mixed infection by moderate-risk human papillomavirus types and human herpesvirus 7 than with these papillomavirus types alone . The presence of human herpesvirus 7 may increase the oncogenic potential of moderate-risk human papillomavirus types.

J Clin Microbiol, 2001 Aug, 39(8), 2928 - 32
Evaluation of COBAS AMPLICOR (Roche): accuracy in detection of Chlamydia trachomatis and Neisseria gonorrhoeae by coamplification of endocervical specimens; Livengood CH 3rd et al.; We evaluated further the accuracy of the COBAS AMPLICOR (Roche) (CA) PCR-based system in detection of Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical specimens . Endocervical specimens collected for any indication for testing for C . trachomatis and N . gonorrhoeae among a university hospital health system population were included . Testing for C . trachomatis was done by two PCR methods, CA and manual microwell AMPLICOR (Roche) (MWA), and by culture; testing for N . gonorrhoeae was done by CA and culture . Discrepancy resolution was performed . Reproducibility testing and hands-on labor time measurements for CA were done . Among 654 C . trachomatis samples, the prevalence of true positivity was 9.2%, and among the 618 N . gonorrhoeae samples, the prevalence of true positivity was 4.4% . For detection of C . trachomatis, the sensitivity, specificity, and negative and positive predictive values were, respectively, as follows for each test: CA, 93.3, 99.7, 99.3, and 96.4%; MWA, 91.7, 99.7, 99.2, and 96.5%; and culture, 65.0, 100, 96.6, and 100% . For detection of N . gonorrhoeae those values were as follows: CA, 96.3, 100, 99.8, and 100%; and culture, 92.6, 100, 99.7, and 100% . Hands-on labor time for each clinical result was estimated to be at 7.5 min . The prevalence of inhibitory specimens was 3.5%, including two positive C . trachomatis samples which would have been missed otherwise . The direct cost of each clinical result with CA was estimated to be $9.09 . Our methods include a diverse range of indications for testing among women, using endocervical swabbing samples, 2 M sucrose phosphate transport medium, and discrepancy resolution for comparison . Under our test conditions, the CA system is an accurate, rapid, and cost- and labor-efficient method for detection of C . trachomatis and N . gonorrhoeae.

Sex Transm Dis, 2001 Aug, 28(8), 468 - 71
Validity of self-reported sexually transmitted diseases among African American female adolescents participating in an HIV/STD prevention intervention trial; Harrington KF et al.; BACKGROUND: Studies assessing the validity attributed to self-reported measures of sexually transmitted diseases (STDs) clearly are needed, particularly those used for high-risk populations such as female adolescents, in whom STD prevention is a priority . GOAL: To determine the accuracy of self-reported STD test results in female adolescents over a relatively brief period ( approximately 28 days) . STUDY DESIGN: A prospective, randomized, controlled clinical trial of STD/HIV prevention for African American females, ages 14 to 18, was conducted . Study participants were recruited from medical clinics and school health classes in low-income neighborhoods of Birmingham, Alabama, that had high rates of unemployment, substance abuse, violence, STDs, and teenage pregnancy . RESULTS: Of the 522 adolescents enrolled in the trial, 92% (n = 479) completed baseline STD testing and follow-up surveys . At baseline, 28% had positive test results for at least one disease: 4.8% for Neisseria gonorrhoeae, 17.1% for Chlamydia trachomatis, and 12.3% for Trichomonas vaginalis . Of the adolescents with negative STD test results, 98.8% were accurate in their self-report of STD status, as compared with 68.7% of the adolescents with positive results . Underreporting varied by type of STD . Adolescents who accurately reported their positive STD status were significantly more likely to report their receipt of treatment accurately (P < 0.001) . CONCLUSIONS: The substantial underreporting of STD incidence in this study suggests that reliance on self-reports of STD history may introduce misclassification bias, potentially leading to false conclusions regarding the efficacy of prevention interventions . This observation highlights the importance of using biologic indicators as outcome measures.

Sex Transm Dis, 2001 Aug, 28(8), 437 - 43
Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae among at-risk women, young sex workers, and street youth attending community organizations in Quebec City, Canada; Poulin C et al.; BACKGROUND: Despite a relatively recent decline in the global incidence of Chlamydia trachomatis and Neisseria gonorrhoeae it seems that some segments of the population such as street youth, sex workers, and individuals with social problems or delinquent behavior could be part of a core group for STDs . These persons may be reluctant to undergo STD diagnosis in traditional medical settings . GOALS: To determine the prevalence of C trachomatis and N gonorrhoeae infection using polymerase chain reaction on urine samples among subjects attending an anonymous HIV testing clinic and four community organizations in Quebec City, and to identify associated risk factors . STUDY DESIGN: A cross-sectional study of 626 street youth, sex workers, and women with social problems or delinquent behavior was conducted . RESULTS: The prevalences of N gonorrhoeae and C trachomatis were, respectively, 1.1% (95% CI, 0.5%--2.3%) and 5.8% (95% CI, 4.1%--7.9%) . No significant difference was found between men and women, but the sexually transmitted disease (STD) prevalence was much higher in subjects younger than 20 years: 11.4% versus 3.6% (P < 0.01) . In a logistic regression model, factors independently associated with STD infection were age younger than 20 years (OR, 2.6; P = 0.007), occasional sex partners (OR, 2.9; P = 0.007), and injection of drugs (OR, 2.8; P = 0.002) in the preceding 6 months . CONCLUSIONS: A moderate STD prevalence was found in the study population . The prevalence, however, can be considered high (>10%) among street youth and young sex workers . Providing community-based STD screening and treatment services appear to be an efficient method for reaching these high-risk groups.

J Infect Dis, 2001 Aug 15, 184(4), 460 - 72 Epub 2001 Jul 25.
Invasion of human fallopian tube epithelium by Escherichia coli expressing combinations of a gonococcal porin, opacity-associated protein, and chimeric lipo-oligosaccharide; Gorby GL et al.; The transepithelial migration of Escherichia coli that expressed all possible combinations of a plasmid-encoded gonococcal porin (Por), opacity-associated protein (Opa), and 3F11 lipo-oligosaccharide (LOS) epitope was investigated . Surface expression of Por mediated selective changes in E . coli antibiotic susceptibility, and coexpression of Opa and the 3F11 LOS epitope mediated bacterial clumping (P<.01) . In the human fallopian tube organ-culture model, Opa-producing variants attached up to 44-fold better than control bacteria (P<.01), and Por-producing variants exceeded submucosal invasion of control bacteria by 500-fold (P<.01) . Opa and Por each facilitated intracellular invasion 20-40-fold (P<.01) . In dual expresser variants, the 3F11 LOS epitope markedly reduced attachment and invasion mediated by Opa or Por . The LOS inhibitory effect was curbed when all 3 factors were expressed, which suggests an additional interaction of the 3 factors at the bacterial surface . Por, Opa, and LOS play important roles in Neisseria gonorrhoeae trafficking across human fallopian tube epithelium.

Melanoma Res, 2001 Jun, 11(3), 219 - 27
A purified GM3 ganglioside conjugated vaccine induces specific, adjuvant-dependent and non-transient antitumour activity against B16 mouse melanoma in vitro and in vivo; Carr A et al.; The presence of substantial amounts of GM3 ganglioside on human melanomas and other tumours, together with its peculiar biological properties, makes this glycolipid a unique target for cancer immunotherapy . B16 mouse melanoma expresses GM3 and constitutes an appropriate model for the development of novel GM3-based vaccines . Recently, we hydrophobically incorporated purified GM3 into the outer membrane protein complex from Neisseria meningitidis to form very small size proteoliposomes (GM3/VSSP) . We have examined the antitumour properties of GM3/VSSP vaccine and compared it with GM3 incorporated in very low density serum lipoproteins (GM3/VLDL) . Immunization with four doses of GM3/VSSP vaccine (120 microg of ganglioside) plus Freund's adjuvant or Montanide ISA 51 significantly increased the overall survival of mice inoculated in the subcutis with 103 B16-F1 cells, whereas the GM3/VLDL immunogen was ineffective . The non-transient character of tumour protection was confirmed in animals surviving the first challenge and re-inoculated with 5 x 103 cells . GM3/VSSP vaccine also reduced the subcutaneous growth of highly aggressive B16-F10 cells . The importance of ganglioside structure in the tumour-protective effect of GM3/VSSP vaccine was confirmed using GM3 containing N-glycolylneuraminic acid, a ganglioside absent in melanoma cells . Immunostaining and enzyme-linked immunosorbent assay (ELISA) experiments showed a high specificity of immune sera against GM3 and the presence of all four IgG subclasses, with a preponderance of IgG2b and IgG3 . In addition, a strong anti-B16 complement-mediated cytotoxicity was induced by vaccination with GM3/VSSP . The present data indicate the molecular specificity of GM3/VSSP vaccine as well as the adjuvant-dependent and non-transient character of tumour protection in the B16 mouse model . These findings suggest that an appropriate GM3 vaccine may be capable of inducing prolonged tumour protection in melanoma patients.

Biochemistry, 2001 Jul 31, 40(30), 9032 - 9
Crystal structures of amylosucrase from Neisseria polysaccharea in complex with D-glucose and the active site mutant Glu328Gln in complex with the natural substrate sucrose; Mirza O et al.; The structure of amylosucrase from Neisseria polysaccharea in complex with beta-D-glucose has been determined by X-ray crystallography at a resolution of 1.66 A . Additionally, the structure of the inactive active site mutant Glu328Gln in complex with sucrose has been determined to a resolution of 2.0 A . The D-glucose complex shows two well-defined D-glucose molecules, one that binds very strongly in the bottom of a pocket that contains the proposed catalytic residues (at the subsite -1), in a nonstrained (4)C(1) conformation, and one that binds in the packing interface to a symmetry-related molecule . A third weaker D-glucose-binding site is located at the surface near the active site pocket entrance . The orientation of the D-glucose in the active site emphasizes the Glu328 role as the general acid/base . The binary sucrose complex shows one molecule bound in the active site, where the glucosyl moiety is located at the alpha-amylase -1 position and the fructosyl ring occupies subsite +1 . Sucrose effectively blocks the only visible access channel to the active site . From analysis of the complex it appears that sucrose binding is primarily obtained through enzyme interactions with the glucosyl ring and that an important part of the enzyme function is a precise alignment of a lone pair of the linking O1 oxygen for hydrogen bond interaction with Glu328 . The sucrose specificity appears to be determined primarily by residues Asp144, Asp394, Arg446, and Arg509 . Both Asp394 and Arg446 are located in an insert connecting beta-strand 7 and alpha-helix 7 that is much longer in amylosucrase compared to other enzymes from the alpha-amylase family (family 13 of the glycoside hydrolases).

Epidemiol Infect, 2001 Jun, 126(3), 373 - 8
A large outbreak of conjunctivitis caused by a single genotype of Neisseria gonorrhoeae distinct from those causing genital tract infections; Mak DB et al.; Several epidemics of gonococcal conjunctivitis have occurred in Aboriginal populations in Central Australia . In 1997, the first outbreak in the Kimberley region of Western Australia occurred, spreading to Central Australia with a total of 447 cases . A genotyping method was applied directly to DNA extracted from patient samples to characterize the gonococcus causing the epidemic and to compare it with contemporaneous genital isolates . Those positive conjunctival specimens from Kimberley and Central Australia that could be genotyped were all indistinguishable, but were distinct from the genital gonococci, even when they shared the same auxotype and serotype . This suggested that the outbreak was due to a single genotype of Neisseria gonorrhoeae that had probably been carried between communities by infected individuals . We did not find evidence to support the existence of a genital reservoir of the types causing epidemic gonococcal conjunctivitis.

J Bacteriol, 2001 Aug, 183(16), 4718 - 26
Insertion-duplication mutagenesis of neisseria: use in characterization of DNA transfer genes in the gonococcal genetic island; Hamilton HL et al.; We created plasmids for use in insertion-duplication mutagenesis (IDM) of Neisseria gonorrhoeae . This mutagenesis method has the advantage that it requires only a single cloning step prior to transformation into gonococci . Chromosomal DNA cloned into the plasmid directs insertion into the chromosome at the site of homology by a single-crossover (Campbell-type) recombination event . Two of the vectors contain an erythromycin resistance gene, ermC, with a strong promoter and in an orientation such that transcription will proceed into the cloned insert . Thus, these plasmids can be used to create insertions that are effectively nonpolar on the transcription of downstream genes . In addition to the improved ermC, the vector contains two copies of the neisserial DNA uptake sequence to facilitate high-frequency DNA uptake during transformation . Using various chromosomal DNA insert sizes, we have determined that even small inserts can target insertion mutation by this method and that the insertions are stably maintained in the gonococcal chromosome . We have used IDM to create knockouts in two genes in the gonococcal genetic island (GGI) and to clone additional regions of the GGI by a chromosome-walking procedure . Phenotypic characterization of traG and traH mutants suggests a role for the encoded proteins in DNA secretion by a novel type IV secretion system.

J Bacteriol, 2001 Aug, 183(16), 4694 - 701
Requirement of novel competence genes pilT and pilU of Pseudomonas stutzeri for natural transformation and suppression of pilT deficiency by a hexahistidine tag on the type IV pilus protein PilAI; Graupner S et al.; The ubiquitous species Pseudomonas stutzeri has type IV pili, and these are essential for the natural transformation of the cells . An absolute transformation-deficient mutant obtained after transposon mutagenesis had an insertion in a gene which was termed pilT . The deduced amino acid sequence has identity with PilT of Pseudomonas aeruginosa (94%), Neisseria gonorrhoeae (67%), and other gram-negative species and it contains a nucleotide-binding motif . The mutant was hyperpiliated but defective for further pilus-associated properties, such as twitching motility and plating of pilus-specific phage PO4 . {(3)H}thymidine-labeled DNA was bound by the mutant but not taken up . Downstream of pilT a gene, termed pilU, coding for a putative protein with 88% amino acid identity with PilU of P . aeruginosa was identified . Insertional inactivation did not affect piliation, twitching motility, or PO4 infection but reduced transformation to about 10% . The defect was fully complemented by PilU of nontransformable P . aeruginosa . When the pilAI gene (coding for the type IV pilus prepilin) was manipulated to code for a protein in which the six C-terminal amino acids were replaced by six histidine residues and then expressed from a plasmid, it gave a nonpiliated and twitching motility-defective phenotype in pilAI::Gm(r) cells but allowed transformability . Moreover, the mutant allele suppressed the absolute transformation deficiency caused by the pilT mutation . Considering the hypothesized role of pilT(+) in pilus retraction and the presumed requirement of retraction for DNA uptake, it is proposed that the pilT-independent transformation is promoted by PilA mutant protein either as single molecules or as minimal pilin assembly structures in the periplasm which may resemble depolymerized pili and that these cause the outer membrane pores to open for DNA entry.

Int J Antimicrob Agents, 2001 Jul, 18(1), 77 - 80
Activity of gatifloxacin against strains resistant to ofloxacin and ciprofloxacin and its ability to select for less susceptible bacterial variants; Fung-Tomc J et al.; Gatifloxacin is an 8-methoxy fluoroquinolone . On quinolones, this side chain imparts increased activity against Gram-positive bacteria and enhanced killing . Gatifloxacin was tested against ofloxacin non-susceptible (ofloxacin MIC>2 mg/l) strains of Streptococcus pneumoniae (gatifloxacin MIC(90), 1 mg/l) and methicillin-resistant Staphylococcus aureus (MRSA, gatifloxacin MIC(90), 4 mg/l), and to ciprofloxacin non-susceptible (ciprofloxacin MIC>1 mg/l) strains of Escherichia coli (gatifloxacin MIC(90),>16 mg/l) and ciprofloxacin non-susceptible (ciprofloxacin MIC>0.06 mg/l) Neisseria gonorrhoeae (gatifloxacin MIC(50), 0.12 mg/l and MIC(90), 0.5 mg/l) . Though gatifloxacin showed some reduced susceptibility to these populations, the MIC(50) and MIC(90) values suggest that gatifloxacin may be useful against pneumococci and some gonococcal strains not susceptible to other fluoroquinolones . Gatifloxacin did not select for less susceptible variants of MRSA and pneumococci, in contrast to the 10- to 100-fold higher selection frequencies with ciprofloxacin and ofloxacin . The single-step E . coli mutants selected by gatifloxacin and the comparator quinolones had quinolone MICs within the susceptible range . These data suggest that gatifloxacin use may hinder the development of quinolone-resistance, particularly in Gram-positive bacteria.

Sex Transm Dis, 2001 Jul, 28(7), 417 - 23
Tests of Buffergel for contraception and prevention of sexually transmitted diseases in animal models; Zeitlin L et al.; BACKGROUND: BufferGel is a novel spermicidal and microbicidal gel formulated to maintain the natural protective acidity of the vagina by acidifying semen, which otherwise alkalinizes the vagina . GOAL: To test the efficacy of BufferGel for preventing sexually transmitted infections and pregnancy in animal models . STUDY DESIGN: Animals were challenged with pathogens or sperm after pretreatment with both test and control agents, or after no pretreatment, then evaluated for infection or pregnancy using standard methods . RESULTS: BufferGel provided significant contraceptive efficacy in the rabbit, and significant protection against vaginal and rectal transmission of herpes simplex virus type 2 (HSV-2) in the mouse, vaginal transmission of Chlamydia trachomatis in the mouse, and skin transmission of cottontail rabbit papillomavirus in the rabbit . It did not protect against vaginal transmission of Neisseria gonorrhoeae in the mouse . CONCLUSIONS: The protective efficacy of BufferGel in five of the six animal models suggests that this microbicide warrants clinical evaluation for both contraception and disease prevention.

Sex Transm Dis, 2001 Jul, 28(7), 372 - 8
Sexually transmitted infections and reproductive health in Azerbaijan; Claeys P et al.; BACKGROUND: Little is known about the prevalence of sexually transmitted infections (STIs) and about sexual and reproductive health in Central and Eastern Europe . However, it is clear that major epidemics of STIs currently exist . GOAL: To provide baseline information for the development of national guidelines on the management of STIs in Azerbaijan . STUDY DESIGN: A prevalence study on STIs, including a questionnaire on sexual and reproductive health, in two regions of Azerbaijan targeted three groups: (1) pregnant women, (2) gynecology patients, and (3) men attending a dermatovenereology clinic . RESULTS: The 407 pregnant women in this study had a mean of 1.47 abortions and 1.40 births per woman . Of these women, 12% reported condom use and 41% previous symptoms of a sexually transmitted infection . Active syphilis was found in 1.7% of the women . The 326 gynecology patients had a mean of 2.54 abortions and 2.63 births per woman . Of these patients, 11% reported use of modern contraceptives, 18.3% previous condom use, and 63% previous symptoms of an STI . The prevalence of active syphilis was 2.2%, Chlamydia trachomatis 3.1%, Neisseria gonorrhoeae 2.8%, Trichomonas vaginalis 7.1%, Candida 33.1%, and bacterial vaginosis 32.5% . Of the 197 male patients, 67% reported multiple partners in the past 3 months, 62% money exchanged for sex, 37% condom use ever, and 40% a history of STIs . Active syphilis was found in 9.5% of the men, C trachomatis in 5.9%, N gonorrhoeae in 17%, and T vaginalis in 4.4% . CONCLUSIONS: The data show high-risk behavior in the men attending STI clinics, poor sexual and reproductive health status in the women, and underreporting of official data.

J Clin Invest, 2001 Jul, 108(2), 233 - 40
A family with complement factor D deficiency; Biesma DH et al.; A complement factor D deficiency was found in a young woman who had experienced a serious Neisseria meningitidis infection, in a deceased family member with a history of meningitis, and in three relatives without a history of serious infections . The patient and these three relatives showed a normal activity of the classical complement pathway, but a very low activity of the alternative complement pathway and a very low capacity to opsonize Escherichia coli and N . meningitidis (isolated from the patient) for phagocytosis by normal human neutrophils . The alternative pathway-dependent hemolytic activity and the opsonizing capacity of these sera were restored by addition of purified factor D . The family had a high degree of consanguinity, and several other family members exhibited decreased levels of factor D . The gene encoding factor D was found to contain a point mutation that changed the TCG codon for serine 42 into a TAG stop codon . This mutation was found in both alleles of the five completely factor D-deficient family members and in one allele of 21 other members of the same family who had decreased or low-normal factor D levels in their serum . The gene sequence of the signal peptide of human factor D was also identified . Our report is the first, to our knowledge, to document a Factor D gene mutation . The mode of inheritance of factor D deficiency is autosomal recessive, in accordance with the localization of the Factor D gene on chromosome 19 . Increased susceptibility for infections in individuals with a partial factor D deficiency is unlikely.

Mol Microbiol, 2001 Jul, 41(1), 263 - 77
A variable genetic island specific for Neisseria gonorrhoeae is involved in providing DNA for natural transformation and is found more often in disseminated infection isolates; Dillard JP et al.; Neisseria gonorrhoeae (the gonococcus) is the causative agent of the sexually transmitted disease gonorrhoea . Most gonococcal infections remain localized to the genital tract but, in a small proportion of untreated cases, the bacterium becomes systemic to produce the serious complication of disseminated gonococcal infection (DGI) . We have identified a large region of chromosomal DNA in N . gonorrhoeae that is not found in a subset of gonococcal isolates (a genetic island), in the closely related pathogen, Neisseria meningitidis or in commensal Neisseria that do not usually cause disease . Certain versions of the island carry a serum resistance locus and a gene for the production of a cytotoxin; these versions of the island are found preferentially in DGI isolates . All versions of the genetic island encode homologues of F factor conjugation proteins, suggesting that, like some other pathogenicity islands, this region encodes a conjugation-like secretion system . Consistent with this hypothesis, a wild-type strain released large amounts of DNA into the medium during exponential growth without cell lysis, whereas an isogenic strain mutated in a peptidoglycan hydrolase gene (atlA) was drastically reduced in its ability to donate DNA for transformation during growth . This genetic island constitutes the first major discriminating factor between the gonococcus and the other Neisseria and carries genes for providing DNA for genetic transformation.

Rev Inst Med Trop Sao Paulo, 2001 May-Jun, 43(3), 119 - 24
Genetic relationships among serogroup B: serotype 4 Neisseria meningitidis strains; Lemos AP et al.; We compared the results obtained by serotyping of PorB epitopes using an expanded panel of monoclonal antibodies (mAb) including mAb 7 and mAb 10, with results obtained by RFLP of rRNA genes (ribotyping) . The purpose of this study was to assess the correlation between phenotypic- and genotypic- methods for typing N . meningitidis . The ribotypes obtained using ClaI or EcoRV endonucleases grouped the strains in seven and two different patterns, respectively . This additional characterization of PorB epitopes improved the correlation between these two methods of typing N . meningitidis.

Arch Pediatr Adolesc Med, 2001 Jun, 155(6), 676 - 9
Self-obtained vaginal swabs for diagnosis of treatable sexually transmitted diseases in adolescent girls; Smith K et al.; OBJECTIVE: To ascertain the acceptability of testing and prevalence of 3 readily treatable sexually transmitted diseases (STDs) (infections with Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis) with the use of patient-obtained vaginal swabs . STUDY DESIGN: Study participants at each initial session were asked to provide self-obtained vaginal swabs for ligase chain reaction testing to detect N gonorrhoeae and C trachomatis, and for culture of T vaginalis . SETTING: Behavioral intervention sessions with African American adolescent girls in a nonclinical program to reduce risk of STDs, human immunodeficiency virus infection, and pregnancy . RESULTS: All study participants were offered their choice of STD screening in the context of a traditional pelvic examination or using self-obtained vaginal swabs . All eligible participants chose self-administered vaginal swabs . Of the 512 participants examined at their initial study visit, 28.7% were found to be infected with 1 or more treatable STDs (5.3% with N gonorrhoeae, 17.8% with C trachomatis, and 12.9% with T vaginalis) . CONCLUSIONS: With the use of newer detection systems, STDs can be readily detected in nonclinical settings with the use of self-obtained vaginal swabs, providing new opportunities for efforts to control STDs.

Am J Emerg Med, 2001 Jul, 19(4), 274 - 5
Empiric treatment of gonorrhea and chlamydia in the ED; Wiest DR et al.; A retrospective study was conducted on female patients who were screened for Chlamydia trachomatis and Neisseria gonorrhea . The purpose of the study was to determine if any factors predict empiric therapy at the index visit . Of 911 patients enrolled in the study, 100 were found to have positive DNA screens and 54 were given empiric therapy . A logistic regression was used to test the ability of age, ethnicity, DNA probe result and provider type (physician or midlevel provider), to predict empiric treatment . DNA probe result and provider type were the only 2 factors that were found to predict empiric therapy . A subsequent analysis using a Cochran-Mantel-Hanszel test to control for DNA probe result revealed that the provider type remained the only variable that predicted empiric treatment.

Mol Microbiol, 2001 Jun, 40(6), 1301 - 10
The recX gene potentiates homologous recombination in Neisseria gonorrhoeae; Stohl EA et al.; In the pathogen Neisseria gonorrhoeae (Gc), the RecA protein is necessary for DNA repair, DNA transformation and pilus antigenic variation . Many bacteria contain a gene, recX, which has been suggested to downregulate recA through an unknown mechanism . To investigate the possible role of recX in Gc, we cloned and insertionally inactivated the recX gene . The recX loss-of-function mutant showed decreases in pilus phase variation, DNA transformation and DNA repair ability compared with wild type . We were able to complement all these deficiencies by supplying a functional copy of recX elsewhere in the chromosome . The recX mutant still showed increases in pilus phase variation under conditions of iron starvation, and the recX mutant showed levels of RecA protein equivalent to wild type . Although the precise role of recX in recombination remains unclear, RecX aids all RecA-related processes in Gc, and this is the first demonstration of a role for recX in homologous recombination in any organism.

Trends Microbiol, 2001 Jul, 9(7), 335 - 43
Detecting anomalous gene clusters and pathogenicity islands in diverse bacterial genomes; Karlin S; A gene in a genome is defined as putative alien (pA) if its codon usage difference from the average gene exceeds a high threshold and codon usage differences from ribosomal protein genes, chaperone genes and protein-synthesis-processing factors are also high . pA gene clusters in bacterial genomes are relevant for detecting genomic islands (GIs), including pathogenicity islands (PAIs) . Four other analyses appropriate to this task are G+C genome variation (the standard method); genomic signature divergences (dinucleotide bias); extremes of codon bias; and anomalies of amino acid usage . For example, the cagA domain of Helicobacter pylori is highly deviant in its genome signature and codon bias from the rest of the genome . Using these methods we can detect two potential PAIs in the Neisseria meningitidis genome, which contain hemagglutinin and/or hemolysin-related genes . Additionally, G+C variation and genome signature differences of the Mycobacterium tuberculosis genome indicate two pA gene clusters.

Pediatrics . 2001 Jul;108(1):E13.
Rapid assessment of sexual behavior, drug use, human immunodeficiency virus, and sexually transmitted diseases in northern thai youth using audio-computer-assisted self-interviewing and noninvasive specimen collection; van Griensven F et al.; BACKGROUND: Drug use, unwanted pregnancy, human immunodeficiency virus (HIV) infection, and sexually transmitted diseases are serious health problems among Thai youth . The gravity of these problems demands high-quality data to direct public health policy and prevention programs . Previous studies of stigmatized behaviors have been hampered by participation bias and underreporting . To evaluate sexual behavior, disease, and drug use, we used audio-computer-assisted self-interviewing (ACASI) and noninvasive specimen collection methods . We also evaluated effectiveness of these methods in minimizing participation bias and underreporting . METHODS: In late 1999, students aged 15 to 21 years attending 3 vocational schools were invited to participate in a cross-sectional survey . Consenting students completed a classroom-based ACASI interview using a confidential code number system . Oral fluid specimens were tested for HIV antibodies, and urine was tested for chlamydial and gonococcal nucleic acids, methamphetamines, and opiates . RESULTS: Of 1736 invited students, 1725 (99%) agreed to participate . Of these, 48% of the male students and 43% of the female students reported ever having had sexual intercourse . Overall, the mean number of lifetime sexual partners was 4.6 among male participants (median: 2) and 2.8 among female participants (median: 1) . Consistent use of condoms with steady partners was reported by 16% of male participants and 11% of female participants who had such partners . Of all male participants, 7% had ever paid for sex, 3% had ever sold sex, and 7% had ever been coerced to have sex . Of all female participants, 3% had ever sold sex and 21% had ever been coerced to have sex . Among women with a history of sexual intercourse, 27% reported at least 1 pregnancy . Of these pregnancies, 83% were terminated . Among those with sexual intercourse experience, the prevalence of HIV infection was 0.5%; of infection with Neisseria gonorrhoeae, 0.4%; and of infection with Chlamydia trachomatis, 5% . Twenty-nine percent of students reported ever having used methamphetamines . Ten percent had a methamphetamine-positive urine test . In the ACASI interview, 16% of these denied ever having used methamphetamines . The prevalence of opiate positive urine tests was low (0.2%) . CONCLUSION: This study shows that adolescents and young adults in Chiang Rai are at high risk for having unprotected intercourse, being coerced to have sex, unwanted pregnancy, sexually transmitted diseases, and drug use . The high enrollment rate demonstrates the feasibility and acceptability of using ACASI and noninvasive specimen collection methods in a developing country . ACASI use may lead to increased, but not to complete, self-reporting of sensitive behaviors.

Commun Dis Intell, 2001 Apr, 25(2), 59 - 63
Annual report of the Australian Gonococcal Surveillance Programme, 2000; Neisseria gonorrhoeae 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase does not catalyze the formation of the ribo analogue; Department of Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan 48109-1065, USA{figure: see text} Neisseria gonorrhoeae 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAH 7-P) synthase catalyzes an aldol-type condensation between D-erythrose 4-phosphate and phosphoenolpyruvate (PEP) to form 3-deoxy-D-arabino-heptulosonate 7-phosphate and not 3-deoxy-D-riboheptulosonate 7-phosphate . Similar to the Escherichia coli enzyme, N . gonorrhoeae DAH 7-P synthase condenses D-arabinose 5-phosphate with PEP to give 3-deoxy-D-manno-octulosonate 8-phosphate . Therefore, the stereochemistry of the reaction catalyzed by N . gonorrhoeae DAH 7-P synthase at C1 of the phosphorylated monosaccharide is the same as that for the E . coli enzyme, namely, re face attack.

J Oral Maxillofac Surg, 2001 Jul, 59(7), 739 - 48; discussion 748-9
The changing face of odontogenic infections; Storoe W et al.; PURPOSE: The purpose of this investigation was to compare characteristics of patients hospitalized with odontogenic infections during the 1980s to those of the 1990s . PATIENTS AND METHODS: This study was a retrospective record review that compared 2 cohorts of patients admitted to the same institution during two 81-month periods, one decade apart . Admission criteria were face or neck swelling suggesting abscess or cellulitis and one or more of the following: temperature above 38 degrees C, white blood cell (WBC) count greater than 10.8 x 10(3)/microL, or concern about airway compromise . Characteristics reviewed were age, gender, race, admission temperature, admission WBC count, fascial space(s) involved, tooth of etiology, duration of hospitalization, and bacteria isolated . Data were compared for statistical significance (P <.05) . RESULTS: No significant differences were found between the 2 cohorts for age, gender, race, admission temperature, admission WBC count, space involvement, or length of stay (P <.05) . One tooth (mandibular left first molar) of 52 was involved more frequently in the 1990 group (P <.03) . Gram-positive cocci were isolated significantly more frequently from the 1990s patients than from the 1980s patients (P <.03) . There were also significant differences (P <.02) between cohorts in the isolation frequency of individual genera, such as alpha-hemolytic Streptococci, coagulase negative staphylococci, Staphylococcus epidermidis, Bacteroides melanogenicus, beta-lactamase positive Bacteroides, Eikenella corrodens, and Neisseria species . Eighty-one percent of the bacteria cultured from the 1990s patients were resistant to one or more common antibiotics; 47% of these organisms were Staphylococcus aureus . CONCLUSION: No clinically significant differences existed in the characteristics of patients hospitalized with odontogenic infections between the 1980s and the 1990s . Although there were differences in the type and prevalence of bacteria isolated, this was probably a result of changes in nomenclature, identification protocols, and isolation techniques .

FEMS Microbiol Lett, 2001 Jun 25, 200(2), 235 - 40
Plasmid-encoded catalase KatA, the main catalase of Pseudomonas fluorescens strain Cb36; Peters M et al.; The plasmid-state catalase gene katA of the phenol gradative Pseudomonas fluorescens isolate Cb36 has been characterized and shown to be the major catalase of this strain . The predicted amino acid sequence of KatA revealed significant similarity with the catalase sequence from Neisseria meningitidis and has probably the non-pseudomonad origin . The specific activity of catalase was investigated and elevated catalase activity was found in stationary phase cells . The consensus sequence for promoters recognized by the stationary phase sigma factor sigma(s) was found 212 bp upstream of the putative ATG start codon . The ability of KatA to detoxify a high concentration of hydrogen peroxide and protect Pseudomonas putida and Escherichia coli cells was shown.

Immunol Rev, 2001 Apr, 180, 86 - 99
Mannose-binding lectin: targeting the microbial world for complement attack and opsonophagocytosis; Jack DL et al.; Mannose-binding lectin (MBL) is an important constituent of the innate immune system . This protein binds through multiple lectin domains to the repeating sugar arrays that decorate many microbial surfaces, and is then able to activate the complement system through a specific protease called MBL-associated protease-2 . We have used flow cytometry to study both the binding of MBL to microorganisms and the subsequent activation of complement . For selected Gram-negative organisms, such as Salmonella and Neisseria, we have examined the relative roles of lipopolysaccharide (LPS) structure and capsule in determining binding and conclude that the LPS is of major importance . Our results from studies with several clinically relevant organisms also show that MBL binding detected by flow cytometry leads to measurable activation of purified C4, suggesting that the bound lectin is capable of initiating opsonophagocytosis and/or bacterial lysis . There is an increasing literature suggesting that MBL deficiency, which mainly results from three relatively common single point mutations in exon 1 of the gene, predisposes both to infection by extracellular pathogens and to autoimmune disease . In addition, the protein also modulates disease severity, at least in part through a complex, dose-dependent influence on cytokine production . The mechanisms and signalling pathways involved in such processes remain to be elucidated.

An Esp Pediatr, 2001 Jul, 55(1), 5 - 10
{The young febrile child . Results of a multicenter survey}; Grupo de Trabajo sobre el Nino Febril de la Sociedad Espanola de Urgencias de Pediatria; OBJECTIVES: To establish the value of clinical and laboratory parameters gathered from a national survey on the management of the young febrile child (0-36 months) in emergency departments and to determine the etiology of potentially serious bacterial infections in Spain in this group of patients . PATIENTS AND METHODS: Prospective survey of febrile children aged less than 36 months who presented to the emergency departments of the participant hospitals and who fulfilled the inclusion criteria . RESULTS: A total of 937 cases were collected . Of these, 43 were excluded because of absence of blood culture and 151 because the patients had not undergone obligatory monitoring until they became afebrile or had no final diagnosis . Consequently, the sample size consisted of 743 patients . Clinical assessment was very important to detect patients with potentially serious bacterial disease . Among the laboratory parameters, a white blood cell count above 15,000/l or below 5,000/l and elevated C-reactive protein levels were the most useful . Urinary tract infection was the most frequent invasive disease . Neisseria meningitidis was the most frequent microorganism isolated in the blood culture . The incidence of occult bacteremia was 3 % with predominance of Streptococcus pneumoniae (74 %) . CONCLUSIONS: The experience of the pediatrician supported by clinical and laboratory parameters are the most effective means of detecting febrile children (0-36 months) with potentially serious bacterial disease.

Diagn Microbiol Infect Dis, 2001 Apr, 39(4), 265 - 6
Neisseria elongata subsp . elongata, as a cause of human endocarditis; Apisarnthanarak A et al.; Neisseria elongata subsp . elongata, previously considered nonpathogenic, is a potential agent of human endocarditis . We report the second case of human endocarditis caused by this organism . The patient was successfully treated with Ceftriaxone alone for a total of six weeks.

J Infect Chemother, 2001 Mar, 7(1), 49 - 50
Emergence of cephem- and aztreonam-high-resistant Neisseria gonorrhoeae that does not produce beta-lactamase; Akasaka S et al.; Regarding Neisseria gonorrhoeae, the National Committee for Clinical Laboratory Standards (NCCLS) has not defined the breakpoint minimum inhibitory concentration (MIC) for expanded spectrum cephems such as cefpodoxime and ceftizoxime, because of the absence of resistant strains to these antibiotics . To date, in gonococcal urethritis, after treatment with third generation cephems and aztreonam, clinical failures caused by resistant N . gonorrhoeae strains have not been reported . However, we experienced two clinical failures in patients with gonococcal urethritis treated with cefdinir and aztreonam . N . gonorrhoeae isolates from these two patients showed high-level MICs to these agents . The MIC of cefdinir was 1 microg/ml for both strains and that of aztreonam was 8 microg/ml for both strains, while the MICs of other beta-lactams were also higher than the NCCLS value, except for ceftriaxone, for which the MIC was 0.125 microg/ml for both strains . Moreover, the MICs of fluoroquinolones, tetracyclines, and erythromycin against these two isolates were higher than the NCCLS susceptibility value . These isolates were susceptible to spectinomycin . In N . gonorrhoeae, the emergence of these beta-lactam-resistant isolates is of serious concern . However, a more serious problem is that these isolates were already resistant to non-beta-lactam antimicrobials . In Japan, ceftriaxone has not been permitted for clinical use against gonococcal infections . Therefore, in Japan, patients with gonococcal urethritis caused by these resistant N . gonorrhoeae strains should be treated with cefodizime or spectinomycin.

Sex Transm Dis, 2001 Jun, 28(6), 356 - 62
Chlamydia, gonorrhea, and HIV-1 prevalence among five populations of women in the Czech and Slovak Republics; Kacena KA et al.; BACKGROUND: Five populations at risk for sexually transmitted diseases (STDs) in the Czech and Slovak Republics were sampled . GOAL: To estimate prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and HIV-1 infections . STUDY DESIGN: Urine specimens were collected serially from women at a Prague prenatal clinic (n = 134), a Prague dermatovenerealogy clinic (n = 91), sex workers from northern and central Bohemia (n = 35), students from a northern Bohemian school (n = 217), and Gypsies from Jarovnice, Slovakia (n = 128) . These specimens were tested for chlamydia and gonorrhea using a ligase chain reaction pooling algorithm, and for HIV using an enzyme immunoassay confirmed by Western blot . RESULTS: The prevalence of chlamydia was 2.2% (95% CI, 0.4-6.4) in the prenatal clinic, 5.5% (95% CI, 1.8-12.4) in the STD clinic, 22.9% (95% CI, 10.4-40.1) among street sex workers, 8.2% (95% CI, 3.6-15.6) among sexually active female high school students, and 3.9% (95% CI, 1.3-8.9) among Gypsy women . Gonorrhea was found in only two populations: 2.2% (95% CI, 0.3-7.7) in the STD clinic, and 2.9% (95% CI, 0.1-14.9) among sex workers . No HIV-1 infection was detected . CONCLUSIONS: Urine screening was an efficient and accurate method for identifying groups at risk for STDs in the Czech Republic and Slovakia because sample collection was fast and noninvasive, and potential participation bias was reduced by high acceptability.

Sex Transm Dis, 2001 Jun, 28(6), 326 - 9
Occurrence of trichomoniasis in women in Denmark, 1967-1997; Dragsted DM et al.; BACKGROUND: The Neisseria Unit at Statens Serum Institut (SSI), Copenhagen, Denmark administers laboratory surveillance systems for gonorrhea and chlamydia, however, no similar system has been available for trichomoniasis . OBJECTIVES: To describe changes in the occurrence of trichomoniasis in women and in the distribution according to age, and to compare these changes with those observed in women with gonorrhea or chlamydia . STUDY DESIGN: Data on the epidemiology of trichomoniasis were based on the annual laboratory records at the Neisseria Unit at SSI . Data on gonorrhea (1967-1997) and chlamydia (1994-1997) originated from the National Laboratory Surveillance Systems . RESULTS: During the period 1967 to 1997, the percentage of Trichomonas vaginalis-positive specimens showed a constant decrease from 19 to fewer than 2 . The annual number of specimens received varied between a maximum of 31,626 in 1975 and a minimum of 3,291 in 1997 . The median age of women with trichomoniasis increased from 31 to 39 years as compared with a change from 21 to 24 years in women with gonorrhea (1967-1997) and a median age of 22 years in women with chlamydia (1994-1997) . CONCLUSIONS: A significant decline in the occurrence of trichomoniasis in women concurrent with a significant rise in the median age of women with the disorder was observed . During the periods investigated, women with trichomoniasis were significantly older than women with gonorrhea or chlamydia.

Sex Transm Dis, 2001 Jun, 28(6), 321 - 5
Self-collection of vaginal swabs for the detection of Chlamydia, gonorrhea, and trichomoniasis: opportunity to encourage sexually transmitted disease testing among adolescents; Wiesenfeld HC et al.; BACKGROUND: Many sexually transmitted diseases (STDs) are prevalent among adolescents, yet compliance to undergo STD testing by this population is suboptimal . Efforts to enhance compliance with testing among at-risk youth are needed . GOAL: To determine the feasibility and acceptability of self-collection of vaginal swabs for the detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis among high-school students attending a school health clinic . STUDY DESIGN: Enrolled in the study were 228 female students between the ages of 15 and 19 years . Each student self-collected a single vaginal swab that was tested for C trachomatis, N gonorrhoeae, and T vaginalis by polymerase chain reaction amplification . Acceptability of self-collection of vaginal swabs was assessed . RESULTS: The prevalence of any STD was 18% . Trichomoniasis, chlamydia, and gonorrhea were diagnosed in 10%, 8%, and 2% of students, respectively . Nearly 13% of females who had never previously had a gynecologic examination tested positive for an STD, and 51% of infected students would not have pursued testing by traditional gynecologic examination if self-collection was not offered . Self-collection of vaginal swabs was almost uniformly reported as easy to perform (99%) and preferable to a gynecologic examination (84%) . Nearly all (97%) stated that they would undergo testing at frequent intervals if self-testing were available . CONCLUSIONS: Self-collected vaginal swabs for STD testing can be easily implemented in a high-school setting with high acceptability among students, enabling the detection of many STDs that would otherwise remain undetected and untreated.

Sex Transm Dis, 2001 Jun, 28(6), 309 - 14
Molecular epidemiology of Neisseria gonorrhoeae exhibiting decreased susceptibility and resistance to ciprofloxacin in Hawaii, 1991-1999; Trees DL et al.; BACKGROUND: Clinically significant resistance to Centers for Disease Control and Prevention (CDC)-recommended doses of fluoroquinolones (ciprofloxacin and ofloxacin) has been reported for Neisseria gonorrhoeae . In Hawaii, fluoroquinolone-resistant gonococcal isolates were first identified in 1991 . GOAL: To assess the diversity, based on phenotypic and genotypic characterization, of gonococcal isolates exhibiting decreased susceptibility (CipI; MICs = 0.125-0.5 microg/ml) or clinically significant resistance (CipR; MICs > or = 1 microg/ml) to ciprofloxacin in Hawaii from 1991 through 1999 . STUDY DESIGN: Antimicrobial susceptibilities, auxotype/serovar (A/S) class, GyrA/ParC alteration patterns, and plasmid profiles were determined for gonococci isolated in Honolulu from 1991 through 1999 that exhibited intermediate or clinically significant resistance to ciprofloxacin . Strain phenotypes were defined by A/S class, GyrA/ParC alteration pattern, and penicillin-tetracycline resistance phenotype supplemented with plasmid profiles for beta-lactamase-producing isolates . RESULTS: Altogether, 68 isolates exhibiting intermediate or clinically significant resistance to ciprofloxacin belonged to 23 and 19 strain phenotypes, respectively . Among the CipI and CipR isolates, 4 and 13 GyrA/ParC alterations patterns were identified, respectively . The 91,95/Asp-86 alteration pattern occurred most frequently among CipR isolates . Forty-four strain phenotypes were represented by only one isolate . In addition, seven pairs and two clusters of isolates were identified . CONCLUSIONS: From 1991 through 1997, few gonococcal strains exhibiting intermediate or clinically significant resistance to CDC-recommended doses of fluoroquinolones were identified from Hawaii . Isolates belonged to a large number of phenotypic and genotypic types, suggesting that most cases were imported, with only a few instances in which isolate pairs indicated that secondary transmission of infections had occurred in Hawaii . Beginning in 1998, the number of CipR isolates increased markedly, and more isolates belonged to fewer phenotypic and genotypic types, suggesting either more frequent importation of fewer strain types or the possibility that the endemic spread of a few strains is beginning to occur.

Microbiology, 2001 Jun, 147(Pt 6), 1473 - 82
An NMR and enzyme study of the carbon metabolism of Neisseria meningitidis; Leighton MP et al.; The pathogenic neisseriae are fastidious bacteria that are only able to grow on a restricted range of carbon sources . The genome sequence of Neisseria meningitidis strain MC58 predicts the presence of a complete citric acid cycle (CAC), but there have been no detailed biochemical studies of carbon metabolism in this important pathogen . In this study, both NMR and conventional enzyme assays were used to investigate the central metabolic pathways of a serogroup B strain (K454) . (13)C-NMR labelling patterns of amino acids from hydrolysed cell proteins after growth with either 2- or 3-{(13)C}pyruvate were consistent with the operation of a complete oxidative CAC . Enzyme assays showed that cell-free extracts contained all the CAC enzymes predicted from the genome sequence, including a membrane-bound malate:quinone oxidoreductase which is present in place of the conventional NAD-linked cytoplasmic malate dehydrogenase . (1)H-NMR studies showed that growth on glucose, lactate and, especially, pyruvate, resulted in the excretion of significant amounts of acetate into the culture supernatant . This occurred via the phosphotransacetylase (PTA)-acetate kinase (ACK) pathway . Extremely high specific activities of PTA (7-14 micromol min(-1) mg(-1)) were detected in cell-free extracts, although ACK activities were much lower (46-298 nmol min(-1) mg(-1)) . Expression of PTA and ACK activities was not co-ordinately regulated during growth on combinations of carbon sources . This may be related to the presence of two ackA paralogues in N . meningitidis which are, unusually, unlinked to the pta gene.

Proc Natl Acad Sci U S A, 2001 Jun 5, 98(12), 6901 - 4 Epub 2001 May 29.
Direct observation of extension and retraction of type IV pili; Skerker JM et al.; Type IV pili are thin filaments that extend from the poles of a diverse group of bacteria, enabling them to move at speeds of a few tenths of a micrometer per second . They are required for twitching motility, e.g., in Pseudomonas aeruginosa and Neisseria gonorrhoeae, and for social gliding motility in Myxococcus xanthus . Here we report direct observation of extension and retraction of type IV pili in P . aeruginosa . Cells without flagellar filaments were labeled with an amino-specific Cy3 fluorescent dye and were visualized on a quartz slide by total internal reflection microscopy . When pili were attached to a cell and their distal ends were free, they extended or retracted at rates of about 0.5 microm s(-1) (29 degrees C) . They also flexed by Brownian motion, exhibiting a persistence length of about 5 microm . Frequently, the distal tip of a filament adsorbed to the substratum and the filament was pulled taut . From the absence of lateral deflections of such filaments, we estimate tensions of at least 10 pN . Occasionally, cell bodies came free and were pulled forward by pilus retraction . Thus, type IV pili are linear actuators that extend, attach at their distal tips, exert substantial force, and retract.

Afr J Med Med Sci, 2000 Jun, 29(2), 171 - 3
E-test method of antimicrobial susceptibility testing of Neisseria gonorrhoeae for routine diagnostic service; Adeyemi-Doro FA et al.; The minimising inhibitory concentrations of four antimicrobial agents for 64 clinical isolates of Neisseria gonorrhoeae including 26 penicillinase producing strains (PPNG) as determined by E test, a recently developed method for sensitivity testing, were compared with those of agar dilution method using isosensitest (IST) agar . The medium was supplemented with either 5% lysed horse blood alone or with both lysed horse blood and 1% vitox defined supplement . The E test MICs compared closely with those obtained by agar dilution with essential agreements within +/- 1 log2 dilution being over 90% with all test antibiotics on medium that did not contain vitox, and between 71 and 93% on medium containing vitox . The Pearson's correlation coefficients ranged from 0.84 to 0.96 on either medium formulation . Excellent categorical agreement was obtained for all isolates with ceftriaxone, ciprofloxacin and tetracycline, while the E test gave a minor categorical discrepancy for two isolates with penicillin . We conclude that the E test is as reliable as conventional agar dilution method for MIC testing of N . gonorrhoeae in a routine laboratory.

FEMS Microbiol Lett, 2001 May 30, 199(2), 203 - 6
Detection of the lst gene in different serogroups and LOS immunotypes of Neisseria meningitidis; Tsang RS et al.; The sialylation of the lipooligosaccharide (LOS) of Neisseria meningitidis is mediated by the LOS sialyltransferase enzyme encoded by the lst gene . PCR using four sets of primers that targeted to different regions of the lst gene was used to survey the distribution of lst in different serogroups and LOS immunotypes of N . meningitidis as well as other Neisseria species . While the lst gene was found in N . meningitidis strains regardless of their capsular serogroup and LOS structures, the gene is also found in N . gonorrhoeae, N . lactamica, N . polysaccharea, and N . subflava biovar subflava . The presence of the lst gene in these organisms and the sialylation of their LOS antigens were discussed.

J Clin Microbiol, 2001 Jun, 39(6), 2335 - 7
Amplified fragment length polymorphism fingerprinting for identification of a core group of Neisseria gonorrhoeae transmitters in the population attending a clinic for treatment of sexually transmitted diseases in Amsterdam, The Netherlands; Spaargaren J et al.; Amplified fragment length polymorphism analysis seems well suited for studying the epidemiology of isolates of Neisseria gonorrhoeae obtained from patients attending the Sexually Transmitted Disease Outpatient Clinic in Amsterdam, The Netherlands . It shows potential to identify the core group of transmitters.

J Clin Microbiol, 2001 Jun, 39(6), 2325 - 9
Genotyping Neisseria gonorrhoeae using fluorescent amplified fragment length polymorphism analysis; Palmer HM et al.; An evaluation of fluorescent amplified fragment length polymorphism (FAFLP) for typing Neisseria gonorrhoeae using 52 isolates revealed that its discriminatory power approached that of Opa-typing in identifying epidemiologically linked isolates . Automated, accurate sizing of FAFLP amplified fragments permits objective data analysis and storage, making it an attractive method for large surveillance projects.

J Clin Microbiol, 2001 Jun, 39(6), 2280 - 2
Evaluation of the Roche Neisseria gonorrhoeae 16S rRNA PCR for confirmation of AMPLICOR PCR-positive samples and comparison of its diagnostic performance according to storage conditions and preparation of endocervical specimens; Van Dyck E et al.; The AMPLICOR PCR was used to detect Neisseria gonorrhoeae in endocervical specimens . A 16S rRNA PCR performed on N . gonorrhoeae-positive samples showed sensitivities of 73.2, 64.3, and 94.4% for samples treated directly with AMPLICOR lysis buffer, samples suspended in 2-sucrose phosphate, and samples suspended in diluted phosphate-buffered saline, respectively.

Int J STD AIDS, 2001 May, 12(5), 342 - 5
Management of epididymo-orchitis in Genitourinary Medicine clinics in the United Kingdom's North Thames region 2000; Dale AW et al.; A questionnaire survey and case notes audit reviewing management of epididymo-orchitis (E-O) by 34 Genitourinary Medicine (GUM) clinics located in the North Thames was undertaken . Twenty-two clinics (65%) returned completed questionnaires and audited a total of 83 newly diagnosed cases . All participating clinics offer microscopy of urethral smears and screening for Neisseria gonorrhoeae and Chlamydia trachomatis to all patients, regardless of age . However, greater numbers of clinics would offer routine microbiology of mid-stream urine (MSU) samples (20/22, 91% versus 16/22, 73%) and scrotal ultrasound (5/22, 23% versus 1/22, 5%) to patients aged over 35, compared with men under 35 . Half of the cases audited were due either to sexually transmitted infections (STIs) (41/83, 49%), or associated with ascending urinary tract infections (4/83, 5%) . No obvious infectious cause was identified for 38/83 cases (46%) . Reported management was appropriate for the causative conditions diagnosed and accorded with the UK National Guidelines for this and related conditions.

Ann Ist Super Sanita, 2000, 36(4), 431 - 5
{Complications of sexually transmitted diseases: clinical course and treatment}; Guaschino S; Sexually transmitted infectious agents cause a wide array of complications in the short-, medium-, and long-term, especially among women . Specifically, infection with these agents can result in sterility, infertility, genital neoplasia, pre-term delivery, and foetal/neonatal pathologies, and the agents most commonly associated with these complications are: Neisseria gonorrhoeae, Chlamydia trachomatis, anaerobic bacteria, Ureaplasma urealyticum, Gardnerella vaginalis and human papilloma virus (HPV), which is involved in the etiology of squamous genital carcinomas . This report describes several of the mechanisms involved in the damage to genital and reproductive organs, focussing on those mechanisms involved in the damage caused by delayed autoimmune response.

Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 1998 Feb, 20(1), 65 - 9
{Identification and study on effect of new stress protein from Neisseria gonorrhoeae}; Zheng H et al.; OBJECTIVE: To investigate the influence of changing environmental growth conditions on the virulence of Neisseria gonorrhoeae . METHODS: The gonococci were subjected to 1 mmol/L hydrogen peroxide (sublethol concentration) for one hour . RESULTS: A novel gonococcal stress protein with molecular weight of 62,000 was identified by radioactive labeling and sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Gonococci exposed to 1.0 mmol/L hydrogen peroxide made it significantly more resistant to higher concentrations of hydrogen peroxide and to neutrophils than control organisms (P < 0.01) . CONCLUSIONS: These results suggest that the 62,000 stress protein might increase the virulence of N . gonorrhoeae and might be an adapted regular protein produced by N . gonorrhoeae survived within human neutrophils.

Int Immunopharmacol, 2001 Mar, 1(3), 423 - 32
C4bp binding to porin mediates stable serum resistance of Neisseria gonorrhoeae; Ram S et al.; Screening of 29 strains of Neisseria gonorrhoeae revealed that 16/21 serum resistant strains and 0/8 serum sensitive strains bound C4bp, suggesting that C4bp binding to gonococci could contribute to serum resistance . C4bp bound to gonococci retained cofactor (C4b-degrading) function . Using allelic exchange to construct strains with hybrid Por1A/B molecules, we demonstrate that the N-terminal loop (loop 1) of Por1A is required for C4bp binding . Serum resistant Por1B gonococcal strains also bind C4bp via their Por molecule . Using allelic exchange and site-directed mutagenesis, we have shown that loops 5 and 7 together form a negatively charged C4bp binding domain . C4bp-Por1B interactions are ionic in nature (inhibited by high salt as well as by heparin), while the C4bp-Por1A bond is hydrophobic . mAbs directed against SCR1 of the alpha-chain of C4bp inhibit C4bp binding to both Por1A and Por1B . Furthermore, only recombinant C4bp mutant molecules that contain alpha-chain SCR1 bind both Por1A and Por1B gonococci, confirming that SCR1 contains Por binding sites . C4bp alpha-chain monomers do not bind strains with either Por molecule, suggesting that the polymeric form of C4bp is required for binding to gonococci . Inhibition of C4bp binding to serum resistant Por1A and Por1B strains in a serum bactericidal assay using fAb fragments against C4bp SCR1 results in complete killing at 30 min of otherwise fully serum resistant strains in only 10% normal serum, underscoring the role of C4bp in mediating gonococcal serum resistance.

Pediatrics . 2001 May;107(5):E73.
Risk assessment for gonococcal and chlamydial infections in young children undergoing evaluation for sexual abuse; Ingram DM et al.; OBJECTIVE: Testing for gonorrhea (GC) and chlamydial (Ct) infection in children who are being evaluated for sexual abuse is invasive and costly . We developed selective criteria to limit unnecessary testing for these infections . METHODS: Over a 10-year period (May 1988 to May 1998), clinical information was collected in a prospectively designed database for all children ages 0 to 12 years by the WakeMed Child Sexual Abuse Team in Raleigh, North Carolina . The study population comprised 3040 (2414 girls and 626 boys) of the 3064 children evaluated for sexual abuse . Children were interviewed, examined, and tested by culture for GC and Ct orally, rectally, and genitally . Information from referral sources, accompanying guardians, and previous recent physical examinations was recorded . Bivariate analyses and logistic regression were used to develop 2 sets of screening criteria to predict children at greatest risk of infection with: 1) GC and/or Ct (GC/Ct) and 2) GC alone . RESULTS: Fifty-eight children were identified with GC/Ct infections (37 with GC, 25 with Ct; 4 children were coinfected) . The proposed algorithm for GC/Ct infections would have identified all children with these infections, while avoiding testing 56% of children without infection . Using genital cultures only, the proposed risk score for GC/Ct infections would have identified 100% of GC/Ct cases with 85% fewer cultures compared with testing all children with oral, rectal, and genital cultures . CONCLUSION: The use of a risk assessment algorithm for GC and Ct infections may reduce the cost and trauma of testing young children who are being evaluated for sexual abuse . sexual abuse, gonorrhea, Neisseria gonorrhoeae, chlamydia, Chlamydia trachomatis, sexually transmitted diseases, children.

AIDS Patient Care STDS, 1998 Jan, 12(1), 29 - 32
Using self-administered tampons to diagnose STDs; Bowden FJ et al.; The new nucleic acid amplification techniques (e.g., LCR and PCR) have allowed the introduction of non- and minimally-invasive techniques for the detection of sexually transmitted diseases (STDs) . In this article, we describe one of these-the self-administrated tampon method-which we have used to detect the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, and human papillomavirus in women.

J Immunol, 2001 Jun 1, 166(11), 6764 - 70
A novel interaction between type IV pili of Neisseria gonorrhoeae and the human complement regulator C4B-binding protein; Blom AM et al.; C4b-binding protein (C4BP) is an important plasma inhibitor of the classical pathway of complement activation . Several bacterial pathogens bind C4BP, which may contribute to their virulence . In the present report we demonstrate that isolated type IV pili from Neisseria gonorrhoeae bind human C4BP in a dose-dependent and saturable manner . C4BP consists of seven identical alpha-chains and one beta-chain linked together with disulfide bridges . We found that pili bind to the alpha-chain of C4BP, which is composed of eight homologous complement control protein (CCP) domains . From the results of an inhibition assay with C4b and a competition assay in which we tested mutants of C4BP lacking individual CCPs, we concluded that the binding area for pili is localized to CCP1 and CCP2 of the alpha-chain . The binding between pili and C4BP was abolished at 0.25 M NaCl, implying that it is based mostly on ionic interactions, similarly to what have been observed for C4b-C4BP binding . Furthermore, the N-terminal part of PilC, a structural component of pili, appeared to be responsible for binding of C4BP . Membrane cofactor protein, previously shown to be a receptor for pathogenic N . gonorrhoeae on the surface of epithelial cells, competed with C4BP for binding to pili only at high concentrations, suggesting that different parts of pili are involved in these two interactions . Accordingly, high concentrations of C4BP were required to inhibit binding of N . gonorrhoeae to Chang conjunctiva cells, and no inhibition of binding was observed with cervical epithelial cells.

Mol Microbiol, 2001 May, 40(3), 645 - 55
Mismatch repair and the regulation of phase variation in Neisseria meningitidis; Richardson AR et al.; Neisseria meningitidis controls the expression of several genes involved in host adaptation by a process known as phase variation . The phase variation frequency of haemoglobin (Hb) receptors among clinical isolates of serogroups A, B and C differed drastically, ranging from approximately 10(-6) to 10(-2) cfu-1 . Frequencies of phase variation are a genetic trait of a particular strain, as two unlinked Hb receptors, hpuAB and hmbR, phase varied with similar frequencies within a given isolate . Based on these frequencies, six Neisserial clinical isolates could be grouped into three distinct classes; slow, medium and fast . An increase in phase variation frequency was accompanied by high rates of spontaneous mutation to rifampicin and nalidixic acid resistance in one medium and one fast strain . The remaining three medium strains displayed elevated levels of phase variation without increases in overall mutability, as they possessed low rates of spontaneous mutation to drug resistance . The mismatch repair system of N . meningitidis was found to play an important role in determining the overall mutability of the clinical isolates . Inactivation of mismatch repair in any strain, regardless of its original phenotype, increased mutability to a level seen in the fast strain . Insertional inactivation of mutS and mutL in the slow strain led to 500- and 250-fold increases in hmbR switching frequency respectively . Concurrently, the frequency of spontaneous point mutations of mutS and mutL mutants from the slow strain was increased 20- to 30-fold to the level seen in the high strain . The status of Dam methylation did not correlate with either the phase variation frequency of Hb receptors or the general mutability of Neisserial strains . Analysis of an expanded set of isolates identified defects in mismatch repair as the genetic basis for strains displaying both the fast Hb switching and high mutation rate phenotypes . In conclusion, elevated frequencies of phase variation were accompanied by increased overall mutability in some N . meningitidis isolates including strains shown to be mismatch repair defective . Other isolates have evolved mechanisms that seem to affect only the switching frequency of phase-variable genes without an accompanied increased accumulation of spontaneous mutations.

Pathology, 2001 May, 33(2), 204 - 5
Urinary screening for Neisseria gonorrhoeae in asymptomatic individuals from Queensland, Australia: an evaluation using three nucleic acid amplification methods; Farrell DJ et al.; A comparison between three nucleic acid amplification methods (COBAS AMPLICOR NG, Abbott LCx NG and in-house cppB gene PCR) for the detection of Neisseria gonorrhoeae was undertaken using non-consecutive urine specimens collected from a high prevalence asymptomatic population (260 patients) from multiple sites in Queensland, Australia . Sensitivity, specificity, positive predictive value and negative predictive value were as follows: COBAS 97.9, 93.9, 78.0, 99.5%; LCx 95.7, 100, 100, 99.1%; cppB 97.9, 100, 100, 99.5% . The results provide further evidence that false-positive COBAS AMPLICOR NG PCR results are a significant problem in our population, and that the Abbott NG LCx is a suitable alternative for screening asymptomatic populations with a high prevalence of N . gonorrhoeae carriage.

Arch Neurol, 2001 May, 58(5), 806 - 10
Acute severe spinal cord dysfunction in bacterial meningitis in adults: MRI findings suggest extensive myelitis; Kastenbauer S et al.; BACKGROUND: Bacterial meningitis is rarely complicated by acute spinal cord involvement (eg, myelitis, ischemic infarction, spinal abscess, or epidural hemorrhage) . In spinal cord dysfunction, magnetic resonance imaging (MRI) is the imaging modality of choice . Still, MRI findings of myelitis due to bacterial meningitis in adults have not been reported . METHODS: Spinal MRIs were obtained during the acute stage of meningitis and on follow-up in 3 adults with bacterial meningitis that was complicated by paraparesis or tetraparesis and bowel and bladder incontinence . The causative pathogens were Streptococcus pneumoniae and Neisseria meningitidis; in 1 patient, the pathogen was not identified . RESULTS: In all cases, spinal MRI ruled out a compression of the cord by an extramedullary mass but demonstrated hyperintensities on T2-weighted images that predominantly involved the gray matter and extended from the cervical to the lumbar cord . Leptomeningeal and discrete nodular intramedullary enhancement on T1-weighted images was detected only in 1 patient . Follow-up examinations revealed that hyperintensities resolved completely in 1 patient, while a central cavitation developed in the cervical spinal cord of another, and the MRI findings were progressive during the first 4 weeks in the third patient . In all cases, severe paresis and bowel and bladder incontinence persisted . CONCLUSION: We demonstrate for the first time the MRI findings of adults with acute spinal cord involvement during bacterial meningitis . Magnetic resonance imaging showed central intramedullary hyperintensities on T2-weighted images that extended from the cervical to the lumbar cord, indicating myelitis . Clinical follow-up examinations suggest that myelitis during bacterial meningitis has an unfavorable prognosis.

Sex Transm Dis, 2001 May, 28(5), 247 - 51
Sexual behavior among adolescent women at high risk for sexually transmitted infections; Katz BP et al.; BACKGROUND: The temporal pattern of partners and sexual encounters may be key factors in the acquisition and transmission of sexually transmitted diseases (STDs) . Behavior among adolescent women is of particular interest because they frequently have the highest prevalence and incidence of infection . GOAL: To examine coital diary data collected during a 7-month longitudinal study of young women at high risk of STDs and to describe their sexual behaviors, with particular attention to issues of partner sequence and overlap . STUDY DESIGN: A 7-month longitudinal study of young women infected with or having a sexual contact infected with Neisseria gonorrhoeae, Chlamydia trachomatis, or Trichomonas vaginalis attending the STD clinic or one of four neighborhood adolescent health clinics . Data were collected at enrollment and at 1, 3, 5, and 7-month follow-up visits . Coital diaries were kept between visits . RESULTS: The average frequency of coital events was 0.94 per week . The median number of sexual partners during the follow-up period was one, and overlapping of partnerships was an uncommon occurrence . The number of days between the last coital event of a current relationship and the first encounter of a new relationship differed for those choosing a new partner (mean, 20.6 days) and those who returned to a previous partner (mean, 7.9 days; P < 0.001) . CONCLUSION: Although at high risk for STDs, high-risk behavior was not common among the study population . Partner choice and the behavior of these partners may be more important elements than personal high-risk behavior in accounting for the high prevalence of sexually transmitted infections among inner-city adolescent women.

Epidemiol Infect, 2001 Apr, 126(2), 219 - 24
Investigation of an outbreak of ciprofloxacin-resistant Neisseria gonorrhoeae using a simplified opa-typing method; Palmer HM et al.; Ciprofloxacin-resistant gonococci have been isolated from patients in the United Kingdom since 1993 . Until recently, evidence has suggested that the majority of infections are not endemic but have been acquired abroad . In October 1999, increasing numbers of ciprofloxacin resistant isolates of the non-requiring auxotype were reported in Oldham and Rochdale (Northwest England) . These and similar isolates from elsewhere in England and Wales were genetically characterized using a simplified opa-typing method (a non-radioactive PCR-RFLP method targeting the opa family of genes) . Of 73 isolates studied, 24 had unique opa-types (10 from infections acquired abroad), whilst the remaining 49 were indistinguishable (none were known to be acquired abroad) . This cluster included 31 isolates from Oldham and Rochdale, 16 from elsewhere in the north of England, and 2 from Southern England and South Wales with known epidemiological links to cases from Manchester and Rochdale respectively . This study illustrates the potential for spread of an antibiotic resistant clone of N . gonorrhoeae both locally and nationally and demonstrates that endemic acquisition of ciprofloxacin-resistant gonococci is now a significant problem in the United Kingdom.

Przegl Epidemiol, 2000, 54(3-4), 351 - 6
{Clinical course and etiologic factors of sepsis in children from different age groups}; Chlebna-Sokol D et al.; The analysis was performed of aetiological factors and clinical course of sepsis in children admitted to the Institute of Pediatrics of Medical University of Lodz in 1995-1998 . Staphyloccocus epidermidis was the most frequently found pathogen in neonates, whereas Neisseria meningitidis in infants and children over 1 year . Candida sp . was responsible for 25% of sepsis in infants . The severity of the disease correlated significantly with increased leucocytosis and shift to the left in the white cell profile as well as elevated levels of bilirubin and transaminases, especially GOT . Lower correlations were found between severity of clinical course and coagulological disturbances or inflammatory changes in the spinal cerebral fluid.

Vaccine, 2001 May 14, 19(25-26), 3568 - 74
Immunogenicity of recombinant class 1 protein from Neisseria meningitidis refolded into phospholipid vesicles and detergent; Niebla O et al.; The possibility of eliciting bactericidal antibodies against a recombinant class 1 protein (P1) from Neisseria meningitidis, joined to the first 45 amino acids of the neisserial LpdA protein (PM82), was examined . P1 was produced in Escherichia coli as intracellular inclusion bodies, from which it was purified and reconstituted by (a) inclusion into phospholipid vesicles and detergent and (b) refolding in 0.1% SDS . When Balb/c mice were immunised, high titres of subtype-specific bactericidal antibodies against P1 were obtained in both cases . These results suggest that in spite of being a denaturing agent, it is possible to use SDS to reconstitute the P1 protein in a conformation that exposes the immunodominat regions.

Vaccine, 2001 May 14, 19(25-26), 3390 - 8
Evaluation of cross-reactive antigens as determinants of cross-bactericidal activity in pathogenic and commensal Neisseria; Sanchez S et al.; Several antisera raised against outer membane vesicles obtained from invasive and carrier Neisseria meningitidis strains and commensal Neisseria and Moraxella catharralis species were assayed to test cross-bactericidal activity on Neisseria meningitidis strains . Results demonstrate that, despite the wide antigenic cross-reactivity previously shown by Western-blotting for the major outer membrane antigenic proteins of all Neisseria species, complement mediated killing shows very variable patterns that can not be predicted on the basis of antigenic cross-reactivity . Results of antibody tritations on homologous and heterologous strains, isotyping, and bactericidal activity of sera raised against denatured purified outer-membrane vesicle proteins, suggest that the responsibility for most of the bactericidal activity of the sera must be conformational and/or shared epitopes not detectable by Western-blotting.

Biochim Biophys Acta, 2001 Jan 12, 1544(1-2), 55 - 63
Cloning, expression and some properties of alpha-carbonic anhydrase from Helicobacter pylori; Chirica LC et al.; The alpha-carbonic anhydrase gene from Helicobacter pylori strain 26695 has been cloned and sequenced . The full-length protein appears to be toxic to Escherichia coli, so we prepared a modified form of the gene lacking a part that presumably encodes a cleavable signal peptide . This truncated gene could be expressed in E . coli yielding an active enzyme comprising 229 amino acid residues . The amino acid sequence shows 36% identity with that of the enzyme from Neisseria gonorrhoeae and 28% with that of human carbonic anhydrase II . The H . pylori enzyme was purified by sulfonamide affinity chromatography and its circular dichroism spectrum and denaturation profile in guanidine hydrochloride have been measured . Kinetic parameters for CO2 hydration catalyzed by the H . pylori enzyme at pH 8.9 and 25 degrees C are kcat=2.4x10(5) s(-1), KM=17 mM and kcat/KM=1.4x10(7) M(-1) x s(-1) . The pH dependence of kcat/KM fits with a simple titration curve with pK(a)=7.5 . Thiocyanate yields an uncompetitive inhibition pattern at pH 9 indicating that the maximal rate of CO2 hydration is limited by proton transfer between a zinc-bound water molecule and the reaction medium in analogy to other forms of the enzyme . The 4-nitrophenyl acetate hydrolase activity of the H . pylori enzyme is quite low with an apparent catalytic second-order rate constant, k(enz), of 24 M(-1) x s(-1) at pH 8.8 and 25 degrees C . However, with 2-nitrophenyl acetate as substrate a k(enz) value of 665 M(-1) x s(-1) was obtained under similar conditions.

Appl Microbiol Biotechnol, 2001 Apr, 55(3), 326 - 32
Proteome and transcriptome based analysis of Bacillus subtilis cells overproducing an insoluble heterologous protein; Jurgen B et al.; Bacillus subtilis and related Bacillus species are frequently used as hosts for the industrial production of recombinant proteins . In this study the cellular response of B . subtilis to the overproduction of an insoluble heterologous protein was investigated . For this purpose PorA, an outer membrane protein from Neisseria meningitidis, which accumulates after overexpression in the cytoplasm of B . subtilis mainly in the form of inclusion bodies, was used . The molecular response to overexpression of porA has been analysed at the transcriptional level using the DNA macro array technique and at the translational level by two-dimensional polyacrylamide gel electrophoresis . It was found that the expression of the heat shock genes of class I (dnaK, groEL and grpE) and class III (clpP and clpC) are increased under overproducing conditions . Furthermore, the protein levels of the two ribosomal proteins RpsB and RplJ are increased in the PorA overproducing cells . The transcriptome analysis indicated that mRNA levels of genes encoding pyrimidine and purine synthesis enzymes but also from ribosomal protein genes have elevated levels under overproducing conditions . Finally, the association of the protease ClpP and its ATPase subunits ClpC and ClpX with the PorA inclusion bodies was demonstrated by means of the immunogold labelling technique.

Enferm Infecc Microbiol Clin, 2001 Mar, 19(3), 99 - 102
{Microbiologic profile in symptomatic pregnant women's genital secretions in Gran Buenos Aires, Argentina}; Di Bartolomeo S et al.; BACKGROUND: Establish the prevalence of microorganisms associated with genital colonization in symptomatic pregnant women . In order to review the evolution of frequent pathogens ecology and adjust the laboratory design, in a population attended at the public health Hospital, in the Great Buenos Aires . METHODS: Vaginal and endocervical samples, were explored for specific detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Streptococcus agalactie, Trichomonas vaginalis, Candida spp., Mycoplasma hominis, Ureaplasma urealyticum and bacterial vaginosis (VB).Direct methods, culture, inmunodetection and PCR, were employed.In 1999; 198 women, with genital discharge, were studied . Age in the group range from 16 to 42 years old (Median 27 years old) . RESULTS: In 51 cases (25.7%) none of the above microorganisms or bacterial vaginosis were detected . In 30 cases (15.1%) bacterial vaginosis was diagnosed . Frequency of detection was: Ureaplasma urealyticum, 49,5%; Candida spp., 34,3%; Mycoplasma hominis, 14.1%; Streptococcus agalactie, 4.5%; Trichomonas vaginalis, 3.5%; Chlamydia trachomatis, 2.5% . No detection of Neisseria gonorrhoeae was demonstrated . CONCLUSIONS: There is a relevant frequency of bacterial vaginosis . On the other hand, lower prevalence of the Trichomonas vaginalis and Chlamydia trachomatis and also the absence of Neisseria gonorrhoeae was demonstrated.Culture for Streptococcus agalactie (at birth) and detection of Chlamydia trachomatis . must be extended to all pregnant women.Study of species and drug sensitivity of Candida spp., and detection of Ureaplasma urealyticum, Mycoplasma hominis and Neisseria gonorrhoeae, have to be explored under specific clinical requirement.

Pediatr Infect Dis J, 2001 Apr, 20(4), 416 - 21
Three-year survey of bacteremia and fungemia in a pediatric intensive care unit; Gray J et al.; BACKGROUND: Knowledge of the pattern of blood stream infection (BSI) in patients in intensive care units (ICUs) can help determine antibiotic prescribing policy and infection control procedures . However, there have been few pediatric-based studies . METHODS: Surveillance of BSI in a pediatric ICU for 3 years, amounting to 131 episodes of significant bacteremia and fungemia . RESULTS: The incidence of BSI was 39.0 per 1,000 admissions (10.6 per 1,000 bed days) . Eighty-four (64.1%) episodes were ICU-acquired, and 27 (20.6%) were community-acquired . Gram-positive, Gram-negative and anaerobic bacteria accounted for 62.2, 30.8 and 1.4%, respectively, of the 143 microorganisms isolated, 5.6% were yeasts . Neisseria meningitidis was the most common species in community-acquired infections, and staphylococci predominated in hospital-acquired episodes . Eighty-seven percent of patients had significant underlying disease, including 60.3% with congenitally acquired conditions . Intravascular devices were the most common source of infection, accounting for 41.2% of all episodes . The crude mortality in children with BSI was 26.5%, compared with 8.1% in those without BSI . CONCLUSIONS: The pattern of BSI in ICUs is partly determined by the type of patient treated . However, some observations are generally applicable, notably the increasing importance of antibiotic-resistant bacteria that are often of low virulence and device-associated . Our experience suggests that universal use of broad spectrum empiric antibiotics to cover these pathogens (which risks further promoting antibiotic resistance) may not improve patient outcome . Our study provides a basis for other pediatric ICUs to evaluate their rates and outcomes of BSI.

J Antimicrob Chemother, 2001 May, 47(5), 651 - 4
Decreased susceptibility to azithromycin and erythromycin mediated by a novel mtr(R) promoter mutation in Neisseria gonorrhoeae; Zarantonelli L et al.; During a screen of Neisseria gonorrhoeae clinical isolates obtained in Uruguay for susceptibility to azithromycin, we noticed that approximately 10% of the strains examined displayed decreased susceptibility to azithromycin and erythromycin due to the mtr(CDE)-encoded efflux pump system, but remained susceptible to Triton X-100 . We now report that the mtr(R) promoter region of one of these isolates contains a dinucleotide insertion (TT) that mediates this resistance phenotype.

J Clin Microbiol, 2001 May, 39(5), 1751 - 6
Detection of Chlamydia trachomatis and Neisseria gonorrhoeae by enzyme immunoassay, culture, and three nucleic acid amplification tests; Van Dyck E et al.; The purpose of this study was to evaluate and compare three commercially available nucleic acid amplification tests (NAATs) for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis . Roche PCR and Becton Dickinson strand displacement amplification (SDA) were performed on 733 endocervical swab specimens from commercial sex workers . Abbott ligase chain reaction (LCR) was performed on a subset of 396 samples . Endocervical specimens from all women were also tested by culture for N . gonorrhoeae and by Syva MicroTrak enzyme immunoassay (EIA) for C . trachomatis . A positive N . gonorrhoeae result was defined as a positive result by culture or by two NAATs, and a positive C . trachomatis result was defined as a positive result by two tests . According to these definitions, the sensitivities and specificities for the subsample of 396 specimens of N . gonorrhoeae culture, PCR, SDA, and LCR were 69.8, 95.2, 88.9, and 88.9% and 100, 99.4, 100, and 99.1%, respectively; the sensitivities and specificities of C . trachomatis EIA, PCR, SDA, and LCR were 42.0, 98.0, 94.0, and 90.0% and 100, 98.0, 100, and 98.6%, respectively . The performance characteristics of N . gonorrhoeae culture, PCR, and SDA and C . trachomatis EIA, PCR, and SDA for all 733 specimens were defined without inclusion of LCR results and by discrepant analysis after resolution of discordant N . gonorrhoeae PCR results and of discordant C . trachomatis EIA and PCR results by LCR testing . The sensitivities of N . gonorrhoeae culture, PCR, and SDA before and after LCR resolution were 67.8, 95.7, and 93.9% and 65, 95.8, and 90.0%, respectively . The sensitivities of C . trachomatis EIA, PCR, and SDA decreased from 39.4, 100, and 100% to 38.7, 98.7, and 94.7%, respectively . All three NAATs proved to be superior to N . gonorrhoeae culture and to C . trachomatis EIA . The accuracies of the different NAATs were quite similar . SDA was the only amplification assay with 100% specificity for detection of both N . gonorrhoeae and C . trachomatis in endocervical specimens.

J Clin Microbiol, 2001 May, 39(5), 1710 - 5
Genetic diversity of Neisseria lactamica strains from epidemiologically defined carriers; Alber D et al.; We assessed the genetic diversity of 26 Neisseria lactamica strains from epidemiologically related sources, i.e., groups of kindergartens and primary schools in three Bavarian towns, by the partial sequencing of the argF, rho, recA, and 16S ribosomal genes . We found a total of 17 genotypes, of which 12 were found only in one strain . The genotypes comprised 5 alleles of the argF gene, 9 of rho, 8 of recA, and 10 of the 16S ribosomal DNA . Sequence analysis by determination of homoplasy ratios and split decomposition analysis revealed abundant recombination within N . lactamica.

J Bacteriol, 2001 May, 183(10), 3160 - 8
ComE, a competence protein from Neisseria gonorrhoeae with DNA-binding activity; Chen I et al.; Neisseria gonorrhoeae is naturally able to take up exogenous DNA and undergo genetic transformation . This ability correlates with the presence of functional type IV pili, and uptake of DNA is dependent on the presence of a specific 10-bp sequence . Among the known competence factors in N . gonorrhoeae, none has been shown to interact with the incoming DNA . Here we describe ComE, a DNA-binding protein involved in neisserial competence . The gene comE was identified through similarity searches in the gonococcal genome sequence, using as the query ComEA, the DNA receptor in competent Bacillus subtilis . The gene comE is present in four identical copies in the genomes of both N . gonorrhoeae and Neisseria meningitidis, located downstream of each of the rRNA operons . Single-copy deletion of comE in N . gonorrhoeae did not have a measurable effect on competence, whereas serial deletions led to gradual decrease in transformation frequencies, reaching a 4 x 10(4)-fold reduction when all copies were deleted . Transformation deficiency correlated with impaired ability to take up exogenous DNA; however, the mutants presented normal piliation and twitching motility phenotype . The product of comE has 99 amino acids, with a predicted signal peptide; by immunodetection, a 8-kDa protein corresponding to processed ComE was observed in different strains of N . gonorrhoeae and N . meningitidis . Recombinant His-tagged ComE showed DNA binding activity, without any detectable sequence specificity . Thus, we identified a novel gonococcal DNA-binding competence factor which is necessary for DNA uptake and does not affect pilus biogenesis or function.

Lancet, 2001 Apr 14, 357(9263), 1161 - 7
The burden of reproductive-organ disease in rural women in The Gambia, West Africa; Walraven G et al.; BACKGROUND: Data on the epidemiology of reproductive-organ morbidity are needed to guide effective interventions, to set health-care priorities, and to target future research . This study aimed to find out the prevalence of reproductive-organ disease in a sample of rural Gambian women . METHODS: A questionnaire on reproductive health was administered by fieldworkers to women aged 15-54 years living in a rural area under demographic surveillance . A female gynaecologist questioned and examined the women (including speculum and bimanual pelvic examinations) . Vaginal swabs were taken to test for Trichomonas vaginalis, Candida albicans, and bacterial vaginosis, cervical smears for cytology, cervical swabs for Chlamydia trachomatis PCR and Neisseria gonorrhoeae culture, and venous blood for haemoglobin, HIV, herpes simplex virus 2, and syphilis serology . FINDINGS: 1348 (72.0%) of 1871 eligible women took part . Reproductive-organ symptoms were more likely to be reported to the gynaecologist (52.7% of women) than to the fieldworker (26.5%) . Menstrual problems, abnormal vaginal discharge, and vaginal itching were the most commonly reported symptoms . A minority of women said they had sought health care for their symptoms . The frequencies of reproductive-organ morbidity were high: menstrual dysfunction 34.1% (95% CI 29.6-39.1), infertility 9.8% (8.2-11.6), reproductive-tract infections 47.3% (43.7-51.0), pelvic tenderness 9.8% ((7.0-13.5), cervical dysplasia 6.7% (5.2-8.4), masses 15.9% (12.5-20.1), and childbirth-related damage to pelvic structures 46.1% (40.1-52.3) . 948 (70.3%) women had at least one reproductive-organ disorder . INTERPRETATION: For these rural women, whose lives depend heavily on their reproductive function, reproductive-organ disease is a large burden . In inadequately resourced rural areas, with poor education, heavy agricultural and domestic labour, and limited access to quality health care, many women are not able to attain and maintain reproductive health and wellbeing.

Sex Transm Dis, 2001 Apr, 28(4), 232 - 5
Management of urethritis in health maintenance organization members receiving care at a multispecialty group practice in Massachusetts; Ratelle S et al.; BACKGROUND: Cost containment has led to a concern that health maintenance organization-insured patients presenting with complaints of urethritis may be treated without being tested . GOAL: To determine the proportion of men presenting with symptoms of urethritis who are tested for Chlamydia trachomatis and Neisseria gonorrhoeae . STUDY DESIGN: Reviews were performed on 196 randomly selected patient records with an outpatient visit and a diagnostic code consistent with urethritis between 1995 and 1997 . Data were collected on demographics, diagnostic testing, and treatment . RESULTS: Diagnostic testing for C trachomatis and N gonorrhoeae was performed, respectively, in 92.3% and 83.2% of the men presenting at an initial visit with complaints of urethritis . Altogether, 98.2% of the patients who met the Centers for Disease Control criteria for documenting urethritis were tested for C trachomatis and N gonorrhoeae . CONCLUSION: Diagnostic testing for C trachomatis and N gonorrhoeae is nearly universal in this multispecialty group practice setting, facilitating surveillance and public health efforts.

J Acquir Immune Defic Syndr, 2001 Apr 1, 26(4), 360 - 4
Selenium deficiency is associated with shedding of HIV-1--infected cells in the female genital tract; Baeten JM et al.; OBJECTIVE: To assess the relation between selenium deficiency and vaginal or cervical shedding of HIV-1-infected cells . DESIGN: Cross-sectional study of 318 HIV-1 seropositive women in Mombasa, Kenya . METHODS: Vaginal and cervical swab specimens were tested for the presence of HIV-1 DNA by polymerase chain reaction . Multivariate logistic regression models, adjusting for CD4 count and vitamin A deficiency, were used . RESULTS: Selenium deficiency (defined as levels <85 microg/L) was observed in 11% of the study population . In unstratified multivariate analyses, there was no significant association between selenium deficiency and vaginal or cervical shedding . In stratified analyses, however, significant associations became apparent after excluding women with predictors of shedding with strong local effects on the genital tract mucosa . Among women who did not use oral contraceptives and who did not have vaginal candidiasis, selenium deficiency was significantly associated with vaginal shedding (adjusted odds ratio {AOR} 2.9, 95% confidence interval {CI} 1.0--8.8, p =.05) . Effect modification was also observed in the relation between selenium deficiency and cervical shedding, with a significant association seen among those women who were not using oral contraceptive pills or depot medroxyprogesterone acetate and who did not have Neisseria gonorrhoeae infection (AOR 2.8, 95% CI 1.1--7.0, p =.02) . CONCLUSIONS: We found selenium deficiency to be associated with a nearly threefold higher likelihood of genital mucosal shedding of HIV-1--infected cells, suggesting that deficiency may increase the infectiousness of women with HIV-1 . Nutritional interventions to prevent HIV-1 transmission warrant investigation.

J Vet Med B Infect Dis Vet Public Health, 2001 Mar, 48(2), 137 - 41
Infectious bovine keratoconjunctivitis and lymphofollicular hyperplasia of the third eyelid in heifers; Yeruham I et al.; On a dairy cattle farm, infectious bovine keratoconjunctivitis was diagnosed in 29 (24%) calves and heifers aged from 2 weeks to 1 year old . The highest infection rate (18%) occurred in animals aged 3-6 months . The bacteriological examination of swabs from the affected animals yielded several species of bacteria: Moraxella bovis, Neisseria ovis, N . cuniculi, plasma coagulase-negative Staphylococcus spp., alpha-haemolytic Streptococcus spp., Arcanobacterium pyogenes and Escherichia coli . Moraxella bovis and N . ovis were the most common isolates . Hyperplasia of the lymphatic tissue of the third eyelid in the form of nodules 7-8 mm in diameter was diagnosed in two heifers aged 8 and 10 months.

J Biol Chem, 2001 Jul 6, 276(27), 25273 - 8 Epub 2001 Apr 16.
Amylosucrase, a glucan-synthesizing enzyme from the alpha-amylase family; Skov LK et al.; Amylosucrase (E.C . 2.4.1.4) is a member of Family 13 of the glycoside hydrolases (the alpha-amylases), although its biological function is the synthesis of amylose-like polymers from sucrose . The structure of amylosucrase from Neisseria polysaccharea is divided into five domains: an all helical N-terminal domain that is not similar to any known fold, a (beta/alpha)(8)-barrel A-domain, B- and B'-domains displaying alpha/beta-structure, and a C-terminal eight-stranded beta-sheet domain . In contrast to other Family 13 hydrolases that have the active site in the bottom of a large cleft, the active site of amylosucrase is at the bottom of a pocket at the molecular surface . A substrate binding site resembling the amylase 2 subsite is not found in amylosucrase . The site is blocked by a salt bridge between residues in the second and eight loops of the (beta/alpha)(8)-barrel . The result is an exo-acting enzyme . Loop 7 in the amylosucrase barrel is prolonged compared with the loop structure found in other hydrolases, and this insertion (forming domain B') is suggested to be important for the polymer synthase activity of the enzyme . The topology of the B'-domain creates an active site entrance with several ravines in the molecular surface that could be used specifically by the substrates/products (sucrose, glucan polymer, and fructose) that have to get in and out of the active site pocket.

J Biol Chem, 2001 Jun 29, 276(26), 24331 - 40 Epub 2001 Apr 16.
Pathogenic Neisseria trigger expression of their carcinoembryonic antigen-related cellular adhesion molecule 1 (CEACAM1; previously CD66a) receptor on primary endothelial cells by activating the immediate early response transcription factor, nuclear factor-kappaB; Muenzner P et al.; Neisseria gonorrhoeae express opacity-associated (Opa) protein adhesins that mediate binding to various members of the carcinoembryonic antigen-related cellular adhesion molecule (CEACAM; previously CD66) receptor family . Although human umbilical vein endothelial cells express little CEACAM receptor in vitro, we found neisserial infection to induce expression of CEACAM1, CEACAM1-3L, and CECAM1-4L splice variants . This mediates an increased Opa(52)-dependent binding of gonococci by these cells . The induced receptor expression did not require bacterial Opa expression, but it was more rapid with adherent bacteria . Because the time course of induction was similar to that seen for induced proinflammatory cytokines, we tested whether CEACAM1 expression could be controlled by a similar mechanism . Gonococcal infection activated a nuclear factor-kappaB (NF-kappaB) heterodimer consisting of p50 and p65, and inhibitors that prevent the nuclear translocation of activated NF-kappaB complex inhibited CEACAM1 transcript expression . Each of these effects could be mimicked by using culture filtrates or purified lipopolysaccharide instead of intact bacteria . Together, our results support a model whereby the outer membrane "blebs" that are actively released by gonococci trigger a Toll-like receptor-4-dependent activation of NF-kappaB, which up-regulates the expression of CEACAM1 to allow Opa(52)-mediated neisserial binding . The regulation of CEACAM1 expression by NF-kappaB also implies a broader role for this receptor in the general inflammatory response to infection.

Clin Infect Dis, 2001 May 1, 32(9), 1313 - 8 Epub 2001 Apr 13.
A randomized trial of ciprofloxacin versus cefixime for treatment of gonorrhea after rapid emergence of gonococcal ciprofloxacin resistance in The Philippines; Aplasca De Los Reyes MR et al.; From 1994 through 1996-1997, high-level ciprofloxacin resistance (minimum inhibitory concentration {MIC}, > or = 4.0 microg/mL) increased from 9% to 49% of gonococcal isolates recovered from consecutive female sex workers in Cebu and Manila, The Philippines (P < .01) . During 1996-1997, 105 female sex workers with gonorrhea were prospectively randomized to receive treatment with oral ciprofloxacin, 500 mg, or cefixime, 400 mg, and followed for test of cure . Neisseria gonorrhoeae was reisolated within 28 days after treatment from 1 (3.8%) of 26 women given cefixime versus 24 (32.3%) of 72 women given ciprofloxacin (P < .01) . Treatment failure (reisolation of pretreatment auxotype/serovar) occurred in 14 (46.7%) of 30 women infected with strains with MICs of ciprofloxacin > or = 4.0 microg/mL versus 1 (3.6%) of 28 infected by strains with MICs < 4.0 microg/mL (P < .01) . High-level, clinically significant gonococcal resistance to ciprofloxacin has rapidly emerged in The Philippines, and spread of fluoroquinolone resistance through commercial sex poses a threat to control of gonorrhea and prevention of human immunodeficiency virus infection and the acquired immunodeficiency syndrome.

Am J Obstet Gynecol, 2001 Apr, 184(5), 856 - 63; discussion 863-4
Clinical predictors of endometritis in women with symptoms and signs of pelvic inflammatory disease; Peipert JF et al.; OBJECTIVE: Careful detection and treatment of pelvic inflammatory disease are essential for the prevention of adverse sequelae . The purpose of this study was to evaluate the diagnostic test characteristics of clinical criteria for the diagnosis of pelvic inflammatory disease . STUDY DESIGN: We performed a cross-sectional analysis of the baseline characteristics of 651 patients enrolled in a multicenter randomized treatment trial for pelvic inflammatory disease . Clinical and laboratory findings were recorded for all patients, and endometrial sampling was performed . We calculated sensitivity and specificity and performed receiver operating characteristic curve analysis and multivariate logistic regression, using histologic endometritis as the criterion standard . RESULTS: The minimal criteria for pelvic inflammatory disease, as recommended by the Centers for Disease Control and Prevention, had a sensitivity of 83%, in comparison with a 95% sensitivity for adnexal tenderness (P =.001) . Of the supportive clinical criteria, the finding most highly associated with endometritis was a positive test result for Chlamydia trachomatis or Neisseria gonorrhoeae (adjusted odds ratio, 4.3; 95% confidence interval, 2.89--6.63) . A multivariate logistic regression model indicated that combinations of criteria significantly improve the prediction of endometritis . CONCLUSION: Sensitivity can be maximized by using the presence of adnexal tenderness as a minimal criterion for the diagnosis of pelvic inflammatory disease, and supportive criteria are helpful in estimating the probability of endometritis.

Contemp Top Lab Anim Sci, 2001 Mar, 40(2), 13 - 7
Standardization of the Whitten Effect to induce susceptibility to Neisseria gonorrhoeae in female mice; Dalal SJ et al.; Female mice (Mus musculus) frequently are used to study hormonally related differences in susceptibility to infectious organisms or response to pharmaceutical agents . Cyclical variation in hormone levels within a group of mice, however, challenges the experimental design of such studies in that it is often difficult to obtain sufficient numbers of mice in the desired phase of the estrous cycle at the time of treatment . The purpose of this work is to provide investigators with a standardized protocol for inducing estrus in mice through exposure to male urine (Whitten Effect) . In addition, we demonstrate how the Whitten Effect can be used to induce susceptibility of mice to Neisseria gonorrhoeae infection . Female BALB/c mice were exposed to male urine via soiled bedding for 0, 24, 48, 72, or 96 h . The effect of exposure on the reproductive cycle was monitored by cytologic examination of vaginal smears and measurement of serum 17-b estradiol levels by using a nonradioactive immunoassay kit . In a separate experiment, mice were exposed to male-urine-soaked bedding for 0, 24, 72, or 96 h prior to intravaginal inoculation with Neisseria gonorrhoeae . Infection was monitored by using vaginal culture for 5 consecutive days . We found that the highest percentage of mice in estrus occurred among mice that were exposed to male-urine-soaked bedding for 96 h . Consistent with this finding was the demonstration that mice were more susceptible to gonococcal infection after exposure to male urine for 3 to 4 days . We conclude that exploitation of this natural murine behavioral response is a simple and inexpensive method by which estrus can be synchronized in a group of mice within a defined period of time . In addition, this protocol can be used to increase mouse susceptibility to experimental gonococcal infection.

Cell Microbiol, 2001 Apr, 3(4), 265 - 75
The pilus-induced Ca2+ flux triggers lysosome exocytosis and increases the amount of Lamp1 accessible to Neisseria IgA1 protease; Ayala BP et al.; The IgA1 protease secreted by the pathogenic Neisseriae cleaves Lamp1, a major integral membrane glycoprotein of lysosomes, and significantly reduces its steady-state levels in an infected cell . IgA1 protease hydrolysis of Lamp1 is inefficient at the low pH of lysosomes, strongly suggesting that the enzyme is unlikely to reduce Lamp1 levels within lysosomes to any appreciable extent . We therefore explored the possibility that the protease may reach Lamp1 through an alternative route . We demonstrate that Neisseria pili induce a transient increase in the levels of cytosolic free Ca2+ in A431 human epithelial cells, as demonstrated previously for ME180 cells . This Ca2+ flux triggers lysosome exocytosis, quickly altering the cellular distribution of Lamp1 and increasing surface Lamp1 levels . Finally, we demonstrate that surface Lamp1 is cleaved by IgA1 protease secreted by adherent bacteria . We conclude that the pilus-induced Ca2+ flux increases the amount of Lamp1 that is cleavable by the IgA1 protease.

Microbios, 2001, 104(408), 87 - 97
Luteolin inhibits the growth and arylamine N-acetyl-transferase activity in Neisseria gonorrhoeae; Tsou MF et al.; Growth inhibition and arylamine N-acetyltransferase (NAT) activity in Neisseria gonorrhoeae were inhibited by luteolin, a drug which originated from herbs . The growth inhibition was based on changes in optical density (OD) using a spectrophotometer, and arylamine NAT activity with 2-aminofluorene (2-AF) was determined using high pressure liquid chromatography . The inhibition of growth in N . gonorrhoeae demonstrated that luteolin elicited a dose-dependent growth inhibition in the N . gonorrhoeae cultures . Suspensions of N . gonorrhoeae with or without specific concentrations of luteolin cotreatment showed different percentages of 2-AF acetylation . The data indicated that there was reduced NAT activity associated with increased levels of luteolin in N . gonorrhoeae suspensions . Time-course experiments showed that NAT activity measured from intact N . gonorrhoeae cells was inhibited by luteolin for at least 4 h . Using standard steady-state kinetic analysis, it was demonstrated that luteolin was a possible uncompetitive inhibitor to NAT activity in N . gonorrhoeae . This report is the first to show that luteolin can inhibit N . gonorrhoeae NAT activity.

Infect Immun, 2001 May, 69(5), 3335 - 42
Monoclonal antibodies specific for Neisseria meningitidis group B polysaccharide and their peptide mimotopes; Shin JS et al.; From five mice immunized with Escherichia coli K1 bacteria, we produced 12 immunoglobulin M hybridomas secreting monoclonal antibodies (MAbs) that bind to Neisseria meningitidis group B (NMGB) . The 12 MAbs also bound the capsular polysaccharide (PS) of E . coli K1 {which, like NMGB, is alpha(2-8)-linked polysialic acid (PSA)} and bound to EV36, a nonpathogenic E . coli K-12 strain producing alpha(2-8) PSA . Except for HmenB5, which cross-reacted with N . meningitidis group C, none of the MAbs bound to N . meningitidis groups A, C, and Y . Of the 12 MAbs, 6 were autoantibodies as defined by binding to CHP-134, a neuroblastoma cell line expressing short-chain alpha(2-8) PSA; five of these MAbs killed NMGB in the presence of rabbit complement, and two also killed NMGB with human complement . The other six MAbs, however, were nonautoreactive; all killed NMGB with rabbit complement, and five killed NMGB with human complement . To obtain peptide mimotopes of NMGB PS, four of the nonautoreactive MAbs (HmenB2, HmenB3, HmenB13, and HmenB14) were used to screen two types of phage libraries, one with a linear peptide of 7 amino acids and the other with a circular peptide of 7 amino acids inserted between two linked cysteines . We obtained 86 phage clones that bound to the screening MAb in the absence but not in the presence of E . coli K1 PSA in solution . The clones contained 31 linear and 4 circular mimotopes expressing unique sequences . These mimotopes nonrandomly expressed amino acids and were different from previously described mimotopes for NMGB PS . The new mimotopes may be useful in producing a vaccine(s) capable of eliciting anti-NMGB antibodies not reactive with neuronal tissue.

Alcohol Clin Exp Res, 2001 Mar, 25(3), 421 - 6
Acetaldehyde production and other ADH-related characteristics of aerobic bacteria isolated from hypochlorhydric human stomach; Vakevainen S et al.; BACKGROUND: Acetaldehyde is a known local carcinogen in the digestive tract in humans . Bacterial overgrowth in the hypochlorhydric stomach enhances production of acetaldehyde from ethanol in vivo after alcohol ingestion . Therefore, microbially produced acetaldehyde may be a potential risk factor for alcohol-related gastric and cardiac cancers . This study was aimed to investigate which bacterial species and/or groups are responsible for acetaldehyde formation in the hypochlorhydric human stomach and to characterize their alcohol dehydrogenase (ADH) enzymes . METHODS: After 7 days of treatment with 30 mg of lansoprazole twice a day, a gastroscopy was performed on eight volunteers to obtain hypochlorhydric gastric juice . Samples were cultured and bacteria were isolated and identified; thereafter, their acetaldehyde production capacity was measured gas chromatographically by incubating intact bacterial suspensions with ethanol at 37 degrees C . Cytosolic ADH activities, Km values, and protein concentration were determined spectrophotometrically . RESULTS: Acetaldehyde production of the isolated bacterial strains (n = 51) varied from less than 1 to 13,690 nmol of acetaldehyde/10(9) colony-forming units/hr . ADH activity of the strains that produced more than 100 nmol of acetaldehyde/10(9) colony-forming units/hr (n = 23) varied from 3.9 to 1253 nmol of nicotinamide adenine dinucleotide per minute per milligram of protein, and Km values for ethanol ranged from 0.65 to 116 mM and from 0.5 to 3.1 M (high Km) . There was a statistically significant correlation (r = 0.64, p < 0.001) between ADH activity and acetaldehyde production from ethanol in the tested strains . The most potent acetaldehyde producers were Neisseria and Rothia species and Streptococcus salivarius, whereas nearly all Stomatococcus, Staphylococcus, and other Streptococcus species had a very low capacity to produce acetaldehyde . CONCLUSIONS: This study demonstrated that certain bacterial species or groups that originate from the oral cavity are responsible for the bulk of acetaldehyde production in the hypochlorhydric stomach . These findings provide new information with the respect to the local production of carcinogenic acetaldehyde in the upper digestive tract of achlorhydric human subjects.

Sex Transm Dis, 2001 Mar, 28(3), 136 - 42
Single-dose gatifloxacin compared with ofloxacin for the treatment of uncomplicated gonorrhea: a randomized, double-blind, multicenter trial; Stoner BP et al.; BACKGROUND: Treatment of gonorrhea is complicated by widespread resistance of Neisseria gonorrhoeae to antimicrobial agents of choice, including decreased susceptibility to ciprofloxacin . GOAL: To demonstrate the efficacy and safety of gatifloxacin, a novel 8-methoxy fluoroquinolone antibiotic, compared with ofloxacin in treating patients with uncomplicated gonococcal infection . STUDY DESIGN: In a double-blind, randomized (2:2:1), controlled trial, 340 men and 388 women with uncomplicated gonorrhea who were 16 years or older received a single oral dose of gatifloxacin (400 mg or 600 mg) or ofloxacin (400 mg) . Primary analysis of efficacy was based on bacteriologic eradication from sites of infection . Secondary analyses examined clinical response and adverse event profiles . RESULTS: Bacteriologic eradication rates for gatifloxacin in evaluable men with urethral gonorrhea were 99% (400 mg) and 100% (600 mg) versus 100% for ofloxacin (n = 117, 122, and 55, respectively; P = ns) . Eradication rates in evaluable women with endocervical gonorrhea were 99% for both 400 mg and 600 mg gatifloxacin versus 100% for ofloxacin (n = 101, 104, and 55, respectively; P = ns) . Eradication rates were 100% for both rectal (n = 43) and pharyngeal (n = 31) infection across all treatment groups . All three drug regimens were well tolerated and exhibited similar clinical response profiles . CONCLUSION: Gatifloxacin is safe and effective as a single 400-mg or 600-mg dose for the treatment of uncomplicated gonorrhea . Similar efficacy rates were observed with the 400-mg and 600-mg doses . A single 400-mg dose can be recommended for treatment of uncomplicated gonorrhea.

Sex Transm Infect, 2001 Apr, 77(2), 125 - 9
Aetiology of urethral discharge in Bangui, Central African Republic; Morency P et al.; OBJECTIVES: To determine the aetiology of urethritis in Bangui, Central African Republic . METHODS: 410 men presenting with urethral discharge and 100 asymptomatic controls were enrolled . Urethral swabs were obtained and processed by gonococcal culture and polymerase chain reaction for the detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Trichomonas vaginalis, and Ureaplasma urealyticum . RESULTS: In multivariate analyses, M genitalium and C trachomatis were significantly associated with urethral discharge when comparing cases of non-gonococcal urethritis (NGU) with controls . T vaginalis was also more common in cases than in controls, but this reached statistical significance only among cases in whom N gonorrhoeae was also detected . U urealyticum was not associated with urethritis . The gonococcus was found in 69% of cases of urethral discharge . M genitalium was the predominant pathogen in patients with NGU, being found in 42% (53/127) of such patients while C trachomatis was found in only 17% (22/127) . T vaginalis was found in 18% (23/127) of patients with NGU, but also in 15% (43/283) of patients with gonococcal urethritis, and two thirds of patients with T vaginalis also had the gonococcus . Multiple infections were common . M genitalium caused a syndrome similar to chlamydial urethritis, with a less severe inflammation than in gonococcal infection . No behavioural or clinical characteristic could discriminate between the various aetiological agents . CONCLUSIONS: M genitalium is more prevalent than C trachomatis and is the most common cause of NGU in BANGUI: It causes a syndrome similar to chlamydial urethritis . T vaginalis is weakly associated with urethritis, and is often found along with other pathogens.

Sex Transm Infect, 2001 Apr, 77(2), 114 - 9
Sexually transmitted infections among married women in Dhaka, Bangladesh: unexpected high prevalence of herpes simplex type 2 infection; Bogaerts J et al.; OBJECTIVES: To document the prevalence of reproductive tract infections (RTI) and sexually transmitted infections (STI) among women attending a basic healthcare clinic in Dhaka, Bangladesh, to identify risk factors associated with the diseases and to estimate the incidence of syphilis, hepatitis C (HCV), hepatitis B (HBV), and herpes simplex type 2 (HSV-2) infection . METHODS: A cross sectional sample of 2335 consecutive women was examined during 1996-8 . Women were interviewed about risk factors for RTI/STI and tested for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Treponema pallidum, HIV, HCV, HBV, HSV-1 and HSV-2 infection as well as vaginal candidosis and bacterial vaginosis . Women with antibodies to T pallidum were retested at regular intervals . One year after ending the study seroconversion for syphilis, HBV, HCV, and HSV-2 infection was detected among women initially negative for the respective diseases . RESULTS: The overall prevalence rate of N gonorrhoeae, C trachomatis, T vaginalis, and T pallidum infection was 0.5%, 1.9%, 2.0%, and 2.9% respectively . Overall, 35% of the women had antibodies to hepatitis B core antigen, 0.9% had HCV, and 12% HSV-2 infection . Risk factors for gonorrhoea/C trachomatis infection were a husband not living at home or suspected of being unfaithful . HSV-2 infection was associated with the same risk factors and with a polygamous marriage . The prevalence of HSV-2 infection among women "at risk" was 23% . HIV infection was not diagnosed . Repeated serological examination indicated that only 32% of women with serological evidence of syphilis had active disease . The seroincidences of HBV, HCV, and HSV-2 were 0.03, 0.007, and 0.009 per person year . Seroconversion for syphilis was not observed.

Sex Transm Infect, 2001 Apr, 77(2), 103 - 6
Endocervical Gram stain smears and their usefulness in the diagnosis of Chlamydia trachomatis; Myziuk L et al.; OBJECTIVE: To evaluate the usefulness of endocervical Gram stain smears in the diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae in a female population attending a STD clinic . METHODS: 494 females attending a STD clinic and undergoing a speculum examination had endocervical specimens submitted for C trachomatis culture, direct fluorescent antibody testing (DFA), and N gonorrhoeae culture . Endocervical smears were also collected for Gram stain . The number of polymorphonuclear leucocytes (PMN) per high power field (HPF), presence of bacteria, yeast, red blood cells, and clue cells were recorded . Clinical signs of cervicitis were also documented . RESULTS: N gonorrhoeae was isolated from one subject who was co-infected with C trachomatis and no further analysis was done regarding N gonorrhoeae . Analysis was performed on 220 participants . The prevalence of C trachomatis infection was 13% . Of the Gram smears collected, 55% were inadequate owing to the presence of vaginal contamination . There were an equal number of C trachomatis isolates in patients with < or = 10 PMN/HPF (48%) and > 10 PMN/HPF (52%) . Endocervical mucopus and erythema were statistically significant for the presence of C trachomatis (p < 0.001 and 0.02 respectively) . The presence of any signs of cervicitis-that is, mucopus, friability, erythema, and ectropion together with > 10 PMN/HPF was statistically significant for the presence of C trachomatis . CONCLUSION: The use of endocervical Gram smear results together with clinical information can be used to identify high risk women for C trachomatis infection.

Proc Natl Acad Sci U S A, 2001 Apr 24, 98(9), 5234 - 9 Epub 2001 Apr 03.
Fit genotypes and escape variants of subgroup III Neisseria meningitidis during three pandemics of epidemic meningitis; Zhu P et al.; The genetic variability at six polymorphic loci was examined within a global collection of 502 isolates of subgroup III, serogroup A Neisseria meningitidis . Nine "genoclouds" were identified, consisting of genotypes that were isolated repeatedly plus 48 descendent genotypes that were isolated rarely . These genoclouds have caused three pandemic waves of disease since the mid-1960s, the most recent of which was imported from East Asia to Europe and Africa in the mid-1990s . Many of the genotypes are escape variants, resulting from positive selection that we attribute to herd immunity . Despite positive selection, most escape variants are less fit than their parents and are lost because of competition and bottlenecks during spread from country to country . Competition between fit genotypes results in dramatic changes in population composition over short time periods.

Curr Infect Dis Rep, 2001 Apr, 3(2), 156 - 161
Quinolone Resistance in Neisseria gonorrhoeae; Tompkins JR et al.; Single-dose oral quinolones have been recommended for gonorrhea treatment since 1989 . The antimicrobial resistance surveillance system has detected several outbreaks of quinolone-resistant gonococcal infections (QRNG), and sporadic treatment failures have been reported from high-incidence areas such as southeast Asia . QRNG may result from mutations that cause structural-functional changes in DNA topoisomerase (the quinolone target enzyme) or by changes in antimicrobial transport into the bacteria . QRNG has occurred sporadically in the United States, predominantly in persons with contact to persons in southeast Asia, and has typically occurred as an epiphenomenon in persons who were treated with other regimens . Nevertheless, this entity warrants close monitoring . The emergence of QRNG is probably related to antimicrobial misuse and overuse, in particular long-term suppressive or subtherapeutic doses.

Microbiology, 2001 Apr, 147(Pt 4), 839 - 49
Antigenic variation of gonococcal pilin expression in vivo: analysis of the strain FA1090 pilin repertoire and identification of the pilS gene copies recombining with pilE during experimental human infection; Hamrick TS et al.; Antigenic variation of gonococcal pilin involves a family of variable genes that undergo homologous recombination, resulting in transfer of variant sequences from the pilS silent gene copies into the complete pilE expression locus . Little is known about the specific recombination events that are involved in assembling new variant pilin genes in vivo . One approach to understanding pilin variation in vivo is to carry out experimental human infections with a gonococcal strain having a fully characterized repertoire of pilin genes, so that the specific recombination events occurring in vivo can be determined . To this end, the authors cloned, sequenced and mapped the pilin genes of strain FA1090 of Neisseria gonorrhoeae . This strain contains one pilE locus and 19 silent gene copies that are arranged in five pilS loci; the pilE locus and four of the pilS loci are clustered in a 35 kb region of the chromosome . The general features of the pilin loci in FA1090 are similar to those in strain MS11, in which the mechanism of pilin variation has been extensively studied . However, none of the silent copy sequences are identical in the two strains, which emphasizes the extreme variability in this gene family among gonococci . Three male volunteers were inoculated with the same variant of strain FA1090 and developed urethritis within 2--4 d . The pilE gene sequences from a total of 23 colonies cultured from the subjects were analysed, determining which pilS silent copy donated each portion of the expressed pilE genes . There were 12 different pilin variants, one of which was the original inoculum variant, among the in vivo-expressed pilE gene sequences . The pilE of the inoculum variant was derived entirely from a single silent copy (pilS6c1) . However, the pilE genes in the majority of the colonies cultured from the infected subjects were chimeras of sequence derived from two or three silent copies . Recombination to generate new pilE sequences involved exchange of single variable minicassettes, multiple minicassettes, entire silent gene copies, or (rarely) recombination within a minicassette.

J Clin Microbiol, 2001 Apr, 39(4), 1429 - 35
Evaluation of the NucliSens Basic Kit for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in genital tract specimens using nucleic acid sequence-based amplification of 16S rRNA; Mahony JB et al.; We evaluated a new RNA amplification and detection kit, the NucliSens Basic Kit (Organon Teknika), for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in genitourinary specimens . The Basic Kit provides an open platform for RNA amplification and detection and contains isolation reagents for nucleic acid extraction, nucleic acid sequence-based amplification (NASBA) reagents (enzymes and buffers), and a generic ruthenium-labeled probe for electrochemiluminescent (ECL) detection of amplified product . Using freshly purified and titrated stocks of C . trachomatis and N . gonorrhoeae and in vitro-generated RNA transcripts for sensitivity determinations, the Basic Kit detected 1 inclusion-forming unit of C . trachomatis, 1 CFU of N . gonorrhoeae, and 100 RNA molecules of 16S rRNA for both bacteria . The clinical performance of the Basic Kit was evaluated by testing a total of 250 specimens for N . gonorrhoeae by culture and NASBA and a total of 96 specimens for C . trachomatis by PCR and NASBA . The Basic Kit detected 139 of 142 N . gonorrhoeae culture-positive specimens and gave a negative result for 73 of 74 culture-negative specimens, for a sensitivity and specificity of 97.9 and 98.7%, respectively . For C . trachomatis, the Basic Kit detected 24 of 24 PCR-positive specimens and gave a negative result for 71 of 72 PCR-negative specimens, for a sensitivity and specificity of 100 and 98.6%, respectively . The Basic Kit also detected specimens containing both N . gonorrhoeae and C . trachomatis, using a multiplex NASBA assay using primers for both bacteria . The NucliSens Basic Kit offers a versatile platform for the development of sensitive RNA detection assays for sexually transmitted diseases.

Genome Res, 2001 Apr, 11(4), 609 - 13
Self-reporting PNA/DNA primers for PCR analysis; Fiandaca MJ et al.; We report a new fluorogenic method for sealed-tube PCR analysis using a quencher-labeled peptide nucleic acid (Q-PNA) probe . The Q-PNA hybridizes to a complementary tag sequence located at the 5' end of a 5' fluorophore-labeled oligonucleotide primer, quenching the primer's fluorescence . Incorporation of the primer into a doublestranded amplicon causes displacement of the Q-PNA such that the fluorescence of the sample is a direct indication of the amplicon concentration . The Q-PNA is able to quench multiple primers bearing distinct 5' fluorophores in a single reaction . We show realtime quantitative detection of a single-copy gene, K-ras, from human genomic DNA, as well as an endpoint multiplex assay for Chlamydia trachomatis and Neisseria gonorrhoeae targets . Because the Q-PNA may be used to quench any primer that contains the 5' tag sequence, it is possible to inexpensively adapt an existing primer set for use in a self-reporting fluorescent assay by including the tag sequence in one of the primers.

Aust Fam Physician, 2001 Feb, 30(2), 177 - 9
Urinary tract infections in men in a primary care population; Russell DB et al.; OBJECTIVE: There have been few studies on urinary tract infections (UTIs) in adult males . This study aimed to look at the clinical features of males presenting with urinary tract infections in a predominantly gay general practice population . METHOD: A retrospective audit and analysis was carried out of all male patients presenting with symptoms suggestive of a UTI and in whom a pathogen was cultured from urine . The subjects were drawn from two urban general practices in Melbourne . Subjects with known pre-existing urinary tract abnormalities, or in whom recent urinary tract instrumentation had been performed, were excluded, as were subjects in whom Neisseria gonorrhoeae was cultured from urine . RESULTS: Thirty-three subjects were identified, with a total of 47 presentations . Six subjects were excluded on the basis of pre-existing known factors which would predispose to UTIs, leaving 27 subjects, with 37 episodes of UTI . The mean age was 43 (range = 28-62) and 25 of the 27 identified as gay . Nine out of the 27 (33%) were HIV positive with a mean CD4 of 574/mm3 (range = 41-1812) . The main presenting symptoms were dysuria/burning on micturition, urinary frequency, fever/sweats and haematuria . Urethral discharge occurred in two episodes . Multiple symptoms on presentation were common . The main organism cultured was E . coli . Fourteen subjects underwent further radiological investigation and two abnormalities were detected . CONCLUSION: Uncomplicated UTIs are uncommon in males presenting to general practice . The symptoms are similar to those in females with UTIs, though fever may be more common . Treatment with oral antibiotics in a primary care setting is generally curative . Further investigation of males with UTIs may be appropriate, but more studies are needed as to the cost-benefit of this.

J Biol Chem, 2001 Apr 20, 276(16), 12785 - 90 Epub 2001 Jan 23.
Dependence of the bi-functional nature of a sialyltransferase from Neisseria meningitidis on a single amino acid substitution; Wakarchuk WW et al.; The L1 immunotype strain 126E of Neisseria meningitidis has been shown to have an N-acetyl-neuraminic acid-containing lipooligosaccharide in which an alpha-linked galactose from a P(k) epitope is substituted at the O6 position (Wakarchuk, W . W., Gilbert, M., Martin, A., Wu, Y., Brisson, J . R., Thibault, P., and Richards, J . C . (1998) Eur . J . Biochem . 254, 626-633) . Using a synthetic P(k)-epitope containing acceptor in glycosyltransferase reactions, we were able to show by NMR analysis of the reaction product that the 126E(L1)-derived sialyltransferase can make both alpha-2,3 and alpha-2,6 linkages to the terminal galactose . Gene disruption experiments showed that the lst gene in 126E(L1) was responsible for the in vivo addition of the alpha-2,6-linked N-acetyl-neuraminic acid residue . By site-directed mutagenesis it was possible to change the MC58(L3)-derived enzyme into a bifunctional enzyme with a single amino acid change at position 168, where a glycine was changed to an isoleucine . We performed a gene replacement experiment where the 126E(L1) alpha-2,3/6-sialyltransferase was replaced by allelic exchange with the monofunctional MC58(L3) alpha-2,3-sialyltransferase and with the mutant MC58(L3) allele G168I . We observed that the level of LOS sialylation with the G168I allele was very similar to that of the wild type 126E(L1), indicating that residue 168 is the critical residue for the alpha-2,6-sialyltransferase activity in vitro as well as in vivo.

J Biol Chem, 2001 May 18, 276(20), 17413 - 9 Epub 2001 Feb 05.
The CGM1a (CEACAM3/CD66d)-mediated phagocytic pathway of Neisseria gonorrhoeae expressing opacity proteins is also the pathway to cell death; Chen T et al.; Phagocytosis of Opa+ Neisseria gonorrhoeae (gonococcus, GC) by neutrophils is in part dependent on the interaction of Opa proteins with CGM1a (CEACAM3/CD66d) antigens, a neutrophil-specific receptor . However, the signaling pathways leading to phagocytosis have not been characterized . Here we show that interaction of OpaI bacteria with neutrophils or CGM1a-transfected DT40 cells induces calcium flux, which correlates with phagocytosis of bacteria . We identified an immunoreceptor tyrosine-based activation motif (ITAM) in CGM1a, and showed that the ability of CGM1a to transduce signals and mediate phagocytosis was abolished by mutation of the ITAM tyrosines . We also demonstrated that CGM1a-ITAM-mediated bacterial phagocytosis is dependent on Syk and phospholipase C activity in DT40 cells . Unexpectedly, the activation of the CGM1a-ITAM phagocytic pathway by Opa+ GC results in induction of cell death.

FEMS Microbiol Lett, 2001 Mar 15, 196(2), 171 - 6
Isolation and characterization of type IIS restriction endonuclease from Neisseria cuniculi ATCC 14688; Furmanek B et al.; Neisseria cuniculi produces the restriction enzyme NcuI which is an isoschizomer of MboII . We have demonstrated that NcuI recognizes a pentanucleotide sequence (5'-GAAGA-3'/3'-CTTCT-5'), and cleaves the DNA 8 and 7 nucleotides downstream from the recognition site leaving a single 3'-protruding nucleotide . We have purified this enzyme to electrophoretic homogeneity using a four-step chromatographic procedure . NcuI endonuclease is a monomeric protein with a M(r)=48,000+/-1000 under denaturing conditions . The properties of NcuI are consistent with those for MboII, the position of the cleavage site being identical and the pH profile and divalent cation requirements being similar . Moreover, NcuI cross-reacts strongly with anti-MboII serum suggesting the presence of similar antigenic determinants . We have determined the sequence of 20 N-terminal amino acids for NcuI and concluded that this sequence is identical to the N-terminal portion of the MboII enzyme.

An Esp Pediatr, 2001 Mar, 54(3), 267 - 71
{Sexual abuse in children: prevention of sexually transmitted diseases}; Garcia Algar O et al.; When a child suffers from sexual abuse clinical guidelines must be established . There is a risk of infection from the following agents responsible for sexually transmitted diseases: the hepatitis B, hepatitis C and human immunodeficiency viruses, Neisseria gonorrhoeae, Chlamydia trachomatis, syphilis, herpes simplex virus, bacterial vaginosis, papillomavirus, Trichomonas vaginalis and Pediculus pubis . Therefore, a follow-up with periodic serological monitoring for 1year and immunoprophylaxis or chemoprophylaxis for some of these diseases should be started . Postpuberal girls should receive emergency contraception.

Can J Microbiol, 2001 Feb, 47(2), 158 - 64
Mapping of monoclonal antibodies specific to P64k: a common antigen of several isolates of Neisseria meningitidis; Nazabal C et al.; P64k is a minor outer membrane protein from Neisseria meningitidis . This protein has been produced at high levels in Escherichia coli . We generated a group of monoclonal antibodies (mAbs) against recombinant P64k, which recognise four non-overlapping epitopes, as shown using competition assays with biotinylated mAbs . The P64k sequences involved in mAbs binding were mapped with synthetic overlapping peptides derived from the P64k protein, and located in the previously determined three-dimensional structure of the protein . These antibodies were also characterised by whole-cell ELISA and bactericidal tests against N . meningitidis . Only two of the recognised epitopes were exposed on the bacterial surface, and none of the mAbs showed bactericidal activity . The relationship between these results and the structural data on the epitopes bound by the mAbs is discussed.

Cell Microbiol, 2001 Mar, 3(3), 133 - 43
Attachment of Neisseria gonorrhoeae to the cellular pilus receptor CD46: identification of domains important for bacterial adherence; Kallstrom H et al.; Pili of Neisseria gonorrhoeae mediate binding of the bacteria to human host cells . Membrane cofactor protein (MCP or CD46), a human cell-surface protein involved in regulation of complement activation, acts as a cellular pilus receptor . In this work, we examined which domains of CD46 mediate bacterial adherence . The CD46 expression was quantified and characterized in human epithelial cell lines . N . gonorrhoeae showed the highest adherence to ME180 cells, which have BC1 as the dominant phenotype . The BC isoforms of CD46 were expressed in all cell lines tested . The adherence was not enhanced by high expression of other isoforms, showing that the BC domain of CD46 is important in adherence of N . gonorrhoeae to human cells . To characterize the pilus-binding site within the CD46 molecule, a set of CD46-BC1 deletion constructs were transfected into COS-7 cells . Piliated N . gonorrhoeae attached well to CD46-BC1-expressing COS-7 cells . We show that the complement control protein repeat 3 (CCP-3) and the serine-threonine-proline (STP)-rich domain of CD46 are important for efficient adherence to host cells . Further, partial deletion of the cytoplasmic tail of CD46 results in low bacterial binding, indicating that the cytoplasmic tail takes part in the process of establishing a stable interaction between N . gonorrhoeae and host cells.

Anaesthesiol Reanim, 2001, 26(1), 11 - 7
{Microbiological contamination of intraoperatively collected erythrocyte concentrate in mechanical autotransfusion in tumor surgery}; Wehner M et al.; Intraoperative autotransfusion is an effective method of the autologous haemotherapy . Gamma irradiation of the salvaged erythrocyte concentrates leads to inactivation of malignant cells and allows retransfusion in oncologic surgery . The risk of microbiological contamination of the autologous blood product has not been systematically studied in this field of surgery . During the evaluation of intraoperative autotransfusion in oncologic surgery at the University Hospital of Leipzig, the salvaged blood of 46 patients who underwent tumour surgery (urology, gynaecology and neurosurgery) was processed to a washed erythrocyte concentrate with a Cell Saver 5 or a Haemolite 2 (Haemonetics) . Specimens for microbiological studies were taken from the area of surgery, the reservoir bottle of the autotransfusion system and the salvaged erythrocyte concentrates . The investigation with plate cultures and the identification of microorganisms were performed according to standard procedures of microbiological diagnosis . The ethics committee of the University Hospital of Leipzig approved the study protocol . We found a microbiological contamination in 10 of 46 salvaged erythrocyte concentrates (22%), above all the growth of saprophytes (Streptococcus, Staphylococcus and Neisseria species) . We also identified bacteria which have their habitats in the intestinal tract (E . coli and Enterococcus) . The highest rate of microbial contaminated erythrocyte concentrates was found in urologic oncosurgery (37%) . In neurosurgery (laminectomy and craniotomy including tumour extirpation), there was no bacterial growth in the salvaged erythrocyte concentrate . Intraoperative autotransfusion should be avoided during surgery touching the urinary tract, e.g . prostatectomy and cystectomy . In neurosurgery, it seems to be safe as far as microbiological contamination is concerned . To assess the real risk of microbiological contamination of the salvaged erythrocytes, more investigations are necessary to determine not only the extent and nature of contamination but also the clinical effects.

Bioorg Khim, 2001 Jan-Feb, 27(1), 21 - 6
{Induction of antimeningitis immunity by synthetic peptides . II.Immunoactive synthetic fragments of OpaB protein from Neisseria meningitidis}; Koroev DO et al.; Mice of various lines were immunized by 11 synthetic peptides that correspond to the sequences of fragments of the OpaB protein from the outer membrane of Neisseria meningitidis involving the known human T-helper epitopes and all the potential mouse T-helper epitopes calculated for the protein . The mice were immunized with the free peptides without their conjugation with a protein carrier . Most of the peptides were found to induce in mice the production of antipeptide antibodies . The mice protection against the experimental infection by a virulent strain of N . meningitidis of the B serotype was studied, and two peptides were shown to exert the most pronounced protective effect.

J Clin Lab Anal . 2000;14(6):336.
Atobe J . H., Hirata M . H., Hoshino-Shimizu S., Schmal M . R., Mamizuka E . M . One-step heminested PCR for amplification of Neisseria meningitidis DNA in cerebrospinal fluid . J Clin Lab Anal 14:193-199, 2000; Malatesta M et al.; On page 195, in Fig . 1, the labels of boxes RW01 and DG74 are switched . The label RW01 in the right box should read DG74 (1540-1521 position), and the label DG74 in the left box should read RW01 (1189-1170 position) . Also in Fig . 1, the primer DG74 listed in the "Primers" box above the legend should read 5'-AGGAGGTGATCCAACCGCA-3', and not 5'-AGGAGGTGATCCAACGCGA-3' as printed . The publisher regrets the error.

Infect Immun, 2001 Apr, 69(4), 2604 - 11
Helicobacter pylori resists phagocytosis by macrophages: quantitative assessment by confocal microscopy and fluorescence-activated cell sorting; Ramarao N et al.; Helicobacter pylori infection of the stomach epithelium is characterized by an infiltration of polymorphonuclear and mononuclear cells . These immune cells contribute to mucosal damage which may eventually lead to gastritis, peptic ulcer, gastric cancer, and/or MALT-associated gastric lymphoma . Here we show that H . pylori inhibits its own uptake, as well as in trans the phagocytosis of Neisseria gonorrhoeae, by human and murine macrophages . This antiphagocytic activity is dependent on the presence of the cag pathogenicity island in the H . pylori genome . We demonstrate that H . pylori also expresses its antiphagocytic activity towards the myelomonocytic cell line JOSKM, thus providing a potent model for the study of the interaction between H . pylori and phagocytes . Our data were obtained using laser confocal microscopy and flow cytometry after quenching the fluorescence of labeled extracellular bacteria . The antiphagocytic activity of H . pylori may explain the persistence of H . pylori and its pathological consequences . The use of cell lines and flow cytometry will hopefully facilitate progress in our understanding of the immune escape of these persistent bacteria.

Infect Immun, 2001 Apr, 69(4), 2230 - 6
Membrane-associated proteins of a lipopolysaccharide-deficient mutant of Neisseria meningitidis activate the inflammatory response through toll-like receptor 2; Ingalls RR et al.; The recent isolation of a lipopolysaccharide (LPS)-deficient mutant of Neisseria meningitidis has allowed us to explore the roles of other gram-negative cell wall components in the host response to infection . The experiments in this study were designed to examine the ability of this mutant strain to activate cells . Although it was clearly less potent than the parental strain, we found the LPS-deficient mutant to be a capable inducer of the inflammatory response in monocytic cells, inducing a response similar to that seen with Staphylococcus aureus . Cellular activation by the LPS mutant was related to expression of CD14, a high-affinity receptor for LPS and other microbial products, as well as Toll-like receptor 2, a member of the Toll family of receptors recently implicated in host responses to gram-positive bacteria . In contrast to the parental strain, the synthetic LPS antagonist E5564 did not inhibit the LPS-deficient mutant . We conclude that even in the absence of LPS, the gram-negative cell wall remains a potent inflammatory stimulant, utilizing signaling pathways independent of those involved in LPS signaling.

Eur J Biochem, 2001 Mar, 268(6), 1613 - 9
Characterization of carbonic anhydrase from Neisseria gonorrhoeae; Elleby B et al.; We have investigated the steady state and equilibrium kinetic properties of carbonic anhydrase from Neisseria gonorrhoeae (NGCA) . Qualitatively, the enzyme shows the same kinetic behaviour as the well studied human carbonic anhydrase II (HCA II) . This is reflected in the similar pH dependencies of the kinetic parameters for CO(2) hydration and the similar behaviour of the kinetics of (18)O exchange between CO(2) and water at chemical equilibrium . The pH profile of the turnover number, k(cat), can be described as a titration curve with an exceptionally high maximal value of 1.7 x 10(6) s(-1) at alkaline pH and a pK(a) of 7.2 . At pH 9, k(cat) is buffer dependent in a saturable manner, suggesting a ping-pong mechanism with buffer as the second substrate . The ratio k(cat)/K(m) is dependent on two ionizations with pK(a) values of 6.4 and 8.2 . However, an (18)O-exchange assay identified only one ionizable group in the pH profile of k(cat)/K(m) with an apparent pK(a) of 6.5 . The results of a kinetic analysis of a His66-->Ala variant of the bacterial enzyme suggest that His66 in NGCA has the same function as a proton shuttle as His64 in HCA II . The kinetic defect in the mutant can partially be overcome by certain buffers, such as imidazole and 1,2-dimethylimidazole . The bacterial enzyme shows similar K(i) values for the inhibitors NCO(-), SCN(-) and N(3)(-) as HCA II, while CN(-) and the sulfonamide ethoxzolamide are considerably weaker inhibitors of the bacterial enzyme than of HCA II . The absorption spectra of the adducts of Co(II)-substituted NGCA with acetazolamide, NCO(-), SCN(-), CN(-) and N(3)(-) resemble the corresponding spectra obtained with human Co(II)-isozymes I and II . Measurements of guanidine hydrochloride (GdnHCl)-induced denaturation reveal a sensitivity of the CO(2) hydration activity to the reducing agent tris(2-carboxyethyl)phosphine (TCEP) . However, the A(292)/A(260) ratio was not affected by the presence of TCEP, and a structural transition at 2.8--2.9 M GdnHCl was observed.

Clin Infect Dis, 2001 Mar 15, 32(6), 884 - 9 Epub 2001 Mar 09.
Treatment failure with the use of ciprofloxacin for gonorrhea correlates with the prevalence of fluoroquinolone-resistant Neisseria gonorrhoeae strains in Bangladesh; Rahman M et al.; Although ciprofloxacin is one of the recommended drugs of choice for the treatment of gonorrhea, in vitro resistance to this drug has been observed in surveillance studies and case reports from many parts of the world, including Bangladesh . However, to our knowledge, there have been no prospective studies of the correlation between in vitro response to the drug and treatment outcome . Therefore, a prospective study of 217 female sex workers in Dhaka, Bangladesh, was conducted to examine the correlation between the in vitro response of Neisseria gonorrhoeae and the outcome of ciprofloxacin treatment . Overall, 37.8% of the gonococcal isolates recovered from female sex workers were resistant to ciprofloxacin, and there was a good correlation between in vitro resistance and treatment failure . These findings suggest that in vitro resistance to ciprofloxacin is predictive of clinical treatment failure in patients with gonorrhea.

Rev Neurol, 2000 Nov 16-30, 31(10), 991 - 3
{Dynamics of intrathecal immunoglobulin synthesis}; Dorta-Contreras AJ; INTRODUCTION: Dynamics of the immune response in Central Nervous System (CSN) is different from the well-known switch of IgM synthesis to IgG synthesis in blood . OBJECTIVE: Broadcast the behavior and the factors involved in the dynamics of the intrathecal immune response in infectious neurological disorders . DEVELOPMENT: The lack of a switch from IgM class response to IgG response could be more related to regulation-modulation mechanisms of the CNS immune response than differs from blood in a different cytokines composition, and the possibility of chemokines synthesis during the neuroinflammatory process, and the neuroimmune-endocrine mechanisms . The immune pattern can be stable like neuroborreliosis, and can be modify like in herpes simplex meningoencephalitis . It could have a typical pattern like in Neisseria meningitidis meningoencephalitis and neurotuberculosis . Also the pattern could be still detectable for many years after sufficient treatment and complete recovery of the symptom-free patients like in neurosyphilis or an advanced precocious response during the childhood . In HIV encephalopathy the pattern remains the same during the evolution but in other virus infections, like Echo 6 or Coxsackie B5, depends on the biological agent . CONCLUSIONS: In order to know the acuity of the disease, we have to know the physiopathologic characteristics of the biological agents, time courses, locations of the pathological processes, and the host age . The main signs in cerebrospinal fluid of an acute, active disease of CNS are the increased of cerebrospinal fluid cell count and the increased of albumin ratio.

J Infect, 2001 Jan, 42(1), 44 - 9
Bacteraemia and mortality among adult medical admissions in Malawi--predominance of non-typhi salmonellae and Streptococcus pneumoniae; Gordon MA et al.; OBJECTIVES: The high seroprevalence of HIV in Malawi might be expected to alter the pattern of pathogens isolated from bacteraemic patients . We aimed to describe the frequency and seasonal pattern of bacterial isolates from blood, their antibiotic susceptibility, and patient outcome, in order to provide data on which to base empirical antibiotic therapy and further studies of pathogenesis . METHODS: Over a 12-month period, blood cultures were taken from all febrile adult medical admissions to Queen Elizabeth Central Hospital, Blantyre . RESULTS: A total of 2789 out of 9298 adult general medical admissions had blood culture performed, of whom 449 (16.1%) grew significant pathogens . Non-typhi salmonellae (NTS) (37%) and Streptococcus pneumoniae (30%) were the two commonest isolates . Mortality was 18% among general medical admissions and 38% among bacteraemic patients . Mortality for individual pathogens was: NTS 33%; S . pneumoniae 36%; Escherichia coli 54%; Klebsiella spp . 58%; Neisseria meningitidis 44%; Salmonella typhi 17% . Despite an overwhelming association between the major pathogens and HIV infection (95% of S . pneumoniae cases and 92% of NTS cases were seropositive for HIV), a seasonal pattern was preserved . Streptococcus pneumoniae was more frequently isolated in the cold dry months, while STM isolates increased following a rise in temperature . A case of bacteraemia with Vibrio cholerae (serotype 01) was detected during a cholera outbreak in the rainy season . Although S . pneumoniae isolates were relatively susceptible to penicillin (88%) and chloramphenicol (74%), S . typhimurium isolates were fully susceptible only to chloramphenicol . CONCLUSIONS: This large study confirms the dominance of NTS and S . pneumoniae in bacteraemia in an area affected by HIV-1 and allows comparison of mortality by individual pathogens . It demonstrates a preserved seasonal pattern of bacteraemia for these major pathogens, despite an overwhelming association with HIV infection .

Chin Med Sci J, 1997 Mar, 12(1), 47 - 9
Analysis of the antigen-antibody specificity in the semen of patients with Neisseria gonorrhoeae; Zheng H; This study was done to define the human genital immune response to infection with Neisseria gonorrhoeae . The semen specimens were obtained from 15 patients with uncomplicated gonococcal infection in the acute and convalescent phases and 15 men with uninfected control . After precipitated with amoniasulfate, the semen was tested against the outer membrane protein of gonococcal isolates from the same patients to examine antigen-antibody interactions by use of the western blot technique . The antibodies in the semen reacted with more gonococcal antigens in the acute phase than in the convalescent phase . IgA in the semen reacted with more antigens than did IgG in the same specimens . The predominant reacted antigens were protein I, protein II, 46-48, kD, 14-16 kD and 88-90 kD protein.

Bull World Health Organ, 2001, 79(2), 118 - 26
Etiology of urethral discharge in West Africa: the role of Mycoplasma genitalium and Trichomonas vaginalis; Pepin J et al.; OBJECTIVE: To determine the etiological role of pathogens other than Neisseria gonorrhoeae and Chlamydia trachomatis in urethral discharge in West African men . METHODS: Urethral swabs were obtained from 659 male patients presenting with urethral discharge in 72 primary health care facilities in seven West African countries, and in 339 controls presenting for complaints unrelated to the genitourinary tract . Polymerase chain reaction analysis was used to detect the presence of N . gonorrhoeae, C . trachomatis, Trichomonas vaginalis, Mycoplasma genitalium, and Ureaplasma urealyticum . FINDINGS: N . gonorrhoeae, T . vaginalis, C . trachomatis, and M . genitalium--but not U . urealyticum--were found more frequently in men with urethral discharge than in asymptomatic controls, being present in 61.9%, 13.8%, 13.4% and 10.0%, respectively, of cases of urethral discharge . Multiple infections were common . Among patients with gonococcal infection, T . vaginalis was as frequent a coinfection as C . trachomatis . M . genitalium, T . vaginalis, and C . trachomatis caused a similar clinical syndrome to that associated with gonococcal infection, but with a less severe urethral discharge . CONCLUSIONS: M . genitalium and T . vaginalis are important etiological agents of urethral discharge in West Africa . The frequent occurrence of multiple infections with any combination of four pathogens strongly supports the syndromic approach . The optimal use of metronidazole in flowcharts for the syndromic management of urethral discharge needs to be explored in therapeutic trials.

Int J Gynecol Cancer, 1999 May, 9(3), 177 - 186
Infection and cervical intraepithelial neoplasia; Boyle DC et al.; The impact of sexually transmitted diseases (STDs) on the development of cervical intraepithelial neoplasia (CIN) has been increasingly recognized over the last 20 years . Much attention has been focused on human papillomavirus (HPV) and the potential for screening for certain HPV types alongside standard cervical cytology in the hope of identifying those females at particular risk of developing high grade CIN or invasive carcinoma . Some infections, for example herpes simplex virus (HSV), have been heavily investigated in the past as they were thought to be involved in the development of CIN but were subsequently discounted . Also discounted as causes of CIN are Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) . These infections were found to be associated with higher rates of CIN in early studies but transpired to be markers for the presence of other infections and pathology and therefore not themselves directly responsible for cytological changes . The role of bacterial vaginosis (BV) is the focus of several current investigations, not only in the genesis of CIN but also in the development of other gynecological and obstetric conditions and complications . Evidence to implicate Trichomonas vaginalis (TV) in the genesis of CIN is conflicting, but there is some evidence that it may exert its influence in a similar way to that hypothesized for BV, ie via abnormal amines . It is well known that there is a high level of concordance of STDs whereby the presence of one infection greatly increases the likelihood of there being one or more others present . There may be a synergism between some infections with regard to the causation of CIN, although the evidence for this is putative . Presented here is an overview of current and previous research in the field of lower genital tract infection as it relates to the development of CIN.

J Infect Dis, 2001 Mar 15, 183(6), 865 - 70 Epub 2001 Feb 21.
Cervical ectopy in adolescent girls with and without human immunodeficiency virus infection; Moscicki AB et al.; The objective of this study was to examine factors, including human immunodeficiency virus (HIV) infection, associated with ectopy among adolescent girls aged 12-20 years who were participating in an ongoing study of HIV infection in adolescents . Samples for detection of bacterial vaginosis, Chlamydia trachomatis, and Neisseria gonorrhoeae and a high-resolution photograph of the cervix for ectopy measurement were collected . Ectopy data for 189 and 92 HIV-positive and -negative adolescents, respectively, were examined . Although univariate analysis found HIV infection and oral contraceptive use to be associated with the amount of ectopy, multivariate logistic regression analysis showed that only number of lifetime sex partners was a significant predictor, with more partners associated with less ectopy (odds ratio, 0.47; 95% confidence interval, 0.22-1.00; P=.05) . In summary, adolescent girls with greater numbers of lifetime sex partners were more likely to have mature cervixes (less ectopy) . HIV infection was not independently associated with ectopy.

Am J Public Health, 2001 Mar, 91(3), 461 - 4
Emergency department screening for asymptomatic sexually transmitted infections; Todd CS et al.; OBJECTIVES: This study assessed the prevalence and correlates of asymptomatic genital tract infection with Neisseria gonorrhoeae and Chlamydia trachomatis among emergency department patients . METHODS: Individuals seeking emergency department evaluation for nongenitourinary complaints provided urine samples for N gonorrhoeae and C trachomatis testing by ligase chain reaction and completed a sociodemographic and behavioral questionnaire . RESULTS: Asymptomatic N gonorrhoeae or C trachomatis was found in 9.7% of persons tested . Correlates of C trachomatis infection included younger age, residence in high-morbidity zip code areas, previous history of N gonorrhoeae or C trachomatis, and number of sex partners in the past year . CONCLUSIONS: Urine-based screening of asymptomatic emergency department patients detected significant numbers of N gonorrhoeae and C trachomatis infections . Targeted screening programs may contribute to community-level prevention and control of sexually transmitted infections.

Int J STD AIDS, 2001 Feb, 12(2), 89 - 93
Antimicrobial susceptibilities and plasmid patterns of Neisseria gonorrhoeae in Bénin; van Dyck E et al.; This study describes antimicrobial susceptibility patterns of Neisseria gonorrhoeae isolates obtained from female sex workers in Cotonou, Benin . All isolates were susceptible to spectinomycin, ceftriaxone and ciprofloxacin, and susceptible to moderately susceptible to kanamycin; 9.8% of isolates were resistant to thiamphenicol; 9%, 87.5% and 3.5% were susceptible, moderately susceptible, resistant to trimethoprim-sulfamethoxazole, respectively; 94.4% and 99.3% were resistant to penicillin and tetracycline, respectively . All isolates with a minimal inhibitory concentration of tetracycline of >8 mg/l carried the 'American type' tetM plasmid; 94% and 6% of penicillinase-producing isolates possessed a 3.2 MDa and a 4.4MDa beta-lactamase plasmid, respectively . Surveillance of antimicrobial susceptibility of N . gonorrhoeae isolates to currently used drugs in Africa should become part of sexually transmitted diseases (STDs) control programmes.

Sex Transm Dis, 2001 Feb, 28(2), 82 - 3
Neisseria gonorrhoeae resistant to ciprofloxacin: first report in Cuba; Llanes R et al.; BACKGROUND AND OBJECTIVES: The Cuban Ministry of Public Health plans to implement the syndromic approach to sexually transmitted diseases in persons with urethral or vaginal syndrome in Cuba using 500 mg ciprofloxacin as therapy . Although the emergence of clinical isolates of Neisseria gonorrhoeae with decreased susceptibility to ciprofloxacin have been sporadically detected in Cuba, there has been no report of isolates that exhibited significant resistance to this drug . This is the first report of the isolation of a N gonorrhoeae strain resistant to ciprofloxacin in Cuba . STUDY DESIGN: Case report . CONCLUSIONS: This case emphasizes the need for awareness regarding the potential emergence of a clinically significant resistance of N gonorrhoeae in Cuba . There is a need for continued antimicrobial susceptibility surveillance of Cuban isolates to ciprofloxacin and other fluoroquinolones.

Sex Transm Dis, 2001 Feb, 28(2), 111 - 4
Epidemiology of the reemergence of gonorrhea in Sweden; Berglund T et al.; BACKGROUND: After many years of decline, the incidence of gonorrhea in Sweden reached an all-time low of 2.4 cases per 100,000 inhabitants in 1996; however, the incidence has been increasing since 1997 . GOAL: To describe the reemergence of gonorrhea in Sweden using clinical epidemiologic data and microbiologic characterization of isolated strains of Neisseria gonorrhoeae . STUDY DESIGN: N gonorrhoeae strains isolated in Sweden from February 1998 to January 1999 were serotyped and an epidemiologic data questionnaire was sent to each clinician reporting a case of gonorrhea . RESULTS: Epidemiologic and microbiologic data were obtained for 357 cases (91% of all cases diagnosed during the period) . Domestic cases had significantly increased since 1997 . Serovar IB-3 was the most common isolate in domestic cases of heterosexually exposed teenagers, and serovar IB-2 was the most frequent isolate in domestic cases of homosexually exposed men . Of the imported cases, 47% were exposed in Asia and 6.5% were exposed in Eastern Europe . CONCLUSION: The increase of gonorrhea in Sweden is due to an increase of domestic cases . Heterosexual teenagers and homosexual men were identified as core groups infected by different serovars of N gonorrhoeae.

J Clin Microbiol, 2001 Mar, 39(3), 1008 - 16
Multicenter evaluation of the BDProbeTec ET System for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine specimens, female endocervical swabs, and male urethral swabs; Van Der Pol B et al.; The performance of the Becton Dickinson BDProbe Tec ET System Chlamydia trachomatis and Neisseria gonorrhoeae Amplified DNA Assays (BD Biosciences, Sparks, Md.) was evaluated in a multicenter study . Specimens were collected from 2,109 men and women, with or without symptoms, attending sexually transmitted disease, family planning, and obstetrics and gynecology clinics . Both swab and urine samples were collected, and the results obtained from 4,131 specimens were compared to those from culture and the LCx nucleic acid amplification test (Abbott Industries, Abbott Park, Ill.) . PCR and cytospin of the culture transport medium with chlamydia direct fluorescent antibody staining were used to adjudicate chlamydia culture-negative results . Sensitivity and specificity were calculated both with and without use of the amplification control (AC), with little apparent difference in the results . Without the AC result, sensitivity for C . trachomatis and N . gonorrhoeae were 92.8 and 96.6%, respectively, for cervical swabs and 80.5 and 84.9% for urine from women . C . trachomatis and N . gonorrhoeae sensitivities were 92.5 and 98.5%, respectively, for male urethral swabs and 93.1 and 97.9% for urine from men . This amplified DNA system for simultaneous detection of chlamydial and gonococcal infections demonstrated superior sensitivity compared to chlamydia culture and has performance characteristics comparable to those of other commercially available nucleic acid-based assays for these organisms.

J Clin Microbiol, 2001 Mar, 39(3), 829 - 35
Comparison between the LCx Probe system and the COBAS AMPLICOR system for detection of Chlamydia trachomatis and Neisseria gonorrhoeae infections in patients attending a clinic for treatment of sexually transmitted diseases in Amsterdam, The Netherlands; van Doornum GJ et al.; Two assays for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae were compared: the LCx Probe system (the LCx system; Abbott Diagnostic Laboratories, North Chicago, Ill.) and the COBAS AMPLICOR C . trachomatis/N . gonorrhoeae system (the COBAS AMPLICOR system; Roche Diagnostic Systems, Branchburg, N.J.) . Endocervical swab specimens, male urethral swab specimens, and female and male urine specimens were collected from 503 female and 498 male visitors attending a sexually transmitted diseases clinic in Amsterdam, The Netherlands . Prevalences for C . trachomatis were 12.5% (63 of 503) and 10.0% (50 of 498) in females and males, respectively . The prevalences for N . gonorrhoeae were 1.2% (6 of 503) and 4.2% (21 of 498) in females and males, respectively . Both assays showed high values for sensitivity and specificity with regard to the detection of C . trachomatis in endocervical swab specimens, male urethral swab specimens, and female and male urine specimens . The sensitivities for the LCx system were 92.1, 90.0, 88.9, and 94.0% for each type of specimen, respectively; and the sensitivies for the COBAS AMPLICOR system were 96.8, 98.0, 82.5, and 92.0% for each type of specimen, respectively . Specificities ranged between 98.4 and 100% . The sensitivity of the LCx system for the detection of N . gonorrhoeae was 100% for female cervical swab and urine specimens and male urethral swab specimens, while for male urine specimens the sensitivity was 95.2%; the specificity was 100% for all types of specimens . For the detection of N . gonorrhoeae by the COBAS AMPLICOR assay, the sensitivity for female cervical swab and male urethral swab specimens was 100%, that for female urine specimens was 66.7%, and that for male urine specimens was 95.2% . However, the predictive values of a positive test for female cervical swab specimens and urine specimens were 31.6 and 36.4%, respectively . Sequence analysis of the amplimers obtained by an in-house 16S rRNA PCR of the solely COBAS AMPLICOR system-positive swab specimens revealed neither N . gonorrhoeae nor other Neisseria spp . The COBAS AMPLICOR assay was considered not suitable for screening for infections with N . gonorrhoeae . If this assay is used for detection of N . gonorrhoeae, confirmation of positive results by a reliable test is mandatory.

Hum Reprod, 2001 Mar, 16(3), 463 - 8
Increased adhesiveness and internalization of Neisseria gonorrhoeae and changes in the expression of epithelial gonococcal receptors in the Fallopian tube of copper T and Norplant users; Fernandez R et al.; Interaction of Neisseria gonorrhoeae with the oviductal epithelium in vitro was examined in 2 cm length segments obtained after surgical sterilization from users of copper T intrauterine device (IUD) or Norplant and control women . Segments perfused with N.gonorrhoeae suspensions were incubated from 30 min up to 4 h, fixed, frozen and cut in 6--10 microm sections . Bacteria were detected immunohistochemically with rabbit anti-gonococcal serum followed by light and confocal microscopy . Adhesion and internalization of gonococci by epithelial cells were observed at all incubation times, and both were higher in explants from users of copper T IUD or Norplant implants than controls . The epithelium of controls expressed CD66 and syndecan-1; but CD46 was found in only one out of six cases . The epithelium of copper T IUD users expressed CD66 but not syndecan-1 or CD46 . Users of Norplant exhibited expression of CD46, CD66 and syndecan-1 . Label was always found along the luminal border of the epithelium . There were more intraepithelial lymphocytes in users of contraceptive methods than in controls . Results indicate that (i) N.gonorrhoeae invade the oviductal epithelium from the first minutes of exposure, (ii) the epithelium is constitutively endowed with two known receptors for the gonococcus, CD66 and syndecan-1, (iii) copper T IUD and Norplant users exhibit higher rates of attachment and internalization of the gonococcus into the oviductal epithelium associated with changes in expression of gonococcal receptors.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Sep-Oct, (5), 15 - 9
{Common epitopes of protein antigens in Neisseria and Moraxella}; Goncharenko AV et al.; Cross reactions between N . meningitidis and M . catarrhalis proteins were studied with the use of a panel of monoclonal antibodies to M . catarrhalis protein antigens . All antigenic preparations under study were shown to give cross reactions between N . meningitidis serotype porin of 39 kD (strain B125) and M . catarrhalis proteins of 40-41 kD . These M . catarrhalis proteins belonged to main proteins of class F and had the function of porins in the cell . In addition, the epitope of 41-kD antigen, detected by monoclonal antibodies 3E10, is common for both N . meningitidis porin and N . meningitidis iron-regulated proteins of 70 and 50 kD . The epitope of M . catarrhalis protein of 67 kD, detected by monoclonal antibodies 1G6, is common for N . meningitidis porin and N . meningitidis iron-regulated proteins of 50 and 55 kD.

Infect Dis Obstet Gynecol, 2000, 8(5-6), 235 - 9
Prevalence of human papillomavirus, Chlamydia trachomatis, and Neisseria gonorrhoeae in commercial sex workers in Japan; Ishi K et al.; OBJECTIVE: We used the hybrid capture assays to investigate the prevalence of human papillomavirus (HPV), Chlamydia trachomatis and Neisseria gonorrhoeae among commercial sex workers in Tokyo . METHODS: Five hundred forty-six consecutive commercial sex workers (CSW) who visited an STD clinic for STD checkup in 1998 and 1999 were studied . A control group consisted of 233 consecutive women who visited a general gynecological clinic for annual checkup . A cervical sample was obtained for hybrid capture assays for HPV-A (low-oncogenic-risk types), HPV-B (intermediate- and high-oncogenic-risk types), C . trachomatis, and N . gonorrhoeae . Fisher's exact test was used for statistical analyses . RESULTS: The positive rate for HPV-B among the CSW was 48.4%, significantly higher than the 6.0% among the control subjects . The positive rates for HPV-A, C . trachomatis, and N . gonorrhoeae were also significantly higher among the CSW than among the control subjects . Among the microorganisms tested, the positive rate for HPV-B was the highest in both the STD and control groups . CONCLUSIONS: The high prevalence of HPV, C . trachomatis, and N . gonorrhoeae infection in CSW poses a risk of further transmission of STD to the general public, suggesting the need for further education and screening for CSW and the general public . We found high- and intermediate-oncogenic-type HPV to be the most prevalent infection among both CSW and control subjects . Screening for HPV may be necessary in STD and general clinics to predict the risk of cervical malignancy . Hybrid capture assays, which permit simultaneous detection of HPV and other STD with high sensitivity, may be a useful diagnostic method.

Pediatr Infect Dis J, 2001 Feb, 20(2), 144 - 7
Comparison of the urine-based ligase chain reaction test to culture for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in pediatric sexual abuse victims; Girardet RG et al.; BACKGROUND: The urine-based ligase chain reaction (LCR) assay for Chlamydia trachomatis and Neisseria gonorrhoeae is an attractive alternative to culture because of the relative ease with which specimens may be collected, transported and processed . In addition LCR offers superior sensitivity while maintaining high specificity when compared with culture in various studies of adolescents and adults . A study comparing LCR to culture has not been published concerning children . METHODS: We conducted a prospective, comparison trial of the urine-based LCR test for Chlamydia trachomatis and Neisseria gonorrhoeae as compared with culture among children at a specialized referral center for evaluation for alleged sexual assault . Of the 1,010 children presenting to the center during the study period, 164 met the study requirements for risk of a sexually transmissible disease and collection of both culture and urine LCR specimens . RESULTS: Eight specimens tested positive by both methods for C . trachomatis . Another 10 specimens tested positive for C . trachomatis by LCR but were negative by culture . No patient with a negative LCR for C . trachomatis had a positive culture . For N . gonorrhoeae 2 specimens tested positive by both methods, and 3 specimens tested positive by LCR but negative by culture . No patient with a negative LCR for N . gonorrhoeae had a positive culture . CONCLUSIONS: The low prevalence of disease in the study population precluded statistical analysis . LCR may prove to be as specific and more sensitive than culture for the detection of C . trachomatis and N . gonorrhoeae in children . Further studies are needed.

Clin Obstet Gynecol, 2001 Mar, 44(1), 122 - 6; quiz 127-8
New contraceptive options; Archer DF; New contraceptive modalities that are being introduced into the US marketplace reflect many changes in the delivery of existing steroidal products . These products are available now not only as oral medications but also as transdermal, intravaginal, intrauterine, and implantable or injectable devices . Each of these has its own unique profile and enhances the ability of consumers to pick and choose the product that is most acceptable to them . However, development of new barrier contraceptives, particularly those that have the potential to reduce the heterosexual transmission of HIV or to reduce a bacterial infection such as Chlamydia or Neisseria gonorrhea, continues to be moving forward in clinical trials . These products can have a lower efficacy compared with the steroidal products but, because of their other benefits, may be of significant medical use.

Mol Gen Genet, 2001 Jan, 264(5), 691 - 701
DNA circle formation in Neisseria gonorrhoeae: a possible intermediate in diverse genomic recombination processes; Barten R et al.; An important attribute that contributes to the virulence of Neisseria gonorrhoeae is its phenotypic variability, which is based on recombination within complex gene families in the genome . In this study we report on the in vivo amplification of large segments of the genome and the existence of circular DNA intermediates in the cell, which might help to explain the evolution of these gene families and provide possible clues as to how genetic variability is maintained . Using an inserted chromosomal marker (cat) in the N . gonorrhoeae MS11 genome that confers low-level resistance to chloramphenicol (Cm), we isolated variants that express resistance to high levels of Cm . Genetic analysis revealed that virtually all variants harboured single or multiple tandem amplifications of the respective genome segments carrying the cat insert . This process occurred independently of both the location of the cat insertion site and of the presence of a functional recA gene . Analysis of the genetically well characterised pilC region revealed a head-to-tail orientation of the amplified segments, with the junctions being located within direct repeats . Identical junctions were detected in extra-chromosomal circular DNA molecules isolated from non-selected wild-type and recA strains, suggesting that both types of structure arise by related processes . The existence of DNA circles was shown by their banding behaviour in caesium chloride/ethidium bromide density centrifugation and their resistance to digestion by exonuclease . The possible roles of such circles in processes such as pilin gene recombination, chromosomal gene amplification and genetic transformation are discussed.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Nov-Dec, (6), 65 - 7
{Computer-controlled maintenance of the dissolved oxygen level in a medium during controlled cultivation of Neisseria meningitidis}; Druzhinin OG et al.; A new method for controlling the processes of N . meningitidis cultivation by the content of oxygen dissolved in the medium with the use of a computer, including the software, technical and methodological provision of the computer-operated control system, has been developed . The processes of the cultivation of N . meningitidis, groups A and B, under fully controlled conditions have been realized . The computer-operated control of the content of oxygen in the medium, the most important parameter of the process, has been shown to be essentially more effective than manual control . Information, accumulated in the course of the experiment, may be used in future for correction of mathematical models and optimization of the process . This will by necessary for obtaining biomass with predetermined properties in the development of new diagnostic and vaccine preparations.

J Reprod Med, 2001 Jan, 46(1), 34 - 8
Risks and adverse outcomes of sexually transmitted diseases . Patients' attitudes and beliefs; Whiteside JL et al.; OBJECTIVE: To determine women's awareness and level of understanding of the risks and sequelae of sexually transmitted diseases (STDs) . STUDY DESIGN: The study was a cross-sectional survey of 103 women seen at an urban center . We recruited women from several clinical sites: (1) patients with signs or symptoms of an STD seen in a busy women's urgent care unit, (2) women enrolled in clinical studies for STDs or bacterial vaginosis, (3) patients admitted to the hospital and found to have an STD, and (4) women at high risk based on age and sexual activity . A trained female interviewer administered the survey to women agreeing to participate . The questionnaire assessed the patient's awareness of various STDs and their association with adverse outcomes . The survey also evaluated women's knowledge regarding STD prevention . Descriptive analyses were performed using SAS (Cary, North Carolina) on the 103 patients surveyed . RESULTS: The median age of our survey population was 23, 66% were nonwhite, and 37% were on public assistance or uninsured . The population was at high risk for STDs based on the reported history of infection with Neisseria gonorrhoeae and Chlamydia trachomatis (6% and 38%, respectively) . Of the 103 women surveyed, 33% had never heard of pelvic inflammatory disease (PID), and 79.6% could not identify any adverse sequelae of PID . Sixty-five percent were unaware that PID increases the risk of ectopic pregnancy, and 56.3% were unaware that it could result in chronic pelvic pain . When asked if they knew of any methods to prevent or reduce their risk of STDs, only 18% mentioned barrier contraception (condoms) . Over 57% of respondents could not name a way to prevent STDs . CONCLUSION: Increased educational efforts targeted at high-risk individuals are necessary to improve knowledge of STDs and their adverse sequelae . With greater knowledge and awareness, individuals and couples may be more likely to practice protective behaviors, such as reducing high-risk exposure, and to increase preventive measures, such as barrier contraception use.

Pharm Res, 2000 Nov, 17(11), 1396 - 401
Application of high performance liquid chromatography coupled with ultraviolet spectroscopy and electrospray mass spectrometry to the characterisation of ellagitannins from Terminalia macroptera roots; Silva O et al.; PURPOSE: Terminalia macroptera roots are used in Guinea-Bissau and other West African countries to treat infectious diseases like gonorrhoea . Previous work showed an ethanol extract of T . macroptera roots (T) to have an in vitro antimicrobial profile against Neisseria gonorrhoae (including resistant strains) and enteropathogenic agents . The most active fractions of this extract were identified as the diethyl ether (T2) and water (T5) fractions . The aim of the present study was the identification of major compounds present in T and simultaneously in T2 or T5 . METHODS: The T extract and T2 and T5 fractions were analysed by high performance liquid chromatography coupled with ultraviolet photodiode array (LC-UV) spectroscopy and electrospray ionization mass spectrometry (ES-MS) . These analyses indicated the presence of ellagitannin derivatives . In order to confirm the identities of the detected compounds, they were isolated from T2 and T5 by preparative chromatographic techniques and identified by spectroscopic methods including tandem mass spectrometry . RESULTS: By using LC-UV-ES-MS, four major compounds (ellagic acid, gallic acid, punicalagin, terchebulin) could be identified in the T extract . Three other compounds (3,3'di-O-methylellagic acid, 3,4,3',4'-tetra-O-methylellagic acid, terflavin A) were also isolated and identified . CONCLUSIONS: LC-UV-ES-MS is a useful technique for the analysis of mixtures containing ellagitannins.

J Mol Microbiol Biotechnol, 2001 Jan, 3(1), 21 - 35
Comparative genomics of prokaryotic GTP-binding proteins (the Era, Obg, EngA, ThdF (TrmE), YchF and YihA families) and their relationship to eukaryotic GTP-binding proteins (the DRG, ARF, RAB, RAN, RAS and RHO families); Mittenhuber G; Several GTP-binding proteins with poorly defined functions were previously identified in Escherichia coli (i.e . Era, ThdF (TrmE)), Bacillus subtilis (i.e . Obg) and Neisseria gonorrhoeae (i.e . EngA) . In these species, every individual protein is encoded by an essential gene . BLAST searches were used to detect orthologs in genomes of various organisms . Alignments of orthologous sequences allowed the construction of phylogenetic trees and the definition of protein families . The BLAST searches also resulted in the identification of two additional families, the YchF and YihA families, named after the ychF and yihA genes of E . coli . Most families are not present in archaeal genomes, but representatives of each family were also detected in eukaryotic genomes . Only representatives of the YchF family are present in every genome sequenced to date, suggesting that YchF-like proteins might be involved in a fundamental life process . The GTP1/DRG family consisting of eukaryotic and archaeal proteins is related to the YchF family of GTP-binding proteins . The relationship of the six prokaryotic families of GTP-binding proteins and the GTP1/DRG family to eukaryotic GTPase families was also investigated: With the exception of the ARF family, a clear separation of the six prokaryotic families and the GTP1/DRG family with respect to eukaryotic (RAB, RAN, RAS and RHO) GTPases was observed.

Sex Transm Dis, 2001 Jan, 28(1), 58 - 61
Curable sexually transmitted infections among the clientele of a family planning clinic in Istanbul, Turkey; Ortayli N et al.; BACKGROUND: Integrating screening and treatment of sexually transmitted infections into existing family planning programs can be a good way to fight this problem . Also, the number of studies on the prevalence of certain sexually transmitted infections in this population in Turkey is limited . GOAL: To find out the prevalence of the most common sexually transmitted infections among the clients of an urban family planning clinic and to evaluate demographic characteristics and clinical signs related to sexually transmitted infections . STUDY DESIGN: Samples for Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis were collected from 569 women who attended the clinic from December 15, 1997 to March 31, 1998 . RESULTS: No case of N gonorrhoeae was found among the samples . The prevalence of C trachomatis infection was 1.9%, whereas the prevalence of of T vaginalis was 2.6% . None of the demographic characteristics or clinical signs was related to these infections . CONCLUSION: Larger studies using more sensitive laboratory tests are needed to determine the prevalence of chlamydial infections in family planning clinics in Turkey.

Sex Transm Dis, 2001 Jan, 28(1), 11 - 3
Motorcycle taxi drivers and sexually transmitted infections in a Peruvian Amazon City; Paris M et al.; BACKGROUND: In Iquitos, Peru, motorcycle taxis are a common form of public transportation used both by residents and visitors . GOAL: To evaluate the prevalence of factors associated with sexually transmitted disease risk and of Neisseria gonorrhoeae and Chlamydia trachomatis infections among motorcycle taxi drivers working in Iquitos, Peru . STUDY DESIGN: A convenience sample of motorcycle taxi drivers was recruited through street outreach . Participants were confidentially interviewed and provided urine specimens for gonococcal and chlamydial infection testing . RESULTS: A history of sexually transmitted disease (62%) or symptoms in the past year (35%) and of referral of clients to commercial sex workers (60%) was common . The prevalence of gonococcal or chlamydial infection was 3.5% . CONCLUSIONS: Motorcycle taxi drivers are a major means of public transportation in Iquitos, Peru, have high personal risks for sexually transmitted disease, and interface often with at-risk persons.

AIDS, 2001 Jan 5, 15(1), 105 - 10
Treatment of cervicitis is associated with decreased cervical shedding of HIV-1; Mcclelland RS et al.; OBJECTIVE: To determine whether cervical mucosal shedding of HIV-1 RNA and HIV-1 infected cells decreases following successful treatment of cervicitis . DESIGN: Prospective interventional study . SETTING: Sexually Transmitted Infections Clinic, Coast Provincial General Hospital, Mombasa, Kenya . PARTICIPANTS: Thirty-six HIV-1 seropositive women with cervicitis: 16 with Neisseria gonorrhoeae, seven with Chlamydia trachomatis, and 13 with non-specific cervicitis . INTERVENTIONS: Treatment of cervicitis . Main outcome measures: Levels of total (cell-free and cell-associated) HIV-1 RNA and presence of HIV-1 DNA (a marker for infected cells) in cervical secretions before and after resolution of cervicitis . RESULTS: After treatment of cervicitis, the median HIV-1 RNA concentration in cervical secretions was reduced from 4.05 to 3.24 log10 copies/swab (P = 0.001) . Significant decreases in cervical HIV-1 RNA occurred in the subgroups with N . gonorrhoeae (3.94 to 3.28 log10 copies/swab; P = 0.02) and C . trachomatis (4.21 to 3.19 log10 copies/swab; P = 0.02) . Overall, the prevalence of HIV-1 infected cells in cervical secretions also decreased after treatment, from 67% to 42% (odds ratio, 2.8; 95% confidence interval, 1.3-6.0; P = 0.009) . Detection of infected cells was associated with higher mean HIV-1 RNA levels (4.04 versus 2.99 log10 copies/swab; P< 0.0001) . CONCLUSIONS: Effective treatment of cervicitis resulted in significant decreases in shedding of HIV-1 virus and infected cells in cervical secretions . Treatment of sexually transmitted diseases may be an important means of decreasing the infectivity of HIV-1 seropositive women by reducing exposure to HIV-1 in genital secretions.

Minerva Ginecol, 2000 Sep, 52(9), 327 - 32
{Gynecologic infections in adolescence}; De Seta F et al.; BACKGROUND: Adolescence in the age bracket with the highest incidence of sexually transmitted diseases (STDs) . An observational study was performed in symptomatic patients attending our Outpatient Clinic for STDs in order to evaluate the diffusion of genital infections in the adolescent population and the different epidemiological distribution compared to adults . METHODS: The incidence of these infections was compared in two groups of patients attending the clinic between April 1995 and April 1999 . The first consisted of 54 teenagers (13-19 years-old) and the second, used as a control, consisted of 917 women of child-bearing age (20-40 years) . All patients underwent clinical examinations and microbiological tests of vaginal secretions (bacterioscopic test on coloured plates using the Gram method, cold tests, cultural assay of bacteria and fungi, assay for (Chlamydia trachomatis, Mycoplasma spp., Neisseria gonorrhoeae, Herpes simplex virus) . RESULTS: The most common genital infection in teenagers was candidosis (35% vs 23.9%), followed by bacterial vaginosis (31.9% vs 25.8%) and mixed flora vaginitis (25.9% vs 22.6%) . HSV was found in 1.8% of adolescents (vs 0.6%), in selected cases, Chlamydia and Mycoplasma were present in 16.6% (vs 1.1%) and 50% (vs 28%) of teenagers respectively . CONCLUSIONS: All genital infections were more frequent in the study population of adolescents compared to the control group . The adolescent population should be the target of a serious and systematic programme of prevention in order to ward off the sequelae of early and often irresponsible sexual activity.

J Bacteriol, 2001 Feb, 183(3), 942 - 50
Structural and immunochemical characterization of the lipooligosaccharides expressed by Neisseria subflava 44; Tong Y et al.; Neisserial lipooligosaccharides (LOSs) are a family of complex cell surface glycolipids . We used mass spectrometry techniques (electrospray ionization, collision-induced dissociation, and multiple step), combined with fluorophore-assisted carbohydrate electrophoresis monosaccharide composition analysis, to determine the structure of the two low-molecular-mass LOS molecules (LOSI and LOSII) expressed by Neisseria subflava 44 . We determined that LOSI contains one glucose on both the alpha and beta chains . LOSII is structurally related to LOSI and differs from it by the addition of a hexose (either glucose or galactose) on the alpha chain . LOSI and LOSII were able to bind monoclonal antibody (MAb) 25-1-LC1 when analyzed by Western blotting experiments . We used a set of genetically defined Neisseria gonorrhoeae mutants that expressed single defined LOS epitopes and a group of Neisseria meningitidis strains that expresses chemically defined LOS components to determine the structures recognized by MAb 25-1-LC1 . We found that extensions onto the beta-chain glucose of LOSI block the recognition by this MAb, as does further elongation from the LOSII alpha chain . The LOSI structure was determined to be the minimum structure that is recognized by MAb 25-1-LC1.

Cell Microbiol, 2001 Jan, 3(1), 33 - 44
Expression of pathogen-like Opa adhesins in commensal Neisseria: genetic and functional analysis; Toleman M et al.; Several species of commensal Neisseriae (Cn) may colonize the human nasopharynx, but little is known about their adhesion mechanisms . We have investigated structural and functional similarities between adhesins of Cn and of Neisseria meningitidis (Nm), also a frequent colonizer of the nasopharynx . In this study, we demonstrate the expression of Opa-like proteins in nine strains of Cn . Phylogenetic analysis segregated the majority of the Cn Opa in a cluster separated from the pathogenic cluster with a few exceptions . One Opa, which located within the pathogenic cluster, was strikingly similar (74%) to an Opa of a Neisseria gonorrhoeae (Ng) strain and, like Ng, it lacked the extra Y11 or the 136DKF138 triplet insert, which are conserved among many N . meningitidis Opa proteins . Most importantly, the majority of the Cn Opa proteins were able to interact with human CEACAM1 (CD66a) molecules, previously identified as receptors for pathogenic Opa proteins . By the use of CEACAM1 N-domain mutants, we demonstrate that Cn Opa target the same region of the N-domain of the receptor as that used by Nm . Furthermore, Cn strains bound to cell-expressed human CEACAM1 . In competition assays, adherent Cn strain C450, exhibiting high affinity for CEACAM1, was not displaced by a Nm isolate and vice versa . But in simultaneous incubation, Nm out-competed the Cn strain . This is the first study to demonstrate the expression of adhesins in Cn that are structurally and functionally closely related to pathogenic adhesins . The studies imply that some Cn have the potential to occupy and thus compete with the pathogens for receptors on human mucosa, their common and exclusive niche.

Cell Microbiol, 2000 Aug, 2(4), 341 - 51
Cholera toxin and extracellular Ca2+ induce adherence of non-piliated Neisseria: evidence for an important role of G-proteins and Rho in the bacteria-cell interaction; Kallstrom H et al.; In this study, we characterize the interaction between non-piliated (P-) Neisseria gonorrhoeae and human epithelial cells . P- mutants lacking the pilus subunit protein PilE attach at low levels to cells . Although the binding may not lead to heavy inflammatory responses, the interaction between P- Neisseria and host cells most probably play a role in colonization and asymptomatic carriage of the pathogen . Here we show that the adherence of P N . gonorrhoeae is blocked by GDP-beta-S {guanosine 5'-O(thio)diphosphate}, a non-hydrolyzable GTP analogue, and by C3 exotoxin, an inhibitor of the small G-protein Rho . G-protein activators such as cholera toxin, that activates Gs, and fluoroaluminate, a general G-protein activator, induced bacterial adherence . Furthermore, increase of the extracellular free {Ca2+} dramatically enhanced adherence of non-piliated Neisseria . The pharynx and the urogenital tract are natural entry sites of the pathogenic Neisseria species, and at both sites the epithelial cells can be exposed to wide variations in Ca2+ concentration . Taken together, these data show the importance of extracellular Ca2+ in the pathogenic Neisseria-host interaction, and reveal a novel function of cholera toxin, namely induction of bacterial adherence.

Cell Microbiol, 2000 Jun, 2(3), 207 - 18
Alteration of epithelial cell transferrin-iron homeostasis by Neisseria meningitidis and Neisseria gonorrhoeae; Bonnah RA et al.; Iron is an essential element for nearly all organisms . In mammals, iron is transported to body tissues by the serum glycoprotein transferrin . Transferrin-iron is internalized by binding to specific receptors followed by endocytosis . In vitro, Neisseria meningitidis and Neisseria gonorrhoeae can use iron from a variety of iron-containing compounds, including human transferrin . In vivo, transferrin is an important source of iron for N . gonorrhoeae: a mutant that is unable to bind and use transferrin-iron is unable to colonize the urethra of men or initiate disease at this site . As pathogenic Neisseria and its human host derive much of their iron from transferrin, we reasoned that a competition may exist between microbe and host epithelial cells for transferrin-iron at certain stages of infection . We therefore tested the hypothesis that N . meningitidis and N . gonorrhoeae may actively interfere with host transferrin-iron metabolism . We report that Neisseria-infected human epithelial cells have reduced levels of transferrin receptor messenger RNA and cycling transferrin receptors . The ability of infected cells to internalize transferrin receptor is also reduced . Finally, the relative distribution of surface and cycling transferrin receptors is altered in an infected cell . We conclude that Neisseria infection alters epithelial cell transferrin-iron homeostasis at multiple levels.

Cell Microbiol, 2000 Feb, 2(1), 69 - 82
Syndecan-1 and syndecan-4 can mediate the invasion of OpaHSPG-expressing Neisseria gonorrhoeae into epithelial cells; Freissler E et al.; Neisseria gonorrhoeae (Ngo) expressing the outer membrane protein OpaHSPG can adhere to and invade epithelial cells via binding to heparan sulphate proteoglycan (HSPG) receptors . In this study, we have investigated the role of syndecan-1 and syndecan-4, two members of the HSPG family, in the uptake of Ngo by epithelial cells . When overexpressed in HeLa cells, both syndecans co-localize with adherent Ngo on the host cell surface . This overexpression of syndecan-1 and syndecan-4 leads to a three- and sevenfold increase in Ngo invasion respectively . In contrast, transfection with the syndecan-1 and syndecan-4 mutant constructs lacking the intracellular domain results in an abrogation of the invasion process, characteristic of a dominant-negative mode of action . A concomitant loss of the capacity to mediate Ngo uptake was also observed with syndecan-4 mutant constructs carrying lesions in the dimerization motif necessary for the binding of protein kinase C (PKC) and phosphatidylinositol 4,5-bisphosphate (PIP2), and mutants that are deficient in a C-terminal EFYA amino acid motif responsible for binding to syntenin or CASK . We conclude that syndecan-1 and syndecan-4 can both mediate Ngo uptake into epithelial cells, and that their intracellular domains play a crucial role in this process, perhaps by mediating signal transduction or anchorage to the cytoskeleton.

Cell Microbiol, 1999 Sep, 1(2), 169 - 81
Molecular analysis of neisserial Opa protein interactions with the CEA family of receptors: identification of determinants contributing to the differential specificities of binding; Popp A et al.; The carcinoembryonic antigen (CEA) gene family members, CEACAM1, CEACAM3, CEACAM5 and CEACAM6, are bound by the Opa outer membrane proteins of pathogenic Neisseria spp., whereas CEACAM8 is not . In this study, we demonstrate that the closely related CEACAM4 and CEACAM7, which are also members of the CEA family, are not Opa receptors . We exploited the high conservation between CEACAM6 and CEACAM8 to generate an extensive set of chimeric receptors in order to delineate the sequences necessary for Opa binding . Using a transfection-based infection system, we showed that binding of Opa52 involves residues 27-42, which are predicted to form beta-strand C and short loops adjacent to it, and residues lying between amino acids 60 and 108 in the amino-terminal domain . The replacement of residues 27-29 in CEACAM6 with the CEACAM1 or CEACAM5 sequences generated recombinant CEACAM6 receptors that are bound by CEACAM1/CEACAM5-specific Opa variants . Together, our data demonstrate that Opa proteins bind to residues exposed on the GFCC' face of the N-terminal domain of CEACAM receptors, and identify an amino acid triplet sequence that is responsible for the differential binding of Opa proteins to CEACAM1, CEACAM5 and CEACAM6.

Antimicrob Agents Chemother, 2001 Mar, 45(3), 734 - 8
Correlation of in vitro susceptibilities to newer quinolones of naturally occurring quinolone-resistant Neisseria gonorrhoeae strains with changes in GyrA and ParC; Shultz TR et al.; The in vitro activities of ciprofloxacin, trovafloxacin, moxifloxacin, and grepafloxacin against 174 strains of Neisseria gonorrhoeae isolated in Sydney, Australia, were determined . The strains included 84 quinolone-less-sensitive and -resistant N . gonorrhoeae (QRNG) strains for which ciprofloxacin MICs were in the range of 0.12 to 16 microg/ml . The QRNG included strains isolated from patients whose infections were acquired in a number of countries, mostly in Southeast Asia . The gyrA and parC quinolone resistance-determining regions (QRDR) of 18 selected QRNG strains were sequenced, and the amino acid mutations observed were related to the MICs obtained . The activities of moxifloxacin and grepafloxacin against QRNG were comparable to that of ciprofloxacin . Trovafloxacin was more active than the other quinolones against some but not all of the QRNG strains . Increments in ciprofloxacin resistance occurred in a step-wise manner with point mutations initiated in gyrA resulting in amino acid alterations Ser91-to-Phe, Ser91-to-Tyr, Asp95-to-Gly, and Asp95-to-Asn . Single gyrA changes correlated with ciprofloxacin MICs in the range 0.12 to 1 microg/ml . The Ser91 changes in GyrA were associated with higher MICs and further QRDR changes . QRNG strains for which ciprofloxacin MICs were greater than 1 microg/ml had both gyrA and parC QRDR point mutations . ParC alterations were seen in these isolates only in the presence of GyrA changes and comprised amino acid changes Asp86-to-Asn, Ser87-to-Asn, Ser87-to-Arg, Ser88-to-Pro, Glu91-to-Lys, and Glu91-to-Gln . QRNG strains for which MICs were in the higher ranges had double GyrA mutations, but again only with accompanying ParC alterations . Not only did the nature and combination of GyrA and ParC changes influence the incremental increases in ciprofloxacin MICs, but they seemingly also altered the differential activity of trovafloxacin . Our findings suggest that the newer quinolones of the type examined are unlikely to be useful replacements for ciprofloxacin in the treatment of gonorrhea, particularly where ciprofloxacin MICs are high or where resistance is widespread.

Infect Immun, 2001 Mar, 69(3), 1909 - 13
Interactions of Neisseria gonorrhoeae with mature human macrophage opacity proteins influence production of proinflammatory cytokines; Makepeace BL et al.; The pathological features of ascending gonococcal infection suggest that proinflammatory mediators secreted by tissue-resident macrophages are important components of the host response . Challenge of fully differentiated, mature macrophages with variants of Neisseria gonorrhoeae strain P9 or purified bacterial surface components (pili, lipooligosaccharide, and outer membrane vesicles) induced the secretion of interleukin 6 (IL-6), tumor necrosis factor alpha, growth-related protein alpha, macrophage inflammatory protein 1alpha (MIP-1alpha), and RANTES cytokines but had no effect on IL-8 production . No secretion of IL-1beta, epithelial-derived neutrophil attractant 78, granulocyte-macrophage colony-stimulating factor, IL-10, or IL-12 cytokines was observed . Notably, the P9-Opa(b) protein, in comparison to P9-Opa(a), increased the association of gonococci with macrophages and elevated the secretion of cytokines . Thus, variation in Opa protein expression by the gonococcus may be a determining factor in the severity of pelvic inflammatory disease.

Int J STD AIDS, 2001 Jan, 12(1), 47 - 9
Reproductive health and STDs among clients of a women's health mobile clinic in rural Bali, Indonesia; Patten JH et al.; Recent studies in Indonesia have reported significant levels of STDs in low-risk urban groups . To gather data on rural women, a prevalence study was undertaken in Bali utilizing a women's health mobile clinic . Rural (n=312) women were tested for vaginal reproductive tract infections (RTIs) by on-site wet mount, for Neisseria gonorrhoeae and Chlamydia trachomatis by unamplified DNA (Gen-Probe) test, and for syphilis by rapid plasma reagin (RPR) . Results were: candidiasis 5.8%; bacterial vaginosis (BV) 37.2%; trichomoniasis 15.1%; gonorrhoea 0.7%; chlamydial infection 5.6%; syphilis 0% . Overall 55.1% had at least one RTI and 19.2% had at least one STD . Reported non-monogamy, pain with urination and genital lesions, observed cervical friability and cervical motion tenderness were associated with cervical STD infection (P<0.05) . We conclude that there is a need for improved services for STD prevention and RTI/STD management in rural Bali, and for condom promotion.

Nat Struct Biol, 2001 Feb, 8(2), 166 - 75
Crystal structure of the retaining galactosyltransferase LgtC from Neisseria meningitidis in complex with donor and acceptor sugar analogs; Persson K et al.; Many bacterial pathogens express lipooligosaccharides that mimic human cell surface glycoconjugates, enabling them to attach to host receptors and to evade the immune response . In Neisseria meningitidis, the galactosyltransferase LgtC catalyzes a key step in the biosynthesis of lipooligosaccharide structure by transferring alpha-d-galactose from UDP-galactose to a terminal lactose . The product retains the configuration of the donor sugar glycosidic bond; LgtC is thus a retaining glycosyltranferase . We report the 2 A crystal structures of the complex of LgtC with manganese and UDP 2-deoxy-2-fluoro-galactose (a donor sugar analog) in the presence and absence of the acceptor sugar analog 4'-deoxylactose . The structures, together with results from site-directed mutagenesis and kinetic analysis, give valuable insights into the unique catalytic mechanism and, as the first structure of a glycosyltransferase in complex with both the donor and acceptor sugars, provide a starting point for inhibitor design.

Curr Opin Microbiol, 2001 Feb, 4(1), 53 - 7
Implications of molecular contacts and signaling initiated by Neisseria gonorrhoeae; Koomey M; Neisseria gonorrhoeae employs diverse strategies with which to adhere to and invade host cells during the course of infection . These primary encounters provide means by which biologically active molecules can be efficiently targeted to disrupt or exploit normal host cell metabolism and immune response elements, which in turn leads to the pathological responses characteristic of gonococcal disease . Current studies have begun to elucidate in detail the molecular interactions orchestrating these processes and the signaling events that they provoke.

Gene, 2000 Dec 23, 259(1-2), 99 - 108
Insertion with long target duplication: a mechanism for gene mobility suggested from comparison of two related bacterial genomes; Nobusato A et al.; The complete genome sequences of two closely related organisms--two Helicobacter pylori strains--have recently become available . Comparison of these genomes at single base pair level has suggested the presence of a mechanism for bacterial gene mobility--insertion with long target duplications . This mechanism is formally similar to classical transposon insertion, but the duplication is much longer, often in the range of 100bp . Restriction and/or modification enzyme genes are often within or adjacent to the insertion . A similar process may have mediated insertion of the cag(+) pathogenicity island in H . pylori . A similar structure was identified in comparisons between Neisseria meningitidis and Neisseria gonorrhoeae genomes . We hypothesize that this mechanism, as well as two other types of polymorphism linked with restriction-modification genes (insertion accompanied by target deletion and a tripartite structure composed of substitution/inversion/deletion), have resulted from attack by restriction enzymes on the chromosome.

Vaccine, 2001 Jan 8, 19(11-12), 1526 - 33
PorB3 outer membrane protein on Neisseria meningitidis is poorly accessible for antibody binding on live bacteria; Michaelsen TE et al.; It is reported here that the PorB3 porin proteins of serotype 4 and 15 are poorly accessible for antibody binding on live Neisseria meningitidis bacteria, whereas the allelic PorB2 and the PorA outer membrane protein appear to be highly accessible . This was revealed by flow cytometry analysis using several mouse monoclonal antibodies (mAbs) as well as PorB3 specific antibodies isolated from post vaccination and patient sera . However, strong antibody binding to the PorB3 protein was observed after killing the bacteria with ethanol . The reason for the lack of epitope exposure could be a shielding effect of the carbohydrate chains of lipopolysaccharides (LPS) possibly combined with short extra-cellular loops in the PorB3 protein . The findings indicate that the PorB3 protein is not an optimal target for protective antibodies induced by vaccination.

Microbiology, 2001 Jan, 147(Pt 1), 225 - 37
Deletion of the cell-division inhibitor MinC results in lysis of Neisseria gonorrhoeae; Ramirez-Arcos S et al.; The minCDE genes involved in division site selection in Neisseria gonorrhoeae were identified using raw data from the N . gonorrhoeae genome project and are part of a cluster of 27 genes . When gonococcal min genes were heterologously expressed as a cluster in Escherichia coli, minicells and filaments were produced, indicating that gonococcal min genes disrupted cell division in other genera . The insertional inactivation of the minC gene of N . gonorrhoeae CH811 resulted in a strain (CSRC1) with decreased viability and grossly abnormal cell division as observed by phase-contrast and electron microscopy analysis . Western blot analysis of N . gonorrhoeae CSRC1 confirmed that MinC(Ng) was not produced . Complementation of CSRC1 by integrating a minC-6xHis tag fusion at the proAB locus by homologous recombination restored viability and 1.9 times wild-type levels of MinC(Ng) expression . This slight increase of expression caused a small percentage of the complemented cells to divide aberrantly . This suggested that the 6xHis tag has partially affected the stability of MinC, or that the chromosomal position of minC is critical to its regulation . Comparison of MinC proteins from different bacteria showed a homologous region corresponding to residues 135-230 with five conserved amino acids . Overexpression of MinC(Ng) in wild-type E . coli cells induced filamentation and an E . coli minC mutant was successfully complemented with minC(Ng) . Therefore, the evidence indicates that MinC from N . gonorrhoeae acts as a cell-division inhibitor and that its role is essential in maintaining proper division in cocci.

J Exp Med, 2001 Feb 5, 193(3), 281 - 95
Binding of C4b-binding protein to porin: a molecular mechanism of serum resistance of Neisseria gonorrhoeae; Ram S et al.; We screened 29 strains of Neisseria gonorrhoeae and found 16/21 strains that resisted killing by normal human serum and 0/8 serum sensitive strains that bound the complement regulator, C4b-binding protein (C4bp) . Microbial surface-bound C4bp demonstrated cofactor activity . We constructed gonococcal strains with hybrid porin (Por) molecules derived from each of the major serogroups (Por1A and Por1B) of N . gonorrhoeae, and showed that the loop 1 of Por1A is required for C4bp binding . Por1B loops 5 and 7 of serum-resistant gonococci together formed a negatively charged C4bp-binding domain . C4bp-Por1B interactions were ionic in nature (inhibited by high salt or by heparin), whereas the C4bp-Por1A bond was hydrophobic . Only recombinant C4bp mutant molecules containing the NH2-terminal alpha-chain short consensus repeat (SCR1) bound to both Por1A and Por1B gonococci, suggesting that SCR1 contained Por binding sites . C4bp alpha-chain monomers did not bind gonococci, indicating that the polymeric form of C4bp was required for binding . Using fAb fragments against C4bp SCR1, C4bp binding to Por1A and Por1B strains was inhibited in a complement-dependent serum bactericidal assay . This resulted in complete killing of these otherwise fully serum resistant strains in only 10% normal serum, underscoring the importance of C4bp in mediating gonococcal serum resistance.

Sex Transm Infect, 1999 Dec, 75(6), 431 - 4
Estimating the prevalence of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and human papillomavirus infection in indigenous women in northern Australia; Bowden FJ et al.; OBJECTIVE: To estimate more accurately the age specific prevalence of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and human papillomavirus infection (HPV) in indigenous women living in urban, rural, and remote areas of the "Top End" of the Northern Territory (NT) . DESIGN: Analysis of data obtained from two community based studies using self administered tampon specimens tested by polymerase chain reaction for sexually transmitted disease (STD) . Data pertaining to the notifiable STDs (N gonorrhoeae and C trachomatis) were obtained from the NT health department . PATIENTS: 1090 indigenous women (age range 12-73 years) were enrolled when they attended local community health centres, family planning clinics, and STD clinics . The majority attended clinics in their home community in the course of "well women's checks" which encourage women to undergo screening for a variety of general medical conditions . RESULTS: The overall prevalence of T vaginalis, C trachomatis, N gonorrhoeae, and HPV was 0.25 (95% CI: 0.22-0.28), 0.11 (0.09-0.13), 0.17 (0.15-0.19), and 0.42 (0.37-0.48) respectively . Of the women found to be infected (excluding HPV), 25.5% had two or more of the above organisms detected . There was a statistically significant increase in the age specific prevalence of T vaginalis but a significant decrease with age for C trachomatis and HPV infection . There was no statistically significant change for N gonorrhoeae with age . CONCLUSIONS: STDs are hyperendemic in this population of indigenous women and the notification data significantly underestimate their prevalence . Distinct patterns of age specific prevalence were demonstrated, highlighting the need to tailor control strategies to specific epidemiological features.

Sex Transm Infect, 1999 Dec, 75(6), 409 - 11
Knowledge of Chlamydia trachomatis genital infection and its consequences in people attending a genitourinary medicine clinic; Devonshire P et al.; OBJECTIVES: To assess knowledge of Chlamydia trachomatis infections, with a comparison of knowledge of Neisseria gonorrhoeae infections . METHODS: A cross sectional survey, by self completed questionnaire, of 200 subjects attending a genitourinary outpatient clinic . RESULTS: The response rate was 82% (90 male and 73 female) . 51% of men (60% of females) had heard of chlamydia . 65 (82%) were unaware of the potential consequences of infection . 66% were unaware that the disease could be asymptomatic . Significantly more men (77%) than women (60%) had heard of gonorrhoea . Most participants (68%-82%) knew little of the possible consequences of this infection, and only 26% were aware that it could be asymptomatic . Knowledge was higher regarding fertility topics . There was no correlation between knowledge and either age or socioeconomic status . However, greater knowledge was displayed by those who read health information leaflets always or often . For both men and women, the preferred source of health information was the doctor . Other popular sources were health information leaflets, women's magazines, and television . CONCLUSIONS: Barely half the participants had heard of chlamydia and gonorrhoea . Further knowledge of either infection was very poor . There are serious implications for public health . The reasons for this are unclear and require exploration before targeted health promotion . Doctors and the popular media are acceptable, and potentially effective, sources of information . Acquisition of knowledge is important, both to reduce sexual risk taking behaviour and its consequences, and to allow for informed consent for chlamydia screening programmes.

Enferm Infecc Microbiol Clin, 2000 Oct, 18(8), 392 - 5
{Incidence of genitourinary infection caused by Chlamydia trachomatis in a STD center calculated by direct antigen detection}; Chavez M et al.; OBJECTIVE: Chlamydia trachomatis is one of the most common sexually transmitted agents which causes a wide spectrum of diseases including urethritis in men and endocervicitis in women . We analyzed patients with genitourinary C . trachomatis infections evaluating risk factors and the association with other sexually transmitted infections . MATERIAL AND METHODS: We processed 1,180 specimens from 913 patients (772 women and 141 men), attended at a Sexually Transmitted Diseases (STD) Center . The diagnostic of C . trachomatis infection was made by an enzyme-linked fluorescent immunoassay, Vidas Chlamydia test (bioMerieux) . RESULTS: The incidence of C . trachomatis infection was 4.8% (57 cases) and was higher in women (70.1%) than in men (29.8%) . The risk groups observed were: 26 prostitute, 7 contact with prostitute or risk partner, 5 homosexual, 5 promiscuous heterosexual and 14 without risk groups . Associated with this infection we observed other: 10 bacterial vaginosis, 8 Papillomavirus infection, 3 Trichomonas vaginalis infections, 2 Neisseria gonorrhoeae infections and 2 Candidiasis . The 53.4% of these patients didn't have any symptomatology at the consult moment . CONCLUSIONS: The control of patients with risk factors is important for the diagnostic of C . trachomatis and other sexually transmitted infections, because most of them were prostitutes and asymptomatic . Within men, homosexuality, contact with prostitute or risk partner were the practices with higher risk.

Sex Transm Infect, 2000 Oct, 76(5), 400 - 2
The sac-4 gene of Neisseria gonorrhoeae and co-existing chlamydial infection; Phillips DJ et al.; BACKGROUND/OBJECTIVES: Recently, the sac-4 gene in Neisseria gonorrhoeae was postulated to increase the risk of developing mixed gonococcal and chlamydial infection . The aims of this study were to determine the frequency of the sac-4 gene in a larger sample of isolates of different serovars and to assess the prevalence of sac-4 in gonococcal isolates from patients with and without coexisting chlamydial infection . METHODS: Isolates from 259 episodes of gonorrhoea were tested by a PCR assay for the sac-4 gene . The presence of co-existing chlamydial infection was determined from both laboratory and GUM clinical records . RESULTS: The overall prevalence of sac-4 was 57.5% (149/259) . The prevalence was not the same in all serovars and ranged from 34.9% in serovar 1B2 to 100% in serovar 1B18 . Exact logistic regression analysis indicated significant differences in sac-4 prevalence in isolates of different serovars . The prevalence of sac-4 was 69.5% (41/59) in gonococcal isolates from patients with co-existing chlamydial infection compared with 57.9% (62/107) for those without chlamydial infection . Exact logistic regression analysis showed that the slightly increased sac-4 prevalence among chlamydia positive patients (p = 0.2) virtually disappeared when serovar status was taken into account (p > 0.9) . CONCLUSION: The sac-4 gene of the gonococcus does not increase the risk for mixed chlamydial infection.

Sex Transm Infect, 2000 Oct, 76(5), 355 - 62
High prevalence of trichomoniasis in rural men in Mwanza, Tanzania: results from a population based study; Watson-Jones D et al.; OBJECTIVES: To measure the prevalence of urethral infections including trichomoniasis in rural Tanzanian men, to assess the prevalence of symptoms and signs among men with Trichomonas vaginalis, and to analyse the risk factors for trichomoniasis . DESIGN: A cross sectional study of 1004 men aged 15-54 years in a rural community in north west Tanzania . METHODS: Participants were interviewed about sexual behaviour and symptoms of sexually transmitted diseases . First fraction urine samples and urethral swabs were collected and used to test for T vaginalis by wet preparation and culture, Neisseria gonorrhoeae by culture, Chlamydia trachomatis by ligase chain reaction and non-specific urethritis by Gram stain . Urine was also tested for the presence of leucocytes using a leucocyte esterase dipstick . Men were re-interviewed 2 weeks later to document new symptoms and signs of urethritis . RESULTS: Complete laboratory results were available on 980 men . One in four men had laboratory evidence of urethritis . T vaginalis was found in 109 individuals (11%), gonorrhoea in eight (0.8%), and chlamydial infection in 15 (1.5%) . Over 50% of men with urethritis were asymptomatic . The prevalence of signs and symptoms was similar among men with T vaginalis alone compared with men with other urethral infections . The sensitivity and specificity of the leucocyte esterase dipstick (LED) test for detecting T vaginalis were 80% and 48% respectively in symptomatic men and 60% and 68% in asymptomatic men . Factors associated with trichomoniasis included religion, type of employment, and marital status . CONCLUSIONS: A high prevalence of urethritis was found in men in this community based study . More than half of the urethral infections detected were asymptomatic . The most prevalent pathogen was T vaginalis . Studies are needed on the prevalence of trichomoniasis in men presenting to health services with complaints suggestive of urethritis since treatment for T vaginalis is not included in the syndromic management of urethritis in most countries . The performance of the LED test as a screening test for trichomoniasis was unsatisfactory in both symptomatic and asymptomatic men . Improved screening tests are urgently needed to identify urethral infections that are asymptomatic and which are not covered by current syndromic management algorithms.

Int J STD AIDS, 2000 Dec, 11(12), 804 - 11
A randomized, placebo-controlled trial of monthly azithromycin prophylaxis to prevent sexually transmitted infections and HIV-1 in Kenyan sex workers: study design and baseline findings; Fonck K et al.; Our objectives were to describe the baseline findings of a trial of antibiotic prophylaxis to prevent sexually transmitted infections (STIs) and HIV-1 in a cohort of Nairobi female sex workers (FSWs) . A questionnaire was administered and a medical examination was performed . HIV-negative women were randomly assigned to either one gram azithromycin or placebo monthly . Mean age of the 318 women was 32 years, mean duration of sex work 7 years and mean number of clients was 4 per day . High-risk behaviour was frequent: 14% practised anal intercourse, 23% sex during menses, and 3% used intravenous drugs . While 20% reported condom use with all clients, 37% never use condoms . However, STI prevalence was relatively low: HIV-1 27%, bacterial vaginosis 46%, Trichomonas vaginalis 13%, Neisseria gonorrhoeae 8%, Chlamydia trachomatis 7%, syphilis 6% and cervical intraepithelial neoplasia (CIN) 3% . It appears feasible to access a population of high-risk FSWs in Nairobi with prevention programmes, including a proposed trial of HIV prevention through STI chemoprophylaxis.

Int J STD AIDS, 2000 Dec, 11(12), 790 - 4
High prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae and Mycoplasma genitalium in female commercial sex workers in Japan; Tsunoe H et al.; Our objectives were to explore the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and Mycoplasma genitalium in Japanese female commercial sex workers (CSWs), in comparison with pregnant women as controls . A high-risk group of 174 female CSWs and 90 asymptomatic pregnant women were enrolled in this study . Detection of C . trachomatis, N . gonorrhoeae, and M . genitalium on the endocervix of the women was performed mainly by using polymerase chain reaction (PCR)-based assays . The prevalence rates of C . trachomatis, N . gonorrhoeae, and M . genitalium were 19.0%, 32.8%, and 12.6%, respectively, in the CSWs, compared with 5.6%, 0%, and 1.1% respectively, in the pregnant women . These results suggest a high prevalence of C . trachomatis, N . gonorrhoeae, and M . genitalium in Japanese CSWs . We conclude that continued close monitoring of the prevalence of C . trachomatis, N . gonorrhoeae, and M . genitalium infection in CSWs is important for preventing the dissemination of these microorganisms, and that further investigation of M . genitalium as a sexually transmitted pathogen in women is needed.

Int J STD AIDS, 2000 Dec, 11(12), 774 - 6
Neisseria gonorrhoeae in a London sexually transmitted infection clinic not fully sensitive to quinolones: are isolates imported and how effective is ciprofloxacin as a first-line therapy?
Ivens D, Martin I, Ison C.
Our objectives were to determine the prevalence of Neisseria gonorrhoeae not fully sensitive to ciprofloxacin from a sexually transmitted infection (STI) clinic in London and where the isolates were acquired from . Data of antibiotic sensitivities of N . gonorrhoeae from 292 patients were reviewed over a 6-month period at St Mary's Genitourinary Medicine (GUM) Clinic, London . Isolates which exhibited reduced susceptibility (minimum inhibitory concentration {MIC} 0.03-0.12 mg/l) and high level resistance (MIC>0.12 mg/l) to ciprofloxacin represented 10% and 1.3% of the total respectively . All patients infected with a high level resistant isolate to ciprofloxacin had had a recent sexual partner from abroad but 18 of the 28 patients infected with a reduced susceptibility isolate denied recent travel . None of the 20 patients with a non-sensitive isolate who re-attended for post treatment cultures had persistant gonococcal infection . From this study we concluded that although N . gonorrhoeae resistant to ciprofloxacin was rare and probably always acquired abroad, isolates exhibiting reduced susceptibility were more common and were mainly as a result of infection from the UK . A stat dose of ciprofloxacin 500 mg and doxycycline 100 mg twice a day for one week was effective treatment.

Proc Natl Acad Sci U S A, 2001 Jan 2, 98(1), 182 - 7
Recombination within natural populations of pathogenic bacteria: short-term empirical estimates and long-term phylogenetic consequences; Feil EJ et al.; The identification of clones within bacterial populations is often taken as evidence for a low rate of recombination, but the validity of this inference is rarely examined . We have used statistical tests of congruence between gene trees to examine the extent and significance of recombination in six bacterial pathogens . For Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus, the congruence between the maximum likelihood trees reconstructed using seven house-keeping genes was in most cases no better than that between each tree and trees of random topology . The lack of congruence between gene trees in these four species, which include both naturally transformable and nontransformable species, is in three cases supported by high ratios of recombination to point mutation during clonal diversification (estimates of this parameter were not possible for Strep . pyogenes) . In contrast, gene trees constructed for Hemophilus influenzae and pathogenic isolates of Escherichia coli showed a higher degree of congruence, suggesting lower rates of recombination . The impact of recombination therefore varies between bacterial species but in many species is sufficient to obliterate the phylogenetic signal in gene trees.

Am J Kidney Dis . 2001 Jan;37(1):E10.
A case of continuous ambulatory peritoneal dialysis peritonitis with an uncommon organism and an atypical clinical course; Konner P et al.; This report describes a 46-year-old patient who experienced an atypical course of peritonitis while undergoing continuous ambulatory peritoneal dialysis (CAPD) . The first sign of peritonitis was progressive impairment of ultrafiltration with increasing fluid absorption . The patient came to the center after 5 days with leg edemas and 645 leukocytes/microL in the first dialysate outflow . On the same day, the dialysate cell count decreased to 208/microL . During the following days, ultrafiltration failure persisted despite spontaneous normalization of PD-fluid leukocytes . No other clinical symptoms were observed, and the serum C-reactive protein (CRP) level remained normal . Magnetic resonance peritoneography and abdominal radiograph did not show dislocation of the catheter, a dialysate leak, or other causes of ultrafiltration failure . At day 14, fever, diarrhea, and an elevated serum CRP level occurred . Dialysate cultures taken on days 8, 11, and 14 showed growth of NEISSERIA: sicca . After initiation of antibiotic therapy with levofloxacine on day 14 ultrafiltration, clinical symptoms and serum CRP normalized within 3 days . In conclusion, Neisseria sicca should be considered as a rare cause of PD peritonitis . Our case report further illustrates the importance of ultrafiltration failure as an early and main symptom of peritoneal inflammation . The frequently used peritonitis criteria may not apply to cases of mild PD peritonitis.

Lancet, 2000 Dec 9, 356(9246), 1984 - 5
Increased ciprofloxacin resistance in gonococci isolated in Scotland; Forsyth A et al.; A review of the susceptibility of Neisseria gonorrhoeae isolated from 4415 episodes of infection in Scotland between 1991 and 1999 showed that the proportion of isolates with lowered susceptibility (ciprofloxacin minimum inhibitory concentration {MIC} > or = 0.05 mg/L) increased from 0.5% in 1991 to 5% in 1999 (p<0.001), whereas the proportion of isolates with clinical resistance (ciprofloxacin MIC > or = 1 mg/L) was significantly higher in 1999 than the average for the preceding 4 years (2.2% vs 0.9%; p=0.02) . Ciprofloxacin is a recommended treatment for gonococcal infection in the UK but if resistance continues to increase at the present rate it might not be suitable as a first-line treatment of gonorrhoea for much longer.

Proc Natl Acad Sci U S A, 2000 Dec 19, 97(26), 14584 - 9
Role of serum amyloid P component in bacterial infection: protection of the host or protection of the pathogen; Noursadeghi M et al.; Serum amyloid P component (SAP) binds to Streptococcus pyogenes, and we show here that it also binds to Neisseria meningitidis, including a lipopolysaccharide (LPS)-negative mutant, and to rough variants of Escherichia coli . Surprisingly, this binding had a powerful antiopsonic effect both in vitro and in vivo, reducing phagocytosis and killing of bacteria . Furthermore, SAP knockout mice survived lethal infection with S . pyogenes and rough E . coli J5, organisms to which SAP binds . The susceptibility of SAP(-/-) mice was fully restored by injection of isolated human SAP . However, SAP(-/-) mice were more susceptible than wild-type animals to lethal infection with E . coli O111:B4, a smooth strain to which SAP does not bind, suggesting that SAP also has some host defense function . Although SAP binds to LPS in vitro, SAP(-/-) mice were only marginally more susceptible to lethal LPS challenge, and injection of large amounts of human SAP into wild-type mice did not affect sensitivity to LPS, indicating that SAP is not a significant modulator of LPS toxicity in vivo . In contrast, the binding of SAP to pathogenic bacteria enabled them to evade neutrophil phagocytosis and display enhanced virulence . Abrogation of this molecular camouflage is thus potentially a novel therapeutic approach, and we show here that administration to wild-type mice of (R)-1-{6-(R)-2-carboxy-pyrrolidin-1-yl}-6-oxo-hexanoyl}pyrrolidine -2- carboxylic acid, a drug that inhibits SAP binding, significantly prolonged survival during lethal infection with E . coli J5.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 359 - 62
In vitro antibiotic susceptibility of Neisseria gonorrhoeae in Jakarta, Indonesia; Lesmana M et al.; Antibiotic susceptibilities were determined for 122 Neisseria gonorrheae isolates obtained from 400 sex workers in Jakarta, Indonesia, and susceptibilities to ciprofloxacin, cefuroxime, cefoxitin, cefotaxime, ceftriaxone, chloramphenicol, and spectinomycin were found . All isolates were resistant to tetracycline . A number of the isolates demonstrated decreased susceptibilities to erythromycin (MIC >/= 1.0 microg/ml), thiamphenicol (MIC >/= 1.0 microg/ml), kanamycin (MIC >/= 16.0 microg/ml), penicillin (MIC >/= 2.0 microg/ml), gentamicin (MIC >/= 16.0 microg/ml), and norfloxacin (MIC = 0.5 microg/ml) . These data showed that certain antibiotics previously used in the treatment of gonorrhea are no longer effective.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 117 - 23
Molecular basis of high-level ciprofloxacin resistance in Neisseria gonorrhoeae strains isolated in Denmark from 1995 to 1998; Su X et al.; In Denmark surveillance of the in vitro susceptibility to ciprofloxacin of Neisseria gonorrhoeae was established in 1990 . The proportion of N . gonorrhoeae strains with decreased susceptibility or resistance to ciprofloxacin (MIC >/= 0.06 microg/ml) was low (0.3 to 2.3%) up to 1995 . Between 1995 and 1998 the rate of less-susceptible and resistant strains rose from 6.9 to 13.2% . Among ciprofloxacin-resistant strains (MIC >/= 1 microg/ml), 81% were highly resistant (MIC >/= 4 microg/ml) . Thirty-five N . gonorrhoeae strains (40 isolates) for which ciprofloxacin MICs were 4 to 32 microg/ml were investigated for the frequency and patterns of mutations within the gyrA and parC genes . The quinolone resistance-determining regions of the gyrA and parC genes were amplified by PCR, and the amplicons were directly sequenced . Alterations at Ser-91 and Asp-95 in GyrA and a single or double alteration in ParC were identified in 32 strains (91%) . Ser-91-to-Phe and Asp-95-to-Gly alterations in GyrA were detected in 28 strains (80%) . The most common ParC alteration, Asp-86 to Asn, was found in 19 strains (54%) . The strains were analyzed for genetic relationship by pulsed-field gel electrophoresis (PFGE) . The analysis showed that nine strains with the same mutation pattern in the gyrA and parC genes, originating from different geographical areas over 3 years, had the same PFGE patterns after SpeI as well as NheI digestion (only one strain with one band difference in the NheI pattern), suggesting that a resistant clone had spread worldwide . The results from this study strongly suggest that double gyrA mutations plus a parC mutation(s) play an important role in the development of high-level fluoroquinolone resistance in N . gonorrhoeae.

J Infect Dis, 2001 Jan 15, 183(2), 269 - 276 Epub 2000 Dec 21.
Association of Mycoplasma genitalium with nongonococcal urethritis in heterosexual men; Totten PA et al.; Chlamydia trachomatis and Neisseria gonorrhoeae are universally acknowledged as urethral pathogens, yet the etiology in the majority of cases of urethritis is unclear . Our case-control study assessed the association of Mycoplasma genitalium, Ureaplasma urealyticum, and other potential pathogens with acute nongonococcal urethritis (NGU) in heterosexual men presenting to an urban sexually transmitted diseases clinic . M . genitalium was detected in 27 (22%) of 121 NGU case patients and in 5 (4%) of 117 control subjects (P<.01) . Although C . trachomatis was detected in 36 (30%) of 121 NGU case patients and in 4 (3%) of 117 control subjects (P<.01), only 3 men with NGU were infected with both C . trachomatis and M . genitalium . U . urealyticum was not associated with NGU . By multivariate analyses, controlling for age, race, history of prior urethritis, and chlamydial infection, M . genitalium was associated with a 6.5-fold increased risk of urethritis (95% confidence interval, 2.1-19.5), which supports a role of this organism in the etiology of NGU.

What Is MIC?, What Is Genome?, What Is Listeria Monocytogenes?, What Is Salmonella?, What Is Molecular Biology?, s, Microorganisms, a, Microbe, n, Bacteria, n, Microbes, o, Bacterium, a, S. cerevisiae, s, Campylobacter, i, Agrobacterium, o, Paracocci, o, Gram negative, o, Antibiotic treatment, a, Escherichia coli, c, Denitrifying, a, Propionibacteria, n, Escherichia coli broth, r, Escherichia coli, c, Escherichia coli, o, Enterobacters, i, Burkholderia, n, Streptococcal, s, Yeasts, n, Paenibacilli, n, Microbial, n, Shigella, e, Bacillus, i, Escherichia coli

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005