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 . 0-3.2