Neisseria

Microbiology, 1999 Aug, 145 ( Pt 8), 2105 - 16
Characterization of apxIVA, a new RTX determinant of Actinobacillus pleuropneumoniae; Schaller A et al.; A fourth type of RTX determinant was identified in Actinobacillus pleuropneumoniae and was designated apxIVA . When expressed in Escherichia coli, recombinant ApxIVA showed a weak haemolytic activity and co-haemolytic synergy with the sphingomyelinase (beta-toxin) of Staphylococcus aureus . These activities required the presence of an additional gene, ORF1, that is located immediately upstream of apxIVA . The apxIVA gene product could not be detected in A . pleuropneumoniae cultures grown under various conditions in vitro; however, pigs experimentally infected with A . pleuropneumoniae serotypes 1, 5 and 7 started to produce antibodies that reacted with recombinant ApxIVA 14 d post-infection, indicating that apxIVA is expressed in vivo . In addition, sera from pigs naturally and experimentally infected with any of the serotypes all reacted with recombinant ApxIVA . The apxIVA gene from the serotype 1 A . pleuropneumoniae type strain Shope 4074T encodes a protein with a predicted molecular mass of 202 kDa which has typical features of RTX proteins including hydrophobic domains in the N-terminal half and 24 glycine-rich nonapeptides in the C-terminal half that bind Ca2+ . The glycine-rich nonapeptides are arranged in a modular structure and there is some variability in the number of modules in the ApxIVA proteins of different serotypes of A . pleuropneumoniae . The deduced amino acid sequences of the ApxIVA proteins have significant similarity with the Neisseria meningitidis iron-regulated RTX proteins FrpA and FrpC, and to a much lesser extent with other RTX proteins . The apxIVA gene could be detected in all A . pleuropneumoniae serotypes and seems to be species-specific . Although the precise role of this new RTX determinant in pathogenesis of porcine pleuropneumonia needs to be determined, apxIVA is the first in vivo induced toxin gene that has been described in A . pleuropneumoniae.

Mol Diagn, 1997 Sep, 2(3), 161 - 168
Detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Ureaplasma urealyticum, and Mycoplasma genitalium in First-void Urine Specimens by Multiplex Polymerase Chain Reaction; Mahony JB et al.; Background: Sexually transmitted diseases are often caused by one or more microorganisms, and asymptomatic carriage and transmission may be of significance . Testing for more than one organism in a single assay could be a useful approach to laboratory diagnosis . Methods and Results: A multiplex polymerase chain reaction (PCR) assay was developed that employed specific primers targeted to the 7.5-kb cryptic plasmid of Chlamydia trachomatis, the cppB gene of the 4.2-kb cryptic plasmid of Neisseria gonorrhoeae, the 140-kd major adhesion protein gene of Mycoplasma genitlium, and the urease gene of Ureaplasma urealyticum . All four polymerase chain reaction products were detectable by agarose gel electorphoresis and were confirmed by Southern hybridization using fluorescein isothiocyanate-labeled oligonucleotide probes and enhanced chemiluminescent detection . Using purified DNA preparations, multiplex PCR had a reproducible detection limit of 1 fg of C . trachomatis DNA, 100 fg of N . gonorrhoeae DNA, and 10 fg U . urealyticum DNA and M . genitalium DNA, which converts to 1-2 genomic equivalents (ge) of C . trachomatis and N . gonorrhoeae, 4 ge of M . genitalium, and 10 ge U . urealyticum . Multiplex PCR was compared with individual uniplex polymerase chian reaction PCR assays by testing 117 first-void urine samples (91 men, 26 women) from Canadian or Kenyan patients . Multiplex PCR detected 45 of 46 (97.8%) urines with C . trachomatis DNA, 42 of 42 (100%) urines with N . gonorrhoeae DNA, 17 of 17 (100%) urines with U . urealyticum DNA, 4 of 4 (100%) urines with M . genitalium DNA, 12 of 12 urines that had DNA from two bacteria, and 2 of 2 urines with DNA from three bacteria . Multiplex PCR correctly identified bacteria in 92 of 93 urines for an overall sensitivity of 98.9% . Specificity calculations were 100% for C . trachomatis (71/71), N . gonorhoeae (75/75), U . urealyticum (100/100), and M . genitalium (113/113) . Conclusions: Multiplex PCR provided a single sensitive and specific test for the detection of four bacteria in first-void urine samples . Testing of first-void urine samples by multiplex PCR could facilitate studies aimed at improving our understanding of the epidemiology of these important sexually transmitted diseases.

Br J Haematol, 1999 Aug, 106(2), 391 - 9
Circulating platelet-neutrophil complexes represent a subpopulation of activated neutrophils primed for adhesion, phagocytosis and intracellular killing; Peters MJ et al.; Platelets play a prominent role in linking the processes of inflammation, haemostasis and thrombosis . Recent studies have shown that platelets form heterotypic aggregates with leucocytes via platelet CD62P and leucocyte beta2 integrins . These interactions have been observed in vitro in blood taken from healthy volunteers and in clinical conditions in which thrombosis and inflammation are prominent . This study investigated the properties of platelet-neutrophil complexes (PNCs) in anticoagulated whole blood . At rest, neutrophils in PNCs exhibit a significantly more activated adhesion molecule profile than free neutrophils with increased CD11b expression and activation (increased binding of the CD11b/CD18 'activation reporter' monoclonal antibody 24) and decreased CD62L expression . In addition, neutrophils in PNCs phagocytosed significantly more Neisseria meningitidis and produced more toxic oxygen metabolites than free neutrophils . Stimulation with the platelet agonist adenosine diphosphate (ADP) led to further increases in CD11b expression and activation, loss of CD62L as well as increased phagocytosis and toxic oxygen metabolite production throughout the whole neutrophil population . When these experiments were repeated with the CD62P blocking antibody G1 the effects were inhibited to a variable extent, dependent upon the parameter under investigation . These results indicate that both soluble and contact-dependent factors contribute to platelet-mediated neutrophil activation . Platelet neutrophil complexes represent a large subpopulation of neutrophils with a more activated adhesion molecule profile, and a greater capacity for phagocytosis and toxic oxygen metabolite production . This study provides further support for a role for PNCs in both health and disease.

Sex Transm Dis, 1999 Aug, 26(7), 410 - 20
Prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and HIV infection among drug users attending an STD/HIV prevention and needle-exchange program in Quebec City, Canada; Poulin C et al.; OBJECTIVES: To determine the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and human immunodeficiency virus (HIV) infection among injection and noninjection drug users in Quebec City and to identify associated risk factors . STUDY DESIGN: Cross-sectional study of 738 drug users attending a sexually transmitted disease/HIV prevention and needle-exchange program in Quebec City, Canada . RESULTS: The prevalences of N . gonorrhoeae and C . trachomatis were, respectively, 0.4% (95% CI: 0.08%-1.2%) and 3.4% (95% Cl: 2.2%-5.0%) . Through multivariate analysis, risk factors associated with sexually transmitted diseases were: among women, being aged between 20 and 24 years and having unprotected sex with commercial partners; in noninjection drug users; cocaine use and first intercourse before 13 years of age; in male noninjection drug users, having regular sexual partners in the previous 6 months . No case of HIV was found in participants who have never injected drugs, but the prevalence was 5.5% (6/110) among the exinjectors and 10.1% (35/347) in current injectors . CONCLUSION: Moderate sexually transmitted disease and HIV prevalences were found, although a high proportion of drug users reported risky behaviors . Needle-exchange program sites may offer a good opportunity to provide sexually transmitted disease/HIV prevention and medical services to drug users.

Infect Immun, 1999 Sep, 67(9), 4974 - 6
Experimental transmission of Neisseria gonorrhoeae from pregnant rat to fetus; Nowicki S et al.; Sprague-Dawley rats were infected on day 20 of pregnancy by intraperitoneal inoculation with Neisseria gonorrhoeae . Disseminated gonococcal infection (DGI) and pelvic inflammatory disease (PID) strains in the presence of C1q but not in the presence of bovine serum albumin (BSA) were able to spread from the pregnant rat to the fetus and resulted in fetal mortality . Transmission of DGI and PID strains that are serum resistant (ser(r)) and sac-4 positive but not of a local infection strain that is ser(s) and sac-4 negative was facilitated by the C1q-dependent mechanism . This study provides the first experimental model that may mimic the transmission of gonococcal infection from mother to the fetus during pregnancy.

Infect Immun, 1999 Sep, 67(9), 4463 - 8
Sulfated polysaccharide-directed recruitment of mammalian host proteins: a novel strategy in microbial pathogenesis; Duensing TD et al.; Fundamental to the virulence of microbial pathogens is their capacity for adaptation and survival within variable, and often hostile, environments encountered in the host . We describe a novel, extragenomic mechanism of surface modulation which may amplify the adaptive and pathogenic potential of numerous bacterial species, including Staphylococcus, Yersinia, and pathogenic Neisseria species, as well as Helicobacter pylori and Streptococcus pyogenes . The mechanism involves specific bacterial recruitment of heparin, glycosaminoglycans, or related sulfated polysaccharides, which in turn serve as universal binding sites for a diverse array of mammalian heparin binding proteins, including adhesive glycoproteins (vitronectin and fibronectin), inflammatory (MCP-3, PF-4, and MIP-1alpha) and immunomodulatory (gamma interferon) intermediates, and fibroblast growth factor . This strategy impacts key aspects of microbial pathogenicity as exemplified by increased bacterial invasion of epithelial cells and inhibition of chemokine-induced chemotaxis . Our findings illustrate a previously unrecognized form of parasitism that complements classical virulence strategies encoded within the microbial genome.

Afr J Med Med Sci, 1997 Sep-Dec, 26(3-4), 185 - 6
Efficacy of pefloxacin in acute gonococcal urethritis; Bakare RA et al.; A clinical trial of pefloxacin as a single oral regimen in the treatment of acute uncomplicated gonococcal infection was evaluated in sixty-four male patients with clinical and laboratory diagnosis of gonococcal urethritis using a single dose of 800 mg pefloxacin . Of the 64 men, 59 (92.2%) were infected by (penicillinase producing Neisseria gonorrhoeae (PPNG) strains, while 5 (7.8%) had non-PPNG strains . The cumulative percentage cure for all infections was 84.4% . Pefloxacin is effective as a single dose therapy for acute gonococcal urethritis in males infected by penicillinase producing and non-penicillinase producing Neisseria gonorrhea in our area of study.

Postgrad Med, 1999 Aug, 106(2), 90 - 2, 97-8, 103-5
Addressing emerging infections . The partnership between public health and primary care physicians; Danila RN et al.; In response to concerns about emerging infections, the Minnesota Department of Health, in conjunction with the Centers for Disease Control and Prevention, developed a model emerging infections program in 1995 . The authors' experience with the program has demonstrated the key role clinicians have as partners with public health agencies in identifying and reporting disease cases, educating patients about infectious risks, and preventing emerging infections . This partnership is well illustrated by two examples from Minnesota: a recent outbreak of Neisseria meningitidis infection in a rural area and a laboratory surveillance study of invasive and drug-resistant Streptococcus pneumoniae infections in a metropolitan area.

J Clin Microbiol, 1999 Sep, 37(9), 3020 - 1
Comparison of commercial Amies transport systems with in-house Amies medium for recovery of Neisseria gonorrhoeae; Thompson DS et al.; Microbiologists are still encumbered by the variable performance of Amies charcoal transport medium in recovery of Neisseria gonorrhoeae . The objective of this study was to evaluate and select a good quality commercial system to replace our in-house preparation . We adsorbed 0.1 ml of a suspension from 30 gonococcal isolates onto each swab type and replaced the swab into the transport medium . We plated the swabs to New York City medium at 0, 24, 48, 72, and 96 h . We compared the survival of each isolate in the commercial Amies transport systems with that in our in-house Amies transport medium . The best recovery was observed with Copan transport systems . Some systems are inadequate and unacceptable for culture of gonococci.

Sex Transm Infect, 1999 Jun, 75(3), 181 - 5
Hydrogels containing monocaprin have potent microbicidal activities against sexually transmitted viruses and bacteria in vitro; Thormar H et al.; OBJECTIVE: To investigate the in vitro microbicidal and cytocidal potency of monocaprin dissolved in pharmaceutical hydrogel formulations and to evaluate their potential use as vaginal microbicides against sexually transmitted pathogens such as herpes simplex virus type 2 (HSV-2), human immunodeficiency virus type 1 (HIV-1), Chlamydia trachomatis, and Neisseria gonorrhoeae . METHODS: Gel formulations were mixed with equal volumes of virus/bacteria suspensions in culture medium and incubated for 1 and 5 minutes . The reduction in virus/bacteria titre was used as a measure of microbicidal activity . Similarly, gels were mixed with human semen to study their effect on leucocytes . The toxicity of the gels was tested in rabbits by the standard vaginal irritation test . RESULTS: Gels containing 20 mM of monocaprin caused a greater than 100,000-fold inactivation of HSV-2 and Neisseria in 1 minute and of Chlamydia in 5 minutes . Similarly, the gels caused a greater than 10,000-fold inactivation of HIV-1 in semen in 1 minute . They caused more than a 10,000-fold reduction in the number of viable leucocytes in semen in 1 minute . No toxic effect on the vaginal mucosa of rabbits was observed after daily exposure for 10 days . CONCLUSIONS: Hydrogels containing monocaprin are potent inactivators of sexually transmitted viruses and bacteria in vitro . This simple lipid seems to be a feasible choice as a mucosal microbicide for prevention of sexually transmitted infections . It is a natural compound found in certain foodstuffs such as milk and is therefore unlikely to cause harmful side effects in the concentrations used.

Sex Transm Infect, 1999 Apr, 75(2), 107 - 11
Susceptibility of gonococci isolated in London to therapeutic antibiotics: establishment of a London surveillance programme . London Gonococcal Working Group; Ison CA et al.; OBJECTIVES: To establish the in vitro susceptibility of gonococci isolated in the London area to antibiotics in current therapeutic use and to establish a sentinel surveillance system for monitoring trends in antibiotic resistant gonorrhoea in London . METHODS: Isolates of Neisseria gonorrhoeae from consecutive patients attending genitourinary medicine clinics at 10 hospitals in the London area were collected over a 3 month period, May to July 1997 . The susceptibility to penicillin, ciprofloxacin, tetracycline, and spectinomycin was determined for each isolate . Isolates exhibiting either plasmid or chromosomally mediated resistance were additionally tested for susceptibility to agents used as alternative treatments including azithromycin, ceftriazone, and ofloxacin . The resistant isolates were also tested for plasmid profiles (penicillinase producing N gonorrhoeae, PPNG), type of tetM determinant (tetracycline resistant N gonorrhoeae, TRNG), and presence of gyrA and parC mutations (quinolone resistant N gonorrhoeae, QRNG) . RESULTS: A total of 1133 isolates were collected which represents > 95% of the total gonococci isolated in the 3 months . Plasmid mediated resistance was exhibited by 48 (4.2%) isolates; six (0.5%) were PPNG, 15 (1.3%) were PP/TRNG, and 27 (2.4%) were TRNG . The majority of PPNG (18 of 20 tested) carried the 3.2 MDa penicillinase plasmid whereas the two types of tetM determinant were more evenly distributed . High level resistance to ciprofloxacin was detected in four (0.4%) isolates and double mutations were found in the quinolone resistance determining region (QRDR) of the gyrA gene in three QRNG with MICs of 16 mg/l and a single mutation in one isolate with a MIC of 1 mg/l to ciprofloxacin . No parC mutations were found . Of the remaining 1081 isolates, 86 (8.0%) were chromosomally mediated resistant N gonorrhoeae (CMRNG) . CONCLUSIONS: A unique collection of gonococcal isolates has been established which can be used as a baseline for surveillance of susceptibility to antibiotics and for epidemiological purposes.

Sex Transm Infect, 1999 Apr, 75(2), 103 - 6
Cervical dysplasia and HIV type 1 infection in African pregnant women: a cross sectional study, Kigali, Rwanda . The Pregnancy and HIV Study Group (EGE); Leroy V et al.; OBJECTIVE: To study the prevalence of cervical squamous intraepithelial lesions (SILs) and their association with HIV-1 infection and immunodeficiency among pregnant women in Kigali, Rwanda . METHODS: As part of a cohort study on the impact of HIV-1 infection on pregnancy outcome, HIV-1 seropositive (HIV+) and seronegative (HIV-) pregnant women were enrolled during the last trimester of pregnancy at the maternity ward of the Centre Hospitalier de Kigali from July 1992 to August 1993 . At inclusion, women were screened for sexually transmitted diseases (STDs)--syphilis, Neisseria gonorrhoeae, chlamydia trachomatis, Trichomonas vaginalis . CD4+ lymphocyte counts were measured and a Papanicolaou smear performed . RESULTS: Papanicolaou smear was interpretable in 103 HIV+ women and 107 HIV- women . Prevalence of SILs was significantly higher in HIV+ women than in HIV- women: 24.3% v 6.5% (odds ratio = 4.6; 95% CI: 1.8-12.3) . SIL+ women (n = 32) tended to have more STDs than SIL- women (n = 178), but this did not reach a statistical difference: 37.5% and 24.7% respectively (p = 0.13) . They also had a mean CD4 count significantly lower than SIL- women (623 and 784 CD4+ cells x 10(6)/l, respectively; p = 0.02) . CONCLUSION: SILs were HIV related and the association with immunosuppression was statistically significant . Prevalence of SILs was high in this population of pregnant women with high HIV/STDs prevalence . Screening policy for STDs and SILs in African women should be assessed in prenatal care.

Sex Transm Infect, 1999 Feb, 75(1), 60 - 6
Molecular epidemiology of tetM genes in Neisseria gonorrhoeae; Turner A et al.; OBJECTIVE: To examine the epidemiology of the tetM gene in Neisseria gonorrhoeae strains with high level resistance to tetracycline (TRNG) using a polymerase chain reaction (PCR) assay . METHODS: A single tube PCR was developed which distinguishes between the American and Dutch variants of the tetM gene . Between 1988 and 1995, 518 strains of TRNG (tetracycline MIC > 8.mg/l) were referred to the Gonococcus Reference Unit by other laboratories or isolated from routine swabs taken at local clinics . The strains were analysed for plasmid content, auxotype, serovar, and the tetM gene type . Travel details of the patients were determined by a questionnaire . RESULTS: A PCR product was obtained from all TRNG examined . 387 TRNG strains produced a 778 bp PCR product (American type tetM) and 131 produced a 443 by PCR product (Dutch type tetM) . Infections acquired in the United Kingdom contributed 57% of the TRNG strains included in this study; 82% of these carried the American type of tetM . The number of UK acquired TRNG received by the GRU increased each year except 1993--from four strains received in 1990 to 92 in 1995 . After the United Kingdom, Caribbean and African countries contributed most strains, with 56 and 60 TRNG acquired in each area respectively . All strains originating in Africa, except one from South Africa, contained the American type tetM . Infections caught in Nigeria and Kenya contributed most strains (15 and 14 respectively) . The TRNG originating from Caribbean countries comprised 36% Dutch tetM type . Infections caught in Jamaica accounted for 82% of the Caribbean strains . All 35 TRNG strains originating in the Far East contained the Dutch type tetM . 25 of the Far East strains were also penicillinase producing (PPNG) . Infections originating in Indonesia accounted for 49% of the Far East strains but these belonged to 12 different auxotype/serovar combinations . A geographical variation in the type of penicillinase coding plasmids found in PPNG/TRNG was also detected . CONCLUSIONS: These data suggest that the Dutch type tetM may have originated in the Far East and the American type in the African continent . Subsequent spread has resulted in a heterogeneous distribution of TRNG types in other parts of the world . At completion of the survey the numbers of TRNG imported each year from the major overseas sources had reached a plateau while UK contracted TRNG continued to rise providing evidence for the establishment of endemic TRNG strains in the United Kingdom.

J Infect Dis, 1999 Sep, 180(3), 762 - 73
T lymphocyte response to Neisseria gonorrhoeae porin in individuals with mucosal gonococcal infections; Simpson SD et al.; T lymphocytes from a majority of patients with urogenital gonococcal disease (67%-80%) proliferated on incubation with gonococcal porin (Por), compared with minimal induced proliferation of T lymphocytes from normal volunteers . A significant increase in Por-specific interleukin (IL)-4-producing CD4+ T helper lymphocytes was seen in patients with mucosal gonococcal disease and not in normal controls . Similar results were observed in CD8+ T lymphocytes from these patients . There was no measured increase in IL-2, IL-10, IL-12, interferon-gamma, or tumor necrosis factor-alpha production by T lymphocytes from infected subjects on incubation with Por . Concomitant increases in IL-4 production in T lymphocytes from infected subjects expressing the mucosal addresin VLAalpha4/beta7 on their surface were also observed on Por incubation, but the increases were similar in T lymphocytes that were VLAalpha4/beta7 negative . In conclusion, mucosal gonococcal disease can induce Por-specific circulating T lymphocytes with a Th2 phenotype, and a portion of these Por-specific T lymphocytes can potentially traffic to mucosal surfaces.

Arch Pediatr Adolesc Med, 1999 Aug, 153(8), 838 - 44
Sexually transmitted diseases in a health maintenance organization teen clinic: associations of race, partner's age, and marijuana use; Boyer CB et al.; OBJECTIVE: To determine the role of sociodemographic risk markers and behavioral risk factors associated with sexually transmitted diseases (STDs) in sexually experienced youth seeking care at an urban, general health maintenance organization teen clinic . DESIGN: Cross-sectional . PARTICIPANTS: A consecutive, racially and ethnically diverse sample of 285 sexually experienced youth who were preponderantly female (58.6%) and who were aged 16.7 years on average . METHODS: All participants completed a self-report questionnaire and were screened for Chlamydia trachomatis using the ligase chain reaction technique and for Neisseria gonorrhoeae and other STDs using conventional methods . RESULTS: Many of these youth were at high risk for STDs, having a self-reported history of sex with multiple partners (49.1%), sex with a new partner (42.5%), inconsistent use of condoms (71.9%), and frequent substance use (24.5% used marijuana 1-2 times per week or more) . Sexually transmitted disease screening revealed that 11.6% of the participants had 1 or more STDs . A logistic regression analysis to determine the best model for predicting STDs indicated that youth who are African American (odds ratio, 3.34; 95% confidence interval, 1.52-7.35), had sexual partners who were 2 or more years older (odds ratio, 2.63, 95% confidence interval, 1.22-5.67), and used marijuana 1 to 2 times or more per week (odds ratio, 2.27; 95% confidence interval, 1.01-5.13) were more likely to have STDs at screening . CONCLUSIONS: A brief sociodemographic and behavioral risk assessment that includes these factors may be useful for clinicians in deciding when to screen for STDs in sexually active youth seeking care for reasons not related to reproductive health.

Urol Int, 1999, 62(1), 64 - 8
Genotypic evolution in a quinolone-resistant Neisseria gonorrhoeae isolate from a patient with clinical failure of levofloxacin treatment; Tanaka M et al.; Recently, a reduction in the antimicrobial susceptibility of clinical isolates of Neisseria gonorrhoeae to newer fluoroquinolones including levofloxacin in vitro has been recognized in Japan . We examined the quinolone resistance mechanisms in N . gonorrhoeae isolates from a patient with clinical failure of levofloxacin treatment . Man with gonococcal urethritis was treated with oral 100 mg levofloxacin 3 times daily for 7 days . However, clinical failure of the treatment was observed . The minimum inhibitory concentration of levofloxacin for the posttreatment isolate (4.0 microg/ml) was 4-fold higher than that for the pretreatment isolate (1.0 microg/ml) . To analyze quinolone resistance mechanisms in the set of isolates, we performed DNA sequencing of the quinolone resistance-determining regions within the gyrA and parC genes . Moreover, we assayed the intracellular levofloxacin and norfloxacin accumulation level in these isolates . The pretreatment isolate contained three substitutions compared to susceptible wild-type isolate, including serine to phenylalanine at position 91 and aspartic acid to asparagine at position 95 in the GyrA protein, and serine to proline at position 88 in the ParC protein . The posttreatment isolate had four substitutions, including the same three substitutions and an additional glutamic acid to glutamine substitution at position 91 in ParC . There was no significant difference in the level of accumulation of levofloxacin and norfloxacin between the pretreatment and posttreatment isolates . Our results indicate that levofloxacin selects a mutant having an additional alteration within the gene cording for the ParC protein during treatment, which may have enhanced quinolone resistance in the organism.

FEMS Immunol Med Microbiol, 1999 Jul 15, 24(4), 447 - 53
Increasing antimicrobial resistance in STDs and the need for surveillance: Neisseria gonorrhoeae as a model; Bal C; Neisseria gonorrhoeae has a rising trend of resistance against antimicrobials . Today, third generation cephalosporins are the only antibiotics for treatment of gonorrhea against which there is no resistance in gonococci . On the other hand, decreased susceptibility against this group, including ceftriaxone, has already been observed . This historically famous pathogen deserves current attention and is reviewed here with respect to its resistance mechanisms and patterns, and the problems concerning standardization of its susceptibility testing are discussed.

FEMS Immunol Med Microbiol, 1999 Jul 15, 24(4), 437 - 46
Nucleic acid tests for the diagnosis of sexually transmitted diseases; Chernesky MA; Nucleic acid (NA) assays have been developed and commercialized for many sexually transmitted diseases (STDs) . Solid phase, liquid phase or in situ hybridization of nucleic acids without amplification procedures have been successfully used for diagnosing Chlamydia trachomatis, Neisseria gonorrhoeae and human papillomaviruses . Tests which use amplification procedures have provided better sensitivity and specificity than traditional tests . With special temperatures and enzymes, the new tests are designed to amplify either the target nucleic acid or the probe after annealing to the target . A third approach uses signal amplification . This article discusses the technology, specimen requirements and the current status of NA assay performance for diagnosing STDs and HIV by traditional and non-invasive clinical specimens.

Gene, 1999 Aug 5, 236(1), 179 - 84
Construction of mutant strains of Neisseria gonorrhoeae lacking new antibiotic resistance markers using a two gene cassette with positive and negative selection; Johnston DM et al.; The pathogenesis of infections caused by Neisseria gonorrhoeae, the causative agent of the sexually transmitted disease gonorrhea, can be studied using experimental infection of human male volunteers . The desire to avoid introducing new antibiotic resistance markers into strains to be used in human experimental infection has complicated the construction of genetically defined mutants in which expression of potential virulence factors is inactivated . To facilitate construction of such mutants, we have used a two-step mutagenesis strategy that allows for gene replacements without introducing new selectable markers into the final strain . The method uses a two-gene cassette containing both a selectable marker (ermC') and a counterselectable marker (rpsL) . The cassette is cloned into the gene of interest and used to replace the wild-type gene on the chromosome by allelic exchange . A second transformation replaces the cassette-containing version of the gene with an engineered version with an unmarked deletion or other mutation . The rpsL gene of Escherichia coli functioned for the counterselection in the gonococcus, albeit with low efficiency . To improve the efficiency of the counterselection, we cloned the gonococcal rpsL gene and incorporated it into the cassette . This technique has been successful in creating defined mutants for human challenge, and also circumvents the limitation in the number of different selectable markers that are useful in Neisseria species.

Am J Reprod Immunol, 1999 Jul, 42(1), 58 - 63
Mucosal immunity in the genital tract: prospects for vaccines against sexually transmitted diseases--a review; Russell MW et al.; PROBLEM: Consistent with the absence of protective immunity resulting from previous infection with Neisseria gonorrhoeae, the genital mucosal immune response in human gonorrhea is weak: only low levels of immunoglobulin G (IgG) and immunoglobulin A (IgA) antibodies are detectable against gonococci, and inflammatory cytokine responses are poor . METHOD OF STUDY: Mucosal immunization strategies designed to induce persisting genital antibody responses might afford protection against infection, if appropriate conserved antigens can also be identified . RESULTS: Intragastric or intranasal immunization with bacterial antigens expressed as recombinant chimeric proteins with cholera toxin A2/B subunits induced persisting IgA antibodies in genital and other secretions, and circulating IgG antibodies . CONCLUSION: Although gonococci may avoid inducing or even suppress immune responses during natural infection, alternative approaches to vaccine development may be successful . However, inadequate understanding of the origins of antibodies in the genital tract, and their effector mechanisms, will need to be rectified to make this possible.

J Exp Med, 1999 Aug 2, 190(3), 331 - 40
Homologue scanning mutagenesis reveals CD66 receptor residues required for neisserial Opa protein binding; Bos MP et al.; The immunoglobulin-like family of CD66 antigens, present on human neutrophils and epithelial cells, are used as receptors for adhesins expressed by the pathogenic Neisseriae . N . gonorrhoeae strain MS11 can express 11 isoforms of these adhesins, called opacity-related (Opa) proteins . Each MS11 Opa protein recognizes a distinct spectrum of CD66 receptors . CD66-Opa binding is mediated by the NH(2)-terminal domain of the receptor and occurs through protein-protein interactions . In this report, we have investigated the molecular basis for the binding between the CD66 and Opa protein families by mapping amino acids in CD66 receptors that determine Opa protein binding . We performed homologue scanning mutagenesis between CD66e, which binds multiple Opa variants, and CD66b, which binds none, and tested both loss-of-function by CD66e and gain-of-function by CD66b in solution assays and in assays involving full-length receptors expressed by epithelial cells . We found that three residues in the CD66e N-domain are required for maximal Opa protein receptor activity . Opa proteins that recognize the same spectrum of native CD66 molecules showed differential binding of receptors with submaximal activity, indicating that the binding characteristics of these Opa proteins are actually slightly different . These data provide a first step toward resolving the structural requirements for Opa-CD66 interaction.

Presse Med, 1999 Jun 26, 28(23), 1227 - 30
{Bacterial concentration in the cerebrospinal fluid in childhood meningitis}; Mariani-Kurkdjian P et al.; OBJECTIVES: The aim of this study was to analyze the epidemiological evolution of causal germs in meningitis in children aged 1 day to 15 years and determine the relationship between pretreatment concentrations of bacteria in cerebral spinal fluid (CSF), patient age, bacterial species and bacteriological eradication . PATIENTS AND METHODS: A quantitative analysis of germs was performed in 212 children with bacterial meningitis (mean age 19.8 months) . RESULTS: Bacterial counts ranged from 2.10(1) to 4.10(9) CFU/ml in CSF . Among the 212 patients, 52 (24.5%) had counts 10(7)/ml . Infants had significantly higher counts than the other age groups . Mean counts for Hoemophilus influenzoe serotype B were not different from those for Streptococcus pneumoniae but were significantly higher than for Neisseria meningitidis . Compared with initial germ counts, 98.5% of the CSF specimens were sterile at 24 and 48 hours and 100% at 72 hours . CONCLUSION: Germ counts were higher in infants.

Mol Microbiol, 1999 Aug, 33(3), 651 - 8
Induction of the mtrCDE-encoded efflux pump system of Neisseria gonorrhoeae requires MtrA, an AraC-like protein; Rouquette C et al.; The mtr (multiple transferable resistance) gene complex in Neisseria gonorrhoeae encodes an energy-dependent efflux pump composed of the MtrC-MtrD-MtrE cell envelope proteins that serves to export structurally diverse antimicrobial, hydrophobic agents (HAs) . Many of these agents have membrane-acting detergent activity . Using Triton X-100 (TX-100) as a representative HA, we found that the mtrCDE efflux pump operon could be induced to higher levels of expression when an HA-sensitive strain was exposed to sublethal concentrations of this non-ionic detergent and the structurally related spermicide, nonoxynol-9 . This induction was at the level of mtrCDE gene transcription and was independent of the MtrR repressor, which normally decreases mtrCDE gene expression . However, the enhanced resistance of gonococci to TX-100 was dependent on the expression of a previously undescribed gonococcal protein that belonged to the AraC/XylS family of transcriptional activators . We have termed this protein MtrA to signify its likely role in the activation of mtrCDE gene expression . Taken together with previous studies dealing with the genetic control of mtrCDE gene expression, we propose that gonococci can modulate their resistance to HAs through both positive and negative transcriptional control processes . The action of these regulatory processes is probably of importance in determining the survival capacity of gonococci at mucosal surfaces that contain detergent-like HAs.

Mol Microbiol, 1999 Aug, 33(3), 635 - 50
The opcA and (psi)opcB regions in Neisseria: genes, pseudogenes, deletions, insertion elements and DNA islands; Zhu P et al.; Previous data have indicated that the opc gene encoding an immunogenic invasin is specific to Neisseria meningitidis (Nm) and is lacking in Neisseria gonorrhoeae (Ng) . The data presented here show that Nm and Ng both contain two paralogous opc-like genes, opcA, corresponding to the former opc gene, and (psi)opcB, a pseudogene . The predicted OpcA and OpcB proteins possess transmembrane regions with conserved non-polar faces but differ extensively in four of the five surface-exposed loops . Gonococcal OpcA was expressed weakly under in vitro conditions, and it is unknown whether these bacteria can express this protein at high levels . Analysis of the sequences flanking opcA and (psi)opcB revealed a framework of conserved housekeeping genes interspersed with DNA islands . These regions also contained several pseudogenes, deletions and IS elements, attesting to considerable genome plasticity . Both opcA and (psi)opcB are located on DNA islands that have probably been imported from unrelated bacteria . A third island encodes the dcmD/dcrD R/M genes in Ng versus a small open reading frame in most strains of Nm . Rare strains of Nm were identified in which the R/M island has been imported . DNA islands in Nm and Ng seem to have been acquired by recombination via conserved flanking housekeeping genes rather than by insertion of mobile genetic elements.

Infect Immun, 1999 Aug, 67(8), 3937 - 46
Limited local and systemic antibody responses to Neisseria gonorrhoeae during uncomplicated genital infections; Hedges SR et al.; Repeated infections with Neisseria gonorrhoeae are common among patients attending sexually transmitted disease clinics . We examined whether previous infections or site of infection altered the local and systemic antigonococcal antibody levels in males and females . Antibodies against N . gonorrhoeae MS11 and the patients' homologous infecting isolates were measured by enzyme-linked immunosorbent assay . In general, the local and systemic immune responses to gonococci were extremely modest . There was a slight increase in serum immunoglobulin G (IgG) against the MS11 strain and the homologous isolates in infected males . Levels of serum IgA1 antibodies against MS11 were slightly higher in infected than in uninfected females . A history of previous infections with N . gonorrhoeae did not alter the antibody levels in patients with a current infection, suggesting that immunological memory is not induced by uncomplicated gonococcal infections . Antibody responses to infected subjects' homologous isolates were observed in cervical mucus; IgA1 levels increased while IgG levels decreased . The decline in mucosal IgG against the homologous isolates was less common in subjects having both rectal and cervical infections; otherwise, no effect of rectal involvement was observed . The absence of substantially higher antibody levels to gonococci where there is infection at a site known to contain organized lymphoid tissue suggests that the low levels of responses to uncomplicated infections may not be due simply to an absence of inductive sites in the genital tract . We propose that in addition to its potential ability to avoid the effects of an immune response, N . gonorrhoeae does not elicit strong humoral immune responses during uncomplicated genital infections.

Sex Transm Dis, 1999 Jul, 26(6), 358 - 63
Association between auxotypes, serogroups, and antibiotic susceptibilities of Neisseria gonorrhoeae isolated from women in Mumbai (formerly Bombay), India; Divekar AA et al.; OBJECTIVES: Gonococcal isolates were differentiated based on susceptibility pattern, penicillinase production (PPNG or non-PPNG), serogroup, auxotype, protein, and plasmid profile . The association between serogroup and auxotype and PPNG was determined . STUDY DESIGN: Women attending tertiary level health centers and the sexually transmitted disease (STD) clinic in Mumbai, India, were screened for Neisseria gonorrhoeae . Minimal inhibitory concentration testing was performed according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines . Auxotypes, serogroups, protein profile, and plasmid content were also studied . RESULTS: Of the 33 isolates, 16 (48.5%) were resistant to penicillin, and 28 (84.8%) showed a chromosomally mediated resistance to tetracycline . Five (15.2%) isolates showed resistance to ciprofloxacin, whereas 12 (36.4%) showed a reduced susceptibility . Twenty-seven (81.8%) isolates belonged to the WI serogroup, and 15 (46.7%) were penicillinase producers (PPNG) . Seventeen (51.5%) isolates were of the nonrequiring auxotype, whereas seven (21.2%) were proline requiring . Fifteen (55.6%) of the isolates belonged to the nonrequiring-WI auxotype/serogroup (A/S) class . Ten of the PPNG isolates possessed the 4.4 MDa plasmid, whereas four had the 3.2 MDa plasmid . Increases in the molecular weight of the major outer membrane protein were observed . CONCLUSION: A high prevalence of chromosomal resistance to penicillin and tetracycline was observed . The 4.4 MDa plasmid was the most prevalent among the PPNG isolates . We observed ciprofloxacin resistance, which has not been reported in previous studies in India . The nonrequiring auxotype was the most prevalent, followed by the proline requiring auxotype . WI serogroup was the most commonly observed among the isolates studied . The nonrequiring/WI A/S class was the most prevalent among the PPNGPIP: This study aims to determine the prevalence of penicillinase-producing Neisseria gonorrhea (PPNG), as well as to differentiate gonococcal isolates based on susceptibility pattern, penicillinase production, serogroup, auxotype, protein and plasmid profile among women attending tertiary level health centers and sexually transmitted disease (STD) clinics in Mumbai (formerly Bombay), India . An association between auxotypes, serogroups and antibiotic susceptibilities of Neisseria gonorrhea was determined . Subjects were screened for Neisseria gonorrhea; the minimal inhibitory concentration testing was performed according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines . The antibiotics tested were penicillin, tetracycline, ciprofloxacin, and spectinomycin . The ability of various typing methods to distinguish different isolates was calculated using the discrimination index . Results showed that in the susceptibility testing group 16 (48.5%) were resistant to penicillin and 28 (84.8%) showed chromosomally mediated resistance to tetracycline . 5 isolates (15.2%) showed resistance to ciprofloxacin, while 12 (36.4%) showed reduced susceptibility . 27 (81.8%) isolates belonged to the WI serogroup, and 15 (46.7%) were penicillinase producers . In auxotyping, 17 (51.5%) isolates were of the nonrequiring auxotype, while 7 (21.2%) were proline requiring . 15 (55.6%) of the isolates belonged to the nonrequiring-WI auxotype/serogroup (A/S) class . In plasmid pattern, 10 of the PPNG isolates possessed the 4.4 MDa plasmid, while 4 had the 3.2 MDa plasmid . Increases in the molecular weight of the major outer membrane protein were observed . In conclusion, the combined use of auxotyping and serogrouping offers a good method for discriminating gonococcal isolates . A high prevalence of chromosomal resistance to penicillin and tetracycline was observed . The 4.4 MDa plasmid was the most prevalent among the PPNG isolates . The nonrequiring auxotype was the most prevalent, followed by the proline requiring auxotype . WI serogroup was commonly observed among the isolates, while the nonrequiring/WI A/S class was the most prevalent among PPNG isolates .

Immunopharmacology, 1999 May, 42(1-3), 23 - 30
Regulation of complement activation by C-reactive protein; Mold C et al.; C-reactive protein (CRP) is an acute-phase serum protein and a mediator of innate immunity . CRP binds to microbial polysaccharides and to ligands exposed on damaged cells . Binding of CRP to these substrates activates the classical complement pathway leading to their uptake by phagocytic cells . Complement activation by CRP is restricted to C1, C4, C2 and C3 with little consumption of C5-9 . Surface bound CRP reduces deposition of and generation of C5b-9 by the alternative pathway and deposition of C3b and lysis by the lectin pathway . These activities of CRP are the result of recruitment of factor H resulting in regulation of C3b on bacteria or erythrocytes . Evidence is presented for direct binding of H to CRP . H binding to CRP or C3b immobilized on microtiter wells was demonstrated by ELISA . Attachment of CRP to a surface was required for H binding . H binding to CRP was not inhibited by EDTA or phosphocholine, which inhibit ligand binding, but was inhibited by a 13 amino acid CRP peptide . The peptide sequence was identical to the region of CRP that showed the best alignment to H binding peptides from Streptococcus pyogenes (M6) and Neisseria gonorrhoeae (Por1A) . The results suggest that CRP bound to a surface provides secondary binding sites for H resulting in greater regulation of alternative pathway amplification and C5 convertases . Complement activation by CRP may help limit the inflammatory response by providing opsonization with minimal generation of C5a and C5b-9.

Mol Immunol, 1999 Mar-Apr, 36(4-5), 241 - 8
Factor H and disease: a complement regulator affects vital body functions; Zipfel PF et al.; Factor H is a multidomain and multifunctional protein . As a complement regulator factor H determines the fate of newly formed C3b and controls formation and stability of C3 convertases both in the fluid phase and on cell surfaces . In addition, this plasma protein displays functions outside complement control as it has been suggested to act as an adhesion protein, to be a ligand for the cellular integrin receptor CR3 (CD11b/CD18) and to display chemotactic activity . Genetic and pathophysiological analyses describe a role for factor H in vital body functions . Depletion or the absence of factor H due to genetic reasons leads to unrestricted C3 consumption . A reduced amount of factor H in plasma or mutations within the factor H gene may lead to glomerulonephritis (type II MPGN) or hemolytic uremic syndrome (HUS) . Certain pathogenic organisms have been shown to evade complement attack by binding factor H from the host . Such specific factor H binding components have been demonstrated on the surface of microbes, e.g., Streptococcus pyogenes and Neisseria gonorrhoeae . Here, we summarize the current knowledge how abnormalities in function of the central complement regulator factor H are associated with human diseases.

Intensive Care Med, 1999 May, 25(5), 475 - 80
Trial of dexamethasone treatment for severe bacterial meningitis in adults . Adult Meningitis Steroid Group; Thomas R et al.; OBJECTIVE: To evaluate the clinical benefit of early adjunctive dexamethasone therapy for severe bacterial meningitis in adults . DESIGN: Multicenter, double-blind, randomized trial initiated in emergency or intensive care units in France and Switzerland . Within 3 h after initiation of an aminopenicillin therapy, patients received dexamethasone (10 mg q.i.d.) or placebo for 3 days . The primary end-point was the rate of patients cured without any neurologic sequelae on day 30 . RESULTS: Sixty patients were enrolled, predominantly with a severe form since 85% required ICU stay and 43% mechanical ventilation . Streptococcus pneumoniae accounted for 31 cases and Neisseria meningitidis for 18 cases . The study had to be stopped prematurely because of a new national recommendation of experts to use third generation cephalosporin and vancomycin as a result of the increasing rate of penicillin-resistant S . pneumoniae in France . After the third sequential analysis by the triangular statistical test, the difference of rate of cured patients without any neurologic sequelae was not statistically significant (p = 0.0711) between the dexamethasone group (74.2%; n = 31) and the placebo group (51.7%; n = 29) . Furthermore, the former group was younger and less sick at inclusion . CONCLUSION: Bacterial meningitis is still a severe disease in adults, since the overall observed rate of death or severe neurologic sequelae was 26.7% . The reported data are inconclusive regarding a systematic use of dexamethasone as an adjunctive therapy for bacterial meningitis in adults . Moreover this treatment impairs antibiotic penetration into the cerebrospinal fluid (CSF) that can lead to therapeutic failure, particularly in areas with high or increasing rates of penicillin-resistant S . pneumoniae.

Pediatr Emerg Care, 1999 Jun, 15(3), 187 - 8
Purpura fulminans associated with Streptococcus pneumoniae infection in a child; Cnota JF et al.; BACKGROUND: Neisseria meningitidis is the most frequent isolate associated with purpura fulminans in children . Although Streptococcus pneumoniae infection has been associated with purpura fulminans, with the exception of one adult, it has only been reported in immunocompromised hosts . PURPOSE: We report an apparently previously healthy child who presented with purpura fulminans associated with pneumococcal meningitis . METHODS: Case report and review of the medical literature from September 1966 to June 1997, using a MEDLINE search . CONCLUSION: While systemic pneumococcal infection is common in childhood, progression to purpura fulminans does not typically occur in overtly healthy children . Our patient illustrates that invasive pneumococcal infection should be considered and empirically treated in a child who presents with purpura fulminans, even in the absence of preexisting functional or anatomic asplenia.

Mol Microbiol, 1999 Jun, 32(6), 1124 - 32
Interactions of pathogenic Neisseria with host cells . Is it possible to assemble the puzzle?
Nassif X, Pujol C, Morand P, Eugene E.
Neisseria meningitidis and Neisseria gonorrhoeae are human pathogens that have to interact with mucosa and/or cellular barriers for their life cycles to progress . Even though they both give rise to dramatically different diseases, the use of in vitro models has shown that most of the mechanisms mediating cellular interactions are common to N . meningitidis and N . gonorrhoeae . This suggests that bacterial cell interactions may be essential not only for pathogenesis but also for other aspects of the bacterial life cycle that are common to both N . meningitidis and N . gonorrhoeae . This manuscript will review the most recent developments concerning the mechanisms mediating cellular interaction of pathogenic Neisseria and will then try to put them into the perspective of pathogenesis and bacterial life cycle.

Mol Microbiol, 1999 Jun, 32(6), 1117 - 23
Iron acquisition systems in the pathogenic Neisseria; Schryvers AB et al.; Pathogenic neisseriae have a repertoire of high-affinity iron uptake systems to facilitate acquisition of this essential element in the human host . They possess surface receptor proteins that directly bind the extracellular host iron-binding proteins transferrin and lactoferrin . Alternatively, they have siderophore receptors capable of scavenging iron when exogenous siderophores are present . Released intracellular haem iron present in the form of haemoglobin, haemoglobin-haptoglobin or free haem can be used directly as a source of iron for growth through direct binding by specific surface receptors . Although these receptors may vary in complexity and composition, the key protein involved in the transport of iron (as iron, haem or iron-siderophore) across the outer membrane is a TonB-dependent receptor with an overall structure presumably similar to that determined recently for Escherichia coli FhuA or FepA . The receptors are potentially ideal vaccine targets in view of their critical role in survival in the host . Preliminary pilot studies indicate that transferrin receptor-based vaccines may be protective in humans.

Hybridoma, 1999 Apr, 18(2), 121 - 9
Human immune response to a peptide mimic of Neisseria meningitidis serogroup C in hu-PBMC-SCID mice; Hutchins WA et al.; An anti-idiotype-based peptide mimic vaccine for Neisseria meningitidis serogroup C polysaccharide (MCPS) has been developed and shown to induce a response in mice that is specific, functional, and T-dependent . In this study, the immunogenicity of the MCPS peptide mimic vaccine preparation, as a potential vaccine for use in humans, is shown using the hu-PBMC-SCID mouse model . The human antibody response to the MCPS peptide mimic vaccine is specific and functional as shown by inhibition enzyme-linked immunoadsorbent assay (ELISA) and bactericidal assay . These data support the usefulness of the peptide mimic vaccine strategy for humans.

Infect Immun, 1999 Jul, 67(7), 3469 - 80
Neisseria gonorrhoeae coordinately uses Pili and Opa to activate HEC-1-B cell microvilli, which causes engulfment of the gonococci; Griffiss JM et al.; This study was undertaken to examine concomitant roles of pili and colony opacity-associated proteins (Opa) in promoting Neisseria gonorrhoeae adherence to and invasion of human endometrial HEC-1-B cells . Adherence of N . gonorrhoeae to cultured HEC-1-B cells was saturable, even though organisms adhered to <50% of the cells . During 4 to 6 h of incubation, adherent mono- and diplococci formed microcolonies on the surfaces of the cells . Microvilli of the HEC-1-B cells adhered by their distal ends to individual cocci within the microcolonies . When the microcolonies grew from isogenic pilus-negative (P-) Opa-, P- Opa+, or P+ Opa- gonococci, microvilli did not elongate, and the colonies were not engulfed . In contrast, the microvilli markedly elongated during exposure to P+ Opa+ gonococci . The microvilli adhered to the organisms along their full lengths and appeared to actively participate in the engulfment of the microcolonies . Internalized microcolonies, with P+ Opa+ gonococci, contained dividing cocci and appeared to be surrounded by cell membrane but were not clearly within vacuoles . In contrast, degenerate individual organisms were within vacuoles . Low doses of chloramphenicol, which inhibits protein synthesis by both prokaryotes and eukaryotes, prevented the microvillar response to and internalization of the P+ Opa+ gonococci; higher doses caused internalization without microvillus activation . Cycloheximide and anisomycin, which inhibit only eukaryotic protein synthesis, caused dose-dependent enhancement of uptake . Cytochalasins reduced engulfment; colchicine had no effect . These results show that gonococci must express both pili and Opa to be engulfed efficiently by HEC-1-B cells.

Med J Aust, 1999 May 17, 170(10), 475 - 8
Bacteraemia in febrile children presenting to a pae3iatric emergency department; Haddon RA et al.; OBJECTIVE: To determine the prevalence of bacteraemia in young febrile children presenting to a paediatric emergency department . DESIGN: Prospective observational case study . SETTING: Emergency Department of the Royal Children's Hospital, Melbourne, between May 1996 and May 1997 . PARTICIPANTS: Patients aged 3-36 months presenting to the Emergency Department with temperature > or = 39 degrees C and without specific viral illnesses (varicella, croup or herpes gingivostomatitis) . OUTCOME MEASURES: Bacteraemia (defined as presence of pathogenic bacteria in a blood culture), white blood cell count (WCC), McCarthy score, and final diagnosis based on clinical features and investigations . RESULTS: Bacteraemia was identified in 18 of 534 patients (3.4%) . Pathogens isolated were Streptococcus pneumoniae (15), Neisseria meningitidis (2) and Klebsiella pneumoniae (1) . Increased WCC counts (P < 0.001) and brief duration of fever (P < 0.001) were associated with bacteraemia . Nevertheless, clinical features, including McCarthy scores, and high WCC counts (> or = 20 x 10(9)/L) had < 10% predictive accuracy for bacteraemia . Overall, final diagnoses in the 534 febrile children included non-specific viral infections (25%), upper respiratory tract infections (24%), otitis media (10%), gastroenteritis (9%), pneumonia (7%), and urinary tract infection (5%) . CONCLUSIONS: Most urban Australian children aged 3-36 months presenting to a paediatric emergency department with temperature > or = 39 degrees C without a clinical focus have a viral infection . However, 3%-4% have occult bacteraemia . Neither clinical features nor high WCC counts reliably identify these patients . As empiric antibiotics may contribute to increasing antibiotic resistance and have not been shown to prevent the rare complication of meningitis, we believe that close contact and regular review of these patients is preferable to empiric antibiotic therapy.

Gene, 1999 Jun 11, 233(1-2), 233 - 40
Identification, expression and chromosome localization of a human gene encoding a novel protein with similarity to the pilB family of transcriptional factors (pilin) and to bacterial peptide methionine sulfoxide reductases; Huang W et al.; Here we report the isolation, characterization and chromosome localization of a subtracted cDNA (CBS-1) isolated from the human ocular ciliary body which encodes a novel protein . As is deduced from the nucleotide sequence of the cDNA, CBS-1 contains an open reading frame consisting of 182 amino acids, with a molecular weight of 19.5kDa . CBS-1 shares significant nucleotide and amino acid sequence identities (residues 51 to 182) with a hypothetical 15.5kDa protein in the ANSA-GAP intergenic region (yeaA) of Escherichia coli, and the carboxyl terminal region of pilB, a transcription factor involved in the regulation of expression of pili, from Neisseria gonorrhoeae . Interestingly, CBS-1 also shares significant identity with the carboxyl terminus of the peptide-methionine sulfoxide reductase (MsrA), a repair enzyme, from Helicobacter pylori and Streptococcus pneumoniae . However, the amino terminal of CBS-1 (residues 23 to 43), which lacks homology to the amino terminal region of gonococcal pilB or pneumococcal MsrA, exhibits significant identity in a stretch of 20 amino acids, with glycine-rich proteins . By Northern blot, CBS-1, hybridized to a 0.6 to 0.7kb transcript in size, is expressed ubiquitously in many tissues, but most abundantly in the retina and ocular ciliary body, skeletal muscle and heart . An epitope-directed antibody to an amino acid sequence at the carboxyl terminus of CBS-1 recognized a main protein of 19.5kDa in ocular ciliary body extracts, and a 23kDa protein in total extracts from E . coli MC1061 cells, which expresses high levels of MsrA . The CBS-1 gene was mapped to human chromosome 10p12 between markers WI-8535 and WI-4724, and is tightly linked to the two STRP markers of D10S1789 and D10S550 . We suggest that the CBS-1 gene encodes a mammalian transcription factor related to the bacterial pilB and certain bacterial MsrA homologues.

Gene, 1999 Jun 11, 233(1-2), 49 - 57
Mutational analysis of the promoter region of the porA gene of Neisseria meningitidis; Sawaya R et al.; The porA gene encodes the class 1 outer membrane protein (OMP1) in Neisseria meningitidis and is under transcriptional control . Promoter regions of porA from different clinical isolates were sequenced and were found to differ in the number of guanosine residues in a poly(G) track located upstream of the -10 region . Isolates that did not express OMP1 had up to nine G residues in the poly(G) track or an adenosine residue within this poly(G) track . Using beta-galactosidase as a reporter gene, the transcriptional activities of the promoter regions of the porA gene from three strains, two of which do not express OMP1, were assayed in both Escherichia coli and N . meningitidis . Mutations in the poly(G) track were created by site-directed mutagenesis and promoter fusions were further analyzed in E . coli and N . meningitidis . The number of nucleotides in the poly(G) track influenced promoter activity: reduction of a poly(G) track of 12nt by one and by two guanosine residues reduced promoter activity . Within the poly(G) track, replacement of an adenosine residue by a guanosine residue increased the promoter activity; replacement of a guanosine residue by an adenosine residue decreased the activity . The similar transcriptional activities for the mutated promoters in E . coli and N . meningitidis are compatible with similar control mechanisms for transcriptional control in both organisms.

Int J Oncol, 1999 Jul, 15(1), 59 - 66
A novel hydrophobized GM3 ganglioside/Neisseria meningitidis outer-membrane-protein complex vaccine induces tumor protection in B16 murine melanoma; Alonso DF et al.; Gangliosides are sialic acid-containing glycosphingolipids that have increased surface membrane expression on cancers of neuroectodermal origin . The present study was designed to investigate at a preclinical level the therapeutic usefulness of a consistently immunogenic and safe conjugate vaccine in melanoma . We have examined a novel vaccine of GM3 monosialoganglioside hydrophobically conjugated with the outer-membrane-protein complex from Neisseria meningitidis plus Montanide ISA 51 in the B16 melanoma mouse model . B16 cell line is characterized by the predominant presence of ganglioside GM3 on the cell surface . Vaccines were administered i.m . in the quadriceps at 14-day intervals and B16 cells were injected in the subcutis of the right flank of C57BL/6 mice, 7 days after the fourth dose . Significant suppression of tumor growth and prolongation of survival were seen by immunization with GM3 vaccine in animals challenged with 5x10(3) or 10(3) live melanoma cells . In addition, vaccination reduced tumor growth in animals challenged with 5x10(4) cells . The reactivity of serum IgG from vaccinated mice was examined by a sensitive immunoperoxidase assay on B16 tumor specimens . Most melanoma cells displayed a distinct positive staining associated with both cell membrane and cytoplasm . In accordance with the immunohistochemical stainings, the antisera of immunized mice reacted brightly against B16 melanoma cells in flow cytometry studies . Anti-sera also mediated complement-mediated cytotoxicity and specific response could be totally ascribed to antibodies of the IgG2b subclass . The present data suggest that GM3 vaccine may provide a useful immunotherapeutic strategy for melanoma.

Infect Dis Obstet Gynecol, 1999, 7(3), 145 - 52
Infertility following pelvic inflammatory disease; Pavletic AJ et al.; OBJECTIVE: To assess the frequency of infertility after pelvic inflammatory disease (PID) and factors important in postinfectious tubal damage in an urban population at high risk for sexually transmitted diseases . METHODS: From a cohort of 213 women with PID documented by laparoscopy and/or endometrial biopsy, 58 women (27% of the initial cohort) were interviewed by phone 2 to 9 years after an index episode of PID . Data regarding the initial history, physical examination, microbiology, laparoscopic, and serologic findings, and data concerning interval contraception, subsequent pregnancy, subsequent infection, and chronic pelvic pain were compared among those with and without infertility at follow up . RESULTS: Nineteen (40%) of the 48 women not using contraception were involuntarily infertile after the index episode of PID . Compared with those who had an interval pregnancy, infertile women were older (P = 0.02), more likely to have a history of infertility prior to the index episode of PID (P = 0.001), and were more likely to have occluded or partially occluded fallopian tubes (P = 0.03), peritubal adhesions (P = 0.007), or perihepatic adhesions (P = 0.02) seen by laparoscopy performed during the index episode . Surprisingly, recovery of Chlamydia trachomatis was negatively related to infertility (P = 0.001), although a similar proportion of both groups had chlamydia immunoglobulin M antibody (40% vs . 31%) . Chlamydia heat shock protein was weakly related to infertility (P = 0.08) . The isolation of Neisseria gonorrhoeae was not significantly different between groups (53% vs . 57%) . CONCLUSIONS: The high rate of postinfection infertility found was probably related to a combination of tubal damage before and during the index episode of PID . Prevention of recurrent PID and better understanding of the pathophysiology of postinfection tubal damage (which may differ between chlamydia and gonorrhea) is needed to develop more effective strategies to reduce permanent tubal damage.

Infect Dis Obstet Gynecol, 1999, 7(3), 138 - 44
Evaluation of ofloxacin in the treatment of laparoscopically documented acute pelvic inflammatory disease (salpingitis); Peipert JF et al.; OBJECTIVE: To evaluate the safety and efficacy of intravenous and oral ofloxacin monotherapy in the treatment of laparoscopically documented acute pelvic inflammatory disease (PID) . METHODS: This study was conducted as an open-label, phase-III, uncontrolled, multicenter study . Patients identified with laparoscopic findings of salpingitis were treated with 400 mg of intravenous ofloxacin every 12 hours followed by 400 mg of oral ofloxacin every 12 hours for 10 to 14 days . Patients were evaluated five times for clinical and microbial efficacy . Since laparoscopy was performed only at admission, pathogens identified laparoscopically were presumed eradicated if they were present on the laparoscopic culture and the patient was clinically cured or improved at final evaluation . RESULTS: Of the 70 patients evaluable for safety (intent-to-treat population), the mean age was 25.6 years . Sixty-one of 70 patients (87%) were cured, one improved, one did not improve, and seven were unevaluable because they discontinued study participation . Fifty-one were evaluable for clinical efficacy: 50 (98%) were cured and one did not improve . Sixteen were evaluable for expanded microbiological efficacy: three had documented Neisseria gonorrhoeae; 12, Chlamydia trachomatis; and one, a mixed infection of both organisms . All cervical, laparoscopic, and endometrial cultured pathogens, including N . gonorrhoeae and C . trachomatis, were eradicated or presumed eradicated at the posttherapy visit . No serious or unexpected adverse events occurred . CONCLUSIONS: Ofloxacin monotherapy was effective and well tolerated in the treatment of laparoscopically proven PID in a geographically diverse population . Future studies are necessary to evaluate long-term outcomes and sequelae of PID treatment with single agent therapy.

Mol Biol Evol, 1999 Jun, 16(6), 773 - 83
Networks and groups within the genus Neisseria: analysis of argF, recA, rho, and 16S rRNA sequences from human Neisseria species; Smith NH et al.; To understand the pattern of nucleotide sequence variation among bacteria that frequently exchange chromosomal genes, we analyzed sequences of the recA, argF, and rho genes, as well as part of the small-subunit (16S) rRNA gene, from about 50 isolates of human commensal Neisseria species and the pathogenic N . meningitidis and N . gonorrhoeae . Almost all isolates of these species could be assigned to five phylogenetic groups that are found for all genes examined and generally are supported by high bootstrap values . In contrast, the phylogenetic relationships among groups varied according to the gene analyzed with notable incongruences involving N . cinerea and N . lactamica . Further analysis using split decomposition showed that for each gene, including 16S rRNA, the patterns of sequence divergence within N . meningitidis and closely related species were inconsistent with a bifurcating treelike phylogeny and better represented by an interconnected network . These data indicate that the human commensal Neisseria species can be separated into discrete groups of related species but that the relationships both within and among these groups, including those reconstructed using 16S rRNA, have been distorted by interspecies recombination events.

J Clin Microbiol, 1999 Jul, 37(7), 2183 - 8
Evaluation of the discriminatory power of typing methods for Neisseria gonorrhoeae; Van Looveren M et al.; A panel of 18 strains of Neisseria gonorrhoeae, known to be temporally and geographically diverse, was used to evaluate a number of typing systems, including conventional auxotyping and serotyping and the molecular methods of arbitrarily primed PCR (AP-PCR), amplified ribosomal-DNA restriction analysis (ARDRA), opa typing, and pulsed-field gel electrophoresis (PFGE) . The discriminatory power of the different typing methods were determined with a collection of 87 clinical isolates from commercial sex workers in Indonesia, and Simpson's index of diversity was calculated . Of the two traditional techniques, auxotyping and serotyping, the latter gives the highest discriminatory index (DI) (DI, 0.846) . The combination of auxotyping and serotyping yields a high DI (DI, 0 . 928) . D11344- and D8635-primed PCR showed low DIs of 0.608 and 0.622, respectively, but a combination of the two primers had a DI of 0 . 849 . The combination of serotyping with D11344-primed or D8635-primed PCR resulted in DIs of 0.936 and 0.937, respectively . ARDRA revealed a low DI of 0.743 alone but a DI of 0.955 in combination with serotyping . PFGE using the restriction enzyme BglII and opa typing produced the highest discrimination (DIs, 0.997 and 0 . 996, respectively) for isolates of N . gonorrhoeae.

Int J STD AIDS, 1999 May, 10(5), 290 - 3
Neisseria gonorrhoeae in Newcastle upon Tyne 1995-1997: increase in ciprofloxacin resistance; Tayal SC et al.; Fluoroquinolones and third generation cephalosporins are the most effective antimicrobial agents for the treatment of gonorrhoea . However, clinically significant resistance to fluoroquinolones in Neisseria gonorrhoeae has been reported worldwide including Britain . The aim of this analysis was to study the factors relating to ciprofloxacin resistance and treatment failure . A total of 201 patients attending the Newcastle Genitourinary Medicine (GUM) clinic from 1995-1997 who were diagnosed with culture positive gonorrhoea was analysed . Treatment failure rates for ciprofloxacin were determined and the minimum inhibitory concentration (MIC) was measured for all cases of treatment failure . The case notes of all patients who had strains with MICs of ciprofloxacin in the resistant range (>0.05 microg/ml) were reviewed to determine the clinical outcome . The ciprofloxacin resistance with treatment failure was seen in 5% (8/160) . All the 8 cases of treatment failure were heterosexual and had isolates resistant to penicillin and 4 cases (50%) were also resistant to tetracycline . All were sensitive to spectinomycin and ceftriaxone . Most of the cases probably acquired their infection from the Far East . As ciprofloxacin resistance seems to be associated with overseas exposure, changes in the standard treatment of gonorrhoea are not justified but consideration should be given to appropriate alternatives when the infection may have arisen from where such resistant strains are endemic . Monitoring fluoroquinolone resistance is now essential for ensuring adequate treatment of infections with resistant strains and for maximizing the time of usage of fluoroquinolones to treat gonorrhoea.

Adolesc Med, 1996 Feb, 7(1), 63 - 82
Nongonococcal Urethritis: Diagnosis, Management, and Complications; Krowchuk DP; Topics covered here include the role of noninfectious and common and uncommon infectious agents in the development of nongonococcal urethritis; the relative usefulness of a variety of laboratory tests for organisms such as Chlamydia trachomatis and Neisseria gonorrhoeae; and complications including acute epididymitis, prostatitis, and Reiter's syndrome.

Clin Chem, 1999 Jun, 45(6 Pt 1), 777 - 84
Strand displacement amplification and homogeneous real-time detection incorporated in a second-generation DNA probe system, BDProbeTecET; Little MC et al.; BACKGROUND: Amplified DNA probes provide powerful tools for the detection of infectious diseases, cancer, and genetic diseases . Commercially available amplification systems suffer from low throughput and require decontamination schemes, significant hands-on time, and specially trained laboratory staff . Our objective was to develop a DNA probe system to overcome these limitations . METHODS: We developed a DNA probe system, the BDProbeTecTMET, based on simultaneous strand displacement amplification and real-time fluorescence detection . The system uses sealed microwells to minimize the release of amplicons to the environment . To avoid the need for specially trained labor, the system uses a simple workflow with predispensed reagent devices; a programmable, expandable-spacing pipettor; and the 96-microwell format . Amplification and detection time was 1 h, with potential throughput up to 564 patient results per shift . We tested 122 total patient specimens obtained from a family practice clinic with the BD ProbeTecET and the Abbott LCx(R) amplified system for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae . Results: Based on reportable results, the BDProbeTecET results for both organisms were 100% sensitive and 100% specific relative to the LCx . Conclusions: The BDProbeTecET is an easy-to-use, high-throughput, closed amplification system for the detection of nucleic acid from C . trachomatis and N . gonorrhoeae and other organisms.

Adolesc Med, 1990 Oct, 1(3), 583 - 596
Office and Laboratory Testing for STDs: Present and Future; Schachter J; This chapter is aimed at the clinician in private practice or in a small clinic setting dealing with sexually transmitted diseases (STDs) but lacking immediate access to larger laboratories . Tests that should be routinely available for patients with STDs are described . Some of these tests may be performed on site, whereas others must be sent out to appropriate laboratories . Among the tests described are Gram stain testing, culture of Neisseria gonorrhoeae, darkfield examination, as well as testing for Chlamydia trachomatis, herpes simplex virus, chancroid, and syphilis.

Epidemiol Mikrobiol Imunol, 1999 Apr, 48(2), 60 - 2
{An unusual isolation of Neisseria meningitidis}; Blahova M et al.; The authors describe a unique case of isolation of Neisseria meningitidis C:2a:P1.2,P1.5 in August 1997 from a 7-year-old female patient from a vaginal smear during purulent inflammation of the external genital following injury . This is the first described case of isolation of N . meningitidis from this site described in our country . The authors draw attention to the necessity to consider uncommon occurrences of N . meningitidis, and to examine atypical materials also under the microscope and to identify by biochemical methods all isolated Neisseriae incl . gonococci.

Antimicrob Agents Chemother, 1999 Jun, 43(6), 1367 - 72
Erythromycin-resistant Neisseria gonorrhoeae and oral commensal Neisseria spp . carry known rRNA methylase genes; Roberts MC et al.; Two Neisseria gonorrhoeae isolates from Seattle and two isolates from Uruguay were resistant to erythromycin (MIC, 4 to 16 microg/ml) and had reduced susceptibility to azithromycin (MIC, 1 to 4 microg/ml) due to the presence of the self-mobile rRNA methylase gene(s) ermF or ermB and ermF . The two Seattle isolates and one isolate from Uruguay were multiresistant, carrying either the 25.2-MDa tetM-containing plasmid (Seattle) or a beta-lactamase plasmid (Uruguay) . Sixteen commensal Neisseria isolates (10 Neisseria perflava-N . sicca, 2 N . flava, and 4 N . mucosa) for which erythromycin MICs were 4 to 16 microg/ml were shown to carry one or more known rRNA methylase genes, including ermB, ermC, and/or ermF . Many of these isolates also were multiresistant and carried the tetM gene . This is the first time that a complete transposon or a complete conjugative transposon carrying an antibiotic resistance gene has been described for the genus Neisseria.

Int J STD AIDS, 1999 Mar, 10(3), 169 - 73
Characterization of an outbreak of tetM-containing Neisseria gonorrhoeae in Argentina; Fernandez Cobo M et al.; Phenotypic and molecular characterization of an outbreak of 9 Neisseria gonorrhoeae (NG) isolates exhibiting high-level plasmid mediated resistance to penicillin and tetracycline (PP-TRNG) that took place in Tandil, Argentina between February and April 1995 . Comparison with the patterns of the 3 PP-TRNG strains previously isolated were made . We determined the following markers for each strain: antimicrobial susceptibility, serogroup, auxotype, plasmid profile, presence of tetM determinant and restriction pattern of the tetM-containing plasmid . Antimicrobial tests values were: tetracycline disk diameter 12-14 mm, minimum inhibitory concentration (MIC) 32 micrograms/ml; penicillin disk diameter 6 mm, MIC 32 micrograms/ml and sensitive by both methods to spectinomycin, cefuroxime, ceftriaxone and ciprofloxacin . All isolates were of the same serogroup (WI) . Ten of the strains, including the 9 from Tandil outbreak, were arginine-requiring, while the other 2 were methionine and arginine-requiring . All of them demonstrate the same plasmid profile (2.6, 3.2, 25.2 MDa) . They were positive for the tetM determinant and the restriction analysis identified it is a Dutch-type plasmid . In spite of the temporal and geographical dispersion, PP-TRNG strains in Argentina seem to be highly homogeneous in terms of antimicrobial susceptibility, serogroup, plasmid profiles and even auxotypePIP: This study determines the phenotypic and molecular characterization of an outbreak of 9 Neisseria gonorrhea isolates exhibiting high-level plasmid-mediated resistance to penicillin and tetracycline (PP-TRNG) that took place in Tandil, Argentina . Subjects included patients attending Tandil's Sanatamarina Hospital . Individuals infected with sexually transmitted disease were examined using a standardized questionnaire . Results show that all isolates produced B-lactamase, were penicillin resistant (disk zone diameter 6 mm, MIC 32 mcg/ml) and high-level tetracycline resistant (disk zone diameter 12-14 mm, MIC 32 mcg/ml) . Moreover, all strains were fully sensitive to cefuroxime, ceftriaxone, ciprofloxacin, and spectinomycin; while autotype and serogroup results showed that all the strains were arginine-requiring and of serogroup WI . In spite of the temporal and geographical dispersion, PP-TRNG strains in Argentina seem to be highly homogeneous in terms of antimicrobial susceptibility, serogroup, plasmid profiles, and even auxotype .

Infect Immun, 1999 Jun, 67(6), 3141 - 5
FbpC is not essential for iron acquisition in Neisseria gonorrhoeae; Sebastian S et al.; The fbpABC locus of Neisseria gonorrhoeae has been proposed to encode a periplasmic protein-dependent iron transport system . Although the function of the gonococcal FbpA protein has been well characterized and its role as a periplasmic binding protein is well defined, little is known about the function of the FbpB and FbpC proteins . To define the function of the gonococcal FbpC protein, an N . gonorrhoeae F62 fbpC mutant was constructed by insertional inactivation with the kanamycin gene . The N . gonorrhoeae F62 fbpC mutant was observed to grow with heme, transferrin, or ferric nitrate as the sole exogenous iron source, indicating that the gonococcal FbpC protein is not absolutely required for growth with these iron sources . In previous studies we were unable to detect fbpB- or fbpC-specific transcripts by Northern analysis . Reverse transcription-PCR analysis with RNA obtained from N . gonorrhoeae F62 grown under iron-replete and -depleted conditions detected fbpA and fbpAB transcripts but failed to detect fbpC or fbpBC transcripts . These results indicate that FbpC does not play a pivotal role in iron transport in N . gonorrhoeae and suggest that additional ABC transport systems are functional in the gonococcus for the acquisition of iron.

Infect Immun, 1999 Jun, 67(6), 3009 - 13
A Neisseria gonorrhoeae immunoglobulin A1 protease mutant is infectious in the human challenge model of urethral infection; Johannsen DB et al.; Many mucosal pathogens, including Neisseria gonorrhoeae, produce proteases that cleave immunoglobulin A (IgA), the predominant immunoglobulin class produced at mucosal surfaces . While considerable circumstantial evidence suggests that IgA1 protease contributes to gonococcal virulence, there is no direct evidence that N . gonorrhoeae requires IgA1 protease activity to infect a human host . We constructed a N . gonorrhoeae iga mutant without introducing new antibiotic resistance markers into the final mutant strain and used human experimental infection to test the ability of the mutant to colonize the male urethra and to cause gonococcal urethritis . Four of the five male volunteers inoculated with the Iga- mutant became infected . In every respect-clinical signs and symptoms, incubation period between inoculation and infection, and the proportion of volunteers infected-the outcome of human experimental infection with FA1090iga was indistinguishable from that previously reported for a variant of parent strain FA1090 matching the mutant in expression of Opa proteins, lipooligosaccharide, and pilin . These results indicate that N . gonorrhoeae does not require IgA1 protease production to cause experimental urethritis in males.

Sex Transm Dis, 1999 May, 26(5), 303 - 8
Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in swab specimens by the Hybrid Capture II and PACE 2 nucleic acid probe tests; Modarress KJ et al.; BACKGROUND AND OBJECTIVES: The Digene Hybrid Capture II (HC II) CT/GC Test (Digene Corp., Beltsville, MD) is a new nucleic acid signal amplification-based test for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in specimens from the genital tract . For optimal results, the HC II CT/GC Test employs a special conical shaped brush for cervical specimen collection from nonpregnant women and swabs from pregnant women . GOALS: To validate a protocol for HC II C . trachomatis and N . gonorrhoeae testing of specimens collected for the GenProbe PACE 2 System . STUDY DESIGN: Specimens were collected from 1,746 patients with a swab and placed in GenProbe transport media according to the manufacturer's recommended procedure . The specimens were first tested at two clinical laboratories by the PACE 2 system, and then blindly tested by HC II CT/GC using an adjusted cutoff value . Discrepant specimens were adjudicated by polymerase chain reaction (PCR), and the result common to two of the three testing methods (HC II, PACE 2, and PCR) was defined as the consensus result . RESULTS: Combining the data from both sites, the relative sensitivity of the HC II Test compared with the consensus result for the detection of 1,761 specimens for C . trachomatis and 1,750 specimens for N . gonorrhoeae was 100% for both organisms . The relative specificities for C . trachomatis and N . gonorrhoeae detection were 99.8% and 99.7%, respectively . The relative sensitivities of the PACE 2 CT and GC Systems were 86.5% and 87.1%, respectively, with relative specificities of 99.9% and 100% . The difference in sensitivity between HC II and PACE 2 for C . trachomatis detection was significant (P < 0.016) . CONCLUSION: The HC II CT/GC Test can be performed using specimens collected in GenProbe transport media and has a significantly greater sensitivity for C . trachomatis detection than the PACE 2 System.

Sex Transm Dis, 1978 Apr-Jun, 5(2), 62 - 4
A single large dose of trimethoprim-sulfamethoxazole fails to cure gonococcal urethritis in men; Prior RB et al.; In a single-blind study, 50 men who had acute gonococcal urethritis were treated with a single oral dose of either 720 mg trimethoprim (TMP) plus 3,600 mg sulfamethoxazole (SMZ) or 3.5 g ampicillin plus 1 g probenecid . Isolates of Neisseria gonorrhoeae were tested for in-vitro susceptibility to the chemotherapeutic agents administered by agar-dilution and disk-diffusion methods, and results were correlated with cure or failure to cure as determined bacteriologically . Among patients returning for follow up, the cure rate after TMP/SMZ was 69% . Cure was predictable when the isolates of N . gonorrhoeae were inhibited by < or = 0.63/11.87 micrograms/ml of TMP/SMZ (fixed ratio, 1:19) or when the zones of inhibition were > or = 23 mm; failure was predictable when > or = 1.25/23.75 micrograms/ml of TMP/SMZ was necessary for inhibition and when zones of inhibition were < or = 21 mm (P < 0.02) . The cure rate after therapy with ampicillin was 100%, a rate significantly higher than that found after TMP/SMZ (P < 0.02); all isolates were inhibited by < or = 0.16 microgram/ml of ampicillin . Adverse reactions were not seen after either TMP/SMZ or ampicillin.

Sex Transm Dis, 1978 Apr-Jun, 5(2), 43 - 50
Comparison of dihydrofolate reductases from trimethoprim- and sulfonamide-resistant strains of Neisseria gonorrhoeae; Ho RI et al.; The dihydrofolate reductases were purified from Neisseria gonorrhoeae strains CDC 9 and 7134, for which MICs of trimethoprim differ twofold and MICs of sulfadiazine differ sixtyfold . Although affinities of the dihydropteroate synthetases from the two strains for sulfonamide were known to differ markedly, no conclusive difference in the extents of binding of trimethoprim by the dihydrofolate reductases from these organisms could be demonstrated . Thus, it was concluded that permeability of the cell envelope or increased production of dihydrofolate reductase may be an important factor among strains of N . gonorrhoeae that have low- and middle-range resistance to trimethoprim.

Sex Transm Dis, 1978 Apr-Jun, 5(2), 39 - 42
Comparison of Gram stain and first-voided urine sediment in the diagnosis of urethritis; Bowie WR; The number of polymorphonuclear leukocytes on gram-stained urethral specimens was compared with the number in the sediment of the first-voided urine to determine what constitutes an abnormal number of polymorphonuclear leukocytes in gram-stained specimens . Seventy-three men attending a clinic for sexually transmitted diseases were studied . An attempt was made to select primarily asymptomatic men and men who had minimal symptoms or signs of urethritis because the diagnosis of urethritis is most difficult in these groups . There was a distinct bimodal distribution of the numbers of polymorphonuclear leukocytes by both techniques . Means of four or fewer polymorphonuclear leukocytes per field in five fields (X 1,000) with gram stain and of < 15 polymorphonuclear leukocytes in all of five fields (X 400) in the first-voided urine sediment were considered normal . The results with the two techniques were highly correlated (P = 1.4 x 10(-10)) . In the absence of cultures for Neisseria gonorrhoeae, Chlamydia trachomatis, and possibly Ureaplasma urealyticum to detect the presence of urethral infection, a mean of more than four polymorphonuclear leukocytes per field (X 1,000) in a Gram-stained urethral specimen indicates the presence of urethritis.

Sex Transm Dis, 1982 Jan-Mar, 9(1), 45 - 7
Evaluation of modified Stuart's medium in Culturettes for transport of Neisseria gonorrhoeae; Ebright JR et al.; A prospective study compared rates of isolation of Neisseria Gonorrhoeae after immediate plating of clinical specimens onto Thayer-Martin medium with isolation rates after initial transport in modified Stuart's medium (MST) contained in Culturettes . Of 75 specimens positive for Neisseria gonorrhoeae after immediate plating onto Thayer-Martin medium, 65 (87%) were also positive after being held in MST medium for < or = 8 hr . Twenty (95%) of 21 clinical specimens remained positive after being held in MST medium for < or = 3 hr . However, the rates of isolation of N . gonorrhoeae were significantly reduced after transport for 24 and 48 hr; 60% and 27% of specimens, respectively, yielded N . gonorrhoeae-positive cultures after these intervals . Therefore, it is concluded the MST medium in Culturettes is an acceptable transport medium for specimens containing N . gonorrhoeae when transport time prior to inoculation of growth medium is < or = 3 hr.

Sex Transm Dis, 1982 Jan-Mar, 9(1), 34 - 6
Evaluation of modified New York City carbohydrate medium for the speciation of Neisseria; Simms DH et al.; A comparative study of modified New York City carbohydrate medium, Minitek (BBL Microbiology Systems, Cockeysville, Md.) and cystine-trypticase agar was conducted for determination of the relative accuracies of these media in the identification of pathogenic Neisseria . Of 170 isolates of Neisseria gonorrhoeae, 169 were correctly identified by New York City carbohydrate media, 131 by Minitek, and 117 by cystine-trypticase agar . Of the 72 isolates of Neisseria meningitidis tested, all were accurately speciated by New York City carbohydrate media, whereas Minitek and cystine-trypticase agar identified 61 and 49 strains, respectively . The greater reliability of New York City carbohydrate media is accompanied by advantages not offered by the other media.

Sex Transm Dis, 1982 Jan-Mar, 9(1), 9 - 14
Treatment of uncomplicated gonococcal infection with trimethoprim-sulfamethoxazole; Brown ST et al.; The efficacy of trimethoprim-sulfamethoxazole (TMP-SMZ; 80 mg of TMP and 400 mg of SMZ per tablet; nine tablets taken once daily for three days; total, 27 tablets) was compared with the U.S . Public Health Service recommended regimen of 2 g of tetracycline daily for five days for the treatment of uncomplicated genital gonorrhea . Fourteen (3%) of the 461 patients treated with tetracycline and 24 (5%) of the 477 patients treated with TMP-SMZ failed to be cured; the difference between the two groups was not significant . Treatment of patients with TMP-SMZ was more likely to fail if the isolates of Neisseria gonorrhoeae had MICs of > or = 0.5 microgram of TMP/ml and > or = 9.5 micrograms of SMZ/ml . Adverse effects were more often reported by patients receiving TMP-SMZ . The results show that TMP-SMZ is an effective therapy for uncomplicated gonococcal infections in men and women and may also eliminate agents causing postgonococcal urethritis . The utility of this drug combination may be limited by the adverse effects that are associated with the large dose used.

Trans R Soc Trop Med Hyg, 1998 Nov-Dec, 92(6), 670 - 4
Antimicrobial resistance of Neisseria gonorrhoeae in Liberia; Guyot A et al.; The prevalence and molecular characteristics of penicillinase-producing Neisseria gonorrhoeae (PPNG) and tetracycline-resistant N . gonorrhoeae (TRNG) were determined in 10 clinics in Monrovia, Liberia, to assess the likely effectiveness of the current standard treatment with penicillin or tetracycline . One hundred gonococcal strains were isolated from 146 urethral swabs and 261 cervical swabs and screened for susceptibility to ceftriaxone, penicillin, spectinomycin and tetracycline by the disk diffusion method; 83% were resistant to penicillin and 63% to tetracycline . Twenty-one strains from 18 men and 3 women with uncomplicated gonorrhoea were subjected to more detailed characterization . These 21 strains belonged to 5 auxotype/serovar classes; 86% were PPNG/TRNG . Three PPNG harboured the 4.4 MDa penicillinase plasmid and 16 the 3.2 MDa plasmid . All TRNG harboured the 25.2 MDa plasmid and their MICs for tetracycline were > 32 mg/L . They gave a PCR product which, according to its restriction pattern, corresponded to the American type tetM gene . By the agar dilution method, all strains exhibited intermediate resistance to sulphamethoxazole-trimethoprim (19:1) (co-trimoxazole) with MICs of 8-32 mg/L . All strains were susceptible to spectinomycin and ciprofloxacin . The MICs for gentamicin were 4-8 mg/L . The use of effective and affordable antimicrobial chemotherapy with either 500 mg ciprofloxacin or a single dose of gentamicin is discussed, with consideration of molecular biological, pharmacological and public health aspects.

J Hum Genet, 1999, 44(3), 215 - 8
A novel nonsense mutation at Glu-631 in a Spanish family with complement component 7 deficiency; Horiuchi T et al.; Deficiency of the seventh component of complement (C7D) is frequently associated with recurrent neisserial infections . We report in the present study the genetic basis for C7D in a Spanish family . We used exon-specific polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) analysis as a screening step for mutations, followed by direct sequencing of the target exon . The mutation in the proband was a homozygous G-to-T transversion at nucleotide 1957, the first nucleotide of the codon GAG for Glu-631, leading to a stop codon TAG (E631X) . Our result provides further evidence that the molecular pathogenesis of C7D is heterogeneous.

Hautarzt, 1999 Mar, 50(3), 186 - 93
{Pathogen spectrum of urethritis in the man}; Gall H et al.; 545 male patients with a tentative diagnosis "urethritis" were examined between November 1984 and December 1994 in the Department of Dermatology and Venerology of the Military Hospital in Ulm . The patients, aged from 18 to 58 years (mean age 24.1 years), were examined according to a standardized diagnostic procedure: Smear preparations from the urethra with subsequent gram staining, bacterial cultures for aerobic bacteria, Neisseria gonorrhoeae (cultures and Phadebact gonococcus test), mycoplasma cultures (Mycoplasma hominis (M . hom.); Ureaplasma urealyticum (U . u), and Chlamydia trachomatis using several methods, primarily DIFT (Syva Micro-Trak) . Trichomonas vaginalis counts in urine sediment 441 patients (81%) had 4 or more leukocytes per high-power (x1000) field in the gram stained specimens . In these 441 urethritis-patients the following germs could be detected: Trichomonas vagin 3 (1%), N . gonorrhoeae 80 (18%), Mycoplasma 94 (21%) {U . u . 59, M . hom . 24, both 11}, C . trachomatis 114 (26%), other pathogenic bacteria 135 (31%) . In 114 patients (26%) no bacteria could be identified . A single infection was diagnosed in a total of 242 patients (55%), a double infection was determined in 71 patients (16%) while a triple infection was found in 14 patients (3%) . The spectrum determined in the single infection included the following: N . gonorrhoeae 41 (9%), Mycoplasma 45 (10%), C . trachomatis 67 (15%), other pathogenic bacteria 89 (20%) (most frequently found germs were Enterococcus, beta-hemolytic Streptococcus, Escherichia coli, Staph . aureus) . In the double infections combinations with aerobic bacteria dominated . In triple infections, mycoplasma were most common . During the investigation period the number of patients with urethritis symptoms declined at a constant rate.

Mol Microbiol, 1999 Apr, 32(2), 233 - 42
Proteoglycan receptor binding by Neisseria gonorrhoeae MS11 is determined by the HV-1 region of OpaA; Grant CC et al.; The interaction of the OpaA protein of Neisseria gonorrhoeae MS11mk with heparan sulphate-containing proteoglycan receptors on Chang conjunctiva epithelial cells was examined using isolated receptor binding and cell adherence/internalization assays . OpaA deletion proteins, in which the four surface-exposed regions of the protein were deleted individually, and chimeric OpaA/B proteins, in which the surface-exposed regions of the OpaA and OpaB proteins were exchanged, were expressed in N . gonorrhoeae . The recombinant deletion proteins and the chimeric OpaA/B proteins were surface exposed in the outer membrane of N . gonorrhoeae . Isolated receptor-binding assays and Chang cell infection assays with OpaA deletion variants indicated that hypervariable region 1 was essential for the interaction of N . gonorrhoeae with the proteoglycan receptor . Expression of chimeric OpaA/B proteins confirmed the central role of hypervariable region 1 in receptor binding and demonstrated that this domain alone confers the invasive biological phenotype in a non-heparan sulphate proteoglycan-binding Opa protein . The other variable regions of OpaA enhanced receptor binding in the presence of region 1, but did not constitute binding domains on their own . The results indicate that proteoglycan receptor binding results from a hierarchical interaction between the variable domains of the OpaA protein of MS11mk.

J Infect Dis, 1999 Jun, 179(6), 1433 - 40
Elderly immune response to a TI-2 antigen: heavy and light chain use and bactericidal activity to Neisseria meningitidis serogroup C polysaccharide; Hutchins WA et al.; Previous studies of the elderly immune response to TI-2 antigens failed to correlate specific antibody levels with function and to compare responses with those of young adults . Neisseria meningitidis serogroup C capsular polysaccharide (MCPS) was used as a model TI-2 antigen . Anti-MCPS antibody levels were determined in elderly individuals and correlated with bactericidal activity . The anti-MCPS response in most persons was characterized by predominant IgG usage, with IgG2>IgG1 . No light chain or IgA subclass predominated, but some responses showed a particular chain type . Bactericidal activity correlated best with IgG2 levels . Elderly subjects had lower anti-MCPS responses than the young adults did in all chain-specific anti-MCPS levels, and levels declined more rapidly . Bactericidal activity following immunization was significantly lower in the elderly persons . These results suggest the anti-MCPS antibody repertoire in the elderly is likely maintained, and the lower level of function is related to the lower antibody levels.

Infect Immun, 1999 May, 67(5), 2406 - 13
Structural and evolutionary inference from molecular variation in Neisseria porins; Derrick JP et al.; The porin proteins of the pathogenic Neisseria species, Neisseria gonorrhoeae and Neisseria meningitidis, are important as serotyping antigens, putative vaccine components, and for their proposed role in the intracellular colonization of humans . A three-dimensional structural homology model for Neisseria porins was generated from Escherichia coli porin structures and N . meningitidis PorA and PorB sequences . The Neisseria sequences were readily assembled into the 16-strand beta-barrel fold characteristic of porins, despite relatively low sequence identity with the Escherichia proteins . The model provided information on the spatial relationships of variable regions of peptide sequences in the PorA and PorB trimers and insights relevant to the use of these proteins in vaccines . The nucleotide sequences of the porin genes from a number of other Neisseria species were obtained by PCR direct sequencing and from GenBank . Alignment and analysis of all available Neisseria porin sequences by use of the structurally conserved regions derived from the PorA and PorB structural models resulted in the recovery of an improved phylogenetic signal . Phylogenetic analyses were consistent with an important role for horizontal genetic exchange in the emergence of different porin classes and confirmed the close evolutionary relationships of the porins from N . meningitidis, N . gonorrhoeae, Neisseria lactamica, and Neisseria polysaccharea . Only members of this group contained three conserved lysine residues which form a potential GTP binding site implicated in pathogenesis . The model placed these residues on the inside of the pore, in close proximity, consistent with their role in regulating pore function when inserted into host cells.

J Acquir Immune Defic Syndr Hum Retrovirol, 1999 Apr 15, 20(5), 502 - 7
Predictors and risk-taking consequences of drug use among HIV-infected women; Novotna L et al.; ObJECTIVE: To determine rates of drug use among women with HIV, and to examine associations between drug use, health, risk behavior, and sexually transmitted diseases (STD) . DESIGN: A longitudinal cohort study of 260 women with confirmed HIV-positive serostatus . METHODS: Each participant contributed a self-report interview, a clinical examination, laboratory testing of cultures for Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and urinalysis for the presence of metabolites of cocaine and opiates . Data were examined on 140 women at 1-year follow-up . Women were defined as drug users if they reported crack, cocaine, or heroin use in the 6 months before the interview or if they had a positive toxicologic test result for cocaine or opiates . RESULTS: 34% of those in the sample were classified as positive for drug use . Drug use was associated with the number of sexual partners, age at first intercourse, prevalence of STDs, and lower quality of life . STDs were present at baseline in 33.7% and 15.5% of drug users and nonusers, respectively . Drug use among this population was also associated at both baseline and follow-up with the likelihood of having a Karnofsky score below 80, and with overall perceived general health . CONCLUSIONS: Drug users in this cohort were more likely to engage in behaviors that place them at risk for STDs, to have elevated STD prevalence, and to have lower perceived health across several indices . Identification of drug use and treatment for it need to be a central component of HIV care for women.

Kansenshogaku Zasshi, 1999 Mar, 73(3), 233 - 8
Prevalence of human papillomavirus infection in women attending a sexually transmitted disease clinic; Kubota T et al.; The purpose of this study was to determine the prevalence of infection due to human papillomavirus (HPV) types of high and intermediate oncogenic risk, which was most frequently associated with uterine cervical neoplasia . The subjects were 236 prostitutes who visited a sexually transmitted diseases (STD) clinic in a metropolitan area in 1998 . Another 95 women who visited a university hospital were selected as a normal control group . A swab sample collected from the uterine cervix and external os was subjected to hybrid capture assays for low-oncogenic-risk HPV types (HPV A; including types 6, 11, 42, 43 and 44) and high- and intermediate-oncogenic-risk HPV types (HPV B; including 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68) . Chlamydia trachomatis and Neisseria gonorrhoeae . Fisher's exact test was used for statistical analyses . Among the microorganisms tested, the positive rate for HPV B was the highest both in the women attending the STD clinic (STD group) and in the control group . The positive rate for HPV B in the STD group was 47.5% (112 of 236), and this was significantly higher than the 5.3% (5 of 95) in the control group (p < 0.0001) . These findings suggest that HPV examination is recommended for women who visit an STD clinic to assess the future risk of cervical neoplasia.

J Bacteriol, 1999 May, 181(9), 2895 - 901
Ferric enterobactin binding and utilization by Neisseria gonorrhoeae; Carson SD et al.; FetA, formerly designated FrpB, an iron-regulated, 76-kDa neisserial outer membrane protein, shows sequence homology to the TonB-dependent family of receptors that transport iron into gram-negative bacteria . Although FetA is commonly expressed by most neisserial strains and is a potential vaccine candidate for both Neisseria gonorrhoeae and Neisseria meningitidis, its function in cell physiology was previously undefined . We now report that FetA functions as an enterobactin receptor . N . gonorrhoeae FA1090 utilized ferric enterobactin as the sole iron source when supplied with ferric enterobactin at approximately 10 microM, but growth stimulation was abolished when an omega (Omega) cassette was inserted within fetA or when tonB was insertionally interrupted . FA1090 FetA specifically bound 59Fe-enterobactin, with a Kd of approximately 5 microM . Monoclonal antibodies raised against the Escherichia coli enterobactin receptor, FepA, recognized FetA in Western blots, and amino acid sequence comparisons revealed that residues previously implicated in ferric enterobactin binding by FepA were partially conserved in FetA . An open reading frame downstream of fetA, designated fetB, predicted a protein with sequence similarity to the family of periplasmic binding proteins necessary for transporting siderophores through the periplasmic space of gram-negative bacteria . An Omega insertion within fetB abolished ferric enterobactin utilization without causing a loss of ferric enterobactin binding . These data show that FetA is a functional homolog of FepA that binds ferric enterobactin and may be part of a system responsible for transporting the siderophore into the cell.

Mol Microbiol, 1999 Apr, 32(1), 111 - 21
Identification of sequences in human transferrin that bind to the bacterial receptor protein, transferrin-binding protein B; Retzer MD et al.; Alignment of amino-acid sequences from the N-terminal and C-terminal halves of transferrin-binding protein B revealed an underlying bilobed nature with several regions of identity . Based on this analysis, purified recombinant fusion proteins of maltose-binding protein (Mbp) with intact TbpB, its N-terminal half or C-terminal half from the human pathogens Neisseria meningitidis and Moraxella catarrhalis were produced . Solid-phase binding assays and affinity isolation assays demonstrated that the N-terminal and C-terminal halves of TbpB could bind independently to human transferrin (hTf) . A solid-phase overlapping synthetic peptide library representing the amino-acid sequence of hTf was probed with soluble, labelled Mbp-TbpB fusions to localize TbpB-binding regions on hTf . An essentially identical series of peptides from domains within both lobes of hTf was recognized by intact TbpB from both organisms, demonstrating a conserved TbpB-hTf interaction . Both halves of TbpB from N . meningitidis bound the same series of peptides, which included peptides from equivalent regions on the two hTf lobes, indicating that TbpB interacts with each lobe of hTf in a similar manner . Mapping of the peptide-binding regions on a molecular model of hTf revealed a series of nearly adjacent surface regions that nearly encircled each lobe . Binding studies with chimeric hTf/bTf transferrins demonstrated that regions in the C-lobe of hTf were preferentially recognized by the N-terminal half of TbpB . Collectively, these results provide evidence that TbpB consists of two lobes, each with distinct yet homologous Tf-binding regions.

Afr J Reprod Health, 1998 Apr, 2(1), 81 - 6
Ophthalmia neonatorum revisited; van Bogaert LJ; The microbiology, epidemiology and pathophysiology of ophthalmia neonatorum are reviewed with special emphasis on its prevention and management in the developing world . Although prophylaxis should be mandatory, no single topical agent is effective to prevent the ocular complications of both Neisseria gonorrhoea and Chlamydia trachomatis . Where levels of resistance to tetracyclines are low, however, tetracycline eye ointment is permissible for ocular prophylaxis . Eye prophylaxis has a relatively low failure rate . Management of ophthalmia neonatorum should be syndromic and systemic . Contact tracing is an integral part of the management.

Afr J Reprod Health, 1998 Apr, 2(1), 32 - 40
Genital infections among antenatal care attendees in Cape Verde; Wessel HF et al.; In a cross-sectional study, 350 pregnant Capeverdian women were examined to assess the prevalence of Chlamydia trachomatis infection (CT), Neisseria gonorrhoeae infection (NG) and Bacterial vaginosis (BV) . Among various analytic methods used, the polymerase chain reaction PCR (for NG, CT) yielded a higher detection rate than did direct microscopy or culture (NG), or direct immuno-fluorescence (CT) . Since the PCR analytic of air-dried specimens is not hampered by harsh storage and transport conditions, it could serve to validate other detection methods where laboratory facilities are suboptimal . Among sociodemographic risk factors young age, and currently living alone, were significantly associated with infection.

Lancet . 1997 Oct 11;350(9084):1076.
Urethral Trichomonas vaginalis infection and HIV-1 transmission; Jackson DJ et al.; PIP: The authors' previous study of 504 male workers in Mombasa, Kenya, provides further documentation of an association between urethral infection and increased shedding of HIV in semen . In this study, Trichomonas vaginalis was isolated by culture in 30 men (6%) and was the most commonly isolated urethral pathogen . Men with Trichomonas vaginalis were significantly older than those with Neisseria gonorrhoeae, suggesting infection may be of long duration . 83% of men with unmixed infections were asymptomatic . Since guidelines for syndromic management of urethral discharge do not include Trichomonas vaginalis, even symptomatic men with this condition who seek treatment are unlikely to be diagnosed and properly treated . More research is needed to confirm whether Trichomonas vaginalis (especially long-standing, low-grade infection) enhances male to female HIV transmission . Since an effective, single-dose, low-cost treatment for Trichomonas vaginalis is available worldwide, mass treatment strategies for this sexually transmitted disease could play an important part in HIV prevention in developing countries .

Glycoconj J, 1998 Oct, 15(10), 1017 - 31
Binding of N-trifluoroacetyl-derivatized natural glycosphingolipids by uropathogenic Escherichia coli and Neisseria subflava; Lanne B et al.; Several neutral glycosphingolipids were hydrogenated and subjected to trifluoroacetylation in trifluoroacetic acid/trifluoroacetic acid anhydride under conditions leading to complete exchange of the N-acetyl groups of GalNAc for N-trifluoroacetyl . The derivatized glycosphingolipids were analyzed for binding by P-fimbriated uropathogenic Escherichia coli, recognizing the globo series of glycolipids (carrying Galalpha1-4Gal) . Using E . coli it was shown that a GalNCO-CF3 next to the minimum binding epitope Galalpha1-4Gal did not substantially influence the binding, as did not a trifluoro acetyl group on the ceramide . Exchange of N-acetyl of GalNAc in the receptor active gangliotetraosylceramide, Galbeta1-3GalNAcbeta1-4Galbeta1-4Glcbeta1-1Cer, for N-trifluoroacetyl, did not change the binding of two out of the three strains tested of the bacterium Neisseria subflava . Discussion concerning the binding epitopes of the bacterial adhesins to carbohydrates is based on these results.

Mol Cell Probes, 1999 Apr, 13(2), 89 - 92
A PCR assay for discriminating Neisseria gonorrhoeaebeta-lactamase-producing plasmids; Dillon JR et al.; Oligonucleotide primers were developed for use in polymerase chain reaction (PCR) assays to differentiate three related, epidemic beta-lactamase-producing plasmids of Neisseria gonorrhoeae-the Asia-(7426 bp), Africa-(5599 bp) and Toronto-(5154 bp) type plasmids . One-hundred and two N . gonorrhoeae isolates with different plasmid profiles were tested-16 isolates carried the Asia plasmid, 41 isolates contained the Africa plasmid, 16 isolates contained the Toronto plasmid and 29 isolates contained no beta-lactamase-producing plasmids . Most (101/102) isolates also carried the gonococcal cryptic plasmid, while 27/102 and 44/102 isolates carried either the transfer plasmid or the tet M-containing plasmids, respectively . The assay was 100% sensitive and specific for identifying the correct plasmid type . This assay is useful for rapidly detecting the presence of gonococcal beta-lactamase-producing plasmids in clinical samples and discriminating them for epidemiological typing .

Am J Obstet Gynecol, 1999 Apr, 180(4), 824 - 36
Prevalence and incidence of gynecologic disorders among women infected with human immunodeficiency virus; Minkoff HL et al.; OBJECTIVE: Our purpose was to ascertain the prevalence, incidence, and predictors of gynecologic disorders among women infected with human immunodeficiency virus . STUDY DESIGN: We serially assessed 292 women infected with human immunodeficiency virus and 681 uninfected women . Outcomes were incidence and prevalence of sexually transmitted diseases, viral shedding, findings of Papanicolaou smears, fungal infections, and menstrual disorders . RESULTS: Women infected with the virus were more likely to have prevalent vulvovaginal candidiasis (odds ratio 1.80, 95% confidence interval 1 . 0-3.25, P =.05), oncogenic human papillomavirus (odds ratio 3.79, 95% confidence interval 2.43-5.91, P =.001), abnormal Papanicolaou smears (odds ratio 5.40, 95% confidence interval 3.35-8.78, P =.001), amenorrhea (4.8% vs 0%, P =.05), positive results on Treponema pallidum hemagglutination assay (odds ratio 1.83, 95% confidence interval 1.16-2.88, P =.01), infection with cytomegalovirus (odds ratio 4.2, 95% confidence interval 1.82-10.62, P =.001), and genital warts (odds ratio 6.93, 95% confidence interval 3.16-16.30, P =.001) but were less likely to have Chlamydia trachomatis infection (odds ratio 0.28, 95% confidence interval 0.10-0.66, P =.01) . Annual incidence rates among women infected with human immunodeficiency virus were 4.0% for candidiasis, 22.0% for oncogenic human papillomavirus, 11.4% for genital warts, 1.7% for infection with C trachomatis, 1.7% for infection with Neisseria gonorrhoeae, 10.3% for Trichomonas vaginalis, 1.1% for positive results on T pallidum hemagglutination assay, 7.4% for an abnormal Papanicolaou smear, and 10.9% for infection with herpes simplex virus . Overall, 46.9% had at least 1 incident condition . Women infected with human immunodeficiency virus were more likely to have incident oncogenic human papillomavirus infection (odds ratio 2.0, 95% confidence interval 1.01-3.8), abnormal Papanicolaou smears (odds ratio 7.76, 95% confidence interval 2.08-42.8), and genital warts (odds ratio 9 . 32, 95% confidence interval 3.04-38.0) . Incidence and prevalence of sexually transmitted diseases and oncogenic human papillomavirus infection increased with increased CD4(+) cell counts . CONCLUSIONS: Women infected with the human immunodeficiency virus are significantly more likely to have prevalent and incident gynecologic disorders but not disorders related to risk taking (eg, incident sexually transmitted diseases) . The latter disorders increased in women with CD4(+) cell counts >500 cells/mm3 . Clinicians should be aware of these patterns so that they can provide appropriate evaluation and treatment of gynecologic disorders.

Mol Microbiol, 1999 Mar, 31(5), 1345 - 57
The comP locus of Neisseria gonorrhoeae encodes a type IV prepilin that is dispensable for pilus biogenesis but essential for natural transformation; Wolfgang M et al.; The expression of type IV pili (Tfp) by Neisseria gonorrhoeae has been shown to be essential for natural genetic transformation at the level of sequence-specific uptake of DNA . All previously characterized mutants defective in this step of transformation either lack Tfp or are altered in the expression of Tfp-associated properties, such as twitching motility, autoagglutination and the ability to bind to human epithelial cells . To examine the basis for this relationship, we identified potential genes encoding polypeptides sharing structural similarities to PilE, the Tfp subunit, within the N . gonorrhoeae genome sequence database . We found that disruption of one such gene, designated comP (for competence-associated prepilin), leads to a severe defect in the capacity to take up DNA in a sequence-specific manner, but does not alter Tfp biogenesis or expression of the Tfp-associated properties of auto-agglutination, twitching motility and human epithelial cell adherence . Indirect evidence based on immunodetection suggests that ComP is expressed at very low levels relative to that of PilE . The process of DNA uptake in gonococci, therefore, is now known to require the expression of at least three distinct components: Tfp, the recently identified PilT protein and ComP.

AIDS, 1999 Feb 25, 13(3), 327 - 32
Increased interleukin-10 in the the endocervical secretions of women with non-ulcerative sexually transmitted diseases: a mechanism for enhanced HIV-1 transmission?
Cohen CR, Plummer FA, Mugo N, Maclean I, Shen C, Bukusi EA, Irungu E, Sinei S, Bwayo J, Brunham RC.
OBJECTIVE: Although non-ulcerative sexually transmitted diseases (STD) and bacterial vaginosis are implicated as cofactors in heterosexual HIV-1 transmission, the mechanisms have not been defined . Recent in vitro data suggest that interleukin (IL)-10 may increase susceptibility of macrophages to HIV-1 infection . Therefore, we performed this study to assess whether non-ulcerative STD are associated with detection of IL-10 in the female genital tract . METHODS: Women with clinical pelvic inflammatory disease with or without cervicovaginal discharge were recruited from an STD clinic in Nairobi, Kenya . Endocervical and endometrial specimens were obtained for Neisseria gonorrhoeae and Chlamydia trachomatis DNA detection, Trichonomas vaginalis culture, and CD4 and CD8 T-cell enumeration . Bacterial vaginosis was diagnosed by Gram stain . IL-10 was detected in endocervical specimens using enzyme-linked immunosorbent assay . Blood was obtained for HIV-1 serology . RESULTS: One hundred and seventy-two women were studied . N . gonorrhoeae, C . trachomatis, bacterial vaginosis, and T . vaginalis were detected in 38 (21%), 17 (9%), 71 (43%), and 22 (12%) women, respectively . Cervical IL-10 was detected more often in women with N . gonorrhoeae {adjusted odds ratio (AOR), 3.4; 95% confidence interval (CI), 1.4-8.4}, C . trachomatis (AOR, 4.4; 95% CI, 1.2-15.6), and bacterial vaginosis (AOR, 3.1; 95% CI, 1.4-6.9) than in women without these infections . CONCLUSIONS: The association of non-ulcerative STD and bacterial vaginosis with increased frequency of IL-10 detection in endocervical secretions suggests a potential mechanism through which these infections may alter susceptibility to HIV-1 infection in women.

Clin Infect Dis, 1999 Mar, 28(3), 491 - 6
Lipopolysaccharide structure influences the macrophage response via CD14-independent and CD14-dependent pathways; Gangloff SC et al.; CD14, a protein expressed on the surface of monocytes and neutrophils, is a major receptor for lipopolysaccharide (LPS) . Studies with normal and CD14-deficient macrophages show that responses to low concentrations of LPS require expression of CD14, whereas responses to high concentrations of LPS are CD14-independent . Since LPS isolated from different bacterial species shows structural variability, studies were performed to determine whether differences in LPS structure influence CD14-dependent and CD14-independent responses . Studies with LPS purified from Escherichia coli, Salmonella abortus subspecies equi, Salmonella minnesota, Pseudomonas aeruginosa, Neisseria meningitidis, Bacteroides fragilis, and Rhodobacter sphaeroides show that the strongest CD14-dependent responses require a typical O-antigen, long carbohydrate chains, at least 6 acyl chains in their lipid A, and 2-phosphorylated Kdo moieties; wild-type LPS lacking a typical O-antigen and containing short carbohydrate chains and 2-phosphorylated Kdo moieties induces the strongest CD14-independent response.

Clin Infect Dis, 1999 Mar, 28(3), 433 - 41
Pathogenic neisseriae: complexity of pathogen-host cell interplay; Meyer TF; Recent studies have provided insight into the function of important neisserial adhesins (pili and Opa) and their interaction with cellular receptors, including members of heparan sulfate proteoglycan, CD66, and integrin receptor families . These interactions not only allow colonization of the human mucosa but also stimulate cellular signaling cascades involving phosphatidylcholine-dependent phospholipase C, acidic sphingomyelinase and protein kinase C in epithelial cells, and Src-related kinases, Rac1, p21-activated kinase, and Jun N-terminal kinase in phagocytic cells . Activation of these pathways is essential for cellular entry and intracellular accommodation of the pathogens but also leads to early induction of cytokine release, thus priming the immune response . Detailed knowledge of the cellular signaling cascades that are activated by infection will aid us in applying both current and novel interfering drugs (in addition to classical antibiotic therapy) as therapy and prophylaxis for persistent or otherwise difficult-to-treat bacterial infections, including periodontal infections.

Arch Pediatr, 1999 Mar, 6(3), 317 - 20
{Chlamydia trachomatis conjunctivitis in the newborn}; Salpietro CD et al.; Infections of the conjunctiva are frequent in the neonatal period . While Neisseria gonorrhoea and chemical agents were considered as the main etiologies of ophtalmiae neonatorum in the past, Chlamydia trachomatis is today a major cause of neonatal conjunctivitis . Thus in a study of 180 uni-or bilateral neonatal conjunctivitis the authors found a prevalence of Chlamydia trachomatis infection of 41% . The importance of the etiological diagnosis of neonatal conjunctivitis is emphasized, in order to define a specific treatment . Etiological diagnosis of Chlamydia trachomatis infection is based upon immunofluorescence and molecular diagnosis techniques (PCR, LCR) . Prevention of neonatal Chlamydia trachomatis conjunctivitis relies upon screening and treatment of Chlamydia trachomatis infections in pregnant women and their partners . Treatment requires oral macrolides, the topical treatment being ineffective.

Cent Afr J Med, 1998 Nov, 44(11), 293 - 6
Syndromic management of sexually transmitted diseases . Part 4--The management of lower abdominal pain in women; Latif AS; The symptom of lower abdominal pain in women is extremely common and does not always indicate the presence of serious illness . However, women with certain serious conditions such as pelvic inflammatory disease (PID), acute appendicitis, ectopic pregnancy and other complications of pregnancy may present initially with this symptom . Therefore, in managing women with lower abdominal pain care should be taken to exclude any serious condition before dismissing the patient . PID is a condition in which there is infection of the reproductive tract of women above the internal os of the cervix . This usually occurs as a result of an ascending cervical infection caused by Neisseria gonorrhoeae, Chlamydia trachomatis and anaerobic bacteria . The immediate and long term effects of PID include salpingitis, pelvic abscess, peritonitis, infertility and predisposition to tubal ectopic pregnancy . Women with lower abdominal pain should be assessed carefully and if PID is the cause they should be treated for gonococcal, chlamydial and anaerobic bacterial infection . Other gynaecological and surgical causes of lower abdominal pain and the immediate complications of PID require urgent referral to a specialist . PID is associated with significant morbidity and mortality.

Antimicrob Agents Chemother, 1999 Apr, 43(4), 983 - 4
In vitro activities of ketolides HMR 3647 {correction of HRM 3647} and HMR 3004 {correction of HRM 3004}, levofloxacin, and other quinolones and macrolides against Neisseria spp . and Moraxella catarrhalis; Saez-Nieto JA et al.; In vitro activities of the ketolides HMR 3647 {corrected} and HMR 3004 {corrected} against pathogenic Neisseria gonorrhoeae and N . meningitidis, saprophytic Neisseria isolates, and Moraxella catarrhalis were determined . The comparison of ketolide activities with those of the other macrolides shows a much better activity in the majority of species, with macrolide MICs at which 90% of the isolates are inhibited between 8- and 10-fold higher.

Antimicrob Agents Chemother, 1999 Apr, 43(4), 745 - 51
Bile salts: natural detergents for the prevention of sexually transmitted diseases; Herold BC et al.; The development of new, safe, topical microbicides for intravaginal use for the prevention of sexually transmitted diseases is imperative . Previous studies have suggested that bile salts may inhibit human immunodeficiency virus infection; however, their activities against other sexually transmitted pathogens have not been reported . To further explore the potential role of bile salts in preventing sexually transmitted diseases, we examined the in vitro activities and cytotoxicities of select bile salts against Chlamydia trachomatis, herpes simplex virus (types 1 and 2), Neisseria gonorrhoeae, and human immunodeficiency virus in comparison to those of nonoxynol-9 and benzalkonium chloride using both primary cells and cell lines derived from the human female genital tract . We found that taurolithocholic acid 3-sulfate and a combination of glycocholic acid and taurolithocholic acid 3-sulfate showed excellent activity against all of the pathogens assayed . Moreover, taurolithocholic acid 3-sulfate alone or in combination was less cytotoxic than nonoxynol-9 and benzalkonium chloride . Thus, taurolithocholic acid 3-sulfate alone or in combination warrants further evaluation as a candidate topical microbicidal agent.

Cent Afr J Med, 1998 Sep, 44(9), 236 - 41
Syndromic management of sexually transmitted diseases . Part 2--The management of genital discharge; Latif AS; In men urethral discharge is commonly caused by Neisseria gonorrhoeae and Chlamydia trachomatis . Both organisms cause an acute anterior urethritis and infected men usually present with symptoms of urethritis . A proportion of men with urethral infection may remain asymptomatic . Amongst women the common cause of vaginal discharge is vaginitis caused by Trichomonas vaginalis, Candida albicans and anaerobic bacterial infection (bacterial vaginosis) . However, cervicitis caused by N . gonorrhoeae and C . trachomatis also causes vaginal discharge . Cervicitis is the more serious cause of vaginal discharge as the aetiologic agents may lead to infection ascending above the internal os of the cervix resulting in pelvic inflammatory disease and other complications.

Yonsei Med J, 1998 Dec, 39(6), 488 - 94
Bacterial resistance to antimicrobial agents: an overview from Korea; Kim WJ et al.; Antimicrobial resistance of bacteria has become a worldwide problem . Available data suggest that the resistance problem is comparatively more serious in Korea . In large hospitals, the proportion of methicillin-resistant Staphylococcus aureus (MRSA) has been reported at over 70%, and of penicillin-nonsusceptible Streptococcus pneumoniae at around 70% . Infection or colonization of vancomycin-resistant enterococci has started to increase . Extended-spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae has become widespread and even carbapenem-resistant Pseudomonas aeruginosa has been increasing . Community-acquired pathogens such as Salmonella, Shigella and Neisseria gonorrhoeae are often resistant to various antimicrobial agents . The prevalence of resistant bacteria can lead to erroneous empirical selection of either noneffective or expensive drugs, prolonging hospitalization and higher mortality . The emergence and spread of resistant bacteria are unavoidable unless antimicrobial agents are not used at all . The high prevalence of resistant bacteria in Korea seems to be related to antibiotic usage: 1) easy availability without prescription at drug stores, 2) injudicious use in hospitals, and 3) uncontrolled use in agriculture, animal husbandry, and fisheries . Nosocomial infection is an important factor in the spread of resistant bacteria . Antimicrobial resistance problems should be regarded as the major public health concern in Korea . It is urgently required to ban the sale of antibiotics without prescription, to use antibiotics more judiciously in hospitals by intensive teaching of the principles of the use of antibiotics, and to establish better control measures of nosocomial infections . Regulation of antimicrobials for other than human use should also be required . These issues are not easy to address and require the collective action of governments, the pharmaceutical industry, health care providers, and consumers.

J Bacteriol, 1999 Apr, 181(7), 2067 - 74
HmbR, a hemoglobin-binding outer membrane protein of Neisseria meningitidis, undergoes phase variation; Richardson AR et al.; Neisseria meningitidis uses hemoglobin (Hb) as an iron source via two TonB-dependent outer membrane receptors, HmbR and HpuB . Analysis of 25 epidemiologically unrelated clinical isolates from serogroups A, B, C, and Y revealed that 64% strains possessed both Hb receptor genes . Examination of the hmbR expression pattern in strains in which the hpuB gene was genetically inactivated revealed two distinct Hb utilization phenotypes . Five strains retained the ability to grow as a confluent lawn, while seven grew only as single colonies around Hb discs . The single-colony phenotype observed for some hpuB mutants is suggestive of phase variation of hmbR . The length of the poly(G) tract starting at position +1164 of hmbR absolutely correlated with the two Hb utilization phenotypes . All five strains that grew as confluent lawns around Hb discs possessed either 9 or 12 consecutive G residues . All seven strains that grew as single colonies around Hb discs had poly(G) tracts of a length other than 9 or 12 . These single-colony variants that arose around the Hb discs had poly(G) tracts with either 9 or 12 consecutive G residues restoring the hmbR reading frame . Inactivation of hmbR in these strains resulted in a loss of Hb utilization, demonstrating that the change in the hmbR gene was responsible for the phenotypic switch . The switching rates from hmbR phase off to phase on were approximately 5 x 10(-4) in four serogroup C strains, 2 x 10(-2) in the serogroup A isolate, and 7 x 10(-6) in the serogroup B isolate.

Mol Microbiol, 1998 Nov, 30(4), 697 - 710
Differential roles of homologous recombination pathways in Neisseria gonorrhoeae pilin antigenic variation, DNA transformation and DNA repair; Mehr IJ et al.; Neisseria gonorrhoeae (Gc) pili undergo antigenic variation when the amino acid sequence of the pilin protein is changed, aiding in immune avoidance and altering pilus expression . Pilin antigenic variation occurs by RecA-dependent unidirectional transfer of DNA sequences from a silent pilin locus to the expressed pilin gene through high-frequency recombination events that occur at limited regions of homology . We show that the Gc recQ and recO genes are essential for pilin antigenic and phase variation and DNA repair but are not involved in natural DNA transformation . This suggests that a RecF-like pathway of recombination exists in Gc . In addition, mutations in the Gc recB, recC or recD genes revealed that a Gc RecBCD pathway also exists and is involved in DNA transformation and DNA repair but not in pilin antigenic variation.

Diagn Microbiol Infect Dis, 1999 Mar, 33(3), 141 - 6
Diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae . Genitourinary infections in males by the Amplicor PCR assay of urine; Palladino S et al.; The Amplicor CT/NG polymerase chain reaction (PCR) test on urine specimens from males was prospectively evaluated against established specimens and laboratory methods for diagnosing Chlamydia trachomatis and Neisseria gonorrhoeae genitourinary infections, in patients from a remote region of Western Australia . Seventy-three males who were tested for both C . trachomatis and N . gonorrhoeae by both conventional methodology and Amplicor PCR on urine were enrolled in the study . Established testing comprised enzyme immunoassay/immunofluorescence antigen testing (EIA/IF) for C . trachomatis and microscopy and/or culture for N . gonorrhoeae on urethral swabs . Positive test results were confirmed using a set of criteria that included supplemental PCR testing and clinical history . Overall, 13.7% of patients were resolved as positive for C . trachomatis and 52.1% as positive for N . gonorrhoeae . The sensitivity and specificity of the Amplicor CT/NG PCR on male urine specimens for C . trachomatis were 80.0% (8/10) and 95.2% (60/63), compared with 60.0% (6/10) and 100.0% (63/63) for EIA/IF on urethral swabs . For N . gonorrhoeae, the sensitivity and specificity of the Amplicor CT/NG PCR on male urine specimens were both 100% (38/38 and 35/35, respectively) compared with 86.8% (33/38) and 100% (35/35) for microscopy and/or culture on urethral swabs . The results of this study indicate that the Amplicor CT/NG multiplex PCR test for C . trachomatis and N . gonorrhoeae performed on urine in males provides a highly sensitive, specific, and robust method for the diagnosis of both C . trachomatis and N . gonorrhoeae, for the early detection of both symptomatic and asymptomatic infected individuals.

J Mol Biol, 1999 Apr 2, 287(3), 499 - 510
Detection of anticodon nuclease residues involved in tRNALys cleavage specificity; Meidler R et al.; The tRNALys-specific anticodon nuclease exists in latent form in Escherichia coli strains containing the optional prr locus . The latency is a result of a masking interaction between the anticodon nuclease core-polypeptide PrrC and the Type IC DNA restriction-modification enzyme EcoprrI . Activation of the latent enzyme by phage T4-infection elicits cleavage of tRNALys 5' to the wobble base, yielding 5'-OH and 2', 3'-cyclic phosphate termini . The N-proximal half of PrrC has been implicated with (A/G) TPase and EcoprrI interfacing activities . Therefore, residues involved in recognition and cleavage of tRNALys were searched for at the C-half . Random mutagenesis of the low-G+C portion encoding PrrC residues 200-313 was performed, followed by selection for loss of anticodon nuclease-dependent lethality and production of full-sized PrrC-like protein . This process yielded a cluster of missense mutations mapping to a region highly conserved between PrrC and two putative Neisseria meningitidis MC58 homologues . This cluster included two adjacent members that relaxed the inherent enzyme's cleavage specificity . We also describe another mode of relaxed specificity, due to mere overexpression of PrrC . This mode was shared by wild-type PrrC and the other mutant alleles . The additional substrates recognised under the promiscuous conditions had, in general, anticodons resembling that of tRNALys . Taken together, the data suggest that the anticodon of tRNALys harbours anticodon nuclease identity elements and implicates a conserved region in PrrC in their recognition .

J Infect, 1999 Jan, 38(1), 45 - 7
Identification of the cause of a brain abscess by direct 16S ribosomal DNA sequencing; Logan JM et al.; We report the case of a young man who apparently suffered successive episodes of meningitis and cerebral abscess over a 1-month period, both of which were diagnosed by two different molecular approaches; PCR for Neisseria meningitidis IS1106 from CSF and 16S rRNA gene sequencing on a specimen of brain pus . In each case, cultures were negative due to prior antibiotic therapy.

J Infect Dis, 1999 Mar, 179 Suppl 2, S375 - 9
Human experimentation with Neisseria gonorrhoeae: progress and goals; Cohen MS et al.; Infection with Neisseria gonorrhoeae has adverse consequences for reproductive health and facilitates the transmission of the human immunodeficiency virus . A major limitation in the development of gonococcal vaccines has been the lack of an animal model . Urethral infection can be initiated in male volunteer subjects through urethral inoculation . Several hundred patients have participated in studies using this experimental infection model . These studies have helped define the natural history of experimental infection and provided a better understanding of phenotypic and genotypic variation of gonococci in vivo . Isogenic molecular mutants can be used to define a role for gonococcal surface structures, including pilin and transferrin-binding proteins; recent results demonstrate that gonococci unable to express transferrin- and lactoferrin-binding proteins cannot cause urethral infection . The experimental model has proven to be an efficient means of studying gonococcal infection and focusing vaccine development . In addition, this model should allow vaccines to be tested quickly and efficiently.

Vaccine, 1999 Jan, 17(1), 72 - 81
Oral immunization with a Streptococcal pneumoniae polysaccharide conjugate vaccine in enterocoated microparticles induces serum antibodies against type specific polysaccharides; Flanagan MP et al.; The authors examined the antibody responses of mice orally immunized with pneumococcal polysaccharide (type 23F) or a pneumococcal polysaccharide conjugated to the outer membrane protein complex of Neisseria meningitides (23F-OMPC) . These antigens were administered either in solution or entrapped within microcapsules . Only the mice receiving the encapsulated conjugate vaccine produced significant levels of anti-polysaccharide serum antibodies . These responses, observed after a second oral immunization with the conjugate, were predominantly IgG . Thus, the conversion from a T-cell-independent to a T-cell dependent response, achieved through conjugation was maintained following oral delivery . However, no local secretory IgA anti-polysaccharide response was detected in these mice indicating that while the OMPC carrier augments orally induced IgG responses, it was insufficient for the induction of secretory IgA.

Microbiology, 1999 Feb, 145 ( Pt 2), 389 - 400
Characterization of the recD gene of Neisseria gonorrhoeae MS11 and the effect of recD inactivation on pilin variation and DNA transformation; Chaussee MS et al.; Pilin antigenic variation in Neisseria gonorrhoeae may result following intrachromosomal recombination between homologous pil genes . Despite extensive study, recA is the only previously characterized gene known to be involved in this process . In this study, the gonococcal recD gene, encoding one subunit of the putative RecBCD holoenzyme, was characterized and its role in pilin variation assessed . The complete recD gene of N . gonorrhoeae MS11 was cloned and its nucleotide sequence determined . The gonococcal recD gene complemented a defined Escherichia coli recD mutant, based on plaque formation of bacteriophage lambda and the restoration of ATP-dependent nuclease activity . Inactivation of the gonococcal recD gene had no measurable effect on cell viability or survival following UV exposure, but did decrease the frequency of DNA transformation approximately threefold . The frequency at which non-parental pilin phenotypes were spawned was 12-fold greater in MS11 recD mutants compared with the parental MS11 rec+ strain . Similar results were obtained using recD mutants that were not competent for DNA transformation . Complementation of the MS11 recD mutant with a wild-type recD gene copy restored the frequency of pilin phenotypic variation to approximately wild-type levels . The nucleotide changes at pilE in the recD mutants were confined to the variable regions of the gene and were similar to changes previously attributed to gene conversion.

Int J Gynaecol Obstet, 1998 Dec, 63 Suppl 1, S183 - 9
Current approach to STD management in women; Amaral E; HIV infection was recognized as a new sexually transmitted disease (STD) at the beginning of the last decade . The knowledge of risk factors for sexual transmission of HIV changed the focus on STD to a broader perspective for prevention and control of HIV infection, and consequently of STD . Barriers to STD control include cultural aspects, difficulties in changing sexual behavior, asymptomatic disease in women and expensive and inaccessible tests for diagnosis . The classical clinical approach based on etiologic treatment has never been achieved by developing countries . The international community has been searching for new approaches . Syndromic management and mass treatment are strategies recently found as useful . Nevertheless the best approach to endocervicitis by Neisseria gonorrhoeae and Chlamydia trachomatis remain problematic . Then, the current approach to STD management must include: prompt attention to every patient seeking care for STD; early diagnosis and treatment; delivery of short term treatment at the clinic; education on STD/HIV; screening for other STDs with pre- and post-test counseling; counseling on risk reduction; provision of condoms; integration of STD services with family planning, prenatal and gynecological services.

J Clin Microbiol, 1999 Apr, 37(4), 1130 - 6
Antimicrobial susceptibilities and plasmid contents of Neisseria gonorrhoeae isolates from commercial sex workers in Dhaka, Bangladesh: emergence of high-level resistance to ciprofloxacin; Bhuiyan BU et al.; Commercial sex workers (CSWs) serve as the most important reservoir of sexually transmitted diseases (STD), including gonorrhea . Periodic monitoring of the antimicrobial susceptibility profile of Neisseria gonorrhoeae in a high-risk population provides essential clues regarding the rapidly changing pattern of antimicrobial susceptibilities . A study concerning the prevalence of gonococcal infection among CSWs was conducted in Bangladesh . The isolates were examined with regards to their antimicrobial susceptibility to, and the MICs of, penicillin, tetracycline, ciprofloxacin, cefuroxime, ceftriaxone, and spectinomycin by disk diffusion and agar dilution methods . The total plasmid profile of the isolates was also analyzed . Of the 224 CSWs, 94 (42%) were culture positive for N . gonorrhoeae . There was a good correlation between the results of the disk diffusion and agar dilution methods . Some 66% of the isolates were resistant to penicillin, and 34% were moderately susceptible to penicillin . Among the resistant isolates, 23.4% were penicillinase-producing N . gonorrhoeae (PPNG) . 60.6% of the isolates were resistant and 38.3% were moderately susceptible to tetracycline, 17.5% were tetracycline-resistant N . gonorrhoeae, 11.7% were resistant and 26.6% had reduced susceptibility to ciprofloxacin, 2.1% were resistant and 11.7% had reduced susceptibility to cefuroxime, and 1% were resistant to ceftriaxone . All PPNG isolates contained a 3.2-MDa African type of plasmid, and a 24.2-MDa conjugative plasmid was present in 34.1% of the isolates . Since quinolones such as ciprofloxacin are recommended as the first line of therapy for gonorrhea, the emergence of significant resistance to ciprofloxacin will limit the usefulness of this drug for treatment of gonorrhea in Bangladesh.

Curr Opin Microbiol, 1998 Feb, 1(1), 75 - 81
Fimbriae-mediated host-pathogen cross-talk; Abraham SN et al.; Recent studies show that the coupling of fimbrial adhesins of uropathogenic Escherichia coli and pathogenic Neisseria species to their complementary receptors on host cells is a dynamic event, involving specific signaling to the bacteria as well as to the host cell . These studies have unveiled intriguing and novel mechanisms by which bacteria utilize their fimbriae to promote virulence at the mucosal surface and in deeper tissue.

East Afr Med J, 1998 Oct, 75(10), 572 - 5
Isolation of Neisseria gonorrhoeae using urine as a holding medium; Ihunwo OA et al.; OBJECTIVE: To utilise urine as a holding medium in isolating Neisseria gonorrhoeae in place of urethral, high-vaginal and endocervical swabs . DESIGN: Screening and diagnostic test . SETTING: A rural community in Jos and Applied Microbiology Research Unit, University of Jos, Jos, Nigeria . SUBJECTS: A random sample of 300 asymptomatic individuals (151 males, 149 females) . MAIN OUTCOME MEASURE: Neisseria gonorrhoeae identification . RESULT: Nineteen Neisseria gonorrhoeae isolates were obtained with the isolation rate to 6.3% . Twelve isolates from males and seven isolates from females representing 63.7% and 36.8% respectively . The peak age group was 21-25 years with 12 positive cases (63.2%) . CONCLUSION: Urine culture proved an effective and painless alternative in screening N . gonorrhoeae in asymptomatic individuals . The age group 21-25 years is suggested for intervention studies . This method is less expensive and easily accepted by subjects as confidentiality is also maintained.

Bull World Health Organ, 1999, 77(1), 22 - 8
Unrecognized sexually transmitted infections in rural South African women: a hidden epidemic; Wilkinson D et al.; Sexually transmitted infections (STIs) are of major public health concern in developing countries, not least because they facilitate transmission of human immunodeficiency virus (HIV) . The present article presents estimates of the prevalence, on any given day, of STIs among women in rural South Africa and the proportion who are asymptomatic, symptomatic but not seeking care, and symptomatic and seeking care . The following data sources from Hlabisa district were used: clinical surveillance for STI syndromes treated in health facilities, microbiological studies among women attending antenatal and family planning clinics, and a community survey . Population census provided denominator data . Adequacy of drug treatment was determined through quality of care surveys . Of 55,974 women aged 15-49 years, a total of 13,943 (24.9%) were infected on any given day with at least one of Trichomonas vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, or Treponema pallidum . Of the women investigated, 6697 (48%) were asymptomatic, 6994 (50%) were symptomatic but not seeking care, 238 (1.7%) were symptomatic and would seek care, and 14 (0.3%) were seeking care on that day . Only 9 of the 14 women (65%) were adequately treated . STIs remained untreated because either women were asymptomatic or the symptoms were not recognized and acted upon . Improved case management alone is therefore unlikely to have a major public health impact . Improving partner treatment and women's awareness of symptoms is essential, while the potential of mass STI treatment needs to be exploredPIP: Estimates of the prevalence, on any given day, of sexually transmitted infections (STIs) among rural South African women and the proportion of these women who are asymptomatic, symptomatic but not seeking care, and symptomatic and seeking care were developed through reference to data from the Hlabisa health district (KwaZulu/Natal) . The prevalence of HIV among women attending antenatal clinics in Hlabisa rose from 4.2% in 1992 to 22% in 1997 . Data sources included clinical surveillance for STI syndromes treated in health facilities, microbiologic studies among women attending antenatal and family planning clinics, and a community survey . The 1991 population census provided denominator data . It was estimated that, on any given day, 13,943 district women 15-49 years of age were infected with at least one STI--a point prevalence of 24.9% of the resident female population in this age group . Of these women, 6697 (48%) were asymptomatic and 7246 (52%) were symptomatic . Only 252 symptomatic women (2%) sought care and, of these, just 164 (65%) received adequate treatment . The majority of women with an STI were in the 15-29 year age group . Pregnant women were significantly less likely than nonpregnant women to have an asymptomatic infection (1% vs . 59%) . Among nonpregnant women, syphilis and Trichomonas vaginalis were more likely to be asymptomatic (67% and 58%, respectively) than infection with Neisseria gonorrhoeae (37%) or Chlamydia trachomatis (36%) . These findings indicate that improved STI case management alone is unlikely to improve STI control . Improving women's recognition of and treatment-seeking behavior for abnormal urogenital symptoms should increase the numbers treated, but a large burden of asymptomatic infection will remain unless screening or mass treatment is considered .

APMIS, 1998 Dec, 106(12), 1181 - 7
Production, isolation and purification of bacteriocins expressed by two strains of Neisseria meningitidis; Allunans J et al.; The systemic Neisseria meningitidis strain P241 and the healthy pharyngeal carrier strain BT878 produce bacteriocin-like substances during growth . A method has been devised for obtaining the active substances in solution . The activity was recovered by freeze-thaw extraction of dialyzed Todd-Hewitt agar medium into which the bacteriocins had diffused during growth of the producer strains . The bacteriocins were purified more than 50-fold by ammonium-sulphate precipitation and hydrophobic interaction chromatography . They are quite stable to heat and remain active 100% after 30 min at 100 degrees C . However, the protein nature of the bacteriocins has been confirmed by their sensitivity to alpha-chymotrypsin . Gel filtration indicated an Mr of 100-110 kDa, whereas SDS-polyacrylamide gel electrophoresis produced a common band by Coomassie staining corresponding to an Mr of 47-48 kDa, suggesting a dimer form of the active protein component.

Proc Natl Acad Sci U S A, 1999 Mar 2, 96(5), 2408 - 13
Pseudomonas aeruginosa killing of Caenorhabditis elegans used to identify P . aeruginosa virulence factors; Tan MW et al.; We reported recently that the human opportunistic pathogen Pseudomonas aeruginosa strain PA14 kills Caenorhabditis elegans and that many P . aeruginosa virulence factors (genes) required for maximum virulence in mouse pathogenicity are also required for maximum killing of C . elegans . Here we report that among eight P . aeruginosa PA14 TnphoA mutants isolated that exhibited reduced killing of C . elegans, at least five also exhibited reduced virulence in mice . Three of the TnphoA mutants corresponded to the known virulence-related genes lasR, gacA, and lemA . Three of the mutants corresponded to known genes (aefA from Escherichia coli, pstP from Azotobacter vinelandii, and mtrR from Neisseria gonorrhoeae) that had not been shown previously to play a role in pathogenesis, and two of the mutants contained TnphoA inserted into novel sequences . These data indicate that the killing of C . elegans by P . aeruginosa can be exploited to identify novel P . aeruginosa virulence factors important for mammalian pathogenesis.

J Bacteriol, 1999 Mar, 181(5), 1395 - 402
Natural competence for DNA transformation by Legionella pneumophila and its association with expression of type IV pili; Stone BJ et al.; We have recently described the expression of two pili of different lengths on the surface of Legionella pneumophila (B . J . Stone and Y . Abu Kwaik, Infect . Immun . 66:1768-1775, 1998) . Production of long pili requires a functional pilEL locus, encoding a type IV pilin protein . Since type IV pili in Neisseria gonorrhoeae are associated with competence for DNA transformation, we examined the competence of L . pneumophila for DNA transformation under conditions that allowed the expression of type IV pili . We show that L . pneumophila is naturally competent for DNA transformation by isogenic chromosomal DNA and by plasmid DNA containing L . pneumophila DNA . Many different L . pneumophila loci are able to transform L . pneumophila after addition of plasmid DNA, including gspA, ppa, asd, and pilEL . The transformation frequency is reduced when competing DNA containing either L . pneumophila DNA or vector sequences is added to the bacteria, suggesting that uptake-specific sequences may not be involved in DNA uptake . Competence for DNA transformation correlates with expression of the type IV pili, and a pilEL mutant defective in expression of type IV pili is not competent for DNA transformation . Complementation of the mutant for competence is restored by the reintroduction of a cosmid that restores production of type IV pili . Minimal competence is restored to the mutant by introduction of pilEL alone . We conclude that competence for DNA transformation in L . pneumophila is associated with expression of the type IV pilus and results in recombination of L . pneumophila DNA into the chromosome . Since expression of type IV pili also facilitates attachment of L . pneumophila to mammalian cells and protozoa, we designated the type IV pili CAP (for competence- and adherence-associated pili).

Mol Microbiol, 1999 Feb, 31(3), 903 - 13
Mutagenesis of the Neisseria gonorrhoeae porin reduces invasion in epithelial cells and enhances phagocyte responsiveness; Bauer FJ et al.; Porin (PorB), the major outer membrane protein of Neisseria gonorrhoeae, has been implicated in pathogenesis previously . However, the fact that porin deletion mutants are not viable has complicated investigations . Here, we describe a method of manipulating the porin gene site-specifically . N . gonorrhoeae MS11, which harbours the porB1B (P.1B) porin allele, was used to generate mutants carrying deletions in the surface loops 1 and 5 . An 11-amino-acid deletion in loop 1 impaired Opa50-dependent invasion into human Chang epithelial cells, whereas loop 5 deletion exhibited no apparent phenotype . In a second approach, the complete gonococcal porB1B was replaced by the porBNia gene of Neisseria lactamica . Such mutants were unable to induce efficient uptake by epithelial cells but induced an enhanced respiratory response in HL60 phagocytic cells . The increased respiratory burst was accompanied by an enhanced phagocytic uptake of the mutant compared with the wild-type strain . Our data extend previous evidence for multiple central functions of PorB in the infection process.

Mol Microbiol, 1999 Feb, 31(3), 743 - 52
Crystallographic structure reveals phosphorylated pilin from Neisseria: phosphoserine sites modify type IV pilus surface chemistry and fibre morphology; Forest KT et al.; Understanding the structural biology of type IV pili, fibres responsible for the virulent attachment and motility of numerous bacterial pathogens, requires a detailed understanding of the three-dimensional structure and chemistry of the constituent pilin subunit . X-ray crystallographic refinement of Neisseria gonorrhoeae pilin against diffraction data to 2.6 A resolution, coupled with mass spectrometry of peptide fragments, reveals phosphoserine at residue 68 . Phosphoserine is exposed on the surface of the modelled type IV pilus at the interface of neighbouring pilin molecules . The site-specific mutation of serine 68 to alanine showed that the loss of the phosphorylation alters the morphology of fibres examined by electron microscopy without a notable effect on adhesion, transformation, piliation or twitching motility . The structural and chemical characterization of protein phosphoserine in type IV pilin subunits is an important indication that this modification, key to numerous regulatory aspects of eukaryotic cell biology, exists in the virulence factor proteins of bacterial pathogens . These O-linked phosphate modifications, unusual in prokaryotes, thus merit study for possible roles in pilus biogenesis and modulation of pilin chemistry for optimal in vivo function.

Curr Opin Microbiol, 1999 Feb, 2(1), 62 - 70
Host cell interactions and signalling with Neisseria gonorrhoeae; Naumann M et al.; Neisseria gonorrhoeae is a highly adapted human pathogen that utilises multiple adhesins to interact with a variety of host cell receptors . Recently, substantial progress has been made in unravelling the signalling events induced by N . gonorrhoae that can lead to cytoskeletal reorganisation, invasion or phagocytic uptake, intraphagosomal accommodation, nuclear signalling, cytokine/chemokine release and apoptosis.

Trends Microbiol, 1998 Dec, 6(12), 489 - 95
The role of neisserial Opa proteins in interactions with host cells; Dehio C et al.; Pathogenic Neisseria spp . possess a repertoire of phase-variable Opa proteins that mediate various pathogen--host cell interactions, including bacterial engulfment by epithelial cells and opsonin-independent phagocytosis by professional phagocytes . Recent studies have identified cellular targets recognized by defined Opa proteins and have begun to reveal host signalling events involved in mediating these Opa-dependent cellular processes.

Infect Immun, 1999 Mar, 67(3), 1149 - 56
Invasion of human mucosal epithelial cells by Neisseria gonorrhoeae upregulates expression of intercellular adhesion molecule 1 (ICAM-1); Jarvis GA et al.; Infection of the mucosa by Neisseria gonorrhoeae involves adherence to and invasion of epithelial cells . Little is known, however, about the expression by mucosal epithelial cells of molecules that mediate cellular interactions between epithelial cells and neutrophils at the site of gonococcal infection . The aim of this study was to determine the expression of intercellular adhesion molecule 1 (ICAM-1) by epithelial cells during the process of gonococcal invasion . The highly invasive strain FA1090 and the poorly invasive strain MS11 were incubated with human endometrial adenocarcinoma (HEC-1-B) or human cervical carcinoma (ME-180) epithelial cells, after which ICAM-1 expression was measured by flow cytometry . After 15 h of infection with FA1090, expression of ICAM-1 increased 4.7- and 2.1-fold for HEC-1-B and ME-180 cells, respectively, whereas 15 h of infection of HEC-1-B cells with MS11 increased ICAM-1 expression only 1.6-fold . ICAM-1 expression was restricted to the cell surface, since no soluble ICAM-1 was detected . The distribution of staining was heterogeneous and mimicked that seen after treatment of HEC-1-B cells with the ICAM-1 agonist tumor necrosis factor alpha (TNF-alpha) in the absence of bacteria . PCR and dot blot analyses of ICAM-1 mRNA showed no change in levels over time in response to infection . Although TNF-alpha was produced by HEC-1-B cells after infection, the extent of ICAM-1 upregulation was not affected by neutralizing anti-TNF-alpha antiserum . Dual-fluorescence flow cytometry showed that the cells with the highest levels of ICAM-1 expression were cells with associated gonococci . We conclude that epithelial cells upregulate the expression of ICAM-1 in response to infection with invasive gonococci . On the mucosa, upregulation of ICAM-1 by infected epithelial cells may function to maintain neutrophils at the site of infection, thereby reducing further invasion of the mucosa by gonococci.

Am J Epidemiol, 1999 Feb 15, 149(4), 353 - 8
Longitudinal evaluation of serovar-specific immunity to Neisseria gonorrhoeae; Fox KK et al.; The serovars of Neisseria gonorrhoeae that are predominant in a community change over time, a phenomenon that may be due to the development of immunity to repeat infection with the same serovar . This study evaluated the epidemiologic evidence for serovar-specific immunity to N . gonorrhoeae . During a 17-month period in 1992-1994, all clients of a sexually transmitted disease clinic in rural North Carolina underwent genital culture for N . gonorrhoeae . Gonococcal isolates were serotyped according to standard methods . Odds ratios for repeat infection with the same serovar versus any different serovar were calculated on the basis of the distribution of serovars in the community at the time of reinfection . Of 2,838 patients, 608 (21.4%; 427 males and 181 females) were found to be infected with N . gonorrhoeae at the initial visit . Ninety patients (14.8% of the 608) had a total of 112 repeat gonococcal infections . Repeat infection with the same serovar occurred slightly more often than would be expected based on the serovars prevalent in the community at the time of reinfection, though the result was marginally nonsignificant (odds ratio = 1.5, 95% confidence interval 1.0-2.4; p = 0.05) . Choosing partners within a sexual network may increase the likelihood of repeat exposure to the same serovar of N . gonorrhoeae . Gonococcal infection did not induce evident immunity to reinfection with the same serovar.

Infect Immun, 1999 Mar, 67(3), 1517 - 20
Characterization of Moraxella (Branhamella) catarrhalis lbpB, lbpA, and lactoferrin receptor orf3 isogenic mutants; Bonnah RA et al.; Pathogenic members of the family Neisseriaceae produce specific receptors to acquire iron from their host's lactoferrin and transferrin . Recently, putative Moraxella catarrhalis lactoferrin receptor genes and a third open reading frame (lbpB, lbpA, and orf3) were cloned and sequenced . We describe the preliminary characterization of isogenic mutants deficient in LbpB, LbpA, or Orf3 protein.

Infect Immun, 1999 Mar, 67(3), 1405 - 14
Relationship between UDP-glucose 4-epimerase activity and oligoglucose glycoforms in two strains of Neisseria meningitidis; Lee FK et al.; Sodium dodecyl sulfate-polyacrylamide gel analysis of lipooligosaccharide (LOS) from Neisseria meningitidis has demonstrated considerable microheterogeneity in the variable region of LOS due to the presence of novel glycoforms . As a step toward understanding the basis for the expression of these novel glycoforms, we have examined the LOS structures and UDP-glucose 4-epimerase (epimerase) activity levels in two strains (NMB and MA-1) and their respective galE mutants . Strain NMB was found to have low epimerase activity and to contain multiple glycoforms, some of which appear to contain only glucose sugars . The galE mutant had only the oligoglucose glycoforms . Strain MA-1 had higher epimerase activity at both log and stationary phases (2- and 12.5-fold, respectively) and one glycoform with a putative lactosyl structure . Strain MA-1 galE had two glycoforms that contained one or two glucose residues . To understand the molecular basis for the different epimerase activities, we examined the predicted amino acid sequences of the respective galE open reading frames and determined the relative amounts of GalE protein . We found no significant differences between the predicted amino acid sequence of the GalE protein in NMB and that in MA-1 . We observed no significant differences in the level of GalE protein between MA-1 and NMB at exponential or stationary phase . We also observed an 8.2-fold drop in epimerase activity in NMB between the log and stationary phases that was not due to the GalE protein level or low glucose levels.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S147 - 52
Screening for sexually transmitted diseases in rural South African women; Schneider H et al.; OBJECTIVE: This paper reports on a study undertaken in a rural area of South Africa, to develop a non-laboratory tool to screen for sexually transmitted diseases (STDs) among family planning clients . METHODS: A cross sectional study was performed of 249 consecutive women attending a family planning service between November and December 1994 . A questionnaire was administered, and a clinical examination and laboratory tests conducted . Sociodemographic, clinical, and other non-laboratory variables that were significantly associated with laboratory evidence of infection were combined to produce non-hierarchical scoring systems for three "syndromes": gonococcal and/or chlamydial cervical infection, trichomoniasis, and cervical infection and/or trichomoniasis combined . The sensitivity, specificity, and predictive values of the scoring systems as a screening tool were assessed against the gold standard of laboratory tests . RESULTS: The prevalence of reproductive tract infections among the study participants was as follows: Chlamydia trachomatis 12%, Neisseria gonorrhoeae 3%, Trichomonas vaginalis 18%, and bacterial vaginosis 29% . Although vaginal discharge and other symptoms were frequently reported, symptoms bore no relation to the presence of infection . The following independent associations with gonococcal/chlamydial cervical infection were found: age less than 25 years and cervical mucopus and/or friability . Abnormal discharge on examination, visible inflammatory changes of the cervix (increased redness), no recent travel, and unemployment were associated with trichomoniasis . The combination of trichomonas and/or cervical infection ("STD syndrome") was associated with cervical mucopus/friability, unemployment, lack of financial support, and increased redness of the cervix . Of the three scoring systems developed on the basis of these associations, that of the "STD syndrome" achieved the best performance characteristics as a screening tool, with a sensitivity of 62%, specificity of 74%, and positive predictive value of 48% . CONCLUSION: STDs are common in a population of rural, sexually active women attending a family planning service . In resource poor settings, non-laboratory screening tools could play some role in identifying and treating infections in these women, especially since the majority would not otherwise have been reached . However, such screening tools cannot be viewed as the only way to identify STDs and should be considered as part of an overall strategy of STD control that includes, for example, good management of symptomatic individuals and their partners.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S139 - 46
Risk scores to detect cervical infections in urban antenatal clinic attenders in Mwanza, Tanzania; Mayaud P et al.; BACKGROUND: Detection and management of gonococcal and/or chlamydial infections in women is a challenge, particularly in developing countries where laboratory tests are not always available . The World Health Organisation (WHO) has developed a risk assessment approach to identify cervical infections among women complaining of vaginal discharge . We have evaluated this approach as a screening strategy among women attending an urban antenatal clinic (ANC) in Tanzania . OBJECTIVES: (i) To measure the prevalence of pathogens associated with sexually transmitted diseases (STD) and reproductive tract infections (RTI) in an urban population of ANC attenders in Tanzania; (ii) to examine characteristics of pregnant women associated with cervical infections; and (iii) to evaluate the performance of a WHO risk assessment algorithm and alternative risk scores for the detection of cervical infections in pregnant women . METHODS: A systematic sample of 660 pregnant women reporting for routine antenatal care at an urban clinic was enrolled . Women were interviewed by a nurse, who applied the WHO risk score . They were referred to a study room for interview about sociodemographic and behavioural factors, examination, and sampling for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Candida albicans, and bacterial vaginosis . Sensitivity, specificity, positive predictive value (PPV), and theoretical cost per true case treated were estimated for the diagnosis of cervical infection with N gonorrhoeae and/or C trachomatis for the WHO and other risk scores . RESULTS: The prevalence of any vaginal or cervical infection was 68% . Prevalence rates of various pathogens were: C albicans 39%, T vaginalis 16%, bacterial vaginosis 24%, N gonorrhoeae 2.3%, C trachomatis 5.9%, any cervical infection (N gonorrhoeae and/or C trachomatis) 7.4% . The WHO score identified only five of 49 women with N gonorrhoeae and/or C trachomatis (sensitivity 10.2%) . The specificity and the PPV were 92% and 9.8% respectively . The theoretical cost per true case treated on the basis of the WHO score was over $18 . Several risk factors were associated with cervical infection on univariate analysis, but only six remained significant at the 10% level after multivariate analysis . These were: never use of contraceptives (OR 3.09), more than one partner in the past 3 months (OR 3.32), partner with symptoms of genital discharge syndrome (GDS) (OR 7.55), frothy vaginal discharge (OR 1.88), 5-19 polymorphonucleocytes per high power field on cervical smear (OR 3.28), or more than 20 polymorphonucleocytes per high power field (OR 16.08), and wet preparation showing evidence of T vaginalis infection (OR 1.96) . Scores based on these variables failed to attain high sensitivities or PPVs (all below 40%) although the costs per true case treated were cheaper than for the WHO score . CONCLUSION: Risk assessment for the screening and management of N gonorrhoeae and/or C trachomatis among women presenting at routine antenatal services appears feasible and acceptable, but of limited value in this population because of its low sensitivity . The optimal risk score may vary considerably from one place to another . The quest for simple, cheap, and reliable tests to diagnose N gonorrhoeae and C trachomatis infections still remains a high priority on the international STD technology research agenda.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S118 - 22
Syndromic approach to detection of gonococcal and chlamydial infections among female sex workers in two Philippine cities; Wi T et al.; BACKGROUND: In many developing countries, STD control efforts often involve registration and periodic examinations of female sex workers (FSW) . Non-availability of sensitive and specific diagnostic tests frequently constrain this approach . METHODS: A model for detection of Chlamydia trachomatis or Neisseria gonorrhoeae in FSW on the basis of risk assessment and examination was developed from data gathered in Manila and evaluated in a second city (Cebu) in the Republic of the Philippines . RESULTS: Gonococcal or chlamydial cervical infection was found in 23.3% of FSW in Manila and 37.0% in Cebu . Unregistered and younger FSW had greatest risk of chlamydial infection and/or gonorrhoea in both cities . In Manila, where gynaecologists performed the pelvic examinations, signs of cervical mucopus or cervical motion, uterine or cervical motion tenderness in women under < 25 years old or unregistered had positive predictive value (PPV) of 0.60 and sensitivity of 42.1% for cervical infection . In Cebu, where women were not examined by gynaecologists, the same model had high PPV, but a sensitivity of only 12.3% . CONCLUSIONS: Experience and training of clinicians undoubtedly can influence the yield of examination in syndromic management of cervical infection . Nevertheless, inexpensive and diagnostic tests are needed for detection of cervical infection in this population.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S106 - 11
Evaluation of simple diagnostic algorithms for Neisseria gonorrhoeae and Chlamydia trachomatis cervical infections in female sex workers in Abidjan, Côte d'Ivoire; Diallo MO et al.; OBJECTIVE: To generate simple algorithms for the diagnosis of cervical infection with Neisseria gonorrhoeae or Chlamydia trachomatis in female sex workers in Abidjan, Cote d'Ivoire and to evaluate their validity . METHODS: From October 1992 to the end of June 1993, female sex workers were interviewed and clinically examined at a confidential clinic . N gonorrhoeae was cultured on modified Thayer-Martin medium and C trachomatis was detected by polymerase chain reaction . The associations of gonococcal or chlamydial cervical infection with sociodemographic, behavioural, clinical, and biological factors were assessed and three algorithms were generated . The validity parameters of these diagnostic algorithms were calculated and compared to those of standard algorithms and mass treatment . RESULTS: Among 683 women, cervical infection was present in 239 (35%) . The sensitivity an algorithm incorporating sociodemographic and behavioural factors and symptoms, of an algorithm incorporating clinical signs and simple laboratory tests, and of a combined algorithm was 83%, 86%, and 79% respectively while the specificity was 32%, 44%, and 54%, and the positive predictive value 40%, 46%, and 48% respectively . A standard algorithm incorporating only the symptom vaginal discharge, and a standard algorithm requiring both the symptom vaginal discharge and the presence of an endocervical mucopurulent discharge on examination had a sensitivity of 44% and 18%, a specificity of 75% and 95%, and a positive predictive value of 49% and 67% respectively . CONCLUSIONS: The algorithms generated in this study may be useful for the control of cervical infections in female sex workers in resource poor settings in the absence of rapid, inexpensive, and accurate laboratory tests for the diagnosis of cervical infections.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S95 - 105
Reproductive tract infections in primary healthcare, family planning, and dermatovenereology clinics: evaluation of syndromic management in Morocco; Ryan CA et al.; OBJECTIVE: To determine where and with what symptoms women seek care for reproductive tract infections (RTI) in Morocco and to guide allocation of resources for training and treatment for RTIs . METHODS: A primary healthcare centre (PHC), a family planning centre (FPC), and a specialty dermatovenereology clinic (SC) were selected in each of three urban areas . Women with symptoms of vaginal discharge, lower abdominal or pelvic pain, or genital lesions (genital ulcer or warts) underwent interviews, physical examinations, serological testing for human immunodeficiency virus (HIV) and syphilis, and collection of vaginal fluid for microscopic examination, and urine for detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) by ligase chain reaction (LCR) . RESULTS: Over 8 months, 1238 women enrolled, including 61.8% at PHCs, 34.8% at FPCs, and 3.4% at SCs . Overall, 54% complained of vaginal discharge, of whom 8.8% had GC or CT infection and 30.1% had trichomoniasis (TV) or bacterial vaginosis (BV); 24.9% complained of lower abdominal pain with or without vaginal discharge, of whom 7.3% had GC or CT and 22.6% had TV or BV . GC or CT infections were found in 10.1% of PHC and 5.4% of FPC patients; while TV and/or BV infections were found in 28.7% and 22.8%, respectively . GC or CT infection was associated with perceived risk behaviours of the male partner (for example, belief partner is unfaithful) more often than with reported risk behaviours of the women themselves . For vaginal infections, a modified World Health Organisation (WHO) test algorithm for vaginal discharge involving risk assessment plus speculum and bimanual examination was 98.0% sensitive at PHCs and 90.8% at FPCs, with positive predictive value (PPV) of 33.4% at PHCs and 26.8% at FPCs . For GC or CT infections this algorithm was 60.6% sensitive at PHCs and 85.7% sensitive at FPCs; but PPV was only 9.9% and 9.0% respectively, little higher than the background prevalence of these infections . An RTI algorithm (Morocco specific) had comparable sensitivity and PPV for vaginal infection, and for cervical infection was less sensitive but had much higher PPV (26.9% for PHCs and 26.7% for FPCs) . CONCLUSION: Women with complaints of vaginal discharge and/or lower abdominal pain presented to PHC and FP clinics, not to SCs . PHCs and FPCs should therefore receive resources for management of vaginal discharge . Both the test algorithm and the new RTI algorithm were useful in allocating treatment for vaginal infection, but only the RTI algorithm discriminated in selecting women with cervical infection . Even with the RTI algorithm, which limited treatment for cervical infection to risk assessment positive patients with signs of cervical infection or PID, the PPV for cervical infection was low, potentially resulting in frequent overtreatment and problems of partner notification.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S85 - 94
Rapid and inexpensive approaches to managing abnormal vaginal discharge or lower abdominal pain: an evaluation in women attending gynaecology and family planning clinics in Peru; Sanchez SE et al.; OBJECTIVES: To assess low abdominal pain, yellow vaginal discharge, other symptoms and signs, and demographic and behavioural variables as predictors for cervical or vaginal infection . METHODS: A cross sectional study of women attending gynaecology and family planning clinics in Lima, Peru was undertaken . 630 consecutive eligible female patients with chief or elicited complaints of yellow vaginal discharge, low abdominal pain, or both were interviewed and examined, together with a comparable reference group without these complaints . Vaginal specimens were tested for trichomoniasis and bacterial vaginosis . Endocervical specimens were tested for Neisseria gonorrhoeae and Chlamydia trachomatis using the ligase chain reaction . RESULTS: Infections found included chlamydial infection in 69 women (10.9%), gonorrhoea in 10 (1.6%), and either infection in 77 (12.2%); trichomoniasis in 46 (7.3%), bacterial vaginosis in 189 (30%), and either infection in 209 (33.2%) . Cervical infection with C trachomatis and/or N gonorrhoeae was independently associated with history of a new sex partner within the last 3 months, more than one sex partner within the last year, use of condoms never or in less than 50% of sex acts, history of sex partner with STD within the last year; with symptoms of persistent low abdominal pain and of yellow vaginal discharge; and with signs of profuse and yellow vaginal discharge, cervical ectopy, easily induced endocervical bleeding, or brown cervical secretion . Using these findings, an algorithm was created that had a positive predictive value (PPV) of 36% for cervical infection among women reporting chief or elicited complaint of this abnormal vaginal discharge and a PPV of 25% among those without a complaint . A chief complaint of yellow vaginal discharge had a PPV of 50% for trichomoniasis or bacterial vaginosis . Among women without a chief complaint of yellow vaginal discharge, clinical findings of yellow vaginal discharge had a PPV of 55% . CONCLUSIONS: Where economic and technical constraints preclude testing, clinical findings and risk assessment are helpful in detecting vaginal and cervical infections . Several demographic, behavioural, clinical, and laboratory variables were predictive of infection in this population.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S77 - 84
Validation of a WHO algorithm with risk assessment for the clinical management of vaginal discharge in Mwanza, Tanzania; Mayaud P et al.; OBJECTIVES: (i) To determine the microbial aetiologies of vaginal discharge in STD clinic and antenatal clinic (ANC) attenders; (ii) to evaluate the performance and costs of a new WHO algorithm for the detection of gonococcal and chlamydial infections in women complaining of vaginal discharge and/or genital itching, using a risk assessment . METHODS: Two groups were enrolled: (i) 395 consecutive female patients attending a hospital outpatient clinic complaining of genital discharge or itching; and (ii) 628 consecutive pregnant women reporting at an urban ANC these symptoms . Patients were interviewed by a nurse, who applied the WHO risk score . They were then referred to the study room for interview concerning the same and other risk factors, examined, and sampled for Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Trichomonas vaginalis (TV), and Candida albicans (CA) . Sensitivity, specificity, positive predictive value, overtreatment and correct treatment rates, and cost of drugs per true case treated were estimated . RESULTS: The prevalence of NG and/or CT infections was 11.4% and 8% at the STD clinic and the ANC respectively . The most prevalent pathogens were CA (38% at both clinics) and TV (25% at the STD clinic and 34% at the ANC) . The sensitivity of the WHO algorithm for NG and/or CT was 62% at the STD clinic and 46% at the ANC, and the specificities were 64% and 84% respectively . The operational feasibility of the method was good . The cost of drugs per true case treated in applying the risk assessment approach was $3.5 among nonpregnant women and $5.0 among pregnant women . This compared favourably with respective costs of $8.8 and $25.0 in applying the syndromic management alone . CONCLUSIONS: The WHO risk assessment algorithm for the diagnosis of NG and/or CT infections among women complaining of genital discharge can considerably reduce overtreatment of NG and/or CT in both pregnant and non-pregnant women, but in this study it failed to identify 38% of non-pregnant and 54% of pregnant women with these infections . The elements of the risk score may need adjustment in different settings.

Sex Transm Infect, 1998 Jun, 74 Suppl 1, S50 - 8
Validation of the WHO diagnostic algorithm and development of an alternative scoring system for the management of women presenting with vaginal discharge in Malawi; Costello Daly C et al.; OBJECTIVE: To evaluate the performance of the WHO algorithm for the detection of cervical infection in women presenting with vaginal discharge and modify the risk assessment score for optimum effectiveness in Malawi . METHODS: 550 consecutive women presenting with non-ulcerative genitourinary complaints were interviewed and examined . Cervical infection was defined as presence of Neisseria gonorrhoeae on culture and/or Chlamydia trachomatis by EIA . Other laboratory investigations included wet mount microscopy, serology for syphilis and HIV, LED testing of cervical and vaginal secretions, and pH testing of vaginal fluid . Sensitivity, specificity, and positive predictive values (PPV) of different algorithms were determined in the analysis . RESULTS: Cervical infection was identified in 19.5% of women (17.1% gonorrhoea, 3.7% chlamydial infection) . The sensitivity/specificity/PPV of the WHO risk assessment were 43%/73%/28%, respectively by history and 62%/61%/27% with the addition of speculum examination . Using Malawi results to modify the risk assessment improved the performance to 61%/68%/31% respectively by history alone, which increased to 73%/64%/33% with bimanual examination and 72%/56%/29% with speculum examination . CONCLUSION: The sensitivity of the WHO risk assessment is low for the detection of cervical infection in Malawi . Although the Malawi risk assessment performed somewhat better on history alone, this study identified external and bimanual examination variables that improved the diagnostic performance of the algorithm in settings where speculum examination is not possible . Although the PPVs of the algorithms are low, country specific risk assessments can provide a framework for management until simple, affordable diagnostic tests for the definitive diagnosis of cervical infection are available.

Arch Pediatr Adolesc Med, 1999 Feb, 153(2), 119 - 25
Is the routine pelvic examination needed with the advent of urine-based screening for sexually transmitted diseases?
Shafer MA, Pantell RH, Schachter J.
OBJECTIVE: To determine the most cost-effective method of screening for chlamydia and gonorrhea to prevent pelvic inflammatory disease (PID) in asymptomatic sexually active adolescent females . DESIGN: Cost-effectiveness decision analysis comparing pelvic examination with cervical screening (the current national standard) with a model of urine screening with ligase chain reaction testing for Chlamydia trachomatis and Neisseria gonorrhoeae . METHODS: Four strategies using decision analysis were compared for a potential cohort of 100000 asymptomatic sexually active young women: (1) pelvic examination screening in 100%; (2) urine screening in 100%; (3) actual predicted pelvic examination screening in 70%; and (4) actual predicted urine screening in 90% . Assumptions and costs were generated from published sources . MAIN OUTCOME MEASURES: Cases of PID prevented per year and cost to prevent a case of PID . RESULTS: A total of 1750 cases of PID would be predicted to occur per year with no screening . Strategy 1 would prevent the most cases of PID (1283) at a mean cost of $10230 . Strategy 2 would prevent 1215 cases of PID at a mean cost of $5093 . The marginal cost to prevent an additional case of PID by strategy 1 is $101454 . Strategy 3 would prevent 898 cases of PID and 1093 cases of PID would be prevented with urine screening in strategy 4 . CONCLUSION: Urine-based ligase chain reaction screening is the most cost-effective strategy to detect chlamydial and gonococcal genital infection in asymptomatic sexually active adolescent females and, owing to ease of implementation, the most likely to prevent the greatest number of cases of PID.

J Infect Dis, 1999 Mar, 179(3), 729 - 33
Correlates of gonococcal infection and of antimicrobial-resistant Neisseria gonorrhoeae among female sex workers, Republic of the Philippines, 1996-1997; Klausner JD et al.; From 1994 to 1997, the proportion of Neisseria gonorrhoeae highly resistant to ciprofloxacin (MIC >/=4 microg/mL) increased substantially among female sex workers (FSWs) in the Philippines . Among 1499 Filipina FSWs, we evaluated factors associated with gonococcal infection and with gonococcal antimicrobial resistance . By multivariate analysis, gonococcal infection was associated with sex with a new client, self-prescribed prophylactic antimicrobial use, work in a brothel, and inconsistent condom use and was negatively associated with registration status and vaginal hygiene practices . Factors associated with ciprofloxacin-resistant gonococci included: marital status, living alone, duration of sex work, and clinic site . Further, gonococci highly resistant to ciprofloxacin were isolated from 10 (11.5%) of 87 FSWs reporting self-prescribed antimicrobial use versus 44 (3.4%) of 1295 reporting no antimicrobial use (P<.001) . Self-prescribed prophylactic antimicrobial use and inconsistent condom use could be important factors in the continued emergence of gonococcal antimicrobial resistance in the Philippines.

Zh Mikrobiol Epidemiol Immunobiol, 1998 Nov-Dec, (6), 33 - 7
{Persistence factors in the biological characterization of Neisseria gonorrhoeae}; Deriabin DG et al.; The study revealed the existence of differences between various representatives of the genus Neisseria gonorrhoeae, as well as intraspecific variations of N.gonorrhoeae by the spectrum of biological characteristics, including antilusozyme and anticomplement activity and resistance to the bactericidal action of blood serum and to the preparation of human leukocyte interferon . In contrast to other Neisseria species, all N.gonorrhoeae were characterized by a high level of anticomplement activity and resistance to the host's bactericidal systems, while a high level of antilysozyme activity, significantly exceeding that of gonococci, was found to be characteristics of other species of the genus Neisseriaceae . Within the species N.gonorrhoeae, the properties under study were associated into a single factor, closely correlated to the duration of the infectious process, which served as the basis for the development of a mathematical algorithm for the identification of persisting gonococcal strains.

Glycobiology, 1999 Feb, 9(2), 157 - 71
An essential saccharide binding domain for the mAb 2C7 established for Neisseria gonorrhoeae LOS by ES-MS and MSn; Muhlecker W et al.; A study of bacterial surface oligosaccharides were investigated among different strains of Neisseria gonorrhoeae to correlate structural features essential for binding to the MAb 2C7 . This epitope is widely expressed and conserved in gonococcal isolates, characteristics essential to an effective candidate vaccine antigen . Sample lipooligosaccharides (LOS), was prepared by a modification of the hot phenol-water method from which de-O-acetylated LOS and oligosaccharide (OS) components were analyzed by ES-MS-CID-MS and ES-MSnin a triple quadrupole and an ion trap mass spectrometer, respectively . Previously documented natural heterogeneity was apparent from both LOS and OS preparations which was admixed with fragments induced by hydrazine and mild acid treatment . Natural heterogeneity was limited to phosphorylation and antenni extensions to the alpha-chain . Mild acid hydrolysis to release OS also hydrolyzed the beta(1-->6) glycosidic linkage of lipid A . OS structures were determined by collisional and resonance excitation combined with MS and multistep MSn which provided sequence information from both neutral loss, and nonreducing terminal fragments . A comparison of OS structures, with earlier knowledge of MAb binding, enzyme treatment, and partial acid hydrolysis indicates a generic overlapping domain for 2C7 binding . Reoccurring structural features include a Hepalpha(1-->3)Hepbeta(1-->5)KDO trisaccharide core branched on the nonreducing terminus (Hep-2) with an alpha(1-->2) linked GlcNAc (gamma-chain), and an alpha-linked lactose (beta-chain) residue . From the central heptose (Hep-1), a beta(1-->4) linked lactose (alpha-chain), moiety is required although extensions to this residue appear unnecessary.

Diagn Microbiol Infect Dis, 1998 Dec, 32(4), 307 - 11
Antimicrobial activity of gatifloxacin tested against Neisseria gonorrhoeae using three methods and a collection of fluoroquinolone-resistant strains; Biedenbach DJ et al.; Gatifloxacin, a new 8-methoxy fluoroquinolone, was tested against 131 strains of Neisseria gonorrhoeae by reference agar dilution, disk diffusion, and Etest (AB BIODISK, Solna, Sweden) methods on supplemented GC agar . Gatifloxacin activity was equal to ciprofloxacin (MIC50, 0.008 microgram/mL) against strains fully susceptible to fluoroquinolones, but was generally four-fold more active (MIC90, 0.064-0.094 microgram/mL) against strains with par C or gyr A mutations and resistance to ciprofloxacin . Etest results were comparable to those generated by the agar dilution test {correlation coefficient (r) = 0.97} . Gatifloxacin zone diameters using 5-microgram disks also correlated well (r = 0.86-0.87) with the agar dilution and Etest MIC results . Breakpoints for laboratory testing of Neisseria gonorrhoeae strains await clinical trial outcome correlations, but susceptibility at < or = 0.125 or < or = 0.25 microgram/mL (> or = 34 mm) seems appropriate . All three tests used in this study seem applicable for laboratory testing of isolates from patients with uncomplicated gonorrhoeae receiving therapy with gatifloxacin.

Sex Transm Infect, 1998 Aug, 74(4), 276 - 8
Periurethral gland abscess: aetiology and treatment; Sanders CJ et al.; OBJECTIVES: To establish some characteristics of patients with periurethral gland abscess, its microbiological profile, and response to treatment . METHOD: The patients were seen at the Khami Road Clinic, Bulawayo, a municipal STD referral clinic, serving an urban population . Twenty consecutive men with periurethral abscesses were studied . Demographic data and a sexual history were obtained from each patient . Aspirates from the abscess cavities and urethral swabs were collected for microbiology, and blood samples taken for syphilis and HIV serology . The patients were treated by aspiration of the abscess cavities, followed by a single injection of kanamycin 2.0 g followed by a 1 week course of oral doxycycline 100 mg twice daily . RESULTS: Neisseria gonorrhoeae was cultured from three aspirates and five urethral specimens . Chlamydia trachomatis was found in two aspirates and three urethral specimens . Other organisms isolated included Gram negative and anaerobic bacilli . HIV antibody was detected in 13 of 18 patients tested . The response to initial treatment was good, but the abscesses ruptured in two patients, one of whom developed a urinary fistula . One patient required treatment with an alternative antimicrobial regimen . CONCLUSION: This study demonstrated a role for N gonorrhoeae and possibly for C trachomatis in the aetiology of periurethral abscess . The prevalence of HIV infection in these patients was high . The results of treatment of periurethral abscess by aspiration of pus and followed by antimicrobial therapy covering both N gonorrhoeae and C trachomatis were acceptable.

Sex Transm Infect, 1998 Aug, 74(4), 256 - 7
Managing STIs identified after testing outside genitourinary medicine departments: one model of care; Haddon L et al.; OBJECTIVES: To develop a local strategy for managing cases of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) which have been identified in the departments of obstetrics and gynaecology (O&G) . METHODS: Weekly notification from the local microbiology laboratory to genitourinary medicine (GUM) departments of all positive CT and GC results generated by tests performed in the two local O&G departments . Direct contact made by GUM departments to index patients identified and "fast track" appointments made . Data recorded for future audit include numbers attending, details of health adviser input, and success of contact tracing . RESULTS: Over 18 months, 294 women were identified and 231 (78%) attended GUM departments; 142 (48%) had received antibiotics before attending GUM departments and of these, 58 (41%) had risked reinfection by an untreated partner and 48 (20%) were found on screening to have a previously undiagnosed genital infection . Over 90% were interviewed by a health adviser . Appropriate follow up was achieved in 87% of index cases . Of the contacts, 194 were treated--150 in the local GUM department . Of these 150 men, 99 (66%) had an identifiable genital infection and 84% of those with CT/non-gonococcal urethritis were asymptomatic . There have been no complaints either formal or informal, by women managed by this system . CONCLUSIONS: GUM clinics are the ideal setting to achieve successful treatment of patients with sexually acquired infections, which must include notification and treatment of their partners if reinfection is to be avoided . For patients with infections diagnosed on other settings, such as O&G, a system of direct notification of results to GUM departments by an agreed protocol can be highly successful . For such a system to work, close cooperation and trust between departments is essential.

Medicina (B Aires), 1998, 58(5 Pt 1), 469 - 73
{Survey of sexually transmitted diseases in the region of Rio Cuarto}; Barberis IL et al.; Sexually transmitted diseases (STD) are acquired mainly through sexual intercourse, being one of the most frequent groups of infectious diseases worldwide and consequently an important public health problem . The aim of this paper was to determine the current state of STD and to compare different diagnostic methods in the population studied . A total of 1060 samples from vaginal flows, endocervical material and urethral discharge were studied during 3 years . Of the total samples, 583 were positive, 493 in women and 90 in men . Microorganisms found in women were: Gardnerella vaginalis (39.3%), Candida albicans (21.1%), Trichomonas vaginalis (17.3%), Candida trachomatis (11.3%), Neisseria gonorrhoeae (3.2%): Mycoplasma hominis and Ureaplasma urelyticum (6.5%) and Treponema pallidum (1.4%), the associations found were, Gardnerella vaginalis with Trichomonas vaginalis 5.5%; Gardnerella vaginalis with Candida albicans 4.9%; Trichomonas vaginalis with Neisseria gonorrhoeae (2.2%) and Gardnerella vaginalis with Chlamydia trachomatis (1.9%) . In men, gonococcal urethritis (UG) represented 37.7% non UG 55.6% and Treponema pallidum 6.7% . These results indicate a decrease in sifilis and in UG when compared to previous studies showing that gonococcal cervicitis had also decreased . We found an important increase in the prevalence of urethritis and non gonococcal cervicitis in agreement with world statistics which consider these diseases as the most common venereal ones . It is necessary to increase the search for Chlamydia trachomatis in pregnant women due to vertical transmission . It should be noted that, in spite of certain fluctuations, the incidence of the STD in our area is still unacceptably high.

J Bacteriol, 1999 Feb, 181(3), 731 - 9
Neisseria gonorrhoeae PilA is an FtsY homolog; Arvidson CG et al.; The pilA gene of Neisseria gonorrhoeae was initially identified in a screen for transcriptional regulators of pilE, the expression locus for pilin, the major structural component of gonococcal pili . The predicted protein sequence for PilA has significant homology to two GTPases of the mammalian signal recognition particle (SRP), SRP54 and SRalpha . Homologs of SRP54 and SRalpha were subsequently identified in bacteria (Ffh and FtsY, respectively) and appear to form an SRP-like apparatus in prokaryotes . Of the two proteins, PilA is the most similar to FtsY (47% identical and 67% similar at the amino acid level) . Like FtsY, PilA is essential for viability and hydrolyzes GTP . The similarities between PilA and the bacterial FtsY led us to ask whether PilA might function as the gonococcal FtsY . In this work, we show that overproduction of PilA in Escherichia coli leads to an accumulation of pre-beta-lactamase, similar to previous observations with other bacterial SRP components . Low-level expression of pilA in an ftsY conditional mutant can complement the ftsY mutation and restore normal growth to this strain under nonpermissive conditions . In addition, purified PilA can replace FtsY in an in vitro translocation assay using purified E . coli SRP components . A PilA mutant that is severely affected in its GTPase activity cannot replace FtsY in vivo or in vitro . However, overexpression of the GTPase mutant leads to the accumulation of pre-beta-lactamase, suggesting that the mutant protein may interact with the SRP apparatus to affect protein maturation . Taken together, these results show that the gonococcal PilA is an FtsY homolog and that the GTPase activity is necessary for its function.

Epidemiol Mikrobiol Imunol, 1998 Dec, 47(4), 145 - 9
{Long-term monitoring of Neisseria meningitidis carrier state in the nasopharynx and seroprevalence of specific antibodies in immunized and non-immunized persons}; Kartusek S et al.; Long-term (three-month and one-year) follow-up of nasopharyngeal carriers of Neisseria meningitidis with a frequency of sample collection of once a week and once a month resp . in the healthy population with a mean age of 43 years revealed a 4.8% positivity (2 of 42 subjects) . Carriership of the same phenotype and ET-genotype of the N . meningitidis group 29E was stable in one subject, while the other long-term carrier of N . meningitidis group B has changes of the phenotype as well as ET-genotype during the follow-up period . Double (after a 3-month interval) verification of the seroprevalence of specific antimeningococcal antibodies revealed in a group of immunized subjects (vaccine A and C) a considerable degree of anti A immunity . Anti A-seropositivity in subjects immunized within a three-year interval since vaccination was 100%, in those immunized within a four-year interval a decline to 66% was recorded . Anti C antibodies after vaccination within a three-year interval were recorded in 75% subjects, in vaccinated subjects the authors detected antibodies during the first year after vaccination in 86% of the subjects . Natural anti A antibodies of the lowest titre 1:6 were recorded in four non-vaccinated subjects (22%) . Naturally acquired anti B antibodies (B:2a:P1.2,P1.5) and anti C (C:2a:P1.2,P1.5) were not detected in any of the subjects.

Sex Transm Dis, 1999 Jan, 26(1), 43 - 8
Epidemiologic aspects of male gonococcal infection in Greece; Kyriakis KP et al.; OBJECTIVE: To report male gonorrhea incidence trends between the years 1974 and 1996 and compare the epidemiologic characteristics of the affected population during periods of high and low incidence of the disease . STUDY DESIGN: Retrospective study of all available data for male gonorrhea cases recorded annually in a major STD hospital in Greece since 1974 . RESULTS: An abrupt drop in gonorrhea relative incidence occurred in 1986, followed by low rates thereafter . In the context of the overall decline in the incidence of the disease, a shift to older ages, longer time elapsed before seeking medical assistance, and higher proportion of disease repeaters was noticed among the patients . Infections caused by penicillinase-producing gonococci were strongly associated with contacts outside the country or with foreigners and their incidence presented erratic time fluctuations, indicating that they have not been established as endemic in the Greek population . CONCLUSION: The core group hypothesis in terms of nighttime sociosexual activity can be used to interpret most of the findingsPIP: Trends in Neisseria gonorrhoeae incidence rates in men presenting to a major sexually transmitted disease hospital (Andreas Sygros) in Athens, Greece, in 1974-96 were evaluated and gonorrhea risk factors during periods of low and high incidence were compared . An abrupt drop in the relative incidence of gonorrhea occurred in 1986, followed by low rates thereafter . A comparison of the periods 1974-85 (high incidence) and 1986-96 (low incidence) revealed a 1.27-fold decrease in total attendance to the hospital and a 5-fold decrease in the number of gonorrhea cases detected . Observed over time was a shift to older ages, longer time elapsed before seeking medical assistance, and a higher proportion of disease repeaters . Infections caused by penicillinase-producing gonococci were strongly associated with contacts outside the country or with foreigners and their incidence was characterized by erratic time fluctuations, suggesting that they have not become endemic in the Greek population . Changes observed in the features of the affected population indicate a trend toward increased risk in sexual behavior .

Sex Transm Dis, 1999 Jan, 26(1), 26 - 32
Subsequent sexually transmitted infections among adolescent women with genital infection due to Chlamydia trachomatis, Neisseria gonorrhoeae, or Trichomonas vaginalis; Fortenberry JD et al.; OBJECTIVE: To identify factors associated with subsequent sexually transmitted infection (STI) (within 1 year of initial infection) due to Chlamydia trachomatis, Neisseria gonorrhoeae or Trichomonas vaginalis . DESIGN: Prospective cohort study . SETTING: A sexually transmitted diseases clinic and four community-based primary care clinics for adolescents . PARTICIPANTS: Female patients (ages 15 to 19 years) with initial diagnosis of chlamydia, gonorrhea, or trichomonas . MAIN OUTCOME MEASURES: Subsequent infection by chlamydia, gonorrhea, or trichomonas . RESULTS: More than 40% of subjects were subsequently infected by at least one STI . Reinfection was common, but infections with sexually transmitted organisms other than the initial infecting organism were also common . Predictors of subsequent infection were black race, gonorrhea as the initial infection, two or more sex partners in the previous 3 months, and inconsistent condom use . CONCLUSIONS: Subsequent STI frequently follow an initial STI, but there is substantial variation in the causal organism . These data suggest the importance of comprehensive STI prevention programs for adolescents rather than organism-specific interventionsPIP: Factors associated with subsequent sexually transmitted infection due to Chlamydia trachomatis, Neisseria gonorrhoeae, or Trichomonas vaginalis were investigated in a prospective study of 236 female adolescents 15-19 years of age who had presented to US sexually transmitted disease (STD) and adolescent health clinics with one of these infections within the previous 12 months . At the second visit, 58 (29.7%) of 195 initially infected women were again infected; 13 were infected with more than one organism . At the third visit, 59 (37.6%) of 157 subjects were reinfected, 20 with more than one organism . Overall, 97 (41.1%) of 236 subjects were again infected within 12 months of an initial STD . Many subsequent infections were due to a sexually transmitted pathogen other than the one causing the index STD . The risk of a subsequent infection was significantly elevated among Blacks, those with gonorrhea at enrollment, and women with 2 or more sex partners in the previous 3 months and significantly reduced among condom users . These findings suggest that organism-specific screening programs would fail to detect a substantial number of subsequent infections due to the presence of other organisms .

Infect Immun, 1999 Feb, 67(2), 834 - 43
Roles of PilC and PilE proteins in pilus-mediated adherence of Neisseria gonorrhoeae and Neisseria meningitidis to human erythrocytes and endothelial and epithelial cells; Scheuerpflug I et al.; Unlike other type 4 pili, the neisserial pili consist of at least two distinct proteins, the highly variable major subunit PilE forming the pilus fiber and the tip-associated adhesin PilC . PilC protein purified either from gonococci or from Escherichia coli interacted with different human epithelial cell lines, primary epithelial and endothelial cells . The binding of PilC protein efficiently prevented the attachment of piliated Neisseria gonorrhoeae and Neisseria meningitidis to these cell types . Fluorescent beads coated with pili prepared from piliated wild-type N . gonorrhoeae also adhered to these cells, in contrast to beads coated with pili prepared from a piliated PilC-deficient mutant . In the latter case, the binding of fluorescent beads was restored after pretreatment of the pilus-loaded beads with purified PilC . Piliated wild-type N . gonorrhoeae, the piliated PilC-deficient mutant, and N . gonorrhoeae pili assembled in Pseudomonas aeruginosa agglutinated human erythrocytes, while nonpiliated gonococci did not . Consistently, purified PilC did not agglutinate or bind to human erythrocytes, suggesting that N . gonorrhoeae PilE is responsible for pilus-mediated hemagglutination.

Infect Immun, 1999 Feb, 67(2), 643 - 52
Neisseria gonorrhoeae mutants altered in toxicity to human fallopian tubes and molecular characterization of the genetic locus involved; Arvidson CG et al.; In an effort to identify potential cytotoxins expressed by Neisseria gonorrhoeae, we have identified a locus that, when mutated in the gonococcus, results in a significant increase in toxicity of the strain to human fallopian tube organ cultures (HFTOC) . This locus, gly1, contains two open reading frames (ORFs) which are likely cotranscribed . ORF1 encodes a polypeptide of 17.8 kDa with a signal sequence that is recognized and processed in Escherichia coli and N . gonorrhoeae . The 15.6-kDa processed polypeptide has been observed in membrane fractions and filtered spent media from cultures of E . coli expressing gly1 and in outer membrane preparations of wild-type N . gonorrhoeae . The gly1 locus is not essential for bacterial survival, and it does not play a detectable role in epithelial cell adhesion, invasion, or intracellular survival . However, a gly1 null mutant causes much more damage to fallopian tube tissues than its isogenic wild-type parent . A strain complemented in trans for the gly1 mutation showed a level of toxicity to HFTOC similar to the level elicited by the wild-type parent . Taken together, these results indicate an involvement of the gly1 locus in the toxicity of N . gonorrhoeae to human fallopian tubes.

Commun Dis Intell, 1998 Dec 24, 22(13), 288 - 91
Resistance in gonococci isolated in the WHO Western Pacific Region to various antimicrobials used in the treatment of gonorrhoea, 1997 . WHO Western Pacific Gonococcal Antimicrobial Surveillance Programme-WHO WPR GASP; Intracellular pathogens and the actin cytoskeleton; Unite des Interactions Bacteries-Cellules, Institut Pasteur, Paris, FranceMany pathogens actively exploit the actin cytoskeleton during infection . This exploitation may take place during entry into mammalian cells after engagement of a receptor and/or as series of signaling events culminating in the engulfment of the microorganism . Although actin rearrangements are a common feature of most internalization events (e.g . entry of Listeria, Salmonella, Shigella, Yersinia, Neisseria, and Bartonella), bacterial and other cellular factors involved in entry are specific to each bacterium . Another step during which pathogens harness the actin cytoskeleton takes place in the cytosol, within which some bacteria (Listeria, Shigella, Rickettsia) or viruses (vaccinia virus) are able to move . Movement is coupled to a polarized actin polymerization process, with the formation of characteristic actin tails . Increasing attention has focused on this phenomenon due to its striking similarity to cellular events occurring at the leading edge of locomoting cells . Thus pathogens are convenient systems in which to study actin cytoskeleton rearrangements in response to stimuli at the plasma membrane or inside cells.

Dermatol Clin, 1998 Oct, 16(4), 747 - 56, xii
Pelvic inflammatory disease . From diagnosis to prevention; Paavonen J; Pelvic inflammatory disease (PID) refers to infection of the uterus, fallopian tubes, and adjacent pelvic structures that is not associated with surgery or pregnancy . PID causes major medical, social, and economic problems worldwide . Long-term sequelae, most notably tubal factor infertility and ectopic pregnancy, are common and extremely costly to the healthcare system . The most important causative micro-organisms are Chlamydia trachomatis, Neisseria gonorrhoeae, and micro-organisms associated with bacterial vaginosis . The clinical spectrum of PID ranges from subclinical endometritis to severe salpingitis, pyosalpinx, tubo-ovarian abscess, pelvic peritonitis, and perihepatitis . Clinical diagnosis of PID has limitations . The clinical diagnostic criteria are insensitive and nonspecific, and false-positive and false-negative diagnosis is common; however, direct visual diagnosis is not always feasible, requires general anesthesia, and is costly . More research is needed of noninvasive diagnosis of PID . Current treatment guidelines call for broad-spectrum antimicrobial coverage . Screening for asymptomatic chlamydial infection is the mainstay of prevention of PID . Emerging evidence from randomized controlled trials provides strong evidence that intervention with selective screening for chlamydial infection effectively reduces the incidence of PID.

J Clin Microbiol, 1999 Feb, 37(2), 386 - 90
Evaluation of AMPLICOR Neisseria gonorrhoeae PCR using cppB nested PCR and 16S rRNA PCR; Farrell DJ; Certain strains of Neisseria subflava and Neisseria cinerea are known to produce false-positive results with the AMPLICOR Neisseria gonorrhoeae PCR (Roche Diagnostic Systems, Branchburg, N.J.) . The analytical sensitivity and analytical specificity of three PCR tests were assessed with 3 geographically diverse N . gonorrhoeae strains and 30 non-N . gonorrhoeae Neisseria spp . The sensitivities of the in-house nested cppB gene and the 16S rRNA PCR methods were greater than that of the AMPLICOR N . gonorrhoeae PCR with purified DNA from all 3 N . gonorrhoeae strains . Six of 14 clinical strains of N . subflava (1 from a vaginal swab, 5 from respiratory sites) produced false-positive AMPLICOR N . gonorrhoeae PCR results and were negative by the two other PCR methods . When applied to 207 clinical specimens selected from a population with a high prevalence ( approximately 9%) of infection, the results for 15 of 96 (15.6%) AMPLICOR-positive specimens and 14 of 17 (82.3%) AMPLICOR-equivocal specimens were not confirmed by the more sensitive nested cppB PCR method . Only 2 of 94 (2.1%) of AMPLICOR N . gonorrhoeae PCR-negative specimens from the same population tested positive by the nested cppB method . These results suggest that for this population the AMPLICOR N . gonorrhoeae PCR test is suitable as a screening test only and all positive results should be confirmed by a PCR method that is more specific and at least as sensitive . This study also illustrates that caution should be used when introducing commercially available nucleic acid amplification-based diagnostic tests into the regimens of tests used for populations not previously tested with these products.

EMBO J, 1999 Jan 15, 18(2), 339 - 52
Neisserial porin (PorB) causes rapid calcium influx in target cells and induces apoptosis by the activation of cysteine proteases; Muller A et al.; The porin (PorB) of Neisseria gonorrhoeae is an intriguing bacterial factor owing to its ability to translocate from the outer bacterial membrane into host cell membranes where it modulates the infection process . Here we report on the induction of programmed cell death after prolonged infection of epithelial cells with pathogenic Neisseria species . The underlying mechanism we propose includes translocation of the porin, a transient increase in cytosolic Ca2+ and subsequent activation of the Ca2+ dependent protease calpain as well as proteases of the caspase family . Blocking the porin channel by ATP eliminates the Ca2+ signal and also abolishes its pro-apoptotic function . The neisserial porins share structural and functional homologies with the mitochondrial voltage-dependent anion channels (VDAC) . The neisserial porin may be an analogue or precursor of the ancient permeability transition pore, the putative central regulator of apoptosis.

N Engl J Med, 1999 Jan 14, 340(2), 85 - 92
Bacteriologic analysis of infected dog and cat bites . Emergency Medicine Animal Bite Infection Study Group; Talan DA et al.; BACKGROUND AND METHODS: To define better the bacteria responsible for infections of dog and cat bites, we conducted a prospective study at 18 emergency departments . To be eligible for enrollment, patients had to meet one of three major criteria for infection of a bite wound (fever, abscess, and lymphangitis) or four of five minor criteria (wound-associated erythema, tenderness at the wound site, swelling at the site, purulent drainage, and leukocytosis) . Wound specimens were cultured for aerobic and anaerobic bacteria at a research microbiology laboratory and, in some cases, at local hospital laboratories . RESULTS: The infected wounds of 50 patients with dog bites and 57 patients with cat bites yielded a median of 5 bacterial isolates per culture (range, 0 to 16) at the reference laboratory . Significantly more isolates grew at the reference laboratory than at the local laboratories (median, 1; range, 0 to 5; P<0.001) . Aerobes and anaerobes were isolated from 56 percent of the wounds, aerobes alone from 36 percent, and anaerobes alone from 1 percent; 7 percent of cultures had no growth . Pasteurella species were the most frequent isolates from both dog bites (50 percent) and cat bites (75 percent) . Pasteurella canis was the most common isolate of dog bites, and Past . multocida subspecies multocida and septica were the most common isolates of cat bites . Other common aerobes included streptococci, staphylococci, moraxella, and neisseria . Common anaerobes included fusobacterium, bacteroides, porphyromonas, and prevotella . Isolates not previously identified as human pathogens included Reimerella anatipestifer from two cat bites and Bacteroides tectum, Prevotella heparinolytica, and several porphyromonas species from dog and cat bites . Erysipelothrix rhusiopathiae was isolated from two cat bites . Patients were most often treated with a combination of a beta-lactam antibiotic and a beta-lactamase inhibitor, which, on the basis of the microbiologic findings, was appropriate therapy . CONCLUSIONS: Infected dog and cat bites have a complex microbiologic mix that usually includes pasteurella species but may also include many other organisms not routinely identified by clinical microbiology laboratories and not previously recognized as bite-wound pathogens.

Microbiology, 1998 Dec, 144 ( Pt 12), 3425 - 36
The Pasteurella haemolytica 35 kDa iron-regulated protein is an FbpA homologue; Kirby SD et al.; In a previous investigation, a 35 kDa iron-regulated protein was identified from total cellular proteins of Pasteurella haemolytica grown under iron-depleted conditions . This study reports identification of the gene (fbpA) encoding the 35 kDa protein based on complementation of an entA Escherichia coli strain transformed with a plasmid derived from a P . haemolytica lambda ZAP II library . Cross-reactivity was demonstrated between an anti-35 kDa mAb and a 35 kDa protein expressed in this strain . Furthermore, a translated ORF identified on the recombinant plasmid corresponded with the N-terminal amino acid sequence of the intact and a CNBr-cleaved fragment of the 35 kDa iron-regulated protein . Nucleotide sequence analysis of the gene encoding the 35 kDa protein demonstrated homology with the cluster 1 group of extracellular solute-binding proteins, especially to the iron-binding proteins of this family . Complete sequence analysis of the recombinant plasmid insert identified three other predominant ORFs, two of which appeared to be in an operonic organization with fbpA . These latter components (fbpB and fbpC) showed homology to the transmembrane and ATPase components of ATP-binding cassette (ABC)-type uptake systems, respectively . Based on amino acid/DNA sequencing, citrate competition assay of iron affinity and visible wavelength spectra, it was concluded that the P . haemolytica 35 kDa protein functions as an FbpA homologue (referred to as PFbpA) and that the gene encoding this protein is part of an operon comprising a member of the FbpABC family of iron uptake systems . Primary sequence analysis revealed rather surprisingly that PFbpA is more closely related to the intracellular Mn/Fe-binding protein IdiA found in cyanobacteria than to any of the homologous FbpA proteins currently known in commensal or pathogenic members of the Pasteurellaceae or Neisseriaceae.

J Bacteriol, 1999 Jan, 181(2), 656 - 61
Role of glycosylation at Ser63 in production of soluble pilin in pathogenic Neisseria; Marceau M et al.; Pilus-mediated adhesion is essential in the pathogenesis of Neisseria meningitidis (MC) and Neisseria gonorrhoeae (GC) . Pili are assembled from a protein subunit called pilin . Pilin is a glycoprotein, and pilin antigenic variation has been shown to be responsible for intrastrain variability with respect to the degree of adhesion in both MC and GC . In MC, high-adhesion pilins are responsible for the formation of bundles of pili which bind bacteria and cause them to grow as colonies on infected monolayers . In this work, we selected MC and GC pilin variants responsible for high and low adhesiveness and introduced them into the other species . Our results demonstrated that a given pilin variant expressed an identical phenotype in either GC or MC with respect to bundling and adhesiveness to epithelial cells . However, the production of truncated soluble pilin (S pilin) was consistently more abundant in GC than in MC . In the latter species, the glycosylation of pilin at Ser63 was shown to be required for the production of a truncated monomer of S pilin . In order to determine whether the same was true for GC, we engineered various pilin derivatives with an altered Ser63 glycosylation site . The results of these experiments demonstrated that the production of S pilin in GC was indeed more abundant when pilin was posttranslationally modified at Ser63 . However, nonglycosylated variants remained capable of producing large amounts of S pilin . These data demonstrated that for GC, unlike for MC, glycosylation at Ser63 is not required for S-pilin production, suggesting that the mechanisms leading to the production of S pilin in GC and MC are different.

J Bacteriol, 1999 Jan, 181(2), 541 - 51
cis- and trans-acting elements involved in regulation of aniA, the gene encoding the major anaerobically induced outer membrane protein in Neisseria gonorrhoeae; Householder TC et al.; AniA (formerly Pan1) is the major anaerobically induced outer membrane protein in Neisseria gonorrhoeae . AniA has been shown to be a major antigen in patients with gonococcal disease, and we have been studying its regulation in order to understand the gonococcal response to anaerobiosis and its potential role in virulence . This study presents a genetic analysis of aniA regulation . Through deletion analysis of the upstream region, we have determined the minimal promoter region necessary for aniA expression . This 130-bp region contains a sigma 70-type promoter and an FNR (fumarate and nitrate reductase regulator protein) binding site, both of which are absolutely required for anaerobic expression . Also located in the minimal promoter region are three T-rich direct repeats and several potential NarP binding sites . This 80-bp region is required for induction by nitrite . By site-directed mutagenesis of promoter sequences, we have determined that the transcription of aniA is initiated only from the sigma 70-type promoter . The gearbox promoter, previously believed to be the major promoter, does not appear to be active during anaerobiosis . The gonococcal FNR and NarP homologs are involved in the regulation of aniA, and we demonstrate that placing aniA under the control of the tac promoter compensates for the inability of a gonococcal fnr mutant to grow anaerobically.

J Bacteriol, 1999 Jan, 181(2), 375 - 81
Sequence analysis of the gene encoding amylosucrase from Neisseria polysaccharea and characterization of the recombinant enzyme; De Montalk GP et al.; The Neisseria polysaccharea gene encoding amylosucrase was subcloned and expressed in Escherichia coli . Sequencing revealed that the deduced amino acid sequence differs significantly from that previously published . Comparison of the sequence with that of enzymes of the alpha-amylase family predicted a (beta/alpha)8-barrel domain . Six of the eight highly conserved regions in amylolytic enzymes are present in amylosucrase . Among them, four constitute the active site in alpha-amylases . These sites were also conserved in the sequence of glucosyltransferases and dextransucrases . Nevertheless, the evolutionary tree does not show strong homology between them . The amylosucrase was purified by affinity chromatography between fusion protein glutathione S-transferase-amylosucrase and glutathione-Sepharose 4B . The pure enzyme linearly elongated some branched chains of glycogen, to an average degree of polymerization of 75.

Microb Pathog, 1998 Nov, 25(5), 237 - 52
Characterization of a sialyltransferase-deficient mutant of Neisseria gonorrhoeae strain F62: instability of transposon Tn1545 delta3 in gonococci and evidence that multiple genetic loci are essential for lipooligosaccharide sialylation; Crooke H et al.; Neisseria gonorrhoeae strain JB1 was previously shown to be defective in the sialylation of lipoologosaccharide (LOS) by exogenous CMP-NANA . The LOS components synthesized by the mutant now have been shown by mass spectrometry to be similar to those in the parental strain, F62, and to include the 4.5 kDa widely conserved lacto-N-neotetraose component that can be sialylated . The same two LOS components could be sialylated on the surface of the mutant and parental strains . One major component was sialylatable after chemical extraction of the LOS from either strain . These data confirm that the mutant, JB1, retains the ability to synthesize the LOS target required for the conversion by sialylation of serum-sensitive gonococci to serum resistance . A single base frame-shift mutation was found in the lst gene from the mutant, resulting in the replacement of the final 61 amino acids at the C-terminus of the sialyltransferase by four residues . Seventeen independent clones of the lst gene were isolated from the parental strain, but none of them complemented the sialyltransferase defect of the mutant and no sialyltransferase activity expressed from the clones could be detected in Escherichia coli . Although the data suggest that the mutant might be defective in genes at more than one chromosomal locus and that multiple loci are essential for sialyltransferase synthesis and activity, the alternative possibility, that DNA adjacent to the lst gene encodes a product which is toxic to E . coli, cannot be excluded . The site of insertion of the transposon Tn1545-Delta3 in strain JB1 was cloned and sequenced . The transposon is located in an intergenic region adjacent to genes for a putative ATP-dependent transport protein, but encoding no recognizable function relevant to LOS sialylation . Evidence that transposon Tn1545-Delta3 is unstable in gonococci is presented .

J Infect Dis, 1999 Feb, 179(2), 371 - 81
Molecular typing of Neisseria gonorrhoeae causing repeated infections: evolution of porin during passage within a community; Hobbs MM et al.; Thirty-three Neisseria gonorrhoeae isolates from 15 persons infected multiple times with the same serovar were compared using por gene sequencing, opa-typing, and arbitrarily primed-polymerase chain reaction . All three molecular techniques were more discriminatory than serotyping and identified differences between some isolates belonging to the same serovar . Although there were differences among Por sequences within some serovars, 10 of 15 subjects became reinfected with gonococci expressing identical Por proteins . Sequence analysis of por genes revealed evidence of horizontal genetic exchange and point mutations in potential surface-exposed regions during passage in the community.

Pediatr Infect Dis J, 1998 Dec, 17(12), 1135 - 40
Evaluation of febrile children with petechial rashes: is there consensus among pediatricians?
Nelson DG, Leake J, Bradley J, Kuppermann N.
BACKGROUND: The evaluation of febrile children with petechial rashes evokes controversy . Although many of these children have viral infections, on occasion such patients may be infected with Neisseria meningitidis . OBJECTIVE: To investigate differences in practice trends for the evaluation and management of non-toxic-appearing febrile children with petechial rashes among pediatric specialty groups . METHODS: We surveyed 833 pediatricians in 4 specialties {community (CGP) and academic (AGP) general pediatrics, emergency medicine (EM) and infectious diseases} regarding 4 hypothetical non-toxic-appearing febrile children ages 1, 2, 5 and 7 years . The patients differed with regard to clinical appearance, distribution of petechiae and complete blood count results . We compared specialty group responses, adjusting for practice setting, population size and years in practice using multiple logistic regression analysis . RESULTS: The survey was completed and returned by 416 (50%) pediatricians . There was substantial variation in the evaluation of the 2 younger febrile children without clear sources for their petechiae . For the 1-year-old the overall blood culture (BCx) rate was 82%, with the EM group (91%) more often requesting BCx than either the CGP (76%) or AGP (73%, P=0.001) groups . The overall hospital admission rate was 31%, with CGP less often requesting admission than infectious disease pediatricians (22% vs . 40%, P=0.007) . In the regression analysis the only significant difference between groups was in BCx rate between the EM and AGP groups . For the 2-year-old the overall rate of BCx was 95%, lumbar puncture was 41% and admission was 44%, with no significant differences among groups . For the scenarios involving the 2 older febrile children with sources for their petechiae, the majority of respondents chose neither lumbar puncture nor admission . There was disagreement regarding BCx, both within and between groups, although most of the between group differences did not persist in the regression analysis . CONCLUSIONS: There are substantial differences among pediatricians in the evaluation of young non-toxic-appearing febrile children with petechial rashes . Although there are some differences between pediatric subspecialties, most of these differences do not persist after adjusting for practice setting, population size and physician experience.

J Biol Chem, 1999 Jan 8, 274(2), 1017 - 25
Neisseria gonorrhoeae that infect men have lipooligosaccharides with terminal N-acetyllactosamine repeats; John CM et al.; Infectious Neisseria gonorrhoeae make relatively large lipooligosaccharides (LOS) that structurally resemble human glycosphingolipids . MS11mkC is an LOS variant of N . gonorrhoeae strain MS11 which was isolated from men at the onset of dysuria (Schneider, H., Griffiss, J . M., Boslego, J . W., Hitchcock, P . J., Zahos, K . M., and Apicella, M . A . (1991) J . Exp . Med . 174, 1601-1605) . Delayed extraction matrix-assisted laser desorption and ionization and electrospray ionization mass spectrometry of O-deacylated MS11mkC LOS produced ions consistent with known LOS which have lacto-N-neotetraose (Galbeta1-->4GlcNAcbeta1-->3Galbeta1-->4Glc; paraglobosyl; monoclonal antibodies (mAbs) 1B2(+) and 06B4(+)) and GalNAc-->lacto-N-neotetraose (gangliosyl; mAb 1-1-M+) oligosaccharides . Ion peaks for a larger LOS which also bound mAb 1B2 indicated the addition of a hexose (+162 Da) to gangliosyl LOS or the addition of a hexose and a N-acetylhexosamine (+365 Da) to paraglobosyl LOS . Analysis of HF-treated and O-deacylated LOS revealed three major components present in a phosphoethanolamine (PEA)0 and a PEA1 series . Digestion of MS11mkC LOS by beta-N-acetylhexosaminidase and beta-galactosidase, alone and sequentially, combined with mAb binding patterns, confirmed the presence of a nonreducing terminal repeating LacNAc ((Galbeta1-->4GlcNAc)2) on the largest LOS, rather than a parallel oligosaccharide structure.

FEMS Microbiol Lett, 1998 Dec 15, 169(2), 375 - 82
Small genes/gene-products in Escherichia coli K-12; Wasinger VC et al.; Forty-two protein spots of observed M(r) 6-15 kDa were resolved by two-dimensional gel electrophoresis, stained by Coomassie blue and subjected to Edman microsequencing . All of the proteins could be related back to their encoding open reading frames, thereby vindicating the bioinformatic tools currently utilised in their identification . However, only 14/42 gene-products were expressed as annotated . Translation was confirmed for 14 open reading frames with no attributed function (EcoGene Y-entries), while N-terminal sequence allowed the start codon to be accurately annotated for the genes yigF, yccU, yqiC, ynfD, and yeeX . The methionine start codon was cleaved in 11 gene-products (AtpE, Hns, RpoZ, RplL, CspC, YccJ, YggX, YjgF, HimA, InfA, RpsQ) and a further five showed loss of a signal peptide (PspE, HdeB, HdeA, YnfD, YkfE) . Internal (Tig, AtpA, TufA) and N-terminal fragmentation (CspD, RpsF, AtcU) of much larger proteins was also detected, which may have resulted from physiological or translational processes . M(r) and pI isoforms were detected respectively for PtsH and GatB, each being phosphoproteins, as well as RplY which manifested differences with respect to predicted M(r) and pI . In addition, YjgF was shown to belong to a small gene family of unknown function with ancient conserved regions across procaryotes and eucaryotes . YgiN was revealed to have a paralogue and orthologues in Bacillus subtilis, Synechocystis sp., Mycobacterium tuberculosis, Neisseria gonorrhoea, and Rhodococcus erythropolis . Orthologues are also reported for YihD, YccU and YeeX . Of the 14 Y-genes, only YkfE possessed no detectable orthologues . These results highlight the need to complement genomic analysis with detailed proteomics in order to gain a better understanding of cellular molecular biology, while the confirmation of the open reading frame start codon using Edman degradation protein microsequencing has yet to be superseded by recent advances in mass spectrometry.

Biol Chem, 1998 Nov, 379(11), 1391 - 5
Cloning and characterization of the gene encoding a new DNA methyltransferase from Neisseria gonorrhoeae; Radlinska M et al.; A HindIII fragment of N . gonorrhoeae MS11 DNA coding for DNA methyltransferase (MTase) activity was cloned and expressed in E . coli AP1-200-9 cells . The sequence of 4681 bp was determined, and its analysis revealed two open reading frames (ORFs) sharing some similarity with known DNA MTases . ORF1 encodes an active N4mC MTase (M.NgoMV) . The enzyme modifies only one strand of double stranded DNA and preferentially recognises the sequence GCCHR although it is able to methylate other sites . The exact recognition sequence cannot be precisely defined due to a relaxed specificity . The second ORF shows high homology to 5mC Mtases, but we were unable to demonstrate DNA methylating activity of its product either in vivo or in vitro.

J Bacteriol, 1999 Jan, 181(1), 24 - 33
The Myxococcus xanthus pilQ (sglA) gene encodes a secretin homolog required for type IV pilus biogenesis, social motility, and development; Wall D et al.; The Myxococcus xanthus sglA1 spontaneous mutation was originally isolated because it allowed dispersed cell growth in liquid yet retained the ability to form fruiting bodies . Consequently, most of today's laboratory strains either contain the sglA1 mutation or were derived from strains that carry it . Subsequent work showed that sglA was a gene for social gliding motility, a process which is mediated by type IV pili . Here sglA is shown to map to the major pil cluster and to encode a 901-amino-acid open reading frame (ORF) that is homologous to the secretin superfamily of proteins . Secretins form a channel in the outer membrane for the transport of macromolecules . The closest homologs found were PilQ proteins from Pseudomonas aeruginosa and Neisseria gonorrhoeae, which are required for type IV pili biogenesis and twitching motility . To signify these molecular and functional similarities, we have changed the name of sglA to pilQ . The hypomorphic pilQ1 (sglA1) allele was sequenced and found to contain two missense mutations at residues 741 (G-->S) and 762 (N-->G) . In addition, 19 independent social (S)-motility mutations are shown to map to the pilQ locus . In-frame deletions of pilQ and its downstream gene, orfL, were constructed . pilQ is shown to be essential for pilus biogenesis, S-motility, rippling, and fruiting body formation, while orfL is dispensable for these processes . The pilQ1 allele, but not the DeltapilQ allele, was found to render cells hypersensitive to vancomycin, suggesting that PilQ1 alters the permeability properties of the outer membrane . Many differences between pilQ1 and pilQ+ strains have been noted in the literature . We discuss some of these observations and how they may be rationalized in the context of our molecular and functional findings.

Infect Immun, 1999 Jan, 67(1), 455 - 9
Identification and functional characterization of the Neisseria gonorrhoeae lbpB gene product; Biswas GD et al.; We cloned lbpB, encoding a predicted 80-kDa lipoprotein, upstream of lbpA . A nonpolar mutant (LbpB- LbpA+) had normal lactoferrin (LF) binding and grew normally with LF as an iron source, whereas LbpB- LbpA- and LbpB+ LbpA- strains had reduced binding of LF and did not grow with LF as an iron source . LbpB bound LF directly in an affinity purification, suggesting that LbpB might play a still-uncharacterized role in the LF iron utilization.

Sex Transm Dis, 1998 Nov, 25(10), 533 - 8
Detection of Neisseria gonorrhoeae infection by ligase chain reaction testing of urine among adolescent women with and without Chlamydia trachomatis infection; Xu K et al.; BACKGROUND AND OBJECTIVES: Culture, the conventional method for detection of Neisseria gonorrhoeae, requires invasive sampling and stringent specimen transport conditions . The recently developed ligase chain reaction test (LCR; Abbott Laboratories; North Chicago, IL) allows noninvasive sampling and stable transport conditions, but has not been evaluated with specimens from adolescent populations . GOAL OF THIS STUDY: To perform a comparative evaluation of a commercial LCR test and culture for the diagnosis of N . gonorrhoeae in adolescent women . STUDY DESIGN: Urine and endocervical swab specimens from 330 teenage women seen in two public health adolescent clinics were tested by LCR and culture . For resolution of discordant results, a polymerase chain reaction (PCR) test was developed that directly amplifies N . gonorrhoeae DNA from urine samples processed for LCR . RESULTS: Thirty-one of 330 (9.4%) cervical specimens were culture-positive for N . gonorrhoeae, and 30 of 330 (9.1%) urine specimens were positive by LCR . After resolution of 13 discordant results, the sensitivity, specificity, and positive and negative predictive values of LCR for urine were 88.2%, 100%, 100%, 98.7%, respectively, and for culture of endocervical specimens were 82.3%, 98.9%, 90.3% and 98%, respectively . CONCLUSIONS: Although more expensive than culture, LCR offers a sensitive means for the detection of N . gonorrhoeae in urine samples and may be useful for this purpose in settings where pelvic examinations are difficult to perform and simultaneous detection of N . gonorrhoeae and Chlamydia trachomatis is advantageous.

Sex Transm Dis, 1998 Nov, 25(10), 522 - 6
Quinolone-resistant Neisseria gonorrhoeae: the beginning of the end? Report of quinolone-resistant isolates and surveillance in the southwestern United States, 1989 to 1997; Ehret JM et al.; BACKGROUND: Fluoroquinolones are one of the most widely used treatments for gonorrhoeae . Changes in the susceptibility of Neisseria gonorrhoeae to these agents may threaten their use . GOAL OF THIS STUDY: To report several resistant strains (>1 mcg/ml) isolated in the western United States and to evaluate the prevalence of strains with reduced susceptibility (ofloxacin 0.25 mcg/ml, ciprofloxacin 0.06 mcg/ml) . STUDY DESIGN: The microbiology and epidemiology of three resistant strains were characterized and 12,761 other strains were evaluated for fluoroquinolone susceptibility as part of the Gonococcal Isolate Surveillance Project of the Centers for Disease Control and Prevention . RESULTS: Fluoroquinolone-resistant strains may appear sporadically . The prevalence of isolates with reduced susceptibility to fluoroquinolones remains low in the Southwest region of the United States . CONCLUSIONS: Continued active surveillance is needed to detect and control the spread of quinolone-resistant Neisseria gonorrhoeae.

Sex Transm Dis, 1998 Nov, 25(10), 501 - 4
Evaluation of the Gen-Probe PACE 2 assay for the detection of asymptomatic Chlamydia trachomatis and Neisseria gonorrhoeae infections in male arrestees; Beltrami JF et al.; BACKGROUND: The LET and Gen-Probe PACE 2 assay are used to screen male arrestees for evidence of infection with chlamydia and gonorrhea in the intake/booking area of the Jefferson Parish Correctional Center . GOAL OF THIS STUDY: To determine the accuracy of the Gen-Probe PACE 2 assay for the detection of asymptomatic infection with Chlamydia trachomatis and Neisseria gonorrhoeae infections in male arrestees . STUDY DESIGN: From December 1993 to March 1994, 196 arrestees younger than 22 years were screened with the Gen-Probe PACE 2 assay and McCoy shell vial culture for Chlamydia trachomatis . From April to October 1994, 444 arrestees of all ages were screened with the Gen-Probe PACE 2 assay and standard culture for Neisseria gonorrhoeae . The sensitivity, specificity, and predictive values of the Gen-Probe PACE 2 assay, compared with culture, were calculated with 95% confidence intervals . RESULTS: The Gen-Probe PACE 2 assay compared with culture had a sensitivity, specificity, and positive predictive value for Chlamydia trachomatis of 84%, 99%, and 93% and for Neisseria gonorrhoeae of 54%, 99.5%, and 78% . CONCLUSION: The Gen-Probe PACE 2 assay is useful for screening young males in this jail setting and is more accurate for detecting Chlamydia trachomatis compared with Neisseria gonorrhoeae.

Ann Acad Med Stetin, 1998, 44, 45 - 64
{An attempt to block histamine release from basophils granulocytes with antibodies obtained as a result of long-term immunization}; Szymaniak L; Pathogenetic mechanisms responsible for efficacy of specific immunotherapy still remain to be fully explained . This concerns both desensitization with classic allergens and very rarely used specific immunotherapy with bacteria . Microbes can play important role as hypersensitivity factor in some allergo-inflammatory processes . Bacterial products may act as basophil histamine liberators through immunological (IgE-mediated) and nonimmunological--particular lectin-sugar way . The aim of study was to verify if histamine release triggered by microbes could be modified (blocked) with specific antibacterial antibodies--taking into consideration both of mechanisms of basophil degranulation . The size of immediate (in healthy persons--Tab . 3, 4) and late as well as delayed (in asthmatic patients--Tab . 8) skin reactivity to examined microorganisms and the degree of basophil histamine release induced with these bacteria were compared . Human basophils were isolated from peripheral blood on Ficoll-Hypaque gradient, next challenged with whole, formalin-killed bacteria and with the same bacteria after incubation with specific and nonspecific sera . To differentiate between IgE-dependent and non-immunological mechanisms of histamine release, the IgE molecules were removed from the surface of the basophils by exposure to pH 3.6 (stripping) . In each experiment histamine release induced by anti-IgE antibodies was used as control of stripping (Tab . 5, 9) . Levels of histamine from the basophils (without and after stripping) incubated with non-coated and specific antibodies coated bacteria were compared . The results were expressed as a percentage of total histamine content in the sample . Histamine release was assayed spectrofluorometrically by using Shore method in Norn modification . The main investigations concerned the basophils from 12 healthy, non-atopic individuals, who had positive immediate skin reactions with at least 1 from 3 microbial strains: Staphylococcus aureus 9615 (unencapsulated), Staphylococcus aureus Smith (encapsulated) and Escherichia coli . Sera containing specific antibodies for these microorganisms were obtained from immunized rabbits . As negative control served sera collected from animals after immunization . Additionally the basophils of 6 asthmatic (intrinsic asthma) patients treated with autovaccines were examined . All patients demonstrated positive late and delayed skin reactions, 3 of them also immediate, to autologous Neisseria and Moraxella species cultured from upper respiratory tract . The bacteria were used as a component of autovaccine and as a basophils stimulating factor in histamine assay . Microbes were incubated with patients own sera before (unspecific serum) and after treatment (source of "specific" antibodies) . CONCLUSIONS: 1 . Bacteria induced basophil histamine release through two ways: immunological (IgE-mediated) and non-immunological (sugar-lectin interactions) . 2 . Non-immunological interactions played the main role in basophil histamine release induced by bacteria--both in normal individuals and asthmatic patients . 3 . Sera of immunized with bacteria animals partially reduced basophil histamine release induced by homologous strains (Tab . 7) . 4 . An incubation of autologous bacterial strains with asthmatic patients's sera collected after autovaccines treatment has no influence on basophil histamine release induced by these microbes (Tab . 9) . 5 . There was no correlation between the skin reactivity to bacteria (both in healthy persons and in asthmatic patients) and the intensity of basophil histamine release induced by microbes.

J Biol Chem, 1998 Dec 25, 273(52), 35332 - 8
Neisseria gonorrhoeae porin modulates phagosome maturation; Mosleh IM et al.; The porin (PorB) of Neisseria gonorrhoeae has been implicated in the pathogenesis of this species . Porin is believed to translocate from the bacterial outer membrane into target cell membranes affecting various cell functions . Here we investigated the effect of porin on phagosome maturation . Phagocytosis of latex beads by human macrophages was allowed in the presence or absence of purified porin . Isolation of latex bead-containing phagosomes and subsequent two-dimensional gel electrophoresis revealed substantial differences in the phagosomal protein composition . Immunoblotting detected higher amounts of annexin II and the early endocytic markers Rab5 and transferrin receptor and decreased levels of the late endocytic markers Rab7 and cathepsin D in phagosomes obtained in the presence of porin compared with those obtained in its absence . Furthermore, association of Rab4 with the latex bead-containing phagosomes was revealed by flow cytometry . The amount of this small GTPase was markedly higher in the phagosomes isolated in the presence of porin . The data thus indicate that neisserial porin is itself able to arrest phagosome maturation within macrophages.

J Protein Chem, 1998 Oct, 17(7), 719 - 28
Characterization of a subunit structure and stability of the recombinant porin from Neisseria gonorrhoeae; Matsuka YV et al.; An outer membrane PIA protein from Neisseria gonorrhoeae strain FA19 was expressed in Escherichia coli and refolded in vitro in the presence of zwitterionic detergent . Its proper folding and subunit organization was confirmed by comparison with the native counterpart . The unfolding of PIA has been investigated using fluorescence spectroscopy and analytical size-exclusion chromatography methods . Analysis of the denaturation pathway of the PIA revealed that it forms an unusually labile quaternary structure . In the presence of 1 M guanidinium chloride (GdmCl) or upon heating up to 50 degrees C, dissociation of the PIA oligomer was observed resulting in the formation of folded monomeric intermediates . Unfolding of monomers occurs at 80 degrees C or in the presence of 4.3 M GdmCl, indicating high intrinsic stability toward both GdmCl and elevated temperatures . Both oligomeric and monomeric forms of PIA exhibited affinity to the hydrophobic probe 1-anilinonaphthalene-8-sulfonic acid (ANS) and bind with Kd=80 and 130 microM, respectively . Denaturation of the PIA completely abolished affinity to ANS, suggesting that hydrophobicity is a property of the folded state of the porin.

APMIS Suppl, 1998, 84, 56 - 61
Competence for natural transformation in Neisseria gonorrhoeae: a model system for studies of horizontal gene transfer; Koomey M; A combined effort integrating studies of gonococcal Tfp biogenesis, the data made available from the gonococcal genome sequence project and applied molecular genetics have been used to identify the fibrillar filaments themselves, the PilT protein and the ComP protein as essential components for the DNA uptake phase of competence for natural transformation . Our ongoing studies are focused on identifying and understanding the complex interactions which exist between these essential constituents . These studies may be relevant not only to the early steps of genetic transformation but also to the two other venues for horizontal gene transfer based on recent findings . First, the thin pili of IncI1 conjugal plasmids required for liquid mating belong to the type IV family of pili (Yoshida et al., 1998) . Secondly, type IV pili are required for lysogenic conversion of Vibrio cholerae by a filamentous phage encoding cholera toxin (Waldor and Mekalanos, 1996) . How these highly conserved surface organelles contribute to such diverse forms of DNA translocation across membranes remains to be seen.

Sex Transm Infect, 1998 Jun, 74(3), 210 - 2
Antimicrobial susceptibility and plasmid profile of Neisseria gonorrhoeae in India (New Delhi); Bhalla P et al.; OBJECTIVES: To determine the antibiotic susceptibility and plasmid profile of all Neisseria gonorrhoeae strains (PPNG and non-PPNG) isolated from May 1995 to March 1996 in Lok Nayak Hospital, New Delhi, India . METHODS: The agar plate dilution method was used to determine the minimum inhibitory concentration of five antimicrobials including norfloxacin and ceftriaxone which are most commonly used for treatment of gonorrhoea in Delhi . Isolates were screened for production of penicillinase by paper acidometric method and plasmid analysis of PPNG and non-PPNG was carried out by agarose gel electrophoresis . RESULTS: 50 consecutive isolates of N gonorrhoeae were studied, 8% among them were found to be PPNG while 28% were highly resistant to tetracycline (TRNG) . Reduced susceptibility to norfloxacin (MIC > or = 1 microgram/ml) was observed in 12% of all isolates . All PPNG harboured the 4.4 MDa beta lactamase plasmid along with the 25.2 MDa tetracycline resistance plasmid . Norfloxacin resistance (MIC > or = 1 microgram/ml) was present in 28.5% of TRNG but only in 5.5% of the other gonococcal isolates . CONCLUSIONS: Results of this study clearly demonstrate that antibiotic resistant gonococcal strains of different clones are frequently found in New Delhi . Continued surveillance of susceptibility to currently prescribed antimicrobials and epidemiological studies are essential to prevent treatment failures leading to further spread of resistant strains.

Sex Transm Infect, 1998 Jun, 74(3), 205 - 9
Auxotypes, serovars, and trends of antimicrobial resistance of Neisseria gonorrhoeae in Kigali, Rwanda (1985-93); Bogaerts J et al.; OBJECTIVE: To investigate the in vitro antimicrobial susceptibility and the auxotype/serovar distribution of Neisseria gonorrhoeae in Kigali, Rwanda, during 1985-93 . METHODS: As part of a monitoring programme the in vitro susceptibility of 1604 isolates of N gonorrhoeae was determined by agar dilution . Auxo- and serotyping was performed on 1350 and 1313 isolates respectively . RESULTS: The prevalence of penicillinase producing N gonorrhoeae (PPNG) remained stable at a rate of 39% during 1985-91 and increased to 61% in 1992-3 . Chromosomal resistance to penicillin was common among non-PPNG and resistance to thiamphenicol and tetracycline was common among both PPNG and non-PPNG . High level, plasmid mediated resistance to tetracycline (TRNG) was observed for the first time at the end of 1989 and increased from 2% of the isolates in 1990 to 50% by 1993 . A trend for increasing resistance to norfloxacin and ofloxacin was observed during 1985-90 but disappeared in 1991-93 . Five isolates with high level resistance to norfloxacin (MIC 2 mg/l) were observed in 1990 . Resistance to trimethoprim-sulphamethoxazole (TMP-SMZ) emerged at the end of 1990 and was observed among 10% of the isolates during 1991-3 . All strains remained susceptible to ofloxacin, ciprofloxacin, spectinomycin, and ceftriaxone . Overall, 75% of the isolates were prototrophic or required proline for their growth and 62% belonged to serovars IA-6 and IB-1 . The prevalence of serovar IB-4 increased strongly during the last 3 years of the study . CONCLUSION: Resistance to penicillin, thiamphenicol, and tetracycline was common in N gonorrhoeae during 1985-1993 . The rapid spread of TRNG after 1989 and the steep increase of PPNG during 1992-3 were the most striking facts of the study period . The auxotype and serovar distribution was comparable with findings from other African countries.

Sex Transm Infect, 1998 Jun, 74(3), 189 - 93
Rapid assessment of sexually transmitted diseases in a sentinel population in Thailand: prevalence of chlamydial infection, gonorrhoea, and syphilis among pregnant women--1996; Kilmarx PH et al.; OBJECTIVE: To determine the prevalence of sexually transmitted diseases (STDs) among pregnant women in Thailand, where case reporting suggests a marked decrease in STDs following a campaign promoting condom use during commercial sex . DESIGN: Cross sectional study of women at their first visit to the study hospitals' antenatal clinics in Chiang Rai (n = 500) and Bangkok (n = 521) . METHODS: First catch urine specimens were tested for Chlamydia trachomatis and Neisseria gonorrhoeae using the Amplicor CT/NG polymerase chain reaction assay . Syphilis and HIV serological testing were performed in the study hospitals' laboratories . RESULTS: The prevalence of chlamydial infection was 5.7%, gonorrhoea 0.2%, and syphilis 0.5% (all VDRL or RPR titres were < or = 1:4) . The prevalence of HIV infection was 7.1% in Chiang Rai and 2.9% in Bangkok . In a multivariate logistic regression analysis, chlamydial infection was associated with younger age and with higher gestational age at first antenatal clinic visit, but was not associated with marital status, gravidity, city of enrollment, or HIV infection status . CONCLUSIONS: There was a low prevalence of gonorrhoea and syphilis among these pregnant women in Thailand . Chlamydial infection was detected at a higher prevalence, especially among younger women and women registering later for antenatal care . Testing of pregnant women using easily collected urine specimens and a sensitive nucleic acid amplification assay is a feasible method of rapidly assessing chlamydial and gonococcal prevalence.

Proc Natl Acad Sci U S A, 1998 Dec 8, 95(25), 14973 - 8
Suppression of an absolute defect in type IV pilus biogenesis by loss-of-function mutations in pilT, a twitching motility gene in Neisseria gonorrhoeae; Wolfgang M et al.; Type IV pili of Neisseria gonorrhoeae, the Gram-negative etiologic agent of gonorrhea, facilitate colonization of the human host . Gonococcal PilT, a protein belonging to a large family of molecules sharing a highly conserved nucleotide binding domain motif, has been shown to be dispensable for organelle biogenesis but essential for twitching motility and competence for genetic transformation . Here, we show that the defect in pilus biogenesis resulting from mutations in the pilC gene, encoding a putative pilus-associated adhesin for human tissue, can be suppressed by the absence of functional PilT . These data conclusively demonstrate that PilT influences the Type IV pilus biogenesis pathway and strongly suggest that organelle expression is a dynamic process . In addition, these findings imply that PilT antagonizes the process of organelle biogenesis and provide the basis for a model for how the counteractive roles of PilT and PilC might relate mechanistically to the phenomenon of twitching motility.

J Infect Dis, 1999 Jan, 179(1), 124 - 35
Complement processing and immunoglobulin binding to Neisseria gonorrhoeae determined in vitro simulates in vivo effects; McQuillen DP et al.; Local inflammation elicited by Neisseria gonorrhoeae correlates closely with sensitivity to killing by normal human serum . Serum-sensitive (SS) isolates are rendered resistant in vitro by lipooligosaccharide sialylation . Differences in C3b processing on N . gonorrhoeae in vitro were found to match findings at the cervical level in vivo . Nonsialylated SS gonococci bound 5-fold more C3b than did stably serum-resistant (SR) gonococci; most was processed to iC3b, yet significant C3b persisted . Sialylated SS gonococci bound 4-fold less total C3 antigen than did SR gonococci, which was promptly converted to iC3b . C3b bound later on stably SR gonococci but again was processed swiftly to iC3b . In vivo, the iC3b/C3 ratio of SS isolates more closely resembled nonsialylated SS isolates in vitro, implying heterogeneous sialylation or desialylation in vivo . In vitro, total IgM bound was unchanged by sialylation of SS isolates, but total C4 bound decreased by 75%, suggesting that sialylation may indirectly regulate the classical complement pathway.

Ann Intern Med, 1998 Nov 15, 129(10), 813 - 28
Infections acquired during cardiopulmonary resuscitation: estimating the risk and defining strategies for prevention; Mejicano GC et al.; PURPOSE: To estimate the risk for acquiring an infectious disease during cardiopulmonary resuscitation (CPR) or CPR training and to identify strategies to minimize that risk . DATA SOURCES: English-language articles published since 1965 were identified through a search of the MEDLINE database and selected bibliographies . STUDY SELECTION: Studies that contained information about transmission of infectious organisms, particularly HIV and other bloodborne viruses that might be transmitted through mouth-to-mouth ventilation, contact exposures, and needlesticks during CPR . DATA EXTRACTION: Descriptive and analytic data from each study . DATA SYNTHESIS: Fear of acquiring infection, especially HIV infection, can delay prompt initiation of mouth-to-mouth ventilation . Although pathogens can be isolated from the saliva of infected persons, salivary transmission of bloodborne viruses is unusual and transmission of infection has been rare: Only 15 documented cases have been reported . Most of these cases involved a bacterial pathogen, such as Neisseria meningitidis . Transmission of hepatitis B virus, hepatitis C virus, or cytomegalovirus during CPR has not been reported; all three reported cases of HIV infection acquired during resuscitation of an infected patient resulted from high-risk cutaneous exposures . There have been no reports of infection acquired during CPR training . Simple infection-control measures, including use of barrier devices, can reduce the risk for acquisition of an infectious disease during CPR and CPR training . Postexposure protocols can further protect potential rescuers and trainees . CONCLUSIONS: The benefit of initiating lifesaving resuscitation in a patient in cardiopulmonary arrest greatly outweighs the risk for secondary infection in the rescuer or the patient . Nevertheless, use of simple infection-control measures during CPR and CPR training can reduce a very low level of risk even further.

Int J Infect Dis, 1998 Jul-Sep, 3(1), 5 - 11
Herpes simplex type II and Mycoplasma genitalium as risk factors for heterosexual HIV transmission: report from the heterosexual HIV transmission study; Perez G et al.; OBJECTIVES: Two hundred twenty-four human immunodeficiency virus (HIV) discordant couples (one HIV negative, one HIV positive) were compared with 78 seroconcordant heterosexually infected couples with HIV with regard to sexually transmitted diseases . METHODS: Serologic testing and cultures were used to determine exposure of participants to sexually transmitted pathogens . These data were compared with HIV concordance of partners to investigate possible risk factors for HIV transmission . RESULTS: Syphilis, chlamydia, and hepatitis B virus (HBV) serologies did not distinguish between concordant and discordant couples nor did cultures for Neisseria gonorrhoeae and Trichomonas or Chlamydia enzyme immunoassay (EIA) . Risk of transmission increased with positive serologies for herpes simplex virus (HSV)-2 (P = 0.002), cytomegalovirus (CMV) (P = 0.04), and Mycoplasma genitalium (P = 0.01), but not with Mycoplasma fermentans or Mycoplasma penetrans . Cytomegalovirus was not a significant risk factor when controlled for HSV-2 status . Examination by partner status showed increased risk of concordance with: HSV-2 positive serology in both partners (odds ratio {OR} = 3.14; confidence interval {CI} = 1.62-6.09; P = 0.007); HSV-2 in female secondary partner (OR = 2.10; CI = 1.12-3.93; P = 0.02) or the male primary partner (OR = 2.15; CI = 1.15-4.02; P = 0.017); M . genitalium antibody in both partners (OR = 3.44; CI = 1.68-7.04; P < 0.001); M . genitalium antibody in the primary male partner (OR = 2.51, CI = 1 . 27-4.91; P = 0.008) and M . genitalium antibody in the secondary female partner (OR = 2.52; CI = 1.21-5.23; P = 0.01) . CONCLUSIONS: These data support the role of HSV-2 in transmission of HIV and, for the first time, suggest a role for M . genitalium as an independent risk factor.

Clin Infect Dis, 1998 Nov, 27(5), 1214 - 20
Epidemic spread of subgroup III of Neisseria meningitidis serogroup A to South Africa in 1996; McGee L et al.; A total of 111 strains of Neisseria meningitidis isolated during a meningitis outbreak in Johannesburg, South Africa, in 1996 were analyzed by serogrouping, arbitrarily primed polymerase chain reaction analysis, ribotyping, and multilocus enzyme electrophoresis . Nineteen different clusters were identified, and typing patterns for the three techniques were comparable . Of the 111 strains analyzed, 55 (49.5%) belonged to the serogroup A clone complex designated internationally as subgroup I-1 . The second largest cluster included 15 isolates (13.5%) that were identified as belonging to the pandemic clone III-1 . A number of unrelated strains were also encountered . Our study provides the first documented evidence of the presence of clone III-1 in South Africa and indicates the continued spread of this clone well outside the African meningitis belt.

Clin Infect Dis, 1998 Nov, 27(5), 1171 - 80
Laboratory testing for Neisseria gonorrhoeae by recently introduced nonculture tests: a performance review with clinical and public health considerations; Koumans EH et al.; Since 1992 two new nucleic acid-based tests (Gen-Probe Pace 2 and Abbott LCR) for the diagnosis of Neisseria gonorrhoeae infection have been approved by the U.S . Food and Drug Administration . We systematically assessed the quality of 21 studies that evaluated these tests' performance compared with that of culture for diagnosis of gonorrhea, on the basis of established criteria . We estimated overall test sensitivity and specificity by the testing method, sex, and anatomic site . None of the studies optimally fulfilled all quality criteria; few studies adequately used reference tests or described blinding . The sensitivity and specificity of nucleic acid hybridization (approximately 85%, approximately 98%) and amplification tests (approximately 95%, approximately 99%) were high and did not appear to differ substantially by sex or anatomic site . When proficiency in the performance of culture is high, the new tests are comparable to culture and may not offer a substantial advantage; in settings where optimization of culture is difficult, nucleic acid amplification may detect more infections than nucleic acid probe or culture.

Dakar Med, 1996, 41(2), 125 - 7
{Bacteriology of vulvo-vaginitis in pediatrics}; Sow AI et al.; Fifty (50) samples of genital secretions are collected from young girls between 1983 and 1994 at Albert Royer Hospital (Dakar, Senegal), to research pathogenic strains . Each sample were studied by microscopic examination and culture in several media after enrichment . 70% of the samples are positive of specific agents (37.25%), particularly Candida albicans (19.6%) . Neisseria gonorrhoeae was found among the elders . This retrospective study shows the rarity of specific agents of genital infections generally isolated among the adults . The authors were not able to research other agents as Chlamydia trachomatis or viruses.

Infect Immun, 1998 Dec, 66(12), 5826 - 32
Evaluation of immunoglobulin A1 (IgA1) protease and IgA1 protease-inhibitory activity in human female genital infection with Neisseria gonorrhoeae; Hedges SR et al.; Immunoglobulin A1 (IgA1) protease, an enzyme that selectively cleaves human IgA1, may be a virulence factor for pathogenic organisms such as Neisseria gonorrhoeae . Host protection from the effects of IgA1 protease includes antibody-mediated inhibition of IgA1 protease activity, and it is believed that the relative balance between IgA1 protease and inhibitory antibodies contributes to the pathogenesis of disease caused by IgA1 protease-producing organisms . We have examined the levels of these two opposing factors in genital tract secretions and sera from women with uncomplicated infection with N . gonorrhoeae . When IgA1 in cervical mucus was examined by Western blotting, no evidence of cleavage fragments characteristic of IgA1 protease activity was seen in gonococcus-infected or control patients . Cleavage fragments typical of IgA1 protease were detected, however, after the addition of exogenous IgA1 protease to cervical mucus . Degraded IgA1 was detected in some vaginal wash samples, but the fragment pattern was not typical of IgA1 protease activity . All N . gonorrhoeae isolates from the infected patients produced IgA1 protease in vitro . All but two serum samples and 16 of 65 cervical mucus samples displayed inhibitory activity against gonococcal IgA1 protease, but there was no significant difference in the level of inhibitory activity between gonococcus-infected and noninfected patients in either cervical mucus or serum . There was no difference in the levels of IgA1 protease-inhibitory activity in serum or cervical mucus collected from patients at recruitment and 2 weeks later . These results suggest that cleavage of IgA1 by gonococcal IgA1 protease within the lumen of the female lower genital tract is unlikely to be a significant factor in the pathogenesis of infections by N . gonorrhoeae.

Mol Microbiol, 1998 Nov, 30(3), 657 - 71
Opa binding to cellular CD66 receptors mediates the transcellular traversal of Neisseria gonorrhoeae across polarized T84 epithelial cell monolayers; Wang J et al.; We have analysed the capacity of the 11 phase-variable, opacity-associated (Opa) proteins encoded by Neisseria gonorrhoeae MS11 to mediate traversal across polarized monolayers of the human colonic carcinoma T84 cell line . Gonococci expressing either the heparan sulphate proteoglycan (HSPG) binding Opa protein (Opa50) or no Opa protein (Opa-) did not interact with the apical pole of T84 monolayers, whereas the 10 variant Opa proteins previously shown to bind CD66 receptors were found to mediate efficient gonococcal adherence and transepithelial traversal . Consistent with this, T84 cells were shown by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting to co-express CD66a (BGP), CD66c (NCA) and CD66e (CEA) . The recruitment of CD66 receptors by Opa-expressing gonococci indicates their involvement in mediating adherence to the surface of T84 cells, and these bacterial interactions could be inhibited completely using polyclonal antibodies cross-reacting with all of the CD66 proteins co-expressed on T84 cells . Consistent results were obtained when Opa proteins were expressed in Escherichia coli, suggesting that the Opa-CD66 interaction is sufficient to mediate bacterial traversal . Transcytosis of Opa-expressing N . gonorrhoeae or E . coli did not disrupt the barrier function of infected monolayers, as indicated by a sustained transepithelial electrical resistance (TEER) throughout the course of infection, and confocal laser scanning and electron microscopy both suggest a transcellular rather than a paracellular route of traversal across the monolayers . Parallels between the results seen here and previous work done with organ cultures confirm that T84 monolayers provide a valid model for studying neisserial interactions with the mucosal surface, and suggest that CD66 receptors contribute to this process in vivo.

J Infect, 1998 Mar, 36(2), 141 - 3
Spread of Neisseria meningitidis group A clone III-I meningitis epidemic into Zambia; Luo N et al.; The epidemic of Neisseria meningitidis group A clone III-I strain began with the 1987 outbreak in Mecca, Saudi Arabia . Intense epidemics in African countries occurred the following year . Clone III-I was first isolated from Africa in 1988 in Chad, followed by epidemics in Ethiopia (1989), Kenya and Tanzania (1989), and The Central African Republic (1992) . We report an epidemic of Neisseria meningitidis group A clone III-I afflicting Zambia during a 2-year period (April 1992-May 1994) . This extension of the 'meningitis belt' southwards into Central Africa is cause for concern . Active surveillance for this disease should be carried out by all African countries, even those outside the well described sub-Saharan 'meningitis belt'.

J Infect, 1998 Jan, 36(1), 49 - 52
Association of Neisseria cinerea with ocular infections in paediatric patients; Dolter J et al.; Twenty-two strains of Neisseria cinerea were recovered from paediatric patients over a 7-year period and forwarded to the Microbial Diseases Laboratory for biochemical identification and/or confirmation . Eighteen of these 22 strains (82%) were recovered from the eyes of very young children (< or = 1 year), > 50% occurring during the neonatal period . The majority of eye isolates were involved in a variety of ocular infections including orbital cellulitis, conjunctivitis, and eye discharge (most common); in four of the 13 instances (31%) where laboratory data was available, Neisseria cinerea was recovered in pure culture . Neisseria cinerea isolates were often submitted to the Microbial Diseases Laboratory as possible 'N . gonorrhoeae' or 'Neisseria species' due to problems resulting from the use of commercial assays or unfamiliarity with the organism . These observations indicate that N . cinerea can produce eye infections in very young children, who presumably acquire this organism vertically from the mother during birth . Accurate identification of N . cinerea in such infants can preclude the social trauma and possible legal ramifications which can initially result from its misidentification as N . gonorrhoeae.

J Clin Microbiol, 1998 Dec, 36(12), 3624 - 8
Pooling of urine samples for screening for Neisseria gonorrhoeae by ligase chain reaction: accuracy and application; Kacena KA et al.; The accuracy of detection of genital Neisseria gonorrhoeae infection in pooled urine samples by ligase chain reaction (LCR) was examined in three populations . Firstly, urine specimens from 300 female military recruits (FMR) were tested by LCR individually and in pools of four and six . Secondly, 300 urine specimens from middle-school students (MSS) were tested individually by LCR, and then the processed specimens were stored frozen for subsequent testing in pools of 4 and 10 . Thirdly, 600 frozen urine specimens from high-school students (HSS) were tested by using the LCR pooling algorithm, i.e., testing processed specimens in pools of four in one test unit dose, and retesting individual specimens from positive pools . Finally, the pooling algorithm results were compared to culture results for a subset of 344 students from the original 600 HSS from whom cervical or urethral samples were taken at the discretion of the school nurse practitioners . Compared to individual testing of specimens by LCR in the FMR population, the pooling-by-four algorithm was 100% sensitive (5 of 5) and 100% pool specific (70 of 70), and the pool-by-six algorithm was 100% sensitive (5 of 5) and 100% pool specific (45 of 45) . In the MSS population, the pool-by-4 algorithm was 95.8% sensitive (23 of 24) and 100% (52 of 52) pool specific, and the pool-by-10 algorithm was 95.8% sensitive (23 of 24) and 100% (17 of 17) pool specific . In the subset of 344 HSS from whom endocervical or urethral specimens were collected for culture, 31 were positive by LCR in urine and 26 were positive by culture . After results discrepant between culture and LCR were adjudicated by a confirmatory LCR test, the pooling algorithm was 93.8% (30 of 32) sensitive and 99.7% (311 of 312) specific . Culture from these 344 HSS was 81.3% (26 of 32) sensitive . The pooling algorithm reduced the cost of the N . gonorrhoeae LCR assay by 60% compared to individual testing of the HSS specimens and was both sensitive and specific.

J Clin Microbiol, 1998 Dec, 36(12), 3549 - 51
Evaluation of the abbott LCx assay for detection of Neisseria gonorrhoeae in endocervical swab specimens from females; Kehl SC et al.; The Abbott LCx Neisseria gonorrhoeae assay (Abbott Laboratories, Abbott Park, Ill.) uses a ligase chain reaction (LCR) amplification in the LCx probe system for detection of a specific nucleotide sequence in the Opa-encoding gene of N . gonorrhoeae . We evaluated the LCx assay in a comparison with conventional culture employing modified Thayer-Martin media for the detection of N . gonorrhoeae from female endocervical specimens obtained from patients attending a sexually transmitted disease clinic . Discordantly LCR-positive and culture-negative specimens were further evaluated by testing with another LCR assay which used an N . gonorrhoeae-specific pilin probe . Specimens positive by both LCR assays were considered confirmed LCx-positive specimens . A specimen was considered to contain N . gonorrhoeae when it was either culture positive or culture negative and confirmed LCx positive . A total of 403 female endocervical specimens were evaluated . The prevalence of N . gonorrhoeae in this population was 8.7% . The sensitivity and specificity of the LCx assay were 94.3 and 99.4%, and those of culture were 77.1 and 100%, respectively . The Abbott LCx assay is a rapid, sensitive method for detection of N . gonorrhoeae in female endocervical specimens.

J Infect Dis, 1998 Dec, 178(6), 1707 - 12
Transmission of Chlamydia trachomatis and Neisseria gonorrhoeae among men with urethritis and their female sex partners; Lin JS et al.; Transmission of Chlamydia trachomatis and Neisseria gonorrhoeae among infected men and their female sex partners was examined using a design enhancing the likelihood that spread was directed from men to women . Chlamydia culture-negative specimens were examined using DNA amplification tests . Infection rates in women exposed to male sex partners with Chlamydia only were 65% (20/31) and with gonorrhea only were 73% (33/45) . Infection of women by either agent was not influenced by the number of sexual exposures to or coinfection in men . There was a 98% (40/41) concordance of N . gonorrhoeae isolates among partners by auxotype and serovar . Chlamydia isolates were serotyped using ELISA and immunofluorescence testing and confirmed by nested polymerase chain reaction: 50% (6/12) of men and 57% (8/14) of women yielded mixed serovars . Sixty-four percent of pairs (9/14) were infected with identical serovars and an additional 28% shared at least one serovar . Multiple serovars of C . trachomatis, but not of N . gonorrhoeae, were common in sex partners and exchanged frequently.

AIDS, 1998 Oct 22, 12(15), 2041 - 8
Incidence of HIV and sexually transmitted diseases (STD) in a cohort of HIV-negative men who have sex with men (MSM); Tabet SR et al.; OBJECTIVE: To determine the prevalence of sexually transmitted diseases (STD) and incidence of and risk factors for STD, including HIV-1, among a cohort of HIV-negative men who have sex with men (MSM) . SETTING: Seattle, Washington, United States . PARTICIPANTS: Prospective cohort of 578 HIV-negative MSM in which risk factors for acquiring a STD over 12 months follow-up were evaluated using a cumulative incidence analysis . MAIN OUTCOME MEASURES: Baseline tests obtained were: herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) Western blots, hepatitis B, and syphilis serologies; anorectal and pharyngeal Neisseria gonorrhoeae (GC) cultures; first-catch urine for leukocyte esterase (LE) and Chlamydia trachomatis (CT) ligase chain reaction (LCR) . Men with a positive urine LE had urethral GC cultures obtained . The following outcomes were measured over 12 months follow-up: incident symptomatic bacterial STD (urethritis, proctitis, epididymitis), HSV-1 and HSV-2 seroconversion, and HIV-1 seroconversion . The 31 incident cases of STD (men with bacterial STD) were compared with those 489 men without symptomatic bacterial STD or seroconversion to HSV-1, HSV-2 or HIV-1 infection . RESULTS: Bacterial STD were found in nine participants at enrollment; there were two cases of nonchlamydial urethritis, two cases of nonchlamydial epididymitis, and five cases of asymptomatic GC infection . At enrollment, HSV-2 antibodies were detected in 149 (26.0%) of 572 men and prior hepatitis B infection in 145 (34.8%) of 417 unvaccinated men . During the 1-year of follow-up, 31 men (5.7/100 person-years) had 34 episodes of a symptomatic bacterial STD syndrome (urethritis, epididymitis or proctitis) . Urethritis was the most common STD syndrome, detected in 29 men, of whom 10 had GC and 19 had nongonococcal urethritis . In the 1-year of follow-up, five participants seroconverted to HIV-1 (1.3/100 person-years), four to HSV-2 (1.0/100 person-years), and seven to HSV-1 (4.3/100 person-years) . Unprotected insertive anal sex {odds ratio (OR) 2.6; 95% confidence interval (CI) 1.2-5.6}; and nitrite inhalant ('poppers') use (OR, 2.3; 95% CI, 1.0-5.0) were independently associated with incident STD . CONCLUSIONS: STD and HIV infection continue to be acquired even in a city with an overall low bacterial STD prevalence and among educated MSM receiving regular HIV screening and risk-reduction . Urethritis was the most common STD detected, and public health messages aimed at MSM need to emphasize safe insertive as well as receptive sexual practices.

Zhonghua Liu Xing Bing Xue Za Zhi, 1997 Apr, 18(2), 86 - 8
{Detection and analysis of mycoplasma in 400 cases of genitourinary infection in Wuhan area}; Liu C et al.; Mycoplasmas were examined by cultured in the urogenital secreta and the midpieceurine from 400 cases (88 men and 312 women) of genitourinary infection and 119 cases (positive rate 29.75%) of genitourinary infection in Wuhan area . The positive rates of ureaplasma urealyticum (Uu) and mycoplasma hominis (Mh) were 24.75% and 8.75% respectively . The authors also studied the infection rates of Neisseria gonorrheae, fungus and trichomonad by cultures and microscopic findings . The positive rates of Neisseria gonorrheae, fungus and trichomonad were 14.50%, 13.50% and 4.50% respectively . Results showed that mycoplasma was one of the common pathogens causing genitourinary infection in Wuhan area.

J Womens Health, 1998 Oct, 7(8), 1033 - 40
Factors predicting upper genital tract inflammation among women with lower genital tract infection; Nelson DB et al.; We sought to identify factors that discriminate between women with a lower genital tract infection and women with a lower genital tract infection and endometritis . This study enrolled women at risk for or having a lower genital tract infection with Chlamydia trachomatis or Neisseria gonorrhoeae and measured behavioral and clinical factors . Women were identified through contact tracing of male partners, presentation with cervicitis, or presentation with symptoms of pelvic inflammatory disease and classified as (1) having a lower genital tract infection without endometritis, (2) having a lower genital tract infection with endometritis, (3) having no lower genital tract infection with endometritis, and (4) having neither a lower genital tract infection nor endometritis . The primary comparison was between women having a lower genital tract infection without endometritis to women having a lower genital tract infection and endometritis . Women with a lower genital tract infection and endometritis were older and reported a history of more sexually transmitted diseases (70.0% vs . 56.7%), abdominal pain (82.2% vs . 60.0%), and use of barrier methods of contraception (28.9% vs . 8.6%) than women with a lower genital tract infection alone . The regression model found that women with a lower genital tract infection and endometritis were 7.1 times (95% CI = 2.2-23.0) more likely to report abdominal pain and 4.6 times (95% CI = 1.5-14.9) more likely to use barrier methods of contraception than women with a lower genital tract infection alone . These results suggest that behavioral factors, in addition to symptoms, can be used to identify women with and without upper genital tract involvement.

S Afr Med J, 1998 Oct, 88(10), 1337 - 40
The sensitivity of a syndromic management approach in detecting sexually transmitted diseases in patients at a public health clinic in Cape Town; Mathews C et al.; OBJECTIVES: To evaluate the sensitivity of a syndromic diagnostic procedure in detecting and treating sexually transmitted diseases (STDs) and genital tract infections (GTIs) . METHODS: All new patients presenting at an STD clinic were sampled systematically by gender over a 6-week period . After the patient's clinical consultation, the clinical records were reviewed . Thereafter all patients were given a thorough genital examination by the research physician, and specimens were collected for laboratory investigations . In a retrospective simulation clinicians' syndromic diagnoses were validated against the laboratory findings, or for genital ulcer syndrome against the findings of the research physician . RESULTS: 170 men and 161 women were included in the sample . Ninety-five per cent of patients were black and the median age was 22 years for women and 26 years for men . In this setting, the Western Cape syndromic diagnostic procedure achieved reasonable levels of sensitivity in detecting Neisseria gonorrhoeae and Chlamydia trachomatis in men and women, and in detecting Trichomonas vaginalis and bacterial vaginosis in women . However, it was estimated to be only 36.4% sensitive in detecting genital ulcers in women, and between 0% and 12.3% sensitive in detecting Candida albicans . With syndromic management 8.2% of men and 32.9% of women would leave the clinic with at least one infection inadequately treated . CONCLUSIONS: Despite the introduction of syndromic protocols, it is likely that a proportion of STDs and GTIs are not being detected and treated owing to the high prevalence of multiple syndromes and mixed infections, both symptomatic and asymptomatic.

Commun Dis Intell, 1998 Oct 1, 22(10), 212 - 6
Annual report of the Australian Gonococcal Surveillance Programme, 1997.
Screening for sexually transmitted diseases in rural women in Papua New Guinea: are WHO therapeutic algorithms appropriate for case detection?
Papua New Guinea Institute of Medical Research, New GuineaThe presence of a large reservoir of untreated sexually transmitted diseases (STDs) in developing countries has prompted a number of suggestions for improving case detection, including the use of clinical algorithms and risk assessments to identify women likely to be infected when they present to clinics for other reasons . We used data from a community-based study of STDs to develop and evaluate algorithms for detection of cervical infection with Chlamydia trachomatis or Neisseria gonorrhoeae, and for detection of vaginal infection with Trichomonas vaginalis or bacterial vaginosis . The algorithms were derived using data from 192 randomly selected women, then evaluated on 200 self-selected women . We evaluated the WHO algorithm for vaginal discharge in both groups . The prevalences of cervical and vaginal infection in the randomly selected group were 27% and 50%, respectively, and 23% and 52%, respectively, in the self-selected group . The derived algorithms had high sensitivities in both groups, but poor specificities in the self-selected women, and the positive predictive values were unacceptably low . The WHO algorithms had extremely low sensitivity for detecting either vaginal or cervical infection because relatively few women reported vaginal discharge . Simple algorithms and risk assessments are not valid for case detection in this population.

Bull World Health Organ, 1998, 76(4), 393 - 400
{Standardization of the Neisseria meningitidis antibiogram . Detection of strains relatively resistant to penicillin}; Nicolas P et al.; Studying the susceptibility of 189 Neisseria meningitidis strains to penicillin, amoxicillin, cefotaxime, ceftriaxone, chloramphenicol and rifampicin by determination of minimum inhibitory concentrations (MICs) by agar dilution (reference method), E-test and disc diffusion method on Mueller-Hinton agar at 37 degrees C with 5% CO2 enabled us to standardize the antibiograms . While MIC determination by agar dilution is still the reference method, it is possible to obtain exact or approximate MIC values using the E-test . For laboratories that cannot determine penicillin MICs, it is impossible to detect strains that are relatively resistant to penicillin (RRP strains: 0.1 < or = MIC < or = 1 mg/l) using a 10-U penicillin disc . A 1 microgram-oxacillin disc allows MIC to be determined in most cases when the oxacillin inhibition zone is < or = 10 mm . Such strains must be sent to a reference laboratory for exact MIC determination . Based on our results and literature data on pharmacokinetics, we propose critical concentrations for these various antibiotics as well as critical diameters for chloramphenicol and rifampicin discs.

Clin Diagn Lab Immunol, 1998 Nov, 5(6), 845 - 55
Proposed standardization of Neisseria meningitidis PorA variable-region typing nomenclature; Sacchi CT et al.; Neisseria meningitidis isolates are conventionally classified by serosubtyping, which characterizes the reactivities of the PorA outer membrane protein variable-region (VR) epitopes with monoclonal antibodies (MAbs) . A newer method (PorA VR typing) uses predicted amino acid sequences derived from DNA sequence analysis . The resulting classification schemes are not standardized, offering conflicting and sometimes irreconcilable data from the two methods . In this paper, we propose a standardization of the PorA VR typing nomenclature that incorporates serologic information from traditional PorA serosubtyping with molecular data from predicted VR sequences . We performed a comprehensive literature and database search, generating a collection of strains and DNA sequences that reflects the diversity within PorA that exists to date . We have arranged this information in a comprehensive logical model that includes both serosubtype and PorA VR type assignments . Our data demonstrate that the current panel of serosubtype-defining MAbs underestimates PorA VR variability by at least 50% . Our proposal for VR typing is informative because amino acid sequence and serologic information, when serosubtype-defining MAbs are available, can be deduced simultaneously from the PorA VR designation . This scheme will be useful in future classification and applied epidemiologic studies of N . meningitidis, being a systematic way of selecting PorA vaccine candidates and analyzing vaccine coverage and failure.

Sex Transm Dis, 1998 Oct, 25(9), 489 - 93
Analysis of quinolone resistance mechanisms in a sparfloxacin-resistant clinical isolate of Neisseria gonorrhoeae; Tanaka M et al.; BACKGROUND AND OBJECTIVES: Recently, a reduction in the susceptibility of clinical isolates of Neisseria gonorrhoeae to newer fluoroquinolones including sparfloxacin in vitro has been recognized in Japan . The quinolone resistance mechanisms in gonococcal isolates from a patient with clinical failure of sparfloxacin treatment was investigated . GOAL: To report a man with gonococcal urethritis in whom clinical failure of sparfloxacin treatment occurred and to examine the quinolone resistance mechanisms in gonococcal isolates from the patient . STUDY DESIGN: A man with gonococcal urethritis was treated with oral 100 mg sparfloxacin three times daily for 5 days . However, clinical failure of the sparfloxacin treatment was observed . The antimicrobial susceptibilities of pretreatment and posttreatment isolates to sparfloxacin and other agents were measured . To analyze quinolone resistance mechanisms in the set of isolates, DNA sequencing of the genes corresponding to the quinolone resistance-determining regions within the GyrA and ParC proteins was performed . We also assayed the intracellular sparfloxacin accumulation level in these gonococcal cells . Moreover, we performed pulsed-field gel electrophoresis analysis to determine whether the pretreatment and posttreatment isolates were isogenic . RESULTS: The minimum inhibitory concentration of sparfloxacin for the posttreatment isolate (4 micrograms/ml) was 16 times higher than that for the pretreatment isolate (0.25 microgram/ml) . The pretreatment isolate contained three mutations, including a Ser-91 to Phe mutation and an Asp-95 to Asn mutation in GyrA and a Ser-88 to Pro mutation in ParC . The posttreatment isolate had four mutations, including the same three mutations and an additional Glu-91 to Gly mutation in ParC . The sparfloxacin accumulation level within 30 minutes in the posttreatment isolate was four times less than that in the pretreatment isolate . There were no differences in the pulsed-field gel electrophoresis patterns between the pretreatment and posttreatment isolates from the patient . CONCLUSIONS: The emergence of a fluoroquinolone-resistant N . gonorrhoeae isolate with multiple mutations involving GyrA and ParC reduced the response to sparfloxacin treatment . Multiple dosing and long-term treatment with sparfloxacin seems to induce a mutation in ParC and an alteration leading to reduced drug accumulation that contribute to increasing the fluoroquinolone resistance level.

Sex Transm Dis, 1998 Oct, 25(9), 464 - 7
Clinical evaluation of ceftibuten in gonorrhea . A pilot study in Hong Kong; Chong LY et al.; BACKGROUND: The escalating rates of gonococcal resistance to quinolone in Hong Kong have prompted a search for an alternative first-line antimicrobial agent for use in treating uncomplicated gonococcal urethritis . Ceftibuten is an orally active third-generation cephalosporin with potent in vitro activity against Neisseria gonorrhoeae . Its pharmacokinetic properties allow single-dose administration . OBJECTIVE: To evaluate the efficacy, safety, and tolerability of ceftibuten in the treatment of uncomplicated gonorrhea in men . STUDY DESIGN: Ceftibuten was evaluated in an open-label, noncomparative, multicenter study . Eligible men with uncomplicated gonococcal urethritis were treated with a single 400-mg oral dose of ceftibuten and reassessed 1 week and 3 weeks after treatment . The main outcome measures were the isolation of N . gonorrhoeae, patient-reported side effects, and other safety parameters (e.g., blood counts and renal and hepatic function tests) . RESULTS: One hundred twenty-five men were enrolled in the study . The overall cure rate was 98.2% (110 of 112 evaluable patients) . Adverse events, which occurred in 4.5% of patients, were all mild, well tolerated, and of short duration . No significant changes in laboratory test results were noted . Of the 125 isolates, 4.8% were -lactamase positive . Susceptibility to ofloxacin was found to be low in 59.2% of isolates (MIC 0.1 to < 1g/mL) and 25.6% of isolates were resistant (MIC1g/mL) to ofloxacin . CONCLUSIONS: A single 400-mg oral dose of ceftibuten is highly effective and well tolerated in the treatment of uncomplicated gonococcal urethritis in men.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2919 - 22
Drift in susceptibility of Neisseria gonorrhoeae to ciprofloxacin and emergence of therapeutic failure; Ison CA et al.; Ciprofloxacin, 500 mg, was introduced as the first-line therapy for gonorrhea at St . Mary's Hospital, London, in 1989, when a surveillance program was initiated to detect the emergence of resistance . Isolates of Neisseria gonorrhoeae from consecutive patients attending the Jefferiss Wing, Genitourinary Medicine Clinic at St . Mary's Hospital, between 1989 and 1997 have been tested for susceptibility to ciprofloxacin by using an agar dilution breakpoint technique . Isolates considered potentially resistant (MIC, >0.12 microg/ml) were further characterized by determination of the MICs of ciprofloxacin, nalidixic acid, and penicillin, auxotyped and serotyped, and screened for mutations in the DNA gyrase gene, gyrA, and the topoisomerase IV gene, parC . A total of 4,875 isolates were tested . While the majority of isolates were highly susceptible (MIC, </=0.008 microg of ciprofloxacin/ml), there was a drift toward reduced susceptibility in N . gonorrhoeae isolated between 1993 and 1996 (P < 0.001) . In 1997 this drift was reduced but remained above pre-1993 levels . Isolates from 18 patients were classed as potentially resistant (MIC, >0.12 microg/ml); all of these belonged to serogroup B, and NR/IB-1 was the most common auxotype/serovar class . The infections in 14 of the 18 patients were known to be acquired abroad, and 5 were known to result in therapeutic failure . The surveillance program has established that ciprofloxacin is still a highly effective antibiotic against N . gonorrhoeae in this population . However, it has identified a drift in susceptibility which may have resulted from increased usage of ciprofloxacin . High-level resistance has now emerged, although treatment failure is still uncommon.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2799 - 803
Gonococcal resistance to beta-lactams and tetracycline involves mutation in loop 3 of the porin encoded at the penB locus; Gill MJ et al.; penB is a chromosomal mutation that confers resistance to beta-lactams and tetracyclines and reduced susceptibility to quinolones in Neisseria gonorrhoeae . It is linked to the porin gene (por) and requires the increased expression of an efflux pump due to mtr . Transformation of a susceptible gonococcus (strain H1) with chromosomal DNA from strain FA140 (penA mtr penB; porin serovar IB1) and conjugal transfer of a beta-lactamase-expressing plasmid was used to produce isogenic strains for determination of equilibrium periplasmic penicillin concentrations by the method of Zimmermann and Rosselet (W . Zimmermann and A . Rosselet, Antimicrob . Agents Chemother . 12:368-372, 1977) . In transformants with the Mtr and PenB phenotypes, equilibrium concentrations of penicillin were reduced . DNA sequence analysis of por from isogenic penB and penB+ transformants revealed 14 sequence differences; nine of these differences resulted in amino acid changes . Three amino acid changes were found in the putative gonococcal equivalent of the pore-constricting loop 3 of Escherichia coli OmpF . Two of these changes (Gly-101-Ala-102-->Asp-Asp) result in an increased negative charge at this position in por loop 3 . PCR products comprising the complete por gene from strain FA140 were transformed into strain H1-2 (penA mtr; porin serovar IB-3), with the resulting transformants having the antibiotic susceptibility phenotype associated with penB . penB-like mutations were found in loop 3 of clinical isolates of gonococci with chromosomally mediated resistance to penicillin . We conclude that penB is a mutation in loop 3 of por that reduces porin permeability to hydrophilic antibiotics and plays an important role in the development of chromosomally mediated resistance to penicillin and tetracycline in gonococci.

Mol Microbiol, 1998 Oct, 30(1), 7 - 17
Type IV pili are involved in plant-microbe and fungus-microbe interactions; Dorr J et al.; Adherence of bacteria to eukaryotic cells is essential for the initiation of infection in many animal and human pathogens, e.g . Neisseria gonorrhoeae and Pseudomonas aeruginosa . Adhesion-mediating type IV pili, filamentous surface appendages formed by pilin subunits, are crucial virulence factors . Here, we report that type IV pilus-dependent adhesion is also involved in plant-bacteria and fungus-bacteria interactions . Nitrogen-fixing, endophytic bacteria, Azoarcus sp., can infect the roots of rice and spread systemically into the shoot without causing symptoms of plant disease . Formation of pili on solid media was dependent on the pilAB locus . PilA encodes an unusually short (6.4 kDa) putative pilin precursor showing 100% homology to the conserved N-terminus of the Pseudomonas aeruginosa type IV pilin . PilB encodes for a 14.2 kDa polypeptide showing similarity to FimF, a component of type I fimbriae of Escherichia coli . It was found to be extruded beyond the cell surface by immunofluorescence studies, and it may, therefore, be part of a pilus assembly complex or the pilus itself . Both genes are involved in the establishment of bacteria on the root surface of rice seedlings, as detected by fluorescence microscopy . Moreover, both genes are necessary for bacterial adhesion to the mycelium of an ascomycete, which was isolated from the same rhizosphere as the bacteria . In co-culture with the fungus, Azoarcus sp . forms complex intracytoplasmic membranes, diazosomes, which are related to efficient nitrogen fixation . Adhesion to the mycelium appears to be crucial for this process, as diazosomes were absent and nitrogen fixation rates were decreased in pilAB mutants in co-culture.

Infect Immun, 1998 Nov, 66(11), 5215 - 23
Neisseria gonorrhoeae heme biosynthetic mutants utilize heme and hemoglobin as a heme source but fail to grow within epithelial cells; Turner PC et al.; Many bacterial pathogens, including pathogenic neisseriae, can use heme as an iron source for growth . To study heme utilization by Neisseria gonorrhoeae, two heme biosynthetic mutants were constructed, one with a mutation in hemH (the gene encoding ferrochelatase) and one with a mutation in hemA (the gene encoding gamma-glutamyl tRNA reductase) . The hemH mutant failed to grow without an exogenous supply of heme or hemoglobin, whereas the hemA mutant failed to grow unless heme, hemoglobin, or heme precursors were present . Growth of the mutants with hemoglobin required expression of the hemoglobin receptor (HpuAB) and was TonB dependent . However, growth with heme required neither HpuAB nor TonB . An fbpA mutant grew normally when either heme or hemoglobin was present in the medium . The heme biosynthetic mutants showed reduced intracellular survival, compared to the parent strain, within A-431 endocervical epithelial cell cultures . These studies demonstrate that in addition to synthesizing their own heme, N . gonorrhoeae strains are able to internalize and utilize exogenous heme independently of FbpA but appear unable to obtain heme from within epithelial cells for growth.

Int J Urol, 1998 Sep, 5(5), 471 - 5
Reduced chlamydial infection and gonorrhea among commercial sex workers in Fukuoka City, Japan; Tanaka M et al.; BACKGROUND: Until recently, epidemiologic studies of sexually transmitted diseases (STDs), including human immunodeficiency virus type-1 (HIV-1) infection, among Japanese female commercial sex workers and their patterns of condom use have been rare . We investigated trends in STDs among female commercial sex workers and their condom use patterns in Fukuoka, Japan, from 1990 through 1995 . METHODS: The study group consisted of 1218 female commercial sex workers who attended an STD clinic to undergo screening for major STDs including chlamydial infection, gonorrhea and HIV-1 infection from 1990 through 1995 . Endocervical smear specimens were taken from the women to detect Chlamydia trachomatis and Neisseria gonorrhoeae, and blood samples were obtained for the serologic diagnosis of HIV-1 . Also, the commercial sex workers were interviewed concerning their condom use patterns . RESULTS: The annual infection and detection rates of C . trachomatis declined significantly from 58.1% in 1990 to 46.2% in 1995 and from 16.3% in 1990 to 10.0% in 1995, respectively, while the annual infection and detection rates of N . gonorrhoeae also declined significantly from 13.2% in 1990 to 3.5% in 1995 and from 1.5% in 1990 to 0.4% in 1995, respectively . None were found to be seropositive for HIV-1 during the 6-year period . The proportion of commercial sex workers using condoms significantly increased during 1992-1993 and 1994-1995 periods, as compared with the 1990-1991 period . CONCLUSION: Reductions in the prevalence of major STDs among female commercial sex workers may be related to an increase in the frequency of condom use.

Adv Exp Med Biol, 1998, 443, 123 - 33
Bacterial lactoferrin receptors; Schryvers AB et al.; Lactoferrin is thought to play a pivotal role in prevention of infection in the host and its ability to sequester iron from potential pathogens has been considered an important component of its antimicrobial function . A number of bacterial species in the Neisseriaceae have developed a mechanism for acquiring iron directly from this host glycoprotein which involves surface receptors capable of specifically binding lactoferrin . Initial attempts at identifying the receptor proteins in Neisseria and Moraxella species using affinity isolation with immobilized lactoferrin under high stringency conditions presumptively identified a single 100 kDa receptor protein, LbpA (formerly Lbp1) . Under modified affinity isolation conditions a second 84 kDa lactoferrin binding protein was isolated and had been presumptively identified as LbpB . This protein was not isolated from a CopB-ve isogenic mutant of Moraxella catarrhalis, indicating that it was in fact CopB . However, another lactoferrin binding protein isolated under high stringency conditions, that comigrated with LbpA in most, but not all, M . catarrhalis strains, was identified by convalescent antisera . Its biochemical properties suggested that it indeed was LbpB . The identity of these proteins was confirmed by preparing isogenic mutants with the lbpA and lbpB genes . Growth studies with isogenic mutants deficient in LbpB, LbpA, CopB or FbpA were performed to evaluate their role in iron acquisition from lactoferrin . LbpA and FbpA were essential for this process, supporting prior models of the iron acquisition pathway . LbpB was not essential which is remniscent of studies with the bacterial transferrin receptors . The isogenic CopB-ve isogenic mutants were deficient in iron acquisition from both transferrin and lactoferrin, suggesting that it is a key component in both pathways . A model providing an alternate explanation of the data is presented . The role and surface accessibility of the lactoferrin receptor proteins suggests that they might be useful vaccine antigens and the preferentially reactivity of convalescent antisera with LbpB suggests that it may be the prime candidate.

Ginekol Pol, 1997 Oct, 68(10), 449 - 58
{The role of lower genital tract infection in PROM}; Blok R et al.; The authors conducted a prospective study of the role of lower genital tract infection in PTL and PROM in high risk obstetric population with special interest in pathogens colonizing the maternal tract . A total amount of 62 women within 28 to 37 weeks of gestation were enrolled and interviewed . Subjects were cultured for Gardnerella vaginalis, group B streptococcus, Trichomonas vaginalis, Neisseria gonorrhoeae, Bacterioides fragilis, M . Hominis, U . Urealyticum, Chlamydia trachomatis, Candida galabrata and they were followed to delivery . Preterm rupture of membranes occurred in 30.6% of these pregnancies.

J Infect Dis, 1998 Nov, 178(5), 1352 - 8
Effect of human immunodeficiency virus type 1 infection upon acute salpingitis: a laparoscopic study; Cohen CR et al.; To determine the effect of human immunodeficiency virus type 1 (HIV-1) infection upon pelvic inflammatory disease (PID), a laparoscopic study of acute PID was conducted in Nairobi, Kenya . Subjects underwent diagnostic laparoscopy, HIV-1 serology, and testing for sexually transmitted diseases . Of the 133 women with laparoscopically verified salpingitis, 52 (39%) were HIV-1-seropositive . Tubo-ovarian abscesses (TOA) were found in 33% of HIV-1-infected and 15% of HIV-1-uninfected women (odds ratio {OR}, 2.8; 95% confidence interval {CI}, 1.2-6.5) . Among seropositive women, TOA was found in 55% of those with CD4 cell percent <14% vs . 28% with CD4 cell percent>14% (OR 3.1, 95% CI 0.6-15.3) . Neisseria gonorrhoeae was detected in 37 women (28%) and Chlamydia trachomatis in 12 (9%); neither was significantly related to HIV-1 seropositivity . Length of hospitalization was not affected by HIV-1 serostatus overall but was prolonged among HIV-1-infected women with CD4 cell percent <14% . Among patients with acute salpingitis, likelihood of TOA was related to HIV-1 infection and advanced immunosuppression . In general, HIV-1-seropositive women with acute salpingitis responded well to treatment.

J Clin Microbiol, 1998 Nov, 36(11), 3205 - 10
Diagnosis of Trichomonas vaginalis infection by PCR using vaginal swab samples; Madico G et al.; Trichomonas vaginalis infection is the most prevalent nonviral sexually transmitted disease (STD) in the world . A PCR test using vaginal swab samples for the detection of T . vaginalis was developed to add T . vaginalis infection to the growing list of STDs that can be detected by DNA amplification techniques . A primer set, BTUB 9/2, was designed to target a well-conserved region in the beta-tubulin genes of T . vaginalis . All strains (15 of 15) of T . vaginalis tested were successfully detected by PCR giving a single predicted product of 112 bp in gel electrophoresis . No such targeted product was amplified with DNA from Trichomonas tenax, Trichomonas gallinae, Chlamydia trachomatis, Neisseria gonorrhoeae, Giardia lamblia, Chilomastix sulcatus, Dientamoeba fragilis, and Entamoeba histolytica . An optimal analytical sensitivity of one T . vaginalis organism per PCR was achieved . Culture, performed with the Inpouch TV culture system, was examined daily with a light microscope to identify T . vaginalis . Twenty-three of 350 (6.6%) vaginal swab samples from women attending an army medical clinic were culture positive for T . vaginalis . Of these culture positive specimens, PCR detected 22 of 23 (96%) with primer set BTUB 9/2, and wet preparation detected only 12 of 23 (52%) . Seventeen specimens were BTUB 9/2-PCR positive and culture negative . Ten of these discordant specimens were determined to be as true positive by PCR using primer sets TVA 5-1/6 and/or AP65 A/B, which target different regions in the T . vaginalis genome, and seven were determined to be false positive . The sensitivity of BTUB 9/2-PCR was 97% and the specificity was 98% . The sensitivities of culture and wet preparation were 70 and 36%, respectively . The diagnosis of T . vaginalis infection by PCR is a sensitive and specific method that could be incorporated into a joint strategy for the screening of multiple STDs by using molecular amplification methods.

Sex Transm Dis, 1998 Sep, 25(8), 415 - 7
Continuing evolution of the pattern of quinolone resistance in Neisseria gonorrhoeae isolated in Sydney, Australia; Tapsall JW et al.; BACKGROUND AND OBJECTIVES: Multiple phenotypes of quinolone-resistant Neisseria gonorrhoeae isolated in Sydney since 1984 originated in Asia and increased in number and level of resistance in 1995 . GOAL: To study the origins, characteristics, and infection pattern of quinolone-resistant Neisseria gonorrhoeae in Sydney from 1995 to 1997 and to compare these results with prior findings . STUDY DESIGN: Quinolone minimal inhibitory concentrations, phenotype, and geographic source of quinolone-resistant Neisseria gonorrhoeae isolated in Sydney from 1995 to 1997 were analyzed . RESULTS: Two hundred nineteen episodes of infection with quinolone-resistant Neisseria gonorrhoeae from 2,236 gonococcal isolates occurred during 1995 through 1997 . The rate of isolation of quinolone-resistant Neisseria gonorrhoeae increased significantly at the end of 1996 and was maintained through 1997 . The increase resulted from sustained domestic transmission of a limited number of phenotypes in heterosexual patients . CONCLUSION: The pattern of isolation of quinolone-resistant Neisseria gonorrhoeae in Sydney changed from the sporadic isolation of multiple phenotypes of imported quinolone-resistant Neisseria gonorrhoeae to a higher rate of endemic disease caused by a few subtypes . Alterations in antibiotic treatment regimens in the affected patient group were required.

Sex Transm Dis, 1998 Sep, 25(8), 395 - 402
Screening for gonorrhea and chlamydia by DNA amplification in adolescents attending middle school health centers . Opportunity for early intervention; Burstein GR et al.; GOAL: To determine prevalence and incidence of Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT) infection and assess risk factors predictive for such infections in a middle school-based clinic sample . STUDY DESIGN: 170 female students and 43 male students making 256 and 47 visits, respectively, > or = 30 days apart, in urban middle school clinics for primary care screening, reproductive health, or illness/injury were routinely asked to provide urine specimens for GC and CT ligase chain reaction testing if sexually active in the preceding 3-month period . Information regarding prior sexually transmitted diseases, reason for visit, and sexual risk behaviors was obtained . RESULTS: GC: 11.4% of female student and 2.1% of male student tests were positive . Incidence was 34.0 cases/1,000 person months (95% Confidence interval {CI}: 19.5-67.5) . Median time to first positive and repeat positive test was 4.6 and 2.6 months, respectively . For CT: 16.4% of female student and 2.1% of male student tests were positive . Incidence was 57.5 cases/1,000 person months (95% CI: 35.2-93.8) . Median time to first positive and repeat positive CT test was 6.0 and 4.8 months, respectively . Assessed risk factors failed to specify a candidate screening population . CONCLUSION: These data suggest that all sexually active adolescent girls in this high risk setting should be offered testing for GC and CT at least twice per year, regardless of age or other sexual risk behaviors and that STD control efforts in high risk middle schools should be encouraged.

Contraception, 1998 Aug, 58(2), 105 - 9
The risk of inadvertent intrauterine device insertion in women carriers of endocervical Chlamydia trachomatis; Faundes A et al.; The most important complication attributed to the use of intrauterine device (IUD) is pelvic inflammatory disease (PID), often associated with Neisseria or Chlamydia infection . Consequently, the IUD should not be inserted in women at risk of infection or with symptoms of endocervicitis . To evaluate the effectiveness of such a policy, a systematic investigation of Chlamydia and Neisseria was carried out among 407 contraceptive acceptors . Twenty-seven cases were positive for Chlamydia and none were positive for Neisseria . There were no statistical differences in the proportion of women with and without Chlamydia who had vulvovaginal or cervical signs or symptoms, although twice as many women had pain at pelvic exam in the Chlamydia-infected group . In 29 women in whom infection was clinically suspected, two were found to be infected with Chlamydia . IUD were not inserted in women suspected of having infection, but 19 of 327 IUD acceptors were subsequently found to have a positive Chlamydia test result . Two women returned with symptoms of PID and 17 were asymptomatic, but all were treated . PID was not suspected in any other subject . These results reinforce the need for careful selection of IUD acceptors and for thorough counseling for symptoms of PID and the need for immediate consultation.

Proc Natl Acad Sci U S A, 1998 Oct 13, 95(21), 12619 - 24
Free recombination within Helicobacter pylori; Suerbaum S et al.; Sequences of three gene fragments (flaA, flaB, and vacA) from Helicobacter pylori strains isolated from patients in Germany, Canada, and South Africa were analyzed for diversity and for linkage equilibrium by using the Homoplasy Test and compatibility matrices . Horizontal genetic exchange in H . pylori is so frequent that different loci and polymorphisms within each locus are all at linkage equilibrium . These results indicate that H . pylori is panmictic . Comparisons with sequences from Escherichia coli, Neisseria meningitidis, and Drosophila melanogaster showed that recombination in H . pylori was much more frequent than in other species . In contrast, when multiple family members infected with H . pylori were investigated, some strains were indistinguishable at all three loci . Thus, H . pylori is clonal over short time periods after natural transmission.

Gene, 1998 Oct 5, 220(1-2), 21 - 9
Neisseria gonorrhoeae contains multiple copies of a gene that may encode a site-specific recombinase and is associated with DNA rearrangements; Carrick CS et al.; A 960-bp ORF potentially encoding a site-specific recombinase has been cloned from Neisseria gonorrhoeae MS11-A . This ORF was designated pivNg on the basis of similarity of the deduced amino acid sequence to the Piv proteins of Moraxella spp . that are site-specific invertases . Southern hybridization and sequence analysis revealed that there were multiple copies of pivNg sequence within the genomes of N . gonorrhoeae strains tested, but not in several other neisserial species . Southern hybridization and sequence analysis further suggested that pivNg sequences may be associated with genomic rearrangements.

Res Microbiol, 1997 May, 148(4), 289 - 303
RegF, an SspA homologue, regulates the expression of the Neisseria gonorrhoeae pilE gene; De Reuse H et al.; The Neisseria gonorrhoeae pilE gene codes for a type IV pilin, the major subunit of pili which constitute an essential virulence factor during gonococcal infection . Expression of pilE seems to be highly regulated, which may allow piliation to adapt to growth conditions . From an N . gonorrhoeae genomic library, we selected plasmid pNG200 encoding a protein (RegF) which caused a 5-fold increase in the expression of pilE::cat fusion in Escherichia coli . This regulation was mediated via the complex pilE promoter region, comprising potential sigma 70- and sigma 54-dependent promoters, and could not be observed in the absence of an active sigma 54 factor . The RegF protein (23,149 Da) showed 42% identity with the E . coli "stringent starvation protein", SspA . This protein was shown to interact with the RNA polymerase holoenzyme and to play a role in the expression of at least 11 proteins in E . coli . In an N . gonorrhoeae strain carrying a regF::mTn3Cm3 mutation constructed by allelic exchange, it was observed that pilin expression was enhanced . Our results were consistent with a model in which (i) in N . gonorrhoeae, RegF acts as a negative regulator of pilE transcription, and (ii) in E . coli, RegF increases pilE transcription by preventing sigma 54-associated steric hindrance at pilE promoters described previously.

Int J STD AIDS, 1998 Sep, 9(9), 548 - 50
HIV infection and asymptomatic sexually transmitted infections in a rural South African community; Colvin M et al.; The objective was to determine the prevalence of HIV and other sexually transmitted infections (STIs) in a rural community . A population-based survey of adults in 110 homesteads was conducted in 1995 . A questionnaire on demographics, sexual practices and history of STDs was administered . Neisseria gonorrhoeae and Chlamydia trachomatis infections were detected using ligase chain reaction (LCR) assay of urine . The seroprevalence of syphilis rapid plasma reagin (RPR) and Treponema pallidum haemagglutination assay (TPHA) and HIV infection (ELISA) was determined . Among 259 subjects the prevalence of HIV was 10.5%, N . gonorrhoeae 4.5%, C . trachomatis 6.1% and active syphilis 8.8% . All infections were asymptomatic . Forty per cent of sexually active men had more than one concurrent sexual partner . Only 14% of subjects had ever used condoms . The STI epidemic is being promoted by high levels of asymptomatic infections, high partner concurrency and low condom usePIP: This study determined the prevalence of HIV and other sexually transmitted infections (STIs) in a rural South African community . A population-based survey of adults in 110 homesteads was conducted in 1995 . A questionnaire on demographics, sexual practices and histories of STIs was administered . Neisseria gonorrhea and Chlamydia trachomatis infections were detected using ligase chain reaction assay of urine . The seroprevalence of syphilis rapid plasma reagin and Treponema pallidum hemagglutination assay and HIV infection (ELISA) was determined . Among 259 subjects, the prevalence of HIV was 10.5%; N . gonorrhea, 4.5%; C . trachomatis, 6.1%; active syphilis, 8.8% . All infections were asymptomatic . About 40% of sexually active men had more than one concurrent sexual partner . Only 14% of subjects had ever used condoms . In general, these findings indicate that high levels of asymptomatic infections, high partner concurrence, and low condom use are promoting the STI epidemic in this community .

Int J STD AIDS, 1998 Sep, 9(9), 543 - 4
Emergency unit care of sexually transmitted infections; Jarvis RR et al.; The aim of the study was to assess provisions for management of sexually transmitted infections (STIs), emergency contraception and pregnancy test in UK emergency departments . Postal questionnaires were sent to all consultant-led emergency departments in the UK in January 1996 . The response rate was 64% . Most departments made direct referrals to genitourinary medicine (GUM) clinics and most had access to appropriate clinics . While 55% had facilities for diagnosis of at least one of the 3 common STIs (Chlamydia trachomatis, Neisseria gonorrhoeae and herpes simplex), only 6.25% had facilities for all 3 . A minority of units provided training in the management of STIs . Emergency physicians should be trained in the early management of STIs and a coordinated working relationship should be developed between emergency and GUM departments to provide optimal sexual health care.

J Exp Med, 1998 Oct 5, 188(7), 1277 - 86
Coordinate activation of activator protein 1 and inflammatory cytokines in response to Neisseria gonorrhoeae epithelial cell contact involves stress response kinases; Naumann M et al.; Neisseria gonorrhoeae (Ngo), the etiologic agent of gonorrhea, induce a number of proinflammatory cytokines by contact to epithelial cells . Cytokine genes and a variety of other immune response genes are activated as a result of the regulatory function of immediate early response transcription factors including activator protein 1 (AP-1) . Since it is established that phosphorylation of c-Jun, the central component of AP-1, by the stress-activated c-Jun NH2-terminal kinase (JNK) increases the transcriptional activity of AP-1, we studied whether Ngo could induce stress response pathways involving JNK . We found that virulent Ngo strains induce phosphorylation and activation of JNK but not of p38 kinase . Analysis of a nonpathogenic Ngo strain revealed only weak JNK activation . In respect to the molecular components upstream of the JNK signaling cascade, we show that a dominant negative mutant of MAP kinase kinase 4 (MKK4) represses transcription of an AP-1-dependent reporter gene . Regarding upstream stress response factors involved in Ngo-induced MKK4/JNK/AP-1 activation, we identified p21-activated kinase (PAK) but not MAPK/ERK kinase kinase (MEKK1) . Inhibition of small GTPases including Rac1 and Cdc42 by Toxin B prevented JNK and AP-1 activation . Our results indicate that Ngo induce the activation of proinflammatory cytokines via a cascade of cellular stress response kinases involving PAK, which directs the signal from the Rho family of small GTPases to JNK/AP-1 activation.

J Mol Biol, 1998, 283(1), 301 - 10
Crystal structure of carbonic anhydrase from Neisseria gonorrhoeae and its complex with the inhibitor acetazolamide; Huang S et al.; The crystal structure of carbonic anhydrase from Neisseria gonorrhoeae has been solved to a resolution of 1.78 A by molecular replacement using human carbonic anhydrase II as a template . After refinement the R factor was 17.8% (Rfree=23.2%) . There are two molecules per asymmetric unit (space group P21), but they have essentially identical structures . The fold of the N . gonorrhoeae enzyme is very similar to that of human isozyme II; 192 residues, 74 of which are identical in the two enzymes, have equivalent positions in the three-dimensional structures . This corresponds to 85% of the entire polypeptide chain of the bacterial enzyme . The only two cysteine residues in the bacterial enzyme, which has a periplasmic location in the cell, are connected by a disulfide bond . Most of the secondary structure elements present in human isozyme II are retained in N . gonorrhoeae carbonic anhydrase, but there are also differences, particularly in the few helical regions . Long deletions in the bacterial enzyme relative to human isozyme II have resulted in a considerable shortening of three surface loops . One of these deletions, corresponding to residues 128 to 139 in the human enzyme, leads to a widening of the entrance to the hydrophobic part of the active site cavity . Practically all the amino acid residues in the active site of human isozyme II are conserved in the N . gonorrhoeae enzyme and have similar structural positions . However, the imidazole ring of a histidine residue, which has been shown to function as a proton shuttle in the catalytic mechanism of the human enzyme, interacts with an extraneous entity, which has tentatively been identified as a 2-mercaptoethanol molecule from the crystallization medium . When this entity is removed by soaking the crystal in a different medium, the side-chain of His66 becomes quite mobile . The structure of a complex with the sulfonamide inhibitor, acetazolamide, has also been determined . Its position in the active site is very similar to that observed in human carbonic anhydrase II .

J Immunol, 1998 Oct 1, 161(7), 3762 - 6
Genetic basis of human complement C8 alpha-gamma deficiency; Kojima T et al.; Deficiency of the alpha-gamma subunit of the eighth component of complement (C8alpha-gammaD) is frequently associated with recurrent neisserial infections, especially meningitis caused by Neisseria meningitidis . We here report the molecular basis of C8alpha-gammaD in two unrelated Japanese subjects . Screening all 11 exons of the C8alpha gene and all 7 exons of the C8gamma gene and their boundaries by exon-specific PCR/single-strand conformation polymorphism demonstrated aberrant single-stranded DNA fragments in exon 2 of C8alpha gene in case 1 and in exons 2 and 9 of C8alpha gene in case 2 . Nucleotide sequencing of the amplified DNA fragments in case 1 revealed a homozygous single-point mutation at the second exon-intron boundary, inactivating the universally conserved 5' splice site consensus sequence of the second intron (IVS2+1G-->T) . Case 2 was a compound heterozygote for the splice junction mutation, IVS2+1G-->T, and a nonsense mutation at Arg394 (R394X) . R394X was caused by a C to T transition at nucleotide 1407, the first nucleotide of the codon CGA for Arg394, leading to a stop codon TGA . No mutations were detected in the C8gamma gene by our method . Our results indicate that the pathogenesis of C8alpha-gammaD might be caused by heterogeneous molecular defects in the C8alpha gene.

J Immunol, 1998 Oct 1, 161(7), 3711 - 8
Membrane cofactor protein: importance of N- and O-glycosylation for complement regulatory function; Liszewski MK et al.; Membrane cofactor protein (MCP; CD46) is a type 1 membrane glycoprotein that inhibits complement activation on host cells . It also is a measles virus (MV) receptor, an adherence factor for group A Streptococcus pyogenes, and a cellular pilus receptor for pathogenic Neisseria . The amino terminus of MCP consists of four complement control protein (CCP) repeats, three of which (CCP-1, -2, and -4) possess N-glycans . Immediately following the CCP modules is an alternatively spliced region for extensive O-glycosylation (termed the STP domain) . Previous studies established that the N-glycan of CCP-2 is essential for MV binding and infection and that the splicing variants of the STP domain not only affect MV binding and fusion, but also differentially protect against complement-mediated cytolysis . In this report, we dissect the role of these carbohydrates on complement regulatory function . We constructed, expressed, and characterized proteins deleting these carbohydrates . For MCP-mediated protection against cytolysis, the N-glycans of CCP-2 and -4 were necessary, the STP segment influenced but was not essential, and the N-glycan of CCP-1 was not required . In addition, the rate and magnitude of cell surface cleavage of C4b to C4c and C4d by MCP and factor I correlated with cytoprotection . These studies expand the structure-function understanding of the active sites of MCP and elucidate an important role for carbohydrates in its function, a finding consistent with their conservation in the MCP of other species.

Orv Hetil, 1998 Sep 6, 139(36), 2115 - 20
{Inflammation of the pelvis minor}; Gardo S; Pelvic inflammatory disease (PID) is the must important gynecologic infectious disease . It causes not only serious clinical symptoms, life threatening complications, but also severe damage to the female upper reproductive tract . Among its important sequale are infertility due to tubal occlusion, ectopic pregnancy, dyspareunia, and chronic pelvic pain . The must important causative organisms are Neisseria gonorrhoeae, Chlamydia trachomatis, as well as anaerobic and facultative bacteria found in the vaginal flora of women with bacterial vaginosis . The author reviews the latest developments regarding the epidemiology, etiology diagnostics, medical and surgical therapy of the disease . The importance and possibilities of prevention are discussed.

Tidsskr Nor Laegeforen, 1998 Aug 10, 118(18), 2783 - 5
{Gonorrhea caused by fluoroquinolone resistant gonococci}; Dosen LK et al.; We describe a case of 4-fluoroquinolone resistant gonorrhoea where treatment failed on several occasions and where there may have been reinfections . Since 1995 peroral single dose 4-fluoroquinolones have been recommended as first preference treatment for uncomplicated gonorrhoea in Norway . However, Neisseria gonorrhoeae has a high ability to create resistance, and the emergence of 4-fluoroquinolone resistant strains is causing worldwide concern . An increasing number of 4-fluoroquinolone resistant gonococcal strains have also been observed in Norway, and the number of failures in the recommended treatment regime are increasing . It is proposed that 4-fluoroquinolone resistant strains be reported nationally.

Biochim Biophys Acta, 1998 Oct 1, 1400(1-3), 45 - 61
Clinical applications of quinolones; Hooper DC; The quinolone antimicrobials are the class of inhibitors of bacterial topoisomerases that has been developed most fully for clinical use in human medicine . Initial members of the class had their greatest potency against Gram-negative bacteria, but newly developed members have exhibited increased potency against Gram-positive bacteria and soon agents will be available with additional activity against anaerobic bacteria, providing a broad spectrum of potency . After nalidixic acid, the earliest member of the class which was used for treatment of urinary tract infections, the later fluoroquinolone congeners have had sufficient potency, absorption, and distribution into tissue for additional uses in treatment of sexually transmitted diseases, infections of the gastrointestinal tract, respiratory tract, skin, and bones and joints . Tolerability of these agents in usual doses has been good . Acquired bacterial resistance resulting from clinical uses has occurred in particular among staphylococci and Pseudomonas aeruginosa . Intense drug use and ability of resistant pathogens to spread have also contributed to development of resistance in initially more susceptible pathogens such as Escherichia coli and Neisseria gonorrhoeae in certain settings . Preservation of the considerable clinical utility of the quinolone class for the long term will be affected by the extent to which their use is judicious.

J Bacteriol, 1998 Oct, 180(19), 5117 - 22
Formation of single-stranded DNA during DNA transformation of Neisseria gonorrhoeae; Chaussee MS et al.; Neisseria gonorrhoeae is naturally competent for DNA transformation . In contrast to other natural prokaryotic DNA transformation systems, single-stranded donor DNA (ssDNA) has not previously been detected during transformation of N . gonorrhoeae . We have reassessed the physical nature of gonococcal transforming DNA by using a sensitive nondenaturing native blotting technique that detects ssDNA . Consistent with previous analyses, we found that the majority of donor DNA remained in the double-stranded form, and only plasmid DNAs that carried the genus-specific DNA uptake sequence were sequestered in a DNase I-resistant state . However, when the DNA was examined under native conditions, S1 nuclease-sensitive ssDNA was identified in all strains tested except for those bacteria that carried the dud-1 mutation . Surprisingly, ssDNA was also found during transformation of N . gonorrhoeae comA mutants, which suggested that ssDNA was initially formed within the periplasm.

Infect Immun, 1998 Oct, 66(10), 5001 - 7
Infection of epithelial cells by pathogenic neisseriae reduces the levels of multiple lysosomal constituents; Ayala P et al.; Members of our group reported recently that neisseria infection of human epithelial cells results in accelerated degradation of the major lysosomal integral membrane protein LAMP1 and that this is due to hydrolysis of this glycoprotein at its immunoglobulin A1 (IgA1)-like hinge by the neisseria type 2 IgA1 protease (L . Lin et al., Mol . Microbiol . 24:1083-1094, 1997) . We also reported that the IgA1 protease plays a major role in the ability of the pathogenic neisseriae to survive within epithelial cells and hypothesized that this is due to alteration of lysosomes as a result of protease-mediated LAMP1 degradation . In this study, we tested the hypothesis that neisseria infection leads to multiple changes in lysosomes . Here, we report that neisseria infection also reduces the levels of three other lysosomal markers: LAMP2, lysosomal acid phosphatase (LAP), and CD63 . In contrast, neither the epidermal growth factor receptor level nor the beta-tubulin level is affected . A detailed examination of LAMP2 indicated that the reduced LAMP2 levels are not the result of an altered biosynthetic rate or of cleavage by the IgA1 protease . Nevertheless, the protease plays a role in reducing LAMP2 and LAP activity levels, as these are partially restored in cells infected with an iga mutant . We conclude that neisseria infection results in multiple changes to the lysosomes of infected epithelial cells and that these changes are likely an indirect result of IgA1 protease-mediated cleavage of LAMP1.

Ginecol Obstet Mex, 1998 Aug, 66, 309 - 15
{Microbial etiology of mild and moderate pelvic inflammatory disease}; Narcio ML et al.; Pelvic inflammatory disease (PID) is one of the most severe complications of sexually transmitted disease (STD) . It can be due to the ascending of normal endogenous microorganisms of the female genital tract or the infection by microorganisms related to STD as Chlamydia trachomatis and Neisseria gonorrhoeae . PID leads to serious gynecoobstetric consequences as infertility and ectopic pregnancy . Clinicians face the problem of knowing the ethiology of PID in order to treat appropriatly patients with this clinical diagnosis . So that, this work pretends to establish what kind of microorganisms are implicated in PID . A proper isolation and identification of microorganisms achieved by culture of lower genital tract samples from endocervix, endometrium and peritoneal fluid, leading to a betther, specific and proper treatment of this disease.

Bull World Health Organ, 1998, 76(3), 277 - 87
Reproductive tract infections, gynaecological morbidity and HIV seroprevalence among women in Mumbai, India; Brabin L et al.; Reported are the prevalence of reproductive tract infections and their contribution to pelvic inflammatory disease (PID), as well as the seroprevalence of human immunodeficiency virus (HIV), among women living in three inner city wards of Mumbai, India . Women aged < or = 35 years were recruited and screened as cases if they had been admitted to hospital for gynaecological investigation for suspected PID (n = 151) or infertility (n = 295); controls were healthy fertile women attending for laparoscopic tubal ligation (n = 2433) . The women were mainly of low socioeconomic status . A total of 59.4% were migrants and 14.9% of these came to Mumbai to seek treatment . Cases reported a history of adverse pregnancy outcomes significantly more often than controls, and 30.5% of suspected PID cases had previously undergone laparoscopic tubal ligation . At examination 24.2% of cases and 8.4% of controls had a vaginal discharge . Pelvic infection was confirmed in 42.0% of suspected PID cases and 14.6% of infertile cases for whom diagnostic laparoscopy was performed . The prevalence of sexually transmitted diseases was low: Chlamydia trachomatis was found in 0.2%; and Neisseria gonorrhoeae was cultured from the cervix in only four cases . Neither of these infections was detected in laparoscopic aspirates . The prevalence of HIV1/2 infections in unlinked samples was 1.9% . Sexually transmitted diseases were not major factors leading to gynaecological morbidity . Heterosexual spread of HIV infection to this population of married women is still relatively low but needs to be carefully monitored . The gynaecological morbidity detected may be a consequence of widespread use of invasive methods of fertility regulationPIP: A case-control study conducted in 1993-95 among women 35 years of age and younger living in three inner-city wards of Mumbai, India, investigated the prevalence of reproductive tract infections and their contribution to pelvic infection . Enrolled as cases were 151 women admitted to the hospital with suspected pelvic inflammatory disease (PID) and 295 infertile women; 2433 healthy fertile women undergoing laparoscopic tubal ligation served as controls . Adverse pregnancy outcomes were reported significantly more often by cases than controls . 31.8% of suspected PID cases, 9.1% of infertile women, and 53.1% of tubal ligation patients reported ever-use of a contraceptive method, primarily a copper IUD . At examination, 24.2% of cases and 8.4% of controls had a vaginal discharge . Pelvic infection was confirmed in 42.0% of suspected PID cases and in 14.6% of infertile women in whom diagnostic laparoscopy was performed . HIV prevalence was 1.9% in unlinked samples . The prevalence of other sexually transmitted diseases (STDs) was under 1% . The gynecologic morbidity recorded in this study is presumed to be a result of widespread use of invasive methods of fertility regulation, not STDs .

J Med Microbiol, 1998 Sep, 47(9), 751 - 5
Resistance to antimicrobial agents: a personal view; Greenwood D; Problems of antimicrobial drug resistance are presently serious, but not yet desperate . The principal areas of concern are two-fold: multiresistant opportunist bacteria that affect vulnerable patients in high dependency areas of hospitals (the most pressing problem for developed countries); and multidrug resistance among classic pathogens like Mycobacterium tuberculosis, Salmonella typhi, Shigella spp., Neisseria gonorrhoeae and Plasmodium falciparum (mainly, although not exclusively, a problem for developing countries) . The first type can be contained to a large extent by good infection control practices and careful prescribing based on agreed policies of antimicrobial drug use . The input of infection control nurses and laboratory-based clinical microbiologists is crucial and these services deserve full support . The second type additionally requires coordinated action to regulate more effectively the manufacture, availability, promotion and use of antimicrobial drugs . In this case the input of governments, international agencies and pharmaceutical companies is essential . Prescription-only status for antimicrobial drugs used in man and animals should be the norm . The number of drugs available for the treatment of viral, fungal and parasitic infections is comparatively small and much less is known about resistance . More research in these areas would be welcome . Teaching good prescribing habits to medical students is presently haphazard and needs to be formalised . Surveillance needs to be improved . The second half of the 20th century has been a golden age of antibiotics, but the outlook is uncertain . If antimicrobial chemotherapy is to have a secure future, prescribers must learn to use these powerful tools with greater discretion and their use worldwide must be regulated effectively.

J Exp Med, 1998 Sep 7, 188(5), 941 - 52
Gonococcal invasion of epithelial cells driven by P.IA, a bacterial ion channel with GTP binding properties; van Putten JP et al.; The neisserial porin P.I is a GTP binding protein that forms a voltage-gated channel that translocates into mammalian cell membranes and modulates host cell signaling events . Here, we report that P.I confers invasion of the bacterial pathogen Neisseria gonorrhoeae into Chang epithelial cells and that this event is controlled by GTP, as well as other phosphorus-containing compounds . Bacterial invasion was observed only for strains carrying the P.IA subtype of porin, which is typically associated with the development of disseminated neisserial disease, and did not require opacity outer membrane proteins, previously recognized as gonococcal invasins . Allelic replacement studies showed that bacterial invasiveness cotransferred with the P.IA (por1A) gene . Mutation of the P.I-associated protein Rmp did not alter the invasive properties . Cross-linking of labeled GTP to the porin revealed more efficient GTP binding to the P.IA than P.IB porin subtype . GTP binding was inhibited by an excess of unlabeled GTP, ATP, and GDP, as well as inorganic phosphate, but not by UTP or beta-glycerophosphate, fully in line with the respective invasion-inhibitory activities observed for these compounds . The P.IA-mediated cellular invasion may explain the more invasive behavior of P.IA strains in the natural infection and may broaden the basis for the development of a P.I-based gonococcal vaccine.

Scand J Infect Dis, 1998, 30(2), 200 - 1
Septicaemia with Neisseria elongata ssp . nitroreducens in a patient with hypertrophic obstructive cardiomyopathia; Hofstad T et al.; Neisseria elongata ssp . nitroreducens, a commensal of the human upper respiratory tract, is a newly recognized cause of endocarditis . We report the isolation of the organism from blood cultures of a 30-y-old man with hypertrophic obstructive cardiomyopathy . The patient was successfully treated with benzylpenicillin and netilmicin in combination, followed by ceftriaxone and amoxicillin.

Vet Immunol Immunopathol, 1998 Jul 31, 64(3), 191 - 205
Occurrence of autoantibodies to cilia in lambs with a 'coughing syndrome'; Niang M et al.; A respiratory disease of lambs that has been termed the 'coughing syndrome' has been observed in the mid-western region of the United States of America . Mycoplasma ovipneumoniae (M . ovipneumoniae) and Mycoplasma arginini (M . arginini) were routinely isolated from the respiratory tract of lambs with this disease . A high level of antibodies reactive with ovine cilia of the upper respiratory tract was detected in the sera from many of the lambs in affected flocks but not in sera of lambs from unaffected flocks . The reactivity of these antibodies with cilia was demonstrated by ELISA and confirmed by indirect immunofluorescent staining and western immunoblotting . These antibodies were predominantly of the IgG isotype . They were distinct from cold or warm agglutinins and could be absorbed from the sera with cilia but not with antigens of common bacterial pathogens of the sheep respiratory tract including M . ovipneumoniae, M . arginini, Pasteurella haemolytica, Pasteurella multocida or Neisseria ovis . In addition, their occurrence appeared to be independent of the specific antibodies to M . ovipneumoniae and M . arginini . Western immunoblotting indicated that the antibodies were directed primarily against an antigen with apparent molecular weight of 50 kDa . In one flock from which serial serum samples were collected from the same lambs over a 10-month period, antibodies to ovine cilia developed before the onset of the clinical disease and persisted for a period of several months until most of the lambs had apparently recovered . However, colonization of the respiratory tract of the lambs by M . ovipneumoniae preceded the production of these antibodies . Sequential serum samples taken from another flock, with no known history of this coughing, showed no such antibodies throughout the sampling period . It is suggested that an immunopathologic mechanism involving production of autoantibodies directed against a ciliary antigen of the lambs could be a contributing factor to the pathogenesis of this clinical disease.

Immunol Invest, 1998 Jul-Sep, 27(4-5), 203 - 20
Immunoassay of human Neisseria meningitidis serogroup A antibody; Diakun KR et al.; An enzyme immunoassay is described which quantitates antibodies to Neisseria meningitidis serogroup A capsular polysaccharide in human sera . Modifications of a previously developed two-day assay by Carlone et al . were made to permit analysis in one day and to be compatible with automation . The allowable variations in assay conditions and the areas in which stringent control must be maintained for consistent assay performance are described . Antigen-coating parameters, the kinetics of primary and secondary antibody incubation steps, the buffer compositions, including detergents, serum requirements, and the need for blocking steps were examined . Our modified one-day assay showed excellent agreement with the standardized method of Carlone, with a correlation coefficient between the two methods of 0.989 . This assay is adaptable within a permissible range of parameters thus facilitating the implementation of the standardized assay . This will maximize the consistency of results from serum analysis for conjugate vaccine trials related to serotype A Neisseria meningitidis.

J Infect Dis, 1998 Sep, 178(3), 742 - 51
Cytokine and antibody responses in women infected with Neisseria gonorrhoeae: effects of concomitant infections; Hedges SR et al.; The levels of interleukin (IL)-1, IL-6, IL-8, IL-10, and transforming growth factor-beta in sera and genital tract secretions from women with gonococcal cervicitis and other genital infections were examined . Cytokines were not elevated in genital secretions from gonococcus-infected compared with uninfected patients . The level of serum IL-6 was higher in gonococcus-infected than in uninfected patients at recruitment . Serum, but not local, IL-1 and IL-6 levels were elevated in patients concomitantly infected with Trichomonas vaginalis or Chlamydia trachomatis in addition to Neisseria gonorrhoeae compared with levels in patients infected with any single organism . Concomitant infection altered neither the total immunoglobulin concentrations nor the levels of antigonococcal antibodies in serum or local secretions . The results suggest that N . gonorrhoeae induces only a limited cytokine and antibody response during uncomplicated cervical infections; however, the presence of other sexually transmitted disease-causing organisms can alter the systemic cytokine but not the antigonococcal antibody levels.

Proc Natl Acad Sci U S A, 1998 Sep 1, 95(18), 10872 - 7
Identification of the gene (lgtG) encoding the lipooligosaccharide beta chain synthesizing glucosyl transferase from Neisseria gonorrhoeae; Banerjee A et al.; The lipooligosaccharide from Neisseria gonorrhoeae (GC), consists of lipid A, an oligosaccharide core and three branches, alpha, beta, and gamma . We report the cloning of the gene (lgtG, lipooligosaccharide glycosyl transferase G) encoding the glucosyl transferase of GC that initiates the beta chain which consists of a lactosyl moiety . This gene contains a homopolymeric tract of cytidine {poly(C)} and we demonstrate that changes in the number of Cs in poly(C) account for the variation of beta chain expression in different GC strains . Biochemical analyses and mass spectrometry clearly attribute the reactivity of mAb 2C7 to the presence of the lactosyl beta chain . In addition, we demonstrate that in the absence of the lactosyl group, a phosphoethanolamine is added to generate a new antigenic epitope as evidenced by the gain of reactivity to mAb 2-L1-8 . These results show that, like the alpha chain, the beta chain of lipooligosaccharide is subject to antigenic variation.

Eur J Clin Microbiol Infect Dis, 1998 May, 17(5), 341 - 3
Neisseria meningitidis serogroup B peritonitis associated with continuous ambulatory peritoneal dialysis; Conrads G et al.; A case of Neisseria meningitidis peritonitis in a 41-year-old female undergoing continuous ambulatory peritoneal dialysis for end-stage renal failure due to insulin-dependent diabetes mellitus is reported . The bacterial strain was serosubtyped as 4:P1.15, a rarely encountered type . Surprisingly, the minimal inhibitory concentration of vancomycin for the isolate was low (16 mg/l) . This may have contributed to the complete recovery of the patient.

Folia Microbiol (Praha), 1998, 43(3), 311 - 9
Pathogenic Neisseria--interplay between pro- and eukaryotic worlds; Meyer TF; The pathogenic Neisseria species constitute a multi-faceted infection model of a highly adapted pathogen-host relationship . Several bacterial and host-cell factors involved in the cellular cross-talk have been recently unraveled . Using Neisseria gonorrhoeae as a prototype, several structurally variable surface proteins, including pili and Opa proteins, have been revealed as adhesins recognizing distinct host-cell receptors . The Opa proteins, in particular, are important in facilitating interaction with heparan sulfate proteoglycan receptors and members of the CD66 and integrin receptor families . These interactions not only enable the pathogens' anchoring, and penetration into, the human mucosa but also stimulate cellular signaling cascades involving the phosphatidylcholine-dependent phospholipase C, acidic sphingomyelinase and protein kinase C in epithelial cells, and Src-related kinases, Rac1, p21-activated kinase and Jun N-terminal kinase in phagocytic cells . Activation of these pathways is essential for the entry and intracellular accommodation of the pathogens but also leads to an early induction of cytokine release, thus priming the immune response . It is believed that detailed knowledge of cellular signaling cascades activated by infection will aid us in applying known and novel interfering drugs, in addition to classical antibiotic therapy, to the therapeutic and prophylactic treatment of persistent or otherwise difficult-to-treat bacterial infections.

Gene, 1998 Jul 30, 215(2), 303 - 10
The ihf mRNA levels decline as Neisseria gonorrhoeae enters the stationary growth phase; Hill SA et al.; Integration host factor (IHF) is a small heterodimeric DNA binding protein found in all Gram-negative bacteria and is implicated as a transcription cofactor of pilE in Neisseria gonorrhoeae (Hill, S.A., Samuels, D.S., Carlson, J.H., Wilson, J., Hogan, D., Lubke, L., Belland, R.J., 1997 . Integration host factor is a transcriptional cofactor of pilE in Neisseria gonorrhoeae . Mol . Microbiol . 23, 649-656) . The ihf genes (ihfA and ihfB) were cloned from N . gonorrhoeae through functional complementation of defined Escherichia coli ihf mutants for plating of phage lambda . The predicted aa sequences of each gonococcal IHF polypeptide showed extensive homology to other reported IHF polypeptide sequences . Northern blotting and primer extension analysis defined the tsp for each gene and indicated a disparity in ihfA and ihfB message levels over time, with ihfB mRNA being more abundant throughout the entire growth cycle . Furthermore, both the ihfA and ihfB message levels declined as cells entered the stationary growth phase . Overall, this study reveals several unique features of ihf transcription in the gonococcus which questions whether certain aspects if ihf transcriptional regulation are universally shared by all Gram-negative bacteria.

Sex Transm Dis, 1998 Aug, 25(7), 346 - 9
Sexually transmitted infections among sex workers in KwaZulu-Natal, South Africa; Ramjee G et al.; OBJECTIVE: The purpose of this study was to investigate the prevalence of sexually transmitted infections, including human immunodeficiency virus (HIV), among female sex workers operating at truckstops in the KwaZulu-Natal midlands of South Africa . METHODS: A total of 145 sex workers were recruited from August 1996 to March 1997 . A gynecologic examination, including a speculum examination, was performed . Investigations were performed to identify gonorrhea, syphilis, chlamydia (infection), candidiasis, trichomoniasis, bacterial vaginosis, and HIV infection . RESULTS: A total of 50.3% of the sex workers were HIV positive . Infection with Trichomonas vaginalis was present in 41.3% of the women, Candida albicans in 40.6%, Neisseria gonorrheae in 14.3%, and Chlamydia trachomatis in 16.4% . Bacterial vaginosis was present in 71% and active syphilis in 42.1% of the women . CONCLUSION: The high prevalence of sexually transmitted infections, including HIV, clearly highlights the urgent need for interventions . In addition to empowering women with methods they can use and control to reduce their risk of infection, an effective approach to control of sexually transmitted infections also needs to recognize the role of the clients, such as truck drivers, so that they also are targeted for interventionsPIP: Findings are reported from a study conducted to assess the prevalence of sexually transmitted infections, including HIV, among female prostitutes working at truckstops in the KwaZulu-Natal midlands of South Africa . The 145 participating prostitutes were recruited from 5 truckstops along the major national road between Durban and Johannesburg from August 1996 to March 1997 . Their mean age was 25 years and they had been prostitutes for an average of 3 years . A gynecologic examination was performed upon each woman, including a speculum examination, and investigations were also conducted to identify gonorrhea, syphilis, chlamydia, candidiasis, trichomoniasis, bacterial vaginosis, and HIV infection . The examinations found that 50.3% of the women were HIV seropositive, 41.3% were infected with Trichomonas vaginalis, 40.6% had Candida albicans, 14.3% had Neisseria gonorrheae, 16.4% had Chlamydia trachomatis, 71% had bacterial vaginosis, and 42.1% had active syphilis . These findings point to the need for HIV/STD prevention interventions among both the prostitutes and their clients .

Mem Inst Oswaldo Cruz, 1998 Jul-Aug, 93(4), 517 - 22
In vitro comparison of disk diffusion and agar dilution antibiotic susceptibility test methods for Neisseria gonorrhoeae; de Castillo MC et al.; At present, most Neisseria gonorrhoeae testing is done with beta-lactamase and agar dilution tests with common therapeutic agents . Generally, in bacteriological diagnosis laboratories in Argentina, study of antibiotic susceptibility of N . gonorrhoeae is based on beta-lactamase determination and agar dilution method with common therapeutic agents . The National Committee for Clinical Laboratory Standards (NCCLS) has recently described a disk diffusion test that produces results comparable to the reference agar dilution method for antibiotic susceptibility of N . gonorrhoeae, using a dispersion diagram for analyzing the correlation between both techniques . We obtained 57 gonococcal isolates from patients attending a clinic for sexually transmitted diseases in Tucuman, Argentina . Antibiotic susceptibility tests using agar dilution and disk diffusion techniques were compared . The established NCCLS interpretive criteria for both susceptibility methods appeared to be applicable to domestic gonococcal strains . The correlation between the MIC's and the zones of inhibition was studied for penicillin, ampicillin, cefoxitin, spectinomycin, cefotaxime, cephaloridine, cephalexin, tetracycline, norfloxacin and kanamycin . Dispersion diagrams showed a high correlation between both methods.

Clin Infect Dis, 1998 Aug, 27 Suppl 1, S12 - 20
Horizontal genetic exchange, evolution, and spread of antibiotic resistance in bacteria; Maiden MC; Some transformable bacteria have acquired target-mediated antibiotic resistance by horizontal genetic exchange of fragments of chromosomal genes . The resistant strains express variants of the antibiotic target that are metabolically active but exhibit a lowered affinity for the antibiotic . The alleles encoding these resistant proteins are mosaics comprising DNA derived from the host and other bacteria, often members of a different species . Examples include penicillin-resistant penicillin-binding proteins (PBPs) in Streptococcus pneumoniae and the pathogenic Neisseria species and sulfonamide-resistant dihydropterate synthase in Neisseria meningitidis . Distinct mosaic alleles encoding antibiotic resistance have arisen on multiple occasions, indicating the mobility of chromosomal genes in these species . Mosaic genes can arise at any chromosomal locus, and S . pneumoniae organisms with high-level penicillin resistance have acquired mosaic PBP genes at three bacterial bpb loci . Furthermore, horizontal genetic exchange permits movement of alleles among bacterial lineages, increasing the opportunities for the spread of antibiotic resistance.

J Exp Med, 1998 Aug 17, 188(4), 671 - 80
Binding of complement factor H to loop 5 of porin protein 1A: a molecular mechanism of serum resistance of nonsialylated Neisseria gonorrhoeae; Ram S et al.; Neisseria gonorrhoeae isolated from patients with disseminated infection are often of the porin (Por1A) serotype and resist killing by nonimmune normal human serum . The molecular basis of this resistance (termed stable serum resistance) in these strains has not been fully defined but is not related to sialylation of lipooligosaccharide . Here we demonstrate that Por1A bearing gonococcal strains bind more factor H, a critical downregulator of the alternative complement pathway, than their Por1B counterparts . This results in a sevenfold reduction in C3b, which is >75% converted to iC3b . Factor H binding to isogenic gonococcal strains that differed only in their porin serotype, confirmed that Por1A was the acceptor molecule for factor H . We identified a surface exposed region on the Por1A molecule that served as the binding site for factor H . We used gonococcal strains with hybrid Por1A/B molecules that differed in their surface exposed domains to localize the factor H binding site to loop 5 of Por1A . This was confirmed by inhibition of factor H binding using synthetic peptides corresponding to the putative exposed regions of the porin loops . The addition of Por1A loop 5 peptide in a serum bactericidal assay, which inhibited binding of factor H to the bacterial surface, permitted 50% killing of an otherwise completely serum resistant gonococcal strain . Collectively, these data provide a molecular basis to explain serum resistance of Por1A strains of N . gonorrhoeae.

J Biol Chem, 1998 Aug 21, 273(34), 21777 - 82
Cell signaling by the type IV pili of pathogenic Neisseria; Kallstrom H et al.; Neisseria gonorrhoeae and Neisseria meningitidis are Gram-negative bacterial pathogens that infect human mucosal epithelia . Type IV pilus-mediated adherence of these bacteria is a crucial early event for establishment of infection . In this work, we show that the type IV pili transduce a signal into the eucaryotic host cell . Purified adherent pili, but not pili from a low binding mutant, trigger an increase in the cytosolic free calcium ({Ca2+}i) in target epithelial cells, a signal known to control many cellular responses . The {Ca2+}i increase was blocked by antibodies against CD46, a putative pilus receptor, suggesting a role for this protein in signal transduction . Pilus-mediated attachment was inhibited by depletion of host cell intracellular Ca2+ stores but not by removal of extracellular Ca2+ . Further, kinase inhibition studies showed that pilus-mediated adherence is dependent on casein kinase II . In summary, these data reveal a novel function of the type IV pili, namely induction of signal transduction pathways in host cells.

Nat Biotechnol, 1998 Aug, 16(8), 769 - 72
The synthesis of sialylated oligosaccharides using a CMP-Neu5Ac synthetase/sialyltransferase fusion; Gilbert M et al.; Large-scale enzymatic synthesis of oligosaccharides, which contain terminal N-acetyl-neuraminic acid residues requires large amounts of the sialyltransferase and the corresponding sugar-nucleotide synthetase, which is required for the synthesis of the sugar-nucleotide donor, CMP-Neu5Ac . Using genes cloned from Neisseria meningitidis, we constructed a fusion protein that has both CMP-Neu5Ac synthetase and alpha-2,3-sialyltransferase activities . The fusion protein was produced in high yields (over 1200 U/L, measured using an alpha-2,3-sialyltransferase assay) in Escherichia coli and functionally pure enzyme could be obtained using a simple protocol . In small-scale enzymatic syntheses, the fusion protein could sialylate various oligosaccharide acceptors (branched and linear) with N-acetyl-neuraminic acid as well as N-glycolyl- and N-propionyl-neuraminic acid in high conversion yield . The fusion protein was also used to produce alpha-2,3-sialyllactose at the 100 g scale using a sugar nucleotide cycle reaction, starting from lactose, sialic acid, phosphoenolpyruvate, and catalytic amounts of ATP and CMP.

Int J STD AIDS, 1998 Aug, 9(8), 435 - 43
The role of effective diagnosis for the control of gonorrhoea in high prevalence populations; Davies PO et al.; Gonorrhoea in the UK is now a highly focal problem that is localized to certain inner city areas where it presents a significant public health concern . However, the majority of men and a proportion of women infested with Neisseria gonorrhoeae do not experience symptoms of sufficient intensity to prompt them to seek medical advice and the current strategy of treating symptomatic individuals in specialist clinics with subsequent tracing of sexual partners is therefore failing to control the spread of infection in these areas . One method for addressing this problem would be the extension of effective diagnostic services into the clinical environments most likely to be accessed by these high-risk individuals . This paper reviews the scientific literature examining the various methodologies available for the identification of N . gonorrhoeae and assesses their suitability for the diagnosis of gonorrhoea in these alternative clinical settings.

Mol Microbiol, 1998 Jul, 29(1), 369 - 79
Entry of OpaA+ gonococci into HEp-2 cells requires concerted action of glycosaminoglycans, fibronectin and integrin receptors; van Putten JP et al.; Heparan sulphate proteoglycans are increasingly implicated as eukaryotic cell surface receptors for bacterial pathogens . Here, we report that Neisseria gonorrhoeae adheres to proteoglycan receptors on HEp-2 epithelial cells but that internalization of the bacterium by this cell type requires the serum glycoprotein fibronectin . Fibronectin was shown to bind specifically to gonococci producing the OpaA adhesin . Binding assays with fibronectin fragments located the bacterial binding site near the N-terminal end of the molecule . However, none of the tested fibronectin fragments supported gonococcal entry into the eukaryotic cells; a 120 kDa fragment carrying the cell adhesion domain with the amino acid sequence RGD even inhibited the fibronectin-mediated uptake of MS11-OpaA . This inhibition could be mimicked by an RGD-containing hexapeptide and by alpha 5 beta 1 integrin-specific antibodies, suggesting that interaction of the central region of fibronectin with integrin receptors facilitated bacterial uptake . Fibronectin was unable to promote gonococcal entry into HEp-2 cells that had been treated with the enzyme heparinase III, which degrades the glycosaminoglycan side-chains of proteoglycan receptors . On the basis of these results, we propose a novel cellular uptake pathway for bacteria, which involves the binding of the pathogen to glycosaminoglycans that, in turn, act as co-receptors facilitating fibronectin-mediated bacterial uptake through integrin receptors . In this scenario, fibronectin would act as a molecular bridge linking to Opa-proteoglycan complex with host cell integrin receptors.

Mol Microbiol, 1998 Jul, 29(1), 321 - 30
PilT mutations lead to simultaneous defects in competence for natural transformation and twitching motility in piliated Neisseria gonorrhoeae; Wolfgang M et al.; Neisseria gonorrhoeae, the Gram-negative aetiological agent of gonorrhoeae, is one of many mucosal pathogens of man that expresses competence for natural transformation . Expression of this phenotype by gonococci appears to rely on the expression of type IV pili (Tfp), but the mechanistic basis for this relationship remains unknown . During studies of gonococcal pilus biogenesis, a homologue of the PilT family of proteins, required for Tfp-dependent twitching motility in Pseudomonas aeruginosa and social gliding motility in Myxococcus xanthus, was discovered . Like the findings in these other species, we show here that gonococcal PilT mutants constructed in vitro no longer display twitching motility . In addition, we demonstrate that they have concurrently lost the ability to undergo natural transformation, despite the expression of structurally and morphologically normal Tpf . These results were confirmed by the findings that two classes of spontaneous mutants that failed to express twitching motility and transformability carried mutations in PilT . Piliated PilT mutants and a panel of pilus assembly mutants were found to be deficient in sequence-specific DNA uptake into the cell, the earliest demonstrable step in neisserial competence . The PilT-deficient strains represent the first genetically defined mutants that are defective in DNA uptake but retain Tfp expression.

Mol Microbiol, 1998 Jul, 29(1), 111 - 24
Structure and function of repetitive sequence elements associated with a highly polymorphic domain of the Neisseria meningitidis PilQ protein; Tonjum T et al.; Secretins are a large family of proteins associated with membrane translocation of macromolecular complexes, and a subset of this family, termed PilQ proteins, is required for type IV pilus biogenesis . We analysed the status of PlIQ expression in Neisseria meningitidis (Mc) and found that PlIQ mutants were non-piliated and deficient in the expression of pilus-associated phenotypes . Sequence analysis of the 5' portion of the pilQ ORF of the serogroup B Mc strain 44/76 showed the presence of seven copies of a repetitive sequence element, in contrast to the situation in N . gonorrhoeae (Gc) strains, which carry either two or three copies of the repeat . The derived amino acid sequence of the consensus nucleotide repeat was an octapeptide PAKQQAAA, designated as the small basic repeat (SBR) . This gene segment was studied in more detail in a collection of 52 Mc strains of diverse origin by screening for variability in the size of the PCR-generated DNA fragments spanning the SBRs . These strains were found to harbour from four to seven copies of the repetitive element . No association between the number of copies and the serogroup, geographic origin or multilocus genotype of the strains was evident . The presence of polymorphic repeat elements in Mc PilQ is unprecedented within the secretin family . To address the potential function of the repeat containing domain, Mc strains were constructed so as to express chimeric PilQ molecules in which the number of SBR repeats was increased or in which the repeat containing domain was replaced in toto by the corresponding region of the Pseudomonas aeruginosa (Pa) PilQ protein . Although the strain expressing PilQ with an increased number of SBRs was identical to the parent strain in pilus phenotypes, a strain expressing PilQ with the equivalent Pa domain had an eightfold reduction in pilus expression level . The findings suggest that the repeat containing domain of PilQ influences Mc pilus expression quantitatively but not qualitatively.

Mem Inst Oswaldo Cruz, 1998 Mar-Apr, 93(2), 237 - 41
Properties of beta-lactamase from Neisseria gonorrhoeae; de Castillo MC et al.; Beta-lactamase activity was studied in Neisseria gonorrhoeae strains . Optimum temperature was found to be 37 degrees C . The enzyme was inactivated at temperatures higher than 60 degrees C, but remained active during storage at low temperatures (4 degrees C, -30 degrees C and -70 degrees C) for two months . Enzyme activity was observed within a pH range of 5.8-8.0, while the optimum pH was 7.0-7.2 . Addition of Ni2+, Fe2+, Mn2+ and p-chloromercurybenzoate to the reaction buffer exerted a negative effect upon the activity, whereas Hg2+ and ethylene diamine tetra-acetic acid produced complete inhibition . These results would indicate the presence of -SH groups at the catalytic site of the enzyme.

Biochim Biophys Acta, 1998 Jun 10, 1365(1-2), 309 - 18
A novel pathway for cytochromes c biogenesis in chloroplasts; Xie Z et al.; The cytochromes c are a useful model for the study of the pathways and mechanisms of assembly of the cofactor-containing components of energy transducing membranes . Genetic analyses have identified proteins that are required for the assembly of c-type cytochromes in mitochondria, bacteria and chloroplasts . The components of the pathway operating in fungal and animal mitochondria, i.e . the cytochrome (cyt) c and c1 heme lyases in the intermembrane space, were identified over a decade ago through the study of cytochrome deficiencies in Neurospora crassa and Saccharomyces cerevisiae . More recently, a large number of membrane or membrane-associated components were identified in various alpha- and gamma-proteobacteria as c-type cytochrome assembly factors; they comprise an assembly pathway that is evolutionarily and mechanistically distinct from that in fungal and animal mitochondria . The components function not only in the lyase reaction but also in the delivery and maintenance of the substrates in a state that is suitable for reaction in the bacterial periplasm . Yet a third pathway is required for cytochrome maturation in chloroplasts . Genetic analyses of Chlamydomonas reinhardtii ccs mutants, which are pleiotropically deficient in both the membrane-anchored cytochrome f and the soluble cytochrome c6, revealed a minimum of six loci, plastid ccsA and nuclear CCS1 through CCS5, that are required for the conversion of the chloroplast apocytochromes to their respective holo forms . Sequence analysis of the cloned ccsA and Ccs1 genes indicates that the predicted protein products are integral membrane proteins with homologues in cyanobacteria, some gram-positive bacteria (Bacillus subtilis, Mycobacterium spp.), beta-proteobacteria (Neisseria spp.) and an epsilon-proteobacterium (Helicobacter pylori) . CcsA and Ccs1 require each other for accumulation in vivo and are therefore proposed to function in a complex, possibly with the products of some of the other CCS loci . A tryptophan-rich motif, which has been proposed to represent a heme binding site in bacterial cytochrome biogenesis proteins (CcmC and CcmF), is functionally important in plastid CcsA . As is the case for CcmC and CcmF, the tryptophan-rich sequence is predicted to occur in a loop on the p-side of the membrane, where the heme attachment reaction occurs . Conserved histidine residues in the CcsA and Ccs1 may serve as ligands to the heme iron . A multiple alignment of the tryptophan-rich regions of the CcsA-, CcmC- and CcmF-like sequences in the genome databases indicates that they represent three different families.

Biochem J, 1998 Aug 15, 334 ( Pt 1), 133 - 9
Vitronectin binds to the gonococcal adhesin OpaA through a glycosaminoglycan molecular bridge; Duensing TD et al.; Several bacterial pathogens including Neisseria gonorrhoeae bind the human serum glycoprotein vitronectin . We aimed at defining the gonococcal receptor for vitronectin . Ligand blots demonstrated that vitronectin bound specifically to the heparin-binding outer-membrane protein OpaA, but that coating OpaA with the sulphated polysaccharide heparin was required for the interaction to occur . Bound vitronectin could be dissociated from OpaA-heparin-vitronectin complexes by the addition of excess heparin, indicating that sulphated polysaccharides provided the main linkage between the two proteins . Binding assays with intact micro-organisms substantiated the requirement of sulphated polysaccharides such as heparin and dextran sulphate for the efficient binding of vitronectin to OpaA+ gonococci . This was underscored by the increased binding of vitronectin to gonococci that had been preincubated with saturating concentrations of dextran sulphate, as opposed to the inhibition of vitronectin binding observed when bacteria were incubated simultaneously with vitronectin and saturating concentrations of dextran sulphate . Binding assays with dextran sulphates of various sizes indicated that vitronectin binding correlated with the size of the polysaccharide rather than with the amount of OpaA produced by the bacteria . The inability of zero-length cross-linking agents to couple vitronectin to OpaA provided further evidence that sulphated polysaccharides formed the linkage between vitronectin and OpaA . Infection experiments demonstrated that proteoglycan-deficient Chinese hamster ovary cells efficiently internalized dextran sulphate/vitronectin-coated gonococci, suggesting that soluble sulphated polysaccharides could substitute for cell surface glycosaminoglycans in the internalization process . On the basis of our results, we propose a novel mechanism of vitronectin binding in which sulphated polysaccharides act as molecular bridges, linking the glycosaminoglycan-binding sites of vitronectin and gonococcal OpaA.

Trans R Soc Trop Med Hyg, 1998 Jan-Feb, 92(1), 105 - 7
Plasmid patterns and antimicrobial susceptibilities of Neisseria gonorrhoeae in Bandung, Indonesia; Djajakusumah T et al.; Antimicrobial susceptibilities of Neisseria gonorrhoeae isolates from female sex workers and from men with urethritis in Bandung, Indonesia, were determined by an agar dilution technique . Typing of the Tet M plasmid in tetracycline-resistant isolates (TRNG) was performed using a polymerase chain reaction (PCR) technique and plasmid profiles of penicillinase-producing isolates (PPNG) were determined . All PPNG possessed the 4.4 MDa beta-lactamase plasmid and all TRNG showed a PCR fragment characteristic of the 'Dutch' type Tet M plasmid . Of the 50 gonococci isolates tested, all were resistant to tetracycline; 47 were TRNG, 26 were PPNG, and 6 were resistant to thiamphenicol . Chromosomal resistance to penicillin was not detected . All isolates were susceptible to ceftriaxone, ciprofloxacin, norfloxacin, ofloxacin, kanamycin, spectinomycin, and trimethoprim/sulfamethoxazole . Spectinomycin and fluoroquinolones are useful primary drugs for treatment of gonococcal infection in Bandung . Continued surveillance of antimicrobial resistance should be part of gonorrhoea control in Indonesia.

Proc Natl Acad Sci U S A, 1998 Aug 4, 95(16), 9584 - 9
CD66 receptor specificity exhibited by neisserial Opa variants is controlled by protein determinants in CD66 N-domains; Bos MP et al.; Neisseria gonorrhoeae strain MS11 is able to express 11 different opacity (Opa) proteins on its outer surface . A number of these Opa proteins have been shown to function as adhesins through binding of CD66 receptors present on human cells . CD66 antigens, or carcinoembryonic antigen family members, constitute a family of glycoproteins belonging to the immunoglobulin superfamily . Opa variants recognize this class of receptors in a differential manner such that certain Opa variants recognize up to four different CD66 receptors (CD66a, -c, -d, and -e), whereas others recognize only two (CD66a and -e) or none . We explored the basis for this receptor tropism in the present study . Our data show that glycoforms of CD66e and deglycosylated CD66e are recognized by gonococci in an Opa-specific manner . Binding by Opa variants of recombinant N-terminal domains of CD66 receptors expressed in Escherichia coli reflected the adherence specificities of Opa variants to HeLa cells expressing native CD66 molecules . These data indicate that recognition of CD66 receptors by Opa variants is mediated by the protein backbone of the CD66 N-domains . Furthermore, by using chimeric constructs between different CD66 N-domains we identified distinct binding regions on the CD66e N-domain for specific groups of Opa variants, suggesting that the differential recognition of CD66 receptors by Opa variants is dictated by the presence of specific binding regions on the N-domain of the receptor.

Eur J Biochem, 1998 Jun 15, 254(3), 626 - 33
Structure of an alpha-2,6-sialylated lipooligosaccharide from Neisseria meningitidis immunotype L1; Wakarchuk WW et al.; The recent cloning of the lipooligosaccharide (LOS) a-2,3-sialyltransferase from Neisseria meningitidis immunotype L3 permitted us to examine other immunotypes for this structural gene . We identified the gene and measured the enzyme activity in the L1 immunotype strain which had previously been reported to lack sialic acid in its LOS because it contains a terminal alpha-linked galactose which was thought not to be an acceptor for the sialyltransferase . This finding prompted us to re-examine the structure of the LOS from the L1 immunotype, which revealed the presence of sialic acid on the terminal alpha-linked galactose . Oligosaccharides derived from the LOS were shown to be sialylated by composition and methylation analysis, mass spectrometry and nuclear magnetic resonance . The detailed structural analysis showed the sialic acid to occur only at 06 of the terminal a-D-galactopyranose residue of the alpha-D-Gal-1,4-beta-D-Gal-1,4-beta-D-glc trisaccharide (Pk epitope) chain of the LOS, in the alpha-D configuration . These data are the first report of a alpha-2,6-linked sialic acid in a bacterial LOS or lipopolysaccharide, and also the first report of a sialylated Pk epitope.

Antimicrob Agents Chemother, 1998 Aug, 42(8), 2103 - 5
Identification of novel mutation patterns in the parC gene of ciprofloxacin-resistant isolates of Neisseria gonorrhoeae; Trees DL et al.; Of 65 ciprofloxacin-resistant, clinical isolates of Neisseria gonorrhoeae, 5 isolates exhibited ParC mutations previously undescribed in the gonococcus . For isolates containing two ParC mutations (the Ser-87-->Ile and Glu-91-->Gly mutations and the Gly-85-->Cys and Arg116-->Leu mutations) the MICs of ciprofloxacin (8.0 to 64.0 microg/ml) were higher than those for the isolate containing the single ParC mutation (Arg-116-->Leu; MIC, 1.0 microg/ml).

Exp Cell Res, 1998 Aug 1, 242(2), 528 - 39
Ligation of cell surface heparan sulfate proteoglycans by antibody-coated beads stimulates phagocytic uptake into epithelial cells: a model for cellular invasion by Neisseria gonorrhoeae; Dehio C et al.; Binding of a particular opacity outer membrane protein (Opa) of Neisseria gonorrhoeae to cell surface heparan sulfate proteoglycans (HSPGs) of epithelial cells results in tight bacterial adherence; however, the role of this ligand-receptor interaction in triggering the subsequent bacterial internalization step is uncertain . Here we have used latex beads coated with HSPG-ligating antibodies as an in vitro model to study the role of HSPGs in gonococcal uptake into epithelial cells . Beads and gonococci showed the same cell line-specified adherence patterns and increase in phagocytic uptake mediated by serum or purified vitronectin (Vn) . Heparitinase digestion as well as antibody competition experiments indicate that a critical level of HSPG ligation is necessary and sufficient to trigger phagocytic uptake into epithelial cells . Vn was found to specifically enhance HSPG-dependent phagocytic uptake while phagocytosis resulting from the ligation of other cell surface receptors was unaffected in the presence of Vn . Pharmacological studies with PKC inhibitors suggest a role for PKC in phagocytic uptake of HSPG-ligating beads . The use of drugs impairing cytoskeletal functions indicates that HSPG-dependent phagocytosis requires actin polymerization by a process distinct from receptor-mediated endocytosis .

J Bacteriol, 1998 Aug, 180(15), 3741 - 9
The regulated outer membrane protein Omp21 from Comamonas acidovorans is identified as a member of a new family of eight-stranded beta-sheet proteins by its sequence and properties; Baldermann C et al.; Omp21, a minor outer membrane protein of the soil bacterium Comamonas acidovorans, was purified from a spontaneous mutant lacking a surface layer and long-chain lipopolysaccharide . Omp21 synthesis is enhanced by oxygen depletion, and the protein has a variable electrophoretic mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis due to its heat-modifiable behavior . The structural gene omp21 encodes a precursor of 204 amino acids with a putative signal peptide of 21 amino acids . Mature Omp21 is a typical outer membrane protein with a high content of beta structure as determined by infrared spectroscopy . Sequence comparisons show that it belongs to a new outer membrane protein family, characterized by eight amphipathic beta strands, which includes virulence proteins, such as the neisserial opacity proteins, Salmonella typhimurium Rck, and Yersinia enterocolitica Ail, as well as the major outer membrane proteins OmpA from Escherichia coli and OprF from Pseudomonas aeruginosa.

Protein Eng, 1998 Apr, 11(4), 295 - 302
Role of paired basic residues in the expression of active recombinant galactosyltransferases from the bacterial pathogen Neisseria meningitidis; Wakarchuk WW et al.; The lgtB gene encoding a beta-1,4-galactosyltransferase gene and the lgtC gene encoding an alpha-1,4-galactosyltransferase from the bacterial pathogen Neisseria meningitidis were cloned into an expression vector and overexpressed in Escherichia coli . Both genes expressed very well, but problems with C-terminal proteolysis were encountered with both proteins . The lgtC protein was initially isolated from extracts of recombinant E.coli as a truncated species that retained enzymatic activity, and was subsequently shown by mass spectrometry to be 19 residues shorter than the expected protein . A specific set of engineered C-terminal deletions was constructed to investigate their effect on the expression of lgtC . As many as 28 residues could be deleted with little effect on activity, and with the concomitant improvement of the overall expression up to fivefold over the full length protein . The lgtB protein was also proteolysed in extracts of normal E.coli strains into enzymatically inactive fragments lacking 28 or 41 C-terminal residues . This degradation could be prevented by expression in an ompT protease deficient strain of E.coli . The full length lgtB protein was not stable in soluble protein extracts from all recombinant strains, however a stable enzyme preparation could be achieved with the membrane fraction from cells of the ompT deficient strain expressing lgtB . Specific deletions of lgtB were also constructed, and 15 residues could be removed without loss of enzyme activity and also with the concomitant improvement of the overall expression up to twofold over the full length protein . Longer deletions produced protein but activity could not be detected in these recombinant strains . Examination of the glycosyltransferase sequences from a wide range of bacteria showed their C-terminal segments of approximately 50 amino acids frequently contained paired basic residues . Engineering of these segments may therefore be required as a general practice to produce these enzymes for use in the large scale chemi-enzymatic synthesis of carbohydrate-based therapeutics.

Int J STD AIDS, 1998 Jun, 9(6), 347 - 9
Comparison of tampon and urine as self-administered methods of specimen collection in the detection of Chlamydia trachomatis, Neisseria gonorrhoeae and Trichomonas vaginalis in women; Tabrizi SN et al.; Self-administered sampling techniques for the detection of sexually transmitted diseases (STDs) are particularly useful due to their ease of collection and better patient compliance . Urine specimens, and recently tampons, have been described as methods of specimen collection for the detection of some STDs in women . In this study, 660 women had both first-void urine (FVU) and tampon specimens analysed by polymerase chain reaction (PCR) for the detection of Chlamydia trachomatis, Neisseria gonorrhoeae and Trichomonas vaginalis . Overall 6.5%, 10.1% and 17.9% of urine samples were positive whereas 7%, 21.2% and 22% of tampon specimens were positive for C . trachomatis, N . gonorrhoeae and T . vaginalis respectively . Tampon-collected specimens tested by PCR were more sensitive than urine specimens for the detection of N . gonorrhoeae and T . vaginalis (P < 0.001) and equally sensitive for the detection of C . trachomatis (P=0.45).

Int J STD AIDS, 1998 Jun, 9(6), 323 - 5
Gonorrhoea treatment failure and ciprofloxacin resistance; Ng PP et al.; Ciprofloxacin-resistant strains of Neisseria gonorrhoeae have been on the increase over the past few years in Singapore and worldwide . The aim of this study is to correlate treatment failures with in vitro ciprofloxacin resistance of N . gonorrhoeae . A total of 694 patients attending the Department of STD Control (DSC) clinic in 1996 who were diagnosed to have gonococcal infection confirmed by culture were analysed . Treatment failure rates for ciprofloxacin were determined and the minimum inhibitory concentration (MIC) of ciprofloxacin were traced for all cases of treatment failure . The case notes of all patients who had strains with MICs of ciprofloxacin in the resistant (> or = 1 microg/ml) and less sensitive (0.125-0.5 microg/ml) range were also reviewed to determine the clinical outcome . Ciprofloxacin treatment failure rate was 1.7% (8/461) which was lower than the percentage of ciprofloxacin-resistant strains isolated in the laboratory . Of these 8 cases, 6 were resistant and 2 were less sensitive to ciprofloxacin . Cure rates with ciprofloxacin for resistant and less sensitive strains were 40% and 92% respectively . In conclusion, in vitro resistance to ciprofloxacin may not translate into clinical treatment failure . Clinical treatment failures, on the other hand, are also seen in less sensitive strains.

Sex Transm Dis, 1998 Jul, 25(6), 322 - 6
Reappraising the value of urine leukocyte esterase testing in the age of nucleic acid amplification; Bowden FJ; BACKGROUND: The leukocyte esterase (LE) test has a limited role in determination of empiric therapy for male patients screened for urethritis because of its poor positive predictive value in low (< 5%) prevalence settings . The recent advent of nucleic acid amplification testing of first-void urine (FVU) has dramatically increased the ease with which widespread screening for Chlamydia trachomatis and Neisseria gonorrhoeae can be performed, but the costs of such testing may be prohibitive . The LE test may therefore have a role in management of urethritis because of its high negative predictive value . OBJECTIVES: To determine the sensitivity, specificity, and positive and negative predictive value of LE testing for the diagnosis of N . gonorrhoeae and C . trachomatis in male FVU specimens in a low-prevalence urban setting using a commercial polymerase chain reaction (PCR) as the gold standard . METHODS: Data were obtained on men presenting to an urban sexually transmitted disease clinic over a 16-month period . Patients were included if an FVU had been tested for the presence of LE using a rapid dipstick, read by an automated urine analyzer, and the sample (either an FVU or urethral swab) had then been processed for the detection of N . gonorrhoeae and C . trachomatis by PCR . RESULTS: Of 301 assessable patients, there were 14 cases of gonorrhoea, 21 cases of chlamydia, and 1 case of dual infection detected by PCR . Most men (245/301; 81.4%) were asymptomatic, of whom 12 of 245 (4.9%) had an infection detected compared with 24 of 56 (42.9%) in the symptomatic men (P < 0.001) . Using a "< or = trace" cutoff, the overall value for the sensitivity of the LE test was 77.8% (95% confidence interval, 60.4-89.3), specificity 80.8% (75.4-85.2), positive predictive value 35.4% (25.2-47.1), and negative predictive value 96.4% (92.8-98.3) . CONCLUSIONS: The negative predictive value of the LE test may be of use in determining which patients should proceed to specific diagnosis by nucleic amplification methods (e.g., PCR or ligase chain reaction) . By limiting testing to patients with positive LE results, cost savings may be made, enabling the technology to be used in a wider community setting . The value of the LE test in higher prevalence populations with access to nucleic amplification testing remains to be established.

Southeast Asian J Trop Med Public Health, 1997 Dec, 28(4), 791 - 800
An assessment and evaluation of methods for diagnosis of chlamydial and gonococcal infections; Chomvarin C et al.; Chlamydia trachomatis and Neisseria gonorrhoeae were studied in 350 females and 140 males attending the sexually transmitted disease clinic and AIDS Center, Khon Kaen zone 6 and the Division of Obstetrics and Gynecology, Khon Kaen Hospital . Chlamydia trachomatis infection was diagnosed by cell culture (CC), enzyme immunoassay (EIA) (Bioquest, NSW, Australia) and nucleic acid hybridization (PACE2 system: Gen-Probe, San Diego, Calif) . It was found that the sensitivity, specificity and positive and negative predictive values in females were 95.7, 100.0, 100.0, 99.7% by the cell culture; 91.3, 99.1, 87.5, 99.4% by the EIA; and 78.3, 99.7, 94.7, 98.5% by the PACE2 respectively . Values of the same parameters in males were 83.3, 100.0, 100.0, 98.5% by the cell culture; 75.0, 99.2, 90.0, 97.7% by the EIA and 91.7, 100.0, 100.0, 99.2% by PACE2 respectively . The methods for detection of Neisseria gonorrhoeae infection were conventional culture, PACE2 test and the direct examination (Gram's stain) . In females, the sensitivity, specificity and positive and negative predictive values of the conventional culture were 85.7, 100.0, 100.0, 99.7% and those of the PACE2 were 85.7, 99.1, 66.7, 99.7% respectively . In males, the values of the same parameters were 81.8, 100.0, 100.0, 100.0% by the conventional culture, 95.5, 100.0, 100.0 and 99.2% by the PACE2 . The prevalence of chlamydial infection in females was 6.6% (23/350) and that in males was 8.6% (12/140) . The prevalence of gonococcal infection in females was 2.0% (7/350) and in males was 15.7% (22/140) . The co-infection of Chlamydia trachomatis and Neisseria gonorrhoeae in females was 0.9% (3/350) and no co-infection was found in males . It is concluded that cell culture is an appropriate method for detection of Chlamydia trachomatis in both genders, particularly in females . PACE 2 test is the best method for such detection in symptomatic males while EIA is a good method in females, particularly in symptomatic females . For gonococcal detection, PACE2 test is a sensitive, specific and alternative method to the conventional culture . It can be appropriately applied for the diagnosis of gonococcal infection, particularly in males.

J Urol, 1998 Feb, 159(2), 405 - 7
Detection of Mycoplasma genitalium in patients with urethritis; Maeda S et al.; PURPOSE: We attempted to detect Mycoplasma genitalium in urethral swab specimens by a polymerase chain reaction based assay to determine the prevalence of M . genitalium in patients with urethritis . MATERIALS AND METHODS: We examined a total of 171 Japanese men who presented to our hospital from February 1995 through January 1997 . Of these men 150 had symptoms and signs compatible with acute urethritis and 21 had no symptoms or signs of urethritis . Urethral swab specimens were used to culture Neisseria gonorrhoeae, to detect Chlamydia trachomatis by an enzyme immunoassay and to detect M . genitalium by a polymerase chain reaction based assay . RESULTS: Gonococcal urethritis was diagnosed in 74 symptomatic men, and nongonococcal urethritis was diagnosed in 76 symptomatic men . Of the 74 cases of gonococcal urethritis 3 (4.1%) were positive for M . genitalium, and 14 (18.9%) were positive for C . trachomatis . Of the 76 cases of nongonococcal urethritis 10 (13.2%) were positive for M . genitalium, and 42 (55.2%) were positive for C . trachomatis . While only 1 of the 42 cases with chlamydial nongonococcal urethritis (2.4%) was positive for M . genitalium, 9 of the 34 chlamydia negative nongonococcal urethritis cases (26.5%) were positive for the mycoplasma . In contrast, all 21 cases men were negative for N . gonorrhoeae, M . genitalium, and C . trachomatis . CONCLUSIONS: The prevalences of M . genitalium in patients with gonococcal urethritis and nongonococcal urethritis who attended our clinic were 4.1 and 13.2%, respectively . M . genitalium was detected significantly more often in men with nongonococcal urethritis than in asymptomatic men . In addition, its prevalence in men with chlamydia negative nongonococcal urethritis (26.5%) was significantly greater than in those with chlamydia positive nongonococcal urethritis (2.4%) . These findings suggest that M . genitalium may be associated with the development of nongonococcal urethritis independent of C . trachomatis.

Br J Obstet Gynaecol, 1998 Jun, 105(6), 599 - 604
A randomised comparison of strategies for reducing infective complications of induced abortion; Penney GC et al.; OBJECTIVES: To determine lower genital tract carriage rates of C . trachomatis, N . gonorrhoeae and bacterial vaginosis among women seeking termination of pregnancy . To compare two clinical management strategies for minimising the risks of infective morbidity after induced abortion . DESIGN: Prevalence of infections was assessed by screening women undergoing abortion . Clinical management strategies were compared by a randomised trial . SETTING: The gynaecology departments of four hospitals in Scotland . PARTICIPANTS: 1672 women undergoing induced abortion . INTERVENTIONS: Women randomised to prophylaxis received metronidazole 1 g rectally before abortion plus doxycycline 100 mg twice daily for seven days . Women randomised to screen-and-treat received appropriate antibiotics only if screening proved positive for one or more infection . MAIN OUTCOME MEASURES: Prevalences of infections; morbidity in the eight weeks following abortion as assessed by reported symptoms, general practitioner consultation and prescription rates and hospital re-attendances; costs to the NHS of alternative managements . RESULTS: Prevalence rates: C . trachomatis 5.6%; N gonorrhoeae 0.19%; bacterial vaginosis 17.5% . Overall, women allocated to receive prophylaxis had lower rates of measures of short term infective morbidity than those allocated to screen-and-treat . These differences only reached statistical significance for women who were reported negative on screening . The direct costs to the NHS of prophylaxis and screen-and-treat were calculated to be 8.17 and 18.34 per woman, respectively . CONCLUSIONS: Prevalences of lower genital tract infections which have been implicated in increased rates of infective morbidity after abortion are similar to those reported elsewhere . Universal antibiotic prophylaxis is at least as effective as a policy of screen-and-treat in minimising the risk of short term infective morbidity and is far more cost efficientPIP: The presence of infection in the lower genital tract at the time of induced abortion has been associated with an increased risk of postabortion pelvic inflammatory disease (PID) . The present study investigated the prevalences of Neisseria gonorrhoeae, Chlamydia trachomatis, and bacterial vaginosis among 1672 women undergoing induced abortion at four Scottish hospitals in 1995-96 . It further compared the effectiveness of two clinical management strategies for minimizing the risk of postabortion infection . Women were randomly assigned to receive either 1 g of metronidazole rectally before abortion and 100 mg/day of doxycycline for 7 days (n = 826) or treatment only if screening was positive for infection (n = 846) . Preabortion lower genital tract screening indicated 3 women (0.2%) were positive for N . gonorrhoeae, 91 (5.6%) for C . trachomatis, and 282 (17.5%) for bacterial vaginosis . A review of the rates of general practitioner consultations, antibiotic prescriptions, and hospital readmissions in the 8 weeks postabortion showed that symptoms were minor and similar in duration and intensity among women in both treatment groups . The postabortion PID/endometriosis rate was 4.6% among women in the prophylaxis group and 6.8% in the screen-and-treat group . Women in these two groups who were initially positive for 1 or more infection had significantly higher rates of postabortion PID/endometriosis (7.7% and 7.4%, respectively) than those who were initially negative (3.1% and 5.7%, respectively) . Antibiotics had to be prescribed postabortion to 13.1% of women initially positive for 1 or more infection compared with 7.8% of those initially negative . The cost of universal prophylaxis (8.17 pounds) was less than half that of screening with treatment and follow up of positive cases .

Wei Sheng Wu Xue Bao, 1996 Oct, 36(5), 379 - 84
{Analysis of surface antigen molecule expression on serovar stains of Neisseria gonorrhoeae by flow cytometry}; Duan Q et al.; In the present experiment, flow cytometry was employed for analysing expression characteristics of the antigen molecules distingushed by 10 monoclonal antibodies against Neisseria gonorrhoeae lipooligosaccharides . Stability of antigen expression and amount of epitope on the surface of N . gonorrhoeae were quantitative determined . Reactivity of the monoclonal antibodies with serovar strains of N . gonorrhoeae were evaluated.

Infect Immun, 1998 Jul, 66(7), 3416 - 9
Neisseria gonorrhoeae induces focal polymerization of actin in primary human urethral epithelium; Giardina PC et al.; The pathogenic Neisseria species induce cytoskeletal reorganization in immortalized cell lines . In Chang conjunctival epithelium and T84 intestinal epithelium, focal cytoskeletal rearrangements in which bacteria contacted the epithelial surface were observed . We show that actin footprints are induced in gonococcus-challenged primary urethral epithelium . Moreover, the microbes induced microvillus extension from the epithelial cell surface . Our results indicate that formation of actin footprints is not an artifact of commonly used immortalized cell lines.

Ann Oncol, 1998 Apr, 9(4), 431 - 5
A novel cancer vaccine composed of human-recombinant epidermal growth factor linked to a carrier protein: report of a pilot clinical trial; Gonzalez G et al.; BACKGROUND: There is evidence of a relationship between epidermal growth factor (EGF) and tumor cell proliferation, such as the overexpression of EGF receptor (EGF-R) in different human tumors, which makes this system an interesting target for cancer treatment . Up to now, passive immunotherapy with monoclonal antibodies against the EGF-R has been assayed in clinics . Our approach consists of active immunotherapy with human EGF (hu-EGF) . We conducted a pilot clinical trial to define the safety, toxicity and immunogenicity of vaccination with hu-EGF coupled to a carrier protein . PATIENTS AND METHODS: Ten patients with histologically-proven malignant carcinomas (colon, lung, stomach and prostate) in advanced clinical stages were enrolled . Patients were immunized twice (on days 0 and 15) with hu-EGF linked to either tetanic toxoid (TT, five patients) or P64K Neisseria Meningitidis recombinant protein (P64k, five patients), intradermically, using aluminium hydroxyde as adjuvant . RESULTS: In both groups 60% of patients developed anti-EGF antibody titers without evidence of toxicity . Secondary reactions were very mild, limited to erythema and itching at the site of injection, which disappeared without medication . CONCLUSIONS: We conclude that the proposed vaccination with hu-EGF was well tolerated and that antibody titers against self EGF were developed . The results of this trial may be useful in the design of new clinical trials with higher dose immunization protocols and using more effective adjuvantsPublication Types:

Clinical Trial

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