Sex Transm Infect, 1998 Apr, 74(2), 120 - 7 Community based study of sexually transmitted diseases in rural women in the highlands of Papua New Guinea: prevalence and risk factors; Passey M et al.; OBJECTIVE: To estimate the prevalence of sexually transmitted diseases (STDs) and determine their risk factors/markers among a rural population of women in the highlands of Papua New Guinea . METHODS: Community based random cluster sample of women of reproductive age were interviewed and examined and had specimens collected for laboratory confirmation of chlamydial and trichomonal infection, gonorrhoea, syphilis, and bacterial vaginosis . RESULTS: Chlamydia trachomatis was detected in 26%, Trichomonas vaginalis in 46%, Neisseria gonorrhoeae in 1%, syphilis in 4%, pelvic inflammatory disease (PID) (diagnosed clinically) in 14%, and bacterial vaginosis in 9% of 201 women . 59% of the women had at least one STD . In a multivariate logistic regression analysis taking the clustered sampling into account, independent risk factors for chlamydial infection were age < or = 25 years, < four living children, visualization of yellow mucopurulent endocervical secretions on a white swab, and bacterial vaginosis . Being married to a man who did not have other wives was protective . For trichomonal infection, independent risk factors were having no formal education, infertility, more than one sexual partner in the previous 12 months, treatment for genital complaints in the previous 3 months, abnormal vaginal discharge detected on examination, and chlamydial infection . Similar levels of trichomonal infection were found in all age groups . Among married women, rates of infection correlated with their perception of their husband having had other sexual partners in the previous 3 months, and this relationship was significant for chlamydial infection among women over 25 . CONCLUSION: STDs are a major problem in this population, with the risk factors varying by outcome . Current treatment regimens are inappropriate given the high prevalence of trichomonal infection, and the available services are inadequate . Effective interventions are required urgently to reduce this burden and to prevent the rapid transmission of HIV. Sex Transm Infect, 1998 Feb, 74(1), 59 - 62 Analysis of quinolone resistance mechanisms in Neisseria gonorrhoeae isolates in vitro; Tanaka M et al.; BACKGROUND AND OBJECTIVES: Gonococcal fluoroquinolone resistance is now a significant problem in Japan . We generated gonococcal mutants resistant to norfloxacin in vitro from norfloxacin sensitive isolates and analysed the contribution of three known mechanisms of quinolone resistance in Neisseria gonorrhoeae . MATERIALS AND METHODS: Three clinical isolates of N gonorrhoeae susceptible to norfloxacin were exposed to increasing concentrations of norfloxacin . To identify mutations in the gyrA and parC genes of the gonococcal mutants, the quinolone resistance determining regions of the gyrA and parC genes were polymerase chain reaction (PCR) amplified and the PCR products were directly sequenced . Norfloxacin accumulation in the gonococcal cells was also measured . RESULTS: The MICs of norfloxacin for three variants containing a single GyrA mutation were 16-fold higher than that for their parent isolates . A variant showing reduced norfloxacin accumulation in the cells, without mutations in the GyrA or ParC proteins, was also less sensitive to norfloxacin, with a 16-fold increase in the MIC, compared with the parent strain . The MIC of norfloxacin for a variant which contained a single GyrA mutation with reduced norfloxacin accumulation in the cells was 128-fold higher than for the parent strain . A variant containing mutations in both GyrA and ParC proteins with reduced accumulation of norfloxacin in the cells showed a 256-fold increase in the norfloxacin MIC compared with the parent strain . There was no variant containing a ParC mutation without the simultaneous presence of a GyrA mutation . CONCLUSIONS: The results from this study suggest that not only a mutation in the gyrA gene but also reduced drug accumulation in cells contributes to the development of fluoroquinolone a mutation in the gyrA gene contributes to a high level of fluoroquinolone resistance in gonococci with decreases in accumulation in cells having an additional but lesser effect. Sex Transm Infect, 1998 Feb, 74(1), 35 - 9 Clinical algorithms for the screening of pregnant women for STDs in Libreville, Gabon: which alternatives? Bourgeois A, Henzel D, Malonga-Mouelet G, Dibanga G, Tsobou C, Peeters M, Delaporte E. OBJECTIVE: Sexually transmitted diseases (STDs) remain one of the major public health problems in the developing world . To implement a systematic screening of STDs among pregnant women in Libreville, Gabon, a preliminary cross sectional study on STD prevalence and risk factors was performed in antenatal clinics . A score, integrating risk factors and elementary clinical signs for the screening of STDs, showed higher performances compared with hierarchical algorithms . The prospective validation of this score based on six criteria (risk factors and simple clinical signs) was done in 1994-5 . The sensitivity (76.7%), compared with results from other studies, was acceptable for diagnosing cervical infection (Neisseria gonorrhoeae and/or Chlamydia trachomatis) but the specificity was low (50.6%) . In addition, the diagnostic values for diagnosing vaginal infection (Trichomonas vaginalis and/or Candida albicans) were poor . We then proposed to evaluate an alternative flowchart for the screening of cervical and vaginal infections . METHODS: In this study, 646 pregnant women were enrolled . Each woman was interviewed and examined by a physician and then was subjected to reference laboratory examinations . An algorithm in two steps, combining a risk assessment score at the beginning of a hierarchical process, followed by a second step more specifically applied to a limited number of women, was developed and evaluated . RESULTS: The prevalence rate was 11.3% for cervical infection and 39.5% for vaginal infection . The first step of the algorithm, applied to all pregnant women, is based on four criteria (age, marital status, dyspareunia, coloured vaginal discharge) . It allows classification of the women into three classes: high, low, and intermediate risk of cervical infection . Only the patients with intermediate risk were submitted to further investigations including speculum and microscopic examination, and subsequently chlamydial antigen detection . This flowchart was 83.6% and 81.2% sensitive and 63.4% and 62.7% specific for predicting cervical infection and vaginal infection, respectively . CONCLUSION: Similar strategies using simple rapid tests for chlamydial and gonococcal infection would certainly constitute a good diagnostic tool . This theoretical model needs to be evaluated prospectively, not only to confirm their diagnostic value but also to evaluate their feasibility, reliability and acceptability, as well as their cost effectiveness. Microb Pathog, 1998 May, 24(5), 299 - 308 Monoclonal antibodies to the epitope alpha-Gal-(1-4)-beta-Gal-(1- of Moraxella catarrhalis LPS react with a similar epitope in type IV pili of Neisseria meningitidis; Rahman M et al.; Murine monoclonal antibodies (MAbs) against the A, B and C LPS serotypes of M . catarrhalis were generated and their binding specificity was examined in an enzyme-linked immunosorbent assay (ELISA) . Two broadly cross-reactive monoclonal antibodies (MCA1 and MCC2) against the outer core region of LPS were further characterized . A panel of synthetic glycoproteins and glycolipids was used to determine the binding specificity of the MAbs . MCA1 and MCC2 bound specifically to alpha-Gal-(1-4)-beta-Gal of galabiose and globotriose glycoconjugates . The reactivity of the MAbs with galabiose was higher than that with globotriose . The MAbs could recognize the alpha-Gal-(1-4)-beta-Gal epitope only when it was in a terminal position . MCA1 was further shown to react with a similar epitope in the glycosylated type IV pili of N . meningitidis, which has been shown to contain a 1-4 linked digalactose at the terminal part of the saccharide present in the pili . MCA1 could efficiently recognize this epitope indicating that it was exposed on the surface of the pili . MMWR Morb Mortal Wkly Rep, 1998 May 29, 47(20), 405 - 8 Fluoroquinolone-resistant Neisseria gonorrhoeae--San Diego, California, 1997. {A comparison of the developmental characteristics of Neisseria gonorrhoeae and Staphylococcus aureus cultures on nutrient media of different compositions} Skliar TV, Krysenko AV, Gavriliuk VG, Vinnikov AI. Dniepropetrovsk State UniversityThe growth parameters of gonococcus and staphylococcus strains in liquid medium were determined . The generation time varied within the limits from 0.8 to 1.3 h for gonococcus and from 0.47 to 0.60 h for staphylococcus, the specific growth rate varied from 0.51 to 0.91 h-1 for gonococcus and from 1.68 to 2.11 h-1 for staphylococcus, depending on the concentration of sodium humate and bovine serum in the medium . The growth characteristics of analysed staphylococcus strains did not depend on the presence of plasmids . The gonococcal cultures grown in the liquid nutrient medium containing 20% of bovine serum for 8-11 h were the most physiologically active . Sodium humate added to the medium for cultivation in concentration 0.01% stimulated the growth of N . gonorrhoeae. J Bacteriol, 1998 Jun, 180(12), 3080 - 90 Preparation and characterization of Neisseria meningitidis mutants deficient in production of the human lactoferrin-binding proteins LbpA and LbpB; Bonnah RA et al.; Pathogenic members of the family Neisseriaceae produce specific receptors facilitating iron acquisition from transferrin (Tf) and lactoferrin (Lf) of their mammalian host . Tf receptors are composed of two outer membrane proteins, Tf-binding proteins A and B (TbpA and TbpB; formerly designated Tbp1 and Tbp2, respectively) . Although only a single Lf-binding protein, LbpA (formerly designated Lbp1), had previously been recognized, we recently identified additional bacterial Lf-binding proteins in the human pathogens Neisseria meningitidis and Moraxella catarrhalis and the bovine pathogen Moraxella bovis by a modified affinity isolation technique (R . A . Bonnah, R.-H . Yu, and A . B . Schryvers, Microb . Pathog . 19:285-297, 1995) . In this report, we characterize an open reading frame (ORF) located immediately upstream of the N . meningitidis B16B6 lbpA gene . Amino acid sequence comparisons of various TbpBs with the product of the translated DNA sequence from the upstream ORF suggests that the region encodes the Lf-binding protein B homolog (LbpB) . The LbpB from strain B16B6 has two large stretches of negatively charged amino acids that are not present in the various transferrin receptor homologs (TbpBs) . Expression of the recombinant LbpB protein as a fusion with maltose binding protein demonstrated functional Lf-binding activity . Studies with N . meningitidis isogenic mutants in which the lbpA gene and the ORF immediately upstream of lbpA (putative lbpB gene) were insertionally inactivated demonstrated that LbpA, but not LbpB, is essential for iron acquisition from Lf in vitro. J Clin Microbiol, 1998 Jun, 36(6), 1765 - 6 Evaluation of silica gel packages for transport of Neisseria meningitidis; Popovic T et al.; Eight Neisseria meningitidis reference strains, representing six different serogroups, were plated on 57 blood agar plates each . The growth was harvested and stored in silica gel packages at different temperatures for up to 90 days . When held at 4 degrees C, all strains were recovered after 90 days of storage . Strains held at room temperature or alternately at 4 degrees C and room temperature survived for at least 10 and 17 days, respectively. J Clin Microbiol, 1998 Jun, 36(6), 1630 - 3 Evaluation of the Abbott LCx ligase chain reaction assay for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine and genital swab specimens from a sexually transmitted disease clinic population; Carroll KC et al.; The Abbott LCx ligase chain reaction (LCR) assay for the simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae was evaluated by using swab and urine specimens from 562 patients . C . trachomatis results by LCR were compared to those by the Gen-Probe PACE 2 assay, whereas N . gonorrhoeae results by LCR were compared to those by culture . The Gen-Probe and LCR assays were performed according to the manufacturers' instructions . Gram-negative diplococci growing on modified Thayer-Martin medium were confirmed as N . gonorrhoeae by the GonoGen II assay . Supplemental data analysis was performed by major outer membrane protein PCR for C . trachomatis and probes for pilin gene detection for N . gonorrhoeae . A true-positive result for each pathogen was defined as a positive result for all three or two of three assays . Overall agreement among the six assays was 94.8% . C . trachomatis prevalence was 16.2%; N . gonorrhoeae prevalence was 5.5% . The overall sensitivity and specificity, respectively, for each test (after supplemental data analysis) were as follows: for C . trachomatis, Gen-Probe, 65.9 and 100%; LCR on urine, 90.1 and 100%; LCR on swab specimens, 96.7 and 100%; and for N . gonorrhoeae, culture, 80.6 and 100%; LCR on urine, 93.5 and 99.8%; and LCR on swab specimens, 96.8 and 100% . For women, the N . gonorrhoeae culture was very insensitive compared to its performance in men (58.3 versus 94.7%, respectively) . For C . trachomatis, the Gen-Probe assay's sensitivity was lower for men than for women (62.3 versus 71.1%, respectively) . The sensitivity for C . trachomatis detection by LCR on urethral and cervical swab specimens was 96.2 and 97.4% for men and women, respectively . For men, swab results were slightly better than urine results for both pathogens (sensitivity for C . trachomatis in swab and urine specimens, 96.2 and 92.5%, respectively; sensitivity for N . gonorrhoeae in swab and urine specimens, 100 and 94.7%, respectively), while for women, cervical swabs were superior in sensitivity to urine samples for detecting C . trachomatis (swab, 97.4%; urine, 81.6%) and equivalent for N . gonorrhoeae (swab, 92.3%; urine, 91.6%) . The LCx LCR appears to be both sensitive and specific for the detection of C . trachomatis and N . gonorrhoeae when performed on urine or genital swab samples . Swab samples had better sensitivity than urine samples for the detection of both pathogens. West Indian Med J, 1998 Mar, 47(1), 23 - 5 Prevalence of viral and bacterial sexually transmitted pathogens in Jamaican pregnant women; Dowe G et al.; In this study we investigated the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum, human immunodeficiency virus type I (HIV-I), human T cell lymphotropic virus type 1 (HTLV-1) and hepatitis B virus (HBV) infections in 200 pregnant women attending antenatal clinics at the University Hospital of the West Indies . 19% of the women had at least one pathogen: C . trachomatis was found in 16%, HTLV-1 in 2%, HIV-1, HBV and N . gonorrhoeae each in 0.5% C . trachomatis infection was more prevalent in women less than 20 years of age (31%) than in those 20 years and older (16%; OR = 0.43; chi 2 = 5.66; p < 0.05) . The study demonstrates the need for identification of sexually transmitted pathogens in antenatal women for syndromic management of genital infections as part of the strategy for prevention and control of HIV/AIDS (acquired immunodeficiency syndrome) in Jamaica. Cell Mol Life Sci, 1998 Apr, 54(4), 332 - 40 Resistant penicillin-binding proteins; Hakenbeck R et al.; Low-affinity penicillin-binding proteins (PBPs), which participate in the beta-lactam resistance of several pathogenic bacteria, have different origins . Natural transformation and recombination events with DNA acquired from neighbouring intrinsically resistant organisms are responsible for the appearance of mosaic genes encoding two or three low-affinity PBPs in highly resistant strains of transformable microorganisms such as Neisseria and Streptococcus pneumoniae . Methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococcal strains possess the mecA determinant gene, which probably evolved within the Staphylococcus genus from a closely related and physiologically functional gene that was modified by point mutations . The expression of mecA is either inducible or constitutive . A stable high-level resistant phenotype requires the synthesis of a normally constituted peptidoglycan . Enterococci have a natural low susceptibility to beta-lactams related to the presence of an intrinsic low-affinity PBP . Highly resistant enterococcal strains overexpress this PBP and/or reduce its affinity. Science, 1998 May 15, 280(5366), 1046 - 8 Chemical amplification: continuous-flow PCR on a chip; Kopp MU et al.; A micromachined chemical amplifier was successfully used to perform the polymerase chain reaction (PCR) in continuous flow at high speed . The device is analogous to an electronic amplifier and relies on the movement of sample through thermostated temperature zones on a glass microchip . Input and output of material (DNA) is continuous, and amplification is independent of input concentration . A 20-cycle PCR amplification of a 176-base pair fragment from the DNA gyrase gene of Neisseria gonorrhoeae was performed at various flow rates, resulting in total reaction times of 90 seconds to 18.7 minutes. Mol Gen Genet, 1998 Apr, 258(1-2), 34 - 44 Cloning and characterisation of the Neisseria gonorrhoeae aroB gene; Barten R et al.; The gene coding for the 3-dehydroquinate synthetase (aroB) of Neisseria gonorrhoeae has been cloned by functional complementation of an Escherichia coli aroB mutant . The aroB gene isolated from a gonococcal plasmid library encodes a 359 amino acid protein with a molecular mass of 38.6 kDa . Alignment of different prokaryotic and eukaryotic aroB gene products reveals an overall identity ranging from 33 to 55% . An open reading frame coding for an aroK homologue is located immediately upstream of aroB . Downstream of aroB a region of inverted repeats and a gene showing high homology to yafJ of E . coli has been identified . Disruption of aroB generates a gonococcal mutant that is unable to grow in the absence of aromatic compounds . Complementation of the mutant with the intact aroB gene in trans indicates that the gene is responsible for the auxotrophic phenotype . In infection assays with AroB-deficient gonococcal strains, binding, entry and short-term survival in epithelial cells is not affected . The aroB gene might be useful as a selectable marker and target for attenuation of a gonococcal live vaccine strain or as a biosafe laboratory strain. Am J Obstet Gynecol, 1998 May, 178(5), 987 - 90 Risk factors for plasma cell endometritis among women with cervical Neisseria gonorrhoeae, cervical Chlamydia trachomatis, or bacterial vaginosis; Korn AP et al.; OBJECTIVE: We sought to determine potential risk factors for upper genital tract inflammation in women with cervical Neisseria gonorrhoeae, Chlamydia trachomatis, or bacterial vaginosis . STUDY DESIGN: In a case-controlled study we compared 111 women with cervical Neisseria gonorrhoeae, Chlamydia trachomatis, or bacterial vaginosis (the study group) with 24 women who had negative tests for each of these infections (the control group) . We evaluated potential risk factors for upper genital tract inflammation by use of bivariate and then logistic regression analysis . RESULTS: We found plasma cell endometritis in 53 of 111 women in the study group and 3 of 24 controls (odds ratio = 6.4, 95% confidence interval 1.7 to 35.0) . On logistic regression, the study group women who were in the proliferative phase had increased likelihood of plasma cell endometritis (odds ratio = 4.5, 95% confidence interval 1.6 to 12.4) . CONCLUSION: The proliferative phase of the menstrual cycle seems to be the primary risk factor for ascending infection by organisms associated with pelvic inflammatory disease . This may be due to a hormonal effect or to the loss of the cervical barrier during menstruation. J Med Chem, 1998 May 7, 41(10), 1688 - 95 Antineoplastic agents . 379 . Synthesis of phenstatin phosphate; Pettit GR et al.; A structure-activity relationship (SAR) study of the South African willow tree (Combretum caffrum) antineoplastic constituent combretastatin A-4 (1b) directed at maintaining the (Z)-stilbene relationship of the olefin diphenyl substituents led to synthesis of a potent cancer cell growth inhibitor designated phenstatin (3b) . Initially phenstatin silyl ether (3a) was unexpectedly obtained by Jacobsen oxidation of combretastatin A-4 silyl ether (1c --> 3a), and the parent phenstatin (3b) was later synthesized (6a --> 3a --> 3b) in quantity . Phenstatin was converted to the sodium phosphate prodrug (3d) by a dibenzyl phosphite phosphorylation and subsequent hydrogenolysis sequence (3b --> 3c --> 3d) . Phenstatin (3b) inhibited growth of the pathogenic bacterium Neisseriagonorrhoeae and was a potent inhibitor of tubulin polymerization and the binding of colchicine to tubulin comparable to combretastatin A-4 (1b) . Interestingly, the prodrugs were found to have reduced activity in these biochemical assays . While no significant tubulin activity was observed with the phosphorylated derivative of combretastatin A-4 (1d), phosphate 3d retained detectable inhibitory effects in both assays. Infect Immun, 1998 Jun, 66(6), 2453 - 9 Age-dependent Neisseria meningitidis serogroup C class-specific antibody concentrations and bactericidal titers in sera from young children from Montana immunized with a licensed polysaccharide vaccine; Maslanka SE et al.; Neisseria meningitidis serogroup C bactericidal titers and class-specific enzyme-linked immunosorbent assay (ELISA) antibody concentrations were measured in sera from 173 children (1 to 5 years old) before and 6 weeks and 7 months following vaccination with a quadrivalent (A/C/Y/W-135) polysaccharide vaccine . The immune responses of the children were compared with those of 40 adults 6 weeks postvaccination . Both bactericidal titers and ELISA antibody concentrations were significantly higher in the adults than in the children (P < 0.05) . In addition, the ratio of immunoglobulin G (IgG) to IgM was higher in the children than in the adults . With an ELISA total antibody concentration of >/=2 microg/ml used as a measure of seroconversion, >/=84% of the individuals from each age group responded to the serogroup C polysaccharide . However, with a >/=4-fold-increase in bactericidal titer used, only 18% of 1-year-olds, 32% of 2-year-olds, and 50 to 60% of 3-, 4-, and 5-year-olds seroconverted . The ELISA results suggest that >50% of all children retained >/=2 microg of total antibody per ml at 7 months postimmunization . However, the bactericidal titers suggest that <10% of children <4 years old retained a >/=4-fold increase at 7 months following vaccination . Of particular note, 59 of 79 sera (75%) from the 1- and 2-year-olds had high ELISA antibody concentrations (2 to 20 microg/ml) with no associated bactericidal titer (<1:8) . Discordant results between bactericidal titers and ELISA antibody concentrations were not explained by the presence of IgA blocking antibody or relative levels of IgG and IgM . The bactericidal results show age-dependent differences in the production and retention of antibody in young children immunized with serogroup C polysaccharide; these differences are not evident with the ELISA data. J Urol, 1998 Jun, 159(6), 2215 - 9 Development of fluoroquinolone resistance and mutations involving GyrA and ParC proteins among Neisseria gonorrhoeae isolates in Japan; Tanaka M et al.; PURPOSE: To investigate the development of fluoroquinolone resistance among Neisseria gonorrhoeae isolates in Japan and the frequency and patterns of mutations involving the GyrA and ParC proteins, which confer quinolone resistance to the bacteria, in isolates . MATERIALS AND METHODS: Antimicrobial susceptibilities of 145 gonococcal isolates, including 79 isolated from February 1992 through February 1993 and 66 isolated from February 1995 through February 1996, to six fluoroquinolones and several other antibiotics were compared with those of 27 isolates obtained from 1981 through 1984 . To identify mutations in gyrA and parC genes of the isolates, the quinolone resistance-determining regions of the gyrA and parC genes were PCR-amplified and the PCR products were directly sequenced . RESULTS: The minimum inhibitory concentration for 90% of strains (MIC90) values of norfloxacin for the isolates from 1992 to 93 (4 microg./ml.) and 1995 to 96 (8 microg./ml.) were 16- and 32-fold, respectively, higher than those for isolates from 1981 to 84 (0.25 microg./ml.) . The MIC90 values of ciprofloxacin for isolates from 1992 to 93 (0.5 microg./ml.) and 1995 to 96 (1 microg./ml.) showed increase of 8- and 16-fold, respectively, in comparison with those from 1981 to 84 (0.063 microg./ml.) . The isolates from 1992 to 93 and 1995 to 96 were also less susceptible to newer fluoroquinolones including levofloxacin, sparfloxacin, DU-6859a and AM-1155, as compared with those from 1981 to 84 . In 46 (67.6%) and 16 (23.5%) of the 68 gonococcal strains sequenced, GyrA and ParC mutations were identified, respectively . No ParC substitutions were identified in any isolates without co-existence of the GyrA mutation . A Ser-91 to Phe mutation, which was detected in 30 (65.2%) of the 46 isolates with GyrA mutations, was the most common GyrA mutation . Mutants with the single Ser-91 to Phe substitution in GyrA were 12-fold and at least 13-fold, respectively, less susceptible to norfloxacin and ciprofloxacin than the wild type . CONCLUSIONS: The results obtained in this study suggest that a high prevalence of gonococcal isolates with the Ser-91 to Phe mutation in GyrA has reduced the susceptibility of this organism to fluoroquinolones in Japan. FEMS Microbiol Lett, 1998 May 1, 162(1), 75 - 82 Naturally occurring isolates of Neisseria gonorrhoeae, which display anomalous serovar properties, express PIA/PIB hybrid porins, deletions in PIB or novel PIA molecules; Cooke SJ et al.; The por gene of Neisseria gonorrhoeae encodes the Protein I porin responsible for serovar specificity . In this study the por genes have been sequenced from clinical isolates which exhibited anomalous serovar reactivity . One group of 'intermediate' strains differed significantly from both Protein IA and IB strains, were more closely related to IA but appeared to represent a distinct class of Protein I . Another strain was closely related to Protein IB of serovar IB-6 but contained a deletion of six amino acids in surface exposed loop 6 which removed epitopes recognized by IB specific monoclonal antibodies . The third group of strains, which reacted with both IA and IB specific monoclonal antibodies, expressed hybrid Protein I molecules containing both IA and IB epitopes . These strains appeared to originate from a double crossover between Proteins IA and IB with the amino and carboxy terminal residues homologous to IB while the surface exposed loop 6 demonstrated close homology to IA . This is the first demonstration of naturally occurring gonococci expressing a hybrid Protein IA/IB. Antimicrob Agents Chemother, 1998 May, 42(5), 1062 - 7 Sulfonamide resistance in Streptococcus pyogenes is associated with differences in the amino acid sequence of its chromosomal dihydropteroate synthase; Swedberg G et al.; Sulfonamide resistance in recent isolates of Streptococcus pyogenes was found to be associated with alterations of the chromosomally encoded dihydropteroate synthase (DHPS) . There were 111 different nucleotides (13.8%) in the genes found in susceptible and resistant isolates, respectively, resulting in 30 amino acid changes (11.3%) . These substantial changes suggested the possibility of a foreign origin of the resistance gene, in parallel to what has already been found for sulfonamide resistance in Neisseria meningitidis . The gene encoding DHPS was linked to at least three other genes encoding enzymes of the folate pathway . These genes were in the order GTP cyclohydrolase, dihydropteroate synthase, dihydroneopterin aldolase, and hydroxymethyldihydropterin pyrophosphokinase . The nucleotide differences in genes from resistant and susceptible strains extended from the beginning of the GTP cyclohydrolase gene to the end of the gene encoding DHPS, an additional indication for gene transfer in the development of resistance . Kinetic measurements established different affinities for sulfathiazole for DHPS enzymes isolated from resistant and susceptible strains. Commun Dis Intell, 1998 Apr 16, 22(4), 52 - 6; discussion 57-8 An outbreak of non-sexually transmitted gonococcal conjunctivitis in Central Australia and the Kimberley region; Matters R et al.; From 13 February to 27 June 1997, 447 cases of gonococcal conjunctivitis were identified by Communicable Disease and Public Health Centres and Community Clinics in the Northern Territory, Western Australia and South Australia . The outbreak involved Aboriginal communities predominantly in Central Australia and the Kimberley region in Western Australia . This was the first outbreak recorded in the Kimberley region . It is not yet known whether the Kimberley cases were part of the larger Central Australian outbreak or whether they represented a separate and unrelated outbreak . Environmental factors associated with this outbreak were similar to those seen in previous outbreaks . Control measures were based on early recognition and treatment of index cases and identifying and treating contacts . Until sexually transmitted Neisseria gonorrhoeae is controlled in communities gonococcal conjunctivitis is likely to appear again . The role of oropharyngeal carriage of N . gonorrhoeae needs to be evaluated further. Genitourin Med, 1997 Dec, 73(6), 564 - 6 Gonorrhoea: auxotypes, serovars, and clinical manifestations among female sex workers from Kinshasa, Zaïre; Mukenge-Tshibaka L et al.; The main question in this paper was to look at the distribution of auxotypes and serovars of Neisseria gonorrhoeae and check whether they correlate with clinical symptoms/signs among female sex workers (FSW) from Kinshasa, Zaire . The subject were 1233 FSW enrolled in a cross sectional study on STDs and HIV infection in 1988; 771 of them were followed prospectively for a median duration of 23 months . At each visit, clinical symptoms and signs of cervicitis were recorded and the subjects were screened for gonococcal and chlamydial infection . The pre-dominant auxotypes were prototrophic (35.2%), proline requiring (29.6%), and proline requiring phenylalanine inhibition (19%) . Serovars 1A-6 (42.5%) and 1B-1 (16.7%) were the commonest . Infection with auxotype prototrophic and phenylalanine inhibition (Proto/Phenali) was significantly associated with both mucopurulent cervicitis and pelvic inflammatory disease; (OR = 8.9; p = 0.002 and OR =19 x9; p = 0.002; respectively) . Despite the few associations found in this study, there was not clear pattern linking clinical manifestations to auxotype/serovar profiles. Genitourin Med, 1997 Dec, 73(6), 558 - 60 Family planning services in developing countries: an opportunity to treat asymptomatic and unrecognised genital tract infections? Wilkinson D, Ndovela N, Harrison A, Lurie M, Connolly C, Sturm AW. OBJECTIVE: To determine the prevalence of asymptomatic and unrecognised genital tract infections among women attending a family planning clinic in rural South Africa . METHODS: 189 consecutive women had genital samples taken to diagnose infection with Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Candida albicans, Treponema pallidum, and HIV, and to diagnose bacterial vaginosis . RESULTS: Mean age was 25 years; 155 (82%) were unmarried, 156 (83%) were currently using contraception, and 41 (22%) reported having an STD treated in the preceding 12 months . Although none volunteered abnormal urogenital symptoms, 74 (39%) had at least one elicited by direct questioning . 119 women (63%) had at least one genital infection: N gonorrhoeae (eight; 4%), C trachomatis (14; 8%), T vaginalis (26; 14%), C albicans (56; 30%), active syphilis (15; 8%), HIV (44; 24%), and bacterial vaginosis (29; 15%) . 49 women (26%) had multiple infections . Most infections (71; 60%) were asymptomatic . Symptomatic women failed to recognise and report their symptoms, and routine services failed to detect the infections . CONCLUSION: Prevalence of genital tract infection is high among these women, most infections are asymptomatic, and symptomatic infections are frequently not recognised . Women attending family planning clinics in such settings should be screened for syphilis and offered testing for HIV infection . Strategies to detect and treat other genital infections need to be developedPIP: This is a study of 189 women attending a family planning clinic in rural South Africa to determine the prevalence of asymptomatic and unrecognized genital tract infections . Genital samples were taken from these women to diagnose infection with Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Candida albicans, Treponema pallidum, and HIV, and to diagnose bacterial vaginosis . Among the 189 women, 41 (22%) reported having had an STD treated in the preceding 12 months . By direct questioning, 74 women stated the following symptoms: genital itch -- 38 (20%); vaginal discharge -- 56 (30%); dysuria -- 33 (18%); dyspareunia -- 22 (12%); and genital ulcers -- 4 (2%) . 45 (24%) women had more than one symptom . 119 (63%) women had at least one genital infection, and 49 (26%) had multiple infections . Most of the infections were asymptomatic; while those that were symptomatic, were unrecognized or not reported . Results showed a high prevalence of genital tract infection among the participating women, with most of their infections remaining asymptomatic or unrecognized . Thus, strategies to detect and treat genital tract infections in rural South Africa need to be developed . Genitourin Med, 1997 Dec, 73(6), 555 - 7 Sexually transmitted disease among married Zambian women: the role of male and female sexual behaviour in prevention and management; Morrison CS et al.; OBJECTIVES: Few studies have evaluated the relation between male and female sexual behaviour and STD among married African women . The objectives of this study were to identify male and female sexual behaviour associated with female STD, and to explore whether incorporating male and female sexual behaviour and male symptoms can improve algorithms for STD management in married African women . METHODS: 99 married couples with one symptomatic member (58 males, 41 females) attending an STD clinic in Lusaka, Zambia were interviewed separately about sexual and contraceptive behaviour, and had physical examinations . Diagnostic tests for Neisseria gonorrhoeae (GC), Trichomonas vaginalis (TV), and HIV were performed . Bivariate and multivariate odds ratios for the association between sexual behaviour and STD were calculated . Predictive algorithms based on current Zambian guidelines for management of STD in women were created . RESULTS: Among women at baseline, 10% were positive for GC, 14% for TV, 52% for HIV . Female alcohol use before sex, a male's paying for sex, and a couple's having sex unprotected by condoms or spermicides were associated with female STD . Incorporation of these behaviours along with symptoms of urethral discharge and dysuria among husbands increased the predictive ability of algorithms for management of STD in women . CONCLUSIONS: The addition of male and female sexual behaviour and male STD symptoms to diagnostic algorithms for female STD should be explored in other settings . Both husbands' and wives' behaviour independently predict STD in these women; risk reduction programmes should target both men's and women's sexual behaviour. Genitourin Med, 1997 Dec, 73(6), 510 - 7 Surveillance of antibiotic resistance in Neisseria gonorrhoeae in The Netherlands, 1977-95; van de Laar MJ et al.; OBJECTIVE: To evaluate the prevalence and epidemiology of penicillinase producing Neisseria gonorrhoeae (PPNG) and tetracycline resistant N gonorrhoeae (TRNG) in the period 1977-95 in the Netherlands . To compare auxotypes, serovars, and antibiograms of PPNG, non-PPNG, and TRNG . To identify determinants in patient characteristics for the epidemic spread of TRNG/PPNG . METHODS: With respect to the national gonococcal surveillance all PPNG isolates from 30 laboratories over the country in 1977-90 and all gonococcal isolates from five sentinel laboratories (during 1 month per quarter) in 1991-5 were collected . Isolates were auxotyped and serotyped, the susceptibility for various antibiotics was tested and plasmid contents were evaluated . Additional data on PPNG infected individuals were collected retrospectively during a microepidemic of TRNG/PPNG . Univariate and multivariate analyses were performed to identify risk factors for TRNG/PPNG infections . RESULTS: In 1995 an overall high prevalence of PPNG infection (27%) and TRNG among PPNG infection (24%) was found in the Netherlands . Importantly, PPNG were found to have higher MICs for ceftriaxone and ciprofloxacin than non-PPNG; clinically relevant resistance to these antibiotics (or related agents) may emerge first among these strains . The observed diversity of strains (123 auxo/serovar classes since 1988) indicates a continuous introduction of new strains into the community . The epidemic increase of TRNG/PPNG was mainly caused by A/S classes NR/1B-6, PRO/1A-3, and PRO/1A-6, suggesting a clonal spread of a few strains; the rapid spread was associated with transmission in high risk individuals (that is, prostitutes and their clients) . CONCLUSION: The prevalence of PPNG in the Netherlands remains high and reduced sensitivity to other antimicrobials was detected among the PPNG strains . This underlines the necessity for a continuous national surveillance of resistance in gonococci including limited epidemiological information. Genitourin Med, 1997 Dec, 73(6), 506 - 9 Cefaclor, an alternative to third generation cephalosporins for the treatment of gonococcal urethritis in the developing world? Crabbe F, Grobbelaar TM, van Dyck E, Dangor Y, Laga M, Ballard RC. OBJECTIVE: To reassess the in vivo and in vitro efficacy of cefaclor for the treatment of uncomplicated gonococcal infection . DESIGN: Open clinical trail conducted in South Africa among consecutive male patients with symptoms and signs of uncomplicated urethritis and laboratory evidence of gonorrhoea . METHODS: Patients were treated with 3 g of cefaclor plus 1 g probenecid as a single dose . Urethral specimens were cultured for Neisseria gonorrhoeae at the initial visit and at follow up . Patients were considered cured if follow up cultures were negative . Treatment was considered to have failed in the patients infected with identical gonococcal strains at the initial and at the control visit . Those with evidence of infection at the follow up visit were administered 400 mg of ofloxacin and doxycycline 100 mg twice daily for 7 days . Minimal inhibitory concentrations (MICs) of cefaclor were determined by an agar dilution technique on the gonococcal isolates from the study subjects . The results were compared with those of isolates from three other African countries . RESULTS: Of 155 patients evaluated, 151 were cured (97%) . Thirty per cent of the patients complained of adverse effects, mainly gastrointestinal . Even though MICs for the isolates from the three other African countries were significantly higher than those for the isolates from the study, none was considered resistant to cefaclor in vitro . MICs were markedly influenced by the type of test medium used . CONCLUSION: The trial demonstrated the efficacy of a single oral dose of cefaclor with probenecid for the treatment of uncomplicated gonococcal urethritis in South Africa . Its potential as an alternative therapy to third generation cephalosporins deserves to be further investigated. Biochim Biophys Acta, 1998 Mar 13, 1370(2), 289 - 98 Successful recovery of the normal electrophysiological properties of PorB (class 3) porin from Neisseria meningitidis after expression in Escherichia coli and renaturation; Song J et al.; Neisseria meningitidis PorB class 3 porins obtained either from native membranes (wild-type) or recovered from inclusion bodies following expression in Escherichia coli (recombinant), have been reconstituted into solvent-free planar phospholipid membranes . The wild-type and recombinant porins exhibited the same single-trimer conductance (1-1.3 nS in 200 mM NaCl), tri-level closure pattern, characteristic of functional channel trimers, and pattern of insertion into planar membranes . Both proteins were open at low voltages and displayed two voltage-dependent closure processes, one at positive and the other at negative potentials . Both showed asymmetric voltage dependence such that one gating process occurred at lower voltages (Vo=15 mV) than the other (Vo=25 mV) . The sign of the potential that resulted in closure at low voltages varied from membrane to membrane indicating that they may have the property of auto-directed insertion (in analogy to the mitochondrial channel, VDAC) . In the case of the recombinant porin, the steepness of the voltage dependence of one gating process was slightly less (n=1.3) than that observed for the other process or for the wild-type channel (n=1.5-1.7) . Both channels have a high (40%) probability of closure even at 0 mV . While both channels show a slight selectivity for Cl- over Na+, the selectivity of the recombinant porin is a bit higher (permeability ratio of 2.8 vs . 1.6) as measured using a 2-fold salt gradient . Thus, the method employed to refold the recombinant porin was successful in not only restoring wild-type structure {H.L . Qi, J.Y . Tai, M.S . Blake, Expression of large amounts of Neisserial porin proteins in Escherichia coli and refolding of the proteins into native trimers, Infect . Immun . 62 (1994) 2432-2439; C.A.S.A . Minetti, J.Y . Tai, M.S . Blake, J.K . Pullen, S.M . Liang, D.P . Remeta, Structural and functional characterization of a recombinant PorB class 2 protein from Neisseria meningitidis . Conformational stability and porin activity, J . Biol . Chem . 272 (1997) 10710-10720} but also the overall electrophysiological function . Acta Med Port, 1997 Oct, 10(10), 637 - 41 {Inflammatory smears in cervicovaginal cytology . A finding meaning infection?}; Ayres de Campos D et al.; INTRODUCTION: Inflammatory changes are relatively frequent findings in cervical smears and are generally believed to be a consequence of genital infection . However, clinical signs of infection are frequently absent and no consensus exists on the management of these patients . The objective of this study was to assess whether or not inflammatory smears are exclusively a consequence of genital infection . For this purpose, the prevalence of genital infection in a group of women with inflammatory cells in the cervical smear and a control group with normal smears was compared . MATERIAL AND METHODS: Sixty-two regularly menstruating women aged 17 to 48 years, attending the outpatient Gynaecology sector of S . Joao Hospital, were prospectively evaluated . Cervical smears were analysed by the same cytologist who chose 10 inflammatory cells per high power field (400x) as the cut-off value for normality . Infection by Candida spp., Trichomonas vaginalis, Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma urealyticum, Chlamydia trachomatis and bacterial vaginosis was investigated . Human Papillomavirus infection was evaluated by colposcopy and biopsy of abnormal colposcopic findings . RESULTS: Thirty-four women and inflammatory cells on their cervical smear and 15 of these (44%) had a genital infection . Of the 28 women with normal smears, 12 (43%) had a genital infection . No statistically significant difference in the prevalence of infection was found between the two groups . CONCLUSIONS: Our results suggest that the presence of inflammatory cells in cervical smears is not necessarily due to infection and other causes may be responsible for their appearance. Infect Immun, 1998 May, 66(5), 2330 - 6 A Neisseria meningitidis fbpABC mutant is incapable of using nonheme iron for growth; Khun HH et al.; The neisserial fbpABC locus has been proposed to act as an iron-specific ABC transporter system . To confirm this assigned function, we constructed an fbpABC mutant in Neisseria meningitidis by insertional inactivation of fbpABC with a selectable antibiotic marker . The mutant was unable to use iron supplied from human transferrin, human lactoferrin, or iron chelates . However, the use of iron from heme and human hemoglobin was unimpaired . These results support the obligatory participation of fbpABC in neisserial periplasmic iron transport and do not indicate a role for this genetic locus in the heme iron pathway. Infect Immun, 1998 May, 66(5), 1918 - 27 Comparisons between colony phase variation of Neisseria gonorrhoeae FA1090 and pilus, pilin, and S-pilin expression; Long CD et al.; The gonococcal pilus is a primary virulence factor, providing the initial attachment of the bacterial cell to human mucosal tissues . Pilin, the major subunit of the pilus, can carry a wide spectrum of primary amino acid sequences which are generated by the action of a complex antigenic variation system . Changes in the pilin amino acid sequence can produce different pilus-dependent colony morphotypes, which have been previously shown to reflect phase variation of pili on the bacterial cell surface . In this study, we further examined the relationships between changes in pilus-dependent colony morphology, pilin sequence, pilus expression, and pilus function in Neisseria gonorrhoeae FA1090 . A group of FA1090 colony variants expressed different pilin sequences and demonstrated different levels of pilin, S-pilin, and pilus expression . The analysis of these colony variants shows that they do not represent two distinct phases of pilus expression, but that changes in pilin protein sequence produce a spectrum of S-pilin production, pilus expression, and pilus aggregation levels . These different levels of pilus expression and aggregation influence not only colony morphology but also DNA transformation efficiency and epithelial cell adherence. J Immunol, 1998 Feb 1, 160(3), 1346 - 53 Activation of complement by mannose-binding lectin on isogenic mutants of Neisseria meningitidis serogroup B; Jack DL et al.; Mannose-binding lectin (MBL) is a serum protein that has been demonstrated to activate the classical complement pathway and to function directly as an opsonin . Although MBL deficiency is associated with a common opsonic defect and a predisposition to infection, the role of the protein in bacterial infection remains unclear . We have investigated MBL binding to Neisseria meningitidis serogroup B1940 and three isogenic mutants, and the subsequent activation of the two major isoforms of C4 (C4A and C4B) by an associated serine protease, MASP . The mutants lacked expression of the capsular polysaccharide (siaD-), the lipo-oligosaccharide (LOS) outer core that prevented LOS sialylation (cpsD-), or both capsule and LOS outer core (cps-) . Using flow cytometry, it was possible to detect strong MBL binding to the cps- and cpsD- mutants over a wide range of concentrations . In contrast, minimal or no MBL binding was detected on the parent organism, with binding to siaD- only at higher MBL concentrations . C4 was activated and bound by mutants that had previously bound MBL/MASP, but there was no significant difference in the amounts of C4A and C4B bound . When sialic acid residues were removed from the parent organism by neuraminidase treatment, the binding of both MBL and C4 increased significantly . Our results suggest that MBL may bind to and activate complement on these encapsulated organisms, and the major determinants of these effects are the LOS structure and sialylation. J Burn Care Rehabil, 1998 Mar-Apr, 19(2), 119 - 23 Purpura fulminans: a disease best managed in a burn center; Brown DL et al.; Victims of purpura fulminans are overcome by a rapidly progressive and sometimes fatal course involving large amounts of tissue loss and multiple organ system failure . From 1986 to 1995, seven children ranging in age from 10 months to 19 years (mean, 6.2 years) were referred to the Shriners Burns Institute in Cincinnati with purpura fulminans . Neisseria meningitidis was identified as the precipitating pathogen in most of the patients . The mean TBSA full-thickness skin loss was 33% . Fourteen extremities were amputated in the seven patients, including three patients with amputations of all four extremities . Transfer to our institution occurred after a mean delay of 20 days, usually after the demarcation of viable tissue . In one patient, however, fasciotomies obviated multiple impending amputations . Monitoring for elevated compartment pressures, early fasciotomies, and expedient transfer to a burn center for a multidiciplinary approach to care should improve the outcome in patients with purpura fulminans. J Bacteriol, 1998 Apr, 180(8), 2152 - 9 An AT-rich tract containing an integration host factor-binding domain and two UP-like elements enhances transcription from the pilEp1 promoter of Neisseria gonorrhoeae; Fyfe JA et al.; The pilE gene of Neisseria gonorrhoeae is transcribed from a sigma70 promoter (pilEp1) with an AT-rich tract extending 65 nucleotides upstream of the -35 box . Within this region is an integration host factor (IHF)-binding core consensus sequence . We have performed a detailed analysis to determine which upstream sequences are required for efficient transcription from pilEp1 in N . gonorrhoeae . Deletion of sequences upstream of the AT-rich tract had no effect on the level of transcription . However, the IHF-binding core consensus sequence and the AT-rich sequence further upstream were both required for enhanced levels of transcription from this promoter in both N . gonorrhoeae and an Escherichia coli strain producing IHF . In addition, an UP-like element positioned between the -35 box and the IHF-binding site was required for maximal transcription . The AT-rich region upstream of the IHF-binding core consensus sequence can also act as an UP-like element when appropriately repositioned upstream of the -35 box. Int J Antimicrob Agents, 1997 Jan, 9(3), 175 - 9 Antimicrobial resistance in gonococci isolated from patients and from commercial sex workers in Harare, Zimbabwe; Mason PR et al.; The objective is to compare antibiotic resistance amongst gonococci isolated from different patient groups in Harare, Zimbabwe . Antimicrobial susceptibilities of Neisseria . gonorrhoeae were determined by disc sensitivity tests . The MICs for penicillin, kanamycin, ceftriaxone, norfloxacin and ciprofloxacin were determined using E-test strips . There were 147 isolates from symptomatic men, 47 isolates from symptomatic women, 29 isolates from asymptomatic women and 41 isolates from female commercial sex workers . A total of 119 (45%) isolates were PPNG and 23 (16%) non-PPNG isolates had a penicillin MIC > 0.64 mg/l . Over 90% of isolates were resistant to TMP/SMX and 16% were resistant to tetracycline . Resistance was uncommon against kanamycin (6%), erythromycin (2%) or ceftriaxone ( < 1%) . For kanamycin, the MIC90 was 32 mg/l, for ceftriaxone the MIC90 was < 0.032 mg/l for non-PPNG and < 0.064 mg/l for PPNG . For norfloxacin and ciprofloxacin the MIC90 was < 0.064 mg/l for both PPNG and non-PPNG . Isolates from the commercial sex workers showed a significantly increased prevalence of PPNG, of penicillin-tolerant non-PPNG and of tetracycline resistance . Four of the 41 isolates from sex workers showed multiple resistance (to penicillin, TMP/SMX, tetracycline and kanamycin) compared to 1/223 isolates from other groups (OR = 24.0) . Antimicrobial resistance is common amongst gonococci in Harare, especially with isolates from commercial sex workers . In order for STD treatment to be implemented as an effective strategy in HIV control, continued monitoring of resistance patterns is essential. FEBS Lett, 1998 Mar 6, 424(1-2), 84 - 8 Vitronectin-dependent invasion of epithelial cells by Neisseria gonorrhoeae involves alpha(v) integrin receptors; Dehio M et al.; Binding of vitronectin (VN) to Neisseria gonorrhoeae expressing the heparan sulfate proteoglycan (HSPG) specific Opa50 protein was recently shown to trigger bacterial internalization into distinct epithelial cell lines . We have investigated the role of VN-binding integrin receptors and protein kinase C (PKC) in VN-triggered bacterial uptake . Blocking integrin function by RGDS peptides or by antibodies specific to alpha(v)beta5 or alpha(v)beta3 resulted in an abrogation of VN-triggered bacterial internalization . Moreover, inhibitors of PKC were found to block VN-triggered uptake . The essential role of alpha(v) integrins and the presumable involvement of PKC in VN-triggered gonococcal uptake are discussed. J Bacteriol, 1998 Apr, 180(7), 1955 - 8 Frequency of pilin antigenic variation in Neisseria gonorrhoeae; Serkin CD et al.; Variation of the pilus of Neisseria gonorrhoeae occurs by the recombination of silent pilin DNA sequences into the pilin expression locus . We have developed a quantitative, competitive reverse transcription-PCR assay which measures the frequency of pilin antigenic variation independently of changes in gonococcal colony morphology and have determined this frequency within a gonococcal population . We have also studied the frequency of antigenic variation during growth and have concluded that growth does not dramatically influence the frequency of pilin antigenic variation, although a reproducible, twofold increase is observed upon the transition into late log/stationary phase. Genitourin Med, 1997 Oct, 73(5), 378 - 82 Characterisation of Neisseria gonorrhoeae in semen during urethral infection in men; Isbey SF et al.; OBJECTIVE: To determine the number of Neisseria gonorrhoeae organisms in urine and semen in men with gonococcal urethritis, and to compare selected phenotypic characteristics of organisms harvested from the urethra and semen . DESIGN: Samples from two groups of subjects were examined . Patients with symptomatic urethritis receiving treatment at an STD clinic, as well as six subjects with experimental urethritis . Semen and urine specimens were obtained after the urethral exudate was sampled . RESULTS: Using quantitative cultures, we found an average of 6 x 10(6) gonococci in urine or semen of 17 men with symptomatic urethritis seeking treatment at an STD clinic, and 2 x 10(4) gonococci in secretions of six male subjects with early experimental infection . Gonococcal outer membrane opacity (Opa) proteins and lipo-oligosaccharide (LOS) recovered from urine and semen of these subjects were very similar . CONCLUSIONS: Men with symptomatic gonorrhoea excrete a large number of gonococci in semen which is not affected by the duration of symptoms . The similar phenotype of organisms in urine and semen suggests the bacteria come from the same compartment . These data help to explain the efficiency of gonococcal transmission from men to their partners, and identify an appropriate target for a preventative vaccine or immunotherapy designed to reduce the inoculum in infected patients. Genitourin Med, 1997 Oct, 73(5), 362 - 4 Genital Chlamydia trachomatis (serotypes D-K) infection in Jamaican commercial street sex workers; Dowe G et al.; OBJECTIVES: To determine the prevalence of genital Chlamydia trachomatis infections in commercial street sex workers (CSSW) in Jamaica . METHODS: The prevalence of C trachomatis infection was determined in 129 Jamaican CSSW using the direct fluorescent antibody (DFA) method and the isolation techniques which utilise fluorescent and iodine staining of endocervical cytobrush specimens cultured in McCoy cells . The seroprevalence of C trachomatis in the CSSW was also compared with that in blood donors (n = 435), using the microimmunofluorescence (MIF) test . RESULTS: The DFA detected C trachomatis in 16% (21/129) of the specimens . The prevalence as determined by the iodine and fluorescein stained cultures was 24% (31/129) and 25% (33/129) respectively . The overall prevalence of current chlamydial infection detected by the isolation techniques used was 25% (33/129) . As determined by the MIF test, a statistically significantly higher seroprevalence rate of C trachomatis (95%, 61/64) was found in CSSW compared with blood donors (53%, 229/435; OR 22.6; chi 2 = 49.8; p < 0.001) . The prevalence of current infection in CSSW as indicated by the isolation of C trachomatis was not influenced by history of previous pelvic inflammatory disease (PID), sexually transmitted disease, or condom use . N gonorrhoeae (9%) and Candida albicans (7%) were found in comparatively low frequencies, while Trichomonas vaginalis (0%) was not found in specimens from the CSSW . CONCLUSIONS: A high seroprevalence rate and a high rate of current infection with C trachomatis occur in Jamaican CSSW . In order to control the spread and prevent the severe clinical complications and sequelae of C trachomatis infection, the diagnosis and treatment in such high risk groups such as CSSW should be optimisedPIP: The prevalence of Chlamydia trachomatis infection was investigated in 129 commercial sex workers (CSWs) recruited on the streets in Kingston, Jamaica . The direct fluorescent antibody method detected C . trachomatis in endocervical cytobrush specimens from 21 women (16%) . When the specimens were cultured, current chlamydial infection was detected by iodine staining in 31 (24%) and by monoclonal antibodies in 33 (25%) . The microimmunofluorescence test for chlamydial antibodies was performed on clotted blood samples obtained from 64 CSWs and, as controls, 435 blood bank donors . A significantly higher seroprevalence rate was found among CSWs (95%) than blood donors (53%) (p 0.001) . Among CSWs, the most common clinical manifestation of C . trachomatis infection was vaginal discharge . The presence of C . trachomatis infection was not related to previous history of pelvic inflammatory disease, sexually transmitted disease (STD), or condom use . Neisseria gonorrhoeae was isolated from 11 (9%) endocervical swabs . This study confirms the predominance of C . trachomatis among the bacterial causes of STDs in high-risk groups in Jamaica, and suggests a need for screening and treatment to control the spread and prevent the severe clinical sequelae of chlamydial infection . Genitourin Med, 1997 Oct, 73(5), 355 - 61 Surveillance of antibiotic susceptibility of Neisseria gonorrhoeae in the WHO western Pacific region 1992-4 . WHO Western Pacific Region Gonococcal Antimicrobial Surveillance Programme; Loss-of-function mutations in the mtr efflux system of Neisseria gonorrhoeae; Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, USAResistance of Neisseria gonorrhoeae to antimicrobial hydrophobic agents (HAs) has been ascribed to the mtr (multiple transferable resistance) operon . This operon is composed of the mtrR gene, which encodes a transcriptional repressor (MtrR), and a three-gene complex (mtrCDE), which encodes cell envelope proteins (MtrC-MtrD-MtrE) that form an energy-dependent efflux pump . HA-hypersusceptible strains are often isolated from patients, but the genetic basis for such hypersusceptibility was heretofore unknown . The genetic basis of HA hypersusceptibility in laboratory-derived strains BR54 and BR87 was studied to learn if this trait could be linked to mutations in the mtr operon . Mutations in the mtrR gene of these strains that could be phenotypically suppressed by mutations in their mtrC or mtrD genes were identified . Thus, small deletions (4-10 bp) in the mtrC or mtrD genes of strains BR87 and BR54 that would result in the production of truncated efflux pump proteins that serve as a membrane fusion protein (MtrC) or transporter of HAs (MtrD) were found to be responsible for their HA-hypersusceptible property. Microb Pathog, 1998 Feb, 24(2), 89 - 100 Biochemical and immunological properties of lactoferrin binding proteins from Moraxella (Branhamella) catarrhalis; Bonnah RA et al.; The Neisseriaceae can acquire iron (Fe) from lactoferrin (Lf) using host-Lf receptors on the bacterial surface . The binding proteins that are proposed to constitute the receptor have been identified by isolation with immobilized Lf . Using CopB-specific monoclonal antibodies and isogenic CopB mutants, we demonstrate that the 84 kDa protein isolated with immobilized human Lf from Moraxella catarrhalis using low stringency conditions is CopB, an 84 kDa membrane-spanning protein with similarities to other TonB-dependent outer membrane proteins . Affinity isolation of Lf receptors from a variety of M . catarrhalis strains using high stringency conditions revealed a 95 kDa protein migrating slightly faster than LbpA on SDS-PAGE in some strains . Convalescent human antisera from patients infected with M . catarrhalis reacted specifically with this protein, but not LbpA . Proteolysis experiments demonstrated that, unlike LbpA, it was rapidly degraded . The 95 kDa protein, but not LbpA, binds labelled Lf after SDS-PAGE and electroblotting, suggesting the 95 kDa protein is LbpB, the homologue of TbpB . This protein comigrates with LbpA in most strains, which may explain why it had not been previously identified. Int J STD AIDS, 1998 Mar, 9(3), 162 - 3 Liaison between gynaecologists, microbiologists and genitourinary medicine clinics in the management of patients with genital chlamydia and gonococcal infections; Huengsberg M et al.; Inadequate treatment and follow-up of women with genital infection with Chlamydia trachomatis and Neisseria gonorrhoeae can cause long-term morbidity . Inadequate contact tracing can predispose to re-infection . As some women with genital infections present to agencies other than genitourinary medicine (GUM) clinics, improved liaison between these and GUM departments are important in safeguarding proper follow-up and contact tracing. Am J Med, 1998 Jan, 104(1), 28 - 32 Randomized trial of trovafloxacin and ofloxacin for single-dose therapy of gonorrhea . Trovafloxacin Gonorrhea Study Group; Jones RB et al.; PURPOSE: To compare trovafloxacin, a new quinolone antibiotic with enhanced activity against Neisseria gonorrhoeae, with ofloxacin as single-dose oral therapy for uncomplicated gonococcal urethritis or cervicitis . PATIENTS AND METHODS: In this multicenter, double-blind trial, 625 patients (270 men, 355 women) with uncomplicated gonococcal urethritis or cervicitis received one 100-mg tablet of trovafloxacin or two 200-mg capsules of ofloxacin as a single dose under direct supervision . RESULTS: Single-dose oral therapy with trovafloxacin was equivalent both bacteriologically and clinically to ofloxacin . Among evaluable patients, N gonorrhoeae was eradicated in 99% of trovafloxacin recipients and in 98% of ofloxacin recipients . Each treatment was well tolerated; vaginitis was the most frequently observed side effect (4% trovafloxacin, 7% ofloxacin) . CONCLUSION: Based on the results presented here, trovafloxacin is a promising agent for single-dose therapy of uncomplicated gonorrhea. Infect Immun, 1998 Apr, 66(4), 1791 - 4 Binding to human extracellular matrix by Neisseria meningitidis; Eberhard T et al.; Adhesion of Neisseria meningitidis strains to extracellular matrix (ECM) and purified matrix components was examined . Most strains bound to subendothelial ECM as well as to immobilized fibronectin and types I, III, and V collagen . Strains from healthy carriers adhered significantly better than isolates from patients . The binding site was localized to the central 75-kDa cell-binding domain of the fibronectin molecule . This domain has not been described previously to interact with bacterial structures. Infect Immun, 1998 Apr, 66(4), 1638 - 47 Nuclear translocation of NF-kappaB in lipopolysaccharide-treated macrophages fails to correspond to endotoxicity: evidence suggesting a requirement for a gamma interferon-like signal; Denlinger LC et al.; Elucidation of a signal transduction pathway essential to lipopolysaccharide (LPS)-induced macrophage activation has the capacity to provide new targets for the treatment of septic shock . In this regard, activation of the transcription factor NF-kappaB is commonly thought to be critical to LPS-stimulated macrophage inflammatory mediator production, although certain immunological, genetic, and molecular evidence suggests that other factors are involved . To address this issue, we hypothesized that the degree of LPS-induced NF-kappaB mobilization should correlate with the murine endotoxicity of the species of LPS used for in vitro study . Therefore, using D-galactosamine-sensitized mice, we assessed the lethal potencies of eight LPS preparations from Escherichia, Salmonella, Klebsiella, Bacteroides, Pseudomonas, Neisseria, and Rhodobacter species as well as that of the endotoxin substructure lipid X . The lethal potencies of these LPS preparations varied by > 160-fold . Treatment of RAW 264.7 cells with the same LPS preparations induced levels of tumor necrosis factor alpha (TNF-alpha) and NO production that correlated with the LPS 50% lethal dose . The combined analysis of the levels of these two mediators produced in response to LPS in RAW cells was found to be a strong predictor of murine endotoxic lethality . Interestingly, while relatively nontoxic in mice, Rhodobacter capsulatus LPS stimulated RAW cell NF-kappaB-like DNA binding protein mobilization and TNF-alpha production to levels comparable to those of more toxic species of LPS but was unable to induce NO generation in RAW cells . These data indicate that neither NF-kappaB activation nor TNF-alpha production alone is a dependable predictor of LPS lethality . Additionally, cotreatment of RAW cells with the potent inflammatory mediator ADP had no effect on the ability of R . capsulatus LPS to stimulate NO production but significantly enhanced induction of NO production by the toxic species of LPS . In contrast, cotreatment of RAW cells or peritoneal macrophages with gamma interferon (IFN-gamma) normalized the abilities of both toxic and nontoxic LPS preparations to induce NO production, suggesting that selected preparations of LPS may preferentially generate an IFN-gamma-like signal that accounts for enhanced toxicity . In sum, the activation of NF-kappaB does not correspond to LPS lethality, thereby complicating models of macrophage activation that highlight NF-kappaB alone as a signal transduction factor necessary for LPS-mediated toxicity. Pediatrics . 1997 Jun;99(6):E8. Vaginal gonococcal cultures in sexual abuse evaluations: evaluation of selective criteria for preteenaged girls; Ingram DL et al.; OBJECTIVE: Accurate selective criteria could limit the number of vaginal cultures for Neisseria gonorrhoeae performed on preteenaged girls as part of their sexual abuse evaluations . This study was performed to determine whether the published selective criteria by the American Academy of Pediatrics (AAP) Committee on Child Abuse and Neglect and by Siegel et al would have accurately detected all cases of vaginal gonococcal infections in our large study population . METHODS: We prospectively studied girls, ages 1 to 12 years, who were referred to our Child Sexual Abuse Team (CSAT) at Wake Medical Center in Raleigh, NC, between July 1, 1976 to July 1, 1996, for sexual abuse evaluations which were performed using a protocol that included collecting historical information, a sexual abuse interview, and a detailed genital examination which included a vaginal culture for N gonorrhoeae . RESULTS: Our study population consisted of 2898 girls of whom 2731 (94%) had vaginal cultures successfully performed for N gonorrhoeae . There were 84 girls with vaginal gonococcal infections, 80 of whom had a vaginal discharge . The four girls without a vaginal discharge included two with a history of having vaginal intercourse with an alleged perpetrator with gonorrhea, one with N gonorrhoeae isolated from a urine culture, and one whose preteenaged sister had gonorrhea . All of the 84 girls would have been identified using the selective culturing criteria of the AAP Committee on Child Abuse and Neglect: culturing when epidemiologically indicated (interpreted as the girl having another sexually transmitted disease {STD}, a child sibling, child household member, a close child associate or a perpetrator with a known STD) or when the history and/or physical findings suggest the possibility of oral, genital, or rectal contact, or Siegel et al's more selective criteria: only culturing prepubertal girls for N gonorrhoeae if there is a vaginal discharge at the time of presentation or if there is a high risk for STD acquisition, defined as having a STD diagnosed, a sibling with a STD, contact with a perpetrator known to have a STD, contact with multiple perpetrators, or Tanner stage III or above . CONCLUSION: Both the selective criteria of the AAP Committee on Child Abuse and Neglect and the more selective criteria of Siegel et al as we interpreted them were accurate when applied to identifying girls with vaginal gonococcal infections in our study population. Ann Acad Med Singapore, 1997 Nov, 26(6), 827 - 33 Sexually-transmitted diseases in Singapore--trends in the last two decades; Ang P et al.; We review the epidemiology and trends of sexually transmitted diseases (STDs) in Singapore from 1977 to 1996 . There has been a progressive decline in the incidence of bacterial STDs over the period of observation . However, viral STDs have not shown the same trends, remaining at a relatively constant level . Early detection and treatment of STDs, and health education and prevention measures targeting sex workers have been a major factor in controlling STDs . Vigilant monitoring of antibiotic resistance patterns in Neisseria gonorrhoeae has allowed timely changes to recommended treatment regimens . The advent of the human immunodeficiency virus (HIV) infection pandemic in the 1980s and public education campaigns have contributed to the success of the STDs control programme . In view of the incurable nature of many viral STDs, prevention and counselling have become a cornerstone of STD control . Surveillance and monitoring of STD trends will continue to provide important information regarding the effectiveness of existing health programmes and help us formulate new plans for the future. Enferm Infecc Microbiol Clin, 1997 Dec, 15(10), 515 - 8 {Usefulness of the disk diffusion method for evaluating the sensitivity of Neisseria meningitidis to penicillin and cefotaxime}; Joyanes P et al.; BACKGROUND: Routine susceptibility testing of Neisseria meningitidis to penicillin and other beta lactams is recommended after the isolation of N . meningitidis of moderately resistant to penicillin (MRP) . We have evaluated the disk-diffusion method to determine susceptibility of N . meningitidis to penicillin (using disks of either penicillin or oxacillin) and to cefotaxime . METHODS: Fifty-four strains of N . meningitidis isolated from clinical samples were studied . MICs of penicillin and cefotaxime were determined by microdilution . Disks of 2 U of penicillin, 1 microgram of oxacillin and 30 micrograms of cefotaxime and two culture media, Mueller-Hinton agar (MHA) and MHA supplemented with 5% sheep blood (MHS) were used in the disk-diffusion assay . RESULTS: For disk of 2 U of penicillin assayed in MHA, 86.4% of the susceptible strains and 20% of MRP strains were considered susceptible when a breakpoint of 28 mm was considered . None of the MRP strains was considered susceptible when using MHS, but only 38.6% of susceptible strains appeared as such on this medium . When a 1 microgram oxacillin disk was used all MRP strains presented an inhibition zone < or = 10 mm on both MHA and MHS, but 54.4 and 4.5% of susceptible strains presented an inhibition zone > or = 11 mm on MHA and MHS, respectively . All strains were susceptible to cefotaxime, showing inhibition zones around a 30 micrograms disk on MHA and MHS of > or = 35 mm and > or = 25 mm, respectively . CONCLUSION: Disk diffusion with cefotaxime (30 micrograms) allows to determine susceptibility of N . meningitidis to this antimicrobial agent . Discs of penicillin (2 U) and oxacillin (1 microgram) are not useful for screening of MRP N . meningitidis. Can J Microbiol, 1998 Jan, 44(1), 56 - 63 Sequencing of porA from clinical isolates of Neisseria meningitidis defines a subtyping scheme and its genetic regulation; Arhin FF et al.; Subtyping Neisseria meningitidis by methods that rely on monoclonal antibody (mAb) reactivity results in an unusually high number of strains that are not subtypeable . To subtype 48 strains isolated (1993-1994) in the province of Quebec that were not subtypeable by mAb-based techniques, we used DNA sequencing of the variable regions of porA, a gene that encodes the class 1 outer membrane protein . We assigned subtypes to all the previously nonserosubtypeable isolates and identified some novel subtypes . Because our sequencing strategy included the promoter region of porA, different isolates were compared in their sequences of the porA promoter region . A poly(G) stretch lies between the -10 and -35 regions of the promoter; replacement of a G residue by an A residue in this region resulted in loss of expression of porA . No correlation was found between the number of G residues in the poly(G) stretch and the level of expression; a minimum of 10 G residues is required in this stretch for expression of porA . One isolate expressed no class 1 outer membrane protein because of the insertion sequence IS1301 in the coding region of porA . Another isolate did not express the protein owing to a frame-shift mutation within the coding region of porA . Sequencing of porA allowed assignments of subtypes to previously uncharacterized isolates and provided insights about the regulation of expression of this gene in N . meningitidis. Sex Transm Dis, 1998 Feb, 25(2), 70 - 5 The relationship of cocaine use and human immunodeficiency virus serostatus to incident sexually transmitted diseases among women; Wilson TE et al.; BACKGROUND AND OBJECTIVES: To assess the incidence of sexually transmitted diseases (STD) in a group of heterosexual women as a function of human immunodeficiency virus (HIV) serostatus and to ascertain the effect of crack cocaine use on these relationships . STUDY DESIGN: At baseline, 445 HIV type 1 (HIV-1) seronegative and 232 seropositive women were provided interviews ascertaining demographic and behavioral risk factors . All participants were tested for Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis at baseline and at 6-month intervals . RESULTS: HIV serostatus was not related to STD incidence . Although HIV-positive women reported more condom use than did HIV-negative women (P < .01), only 65% reported using them consistently . Increases in the frequency of crack cocaine use, measured on a 4-point scale, were positively associated with rates of new STDs (relative risk {RR} = 1.23, P < .01) . Crack cocaine use was also associated with greater numbers of sexual partners and less consistent condom use . The relationship between HIV status and the probability of acquiring an STD was not influenced by frequency of crack use . CONCLUSION: Women infected with HIV or who use crack cocaine are at risk for transmitting HIV and acquiring other STDs . Whether women are infected with or at risk for HIV, programs are needed to prevent and treat crack addiction . Interventions should target high-risk sexual practices among both female crack users and women living with HIV. Int J STD AIDS, 1998 Jan, 9(1), 21 - 4 Detection of male genital infection with Chlamydia trachomatis and Neisseria gonorrhoeae using an automated multiplex PCR system (Cobas Amplicor); Higgins SP et al.; We evaluated Cobas Amplicor, a highly automated polymerase chain reaction (PCR) system, to test first-void urine (FVU) and urethral swab specimens for Chlamydia trachomatis and Neisseria gonorrhoeae in men attending a sexually transmitted infection (STI) clinic . Results were compared against an in-house radioimmune dot blot (DB) test for C . trachomatis and selective culture for N . gonorrhoeae . Three hundred and ninety sets of specimens were obtained from 378 consecutive new and returned-new patients . Gonorrhoea prevalence was 9.49%, with no significant difference in sensitivity or specificity between culture and PCR . Chlamydia prevalence was 15.4%, with sensitivities of: DB 55%, PCR of FVU 86.7%, urethral swab PCR 90% . The specificity of PCR on FVU and urethral swabs was 100% . We have shown that Cobas Amplicor PCR is highly sensitive and specific in the diagnosis of chlamydia and gonorrhoea in men attending an STI clinic . Further economic and scientific studies are needed to determine the cost-effectiveness of this technique for screening in primary care settings. Antimicrob Agents Chemother, 1998 Mar, 42(3), 579 - 82 Reduced clinical efficacy of pazufloxacin against gonorrhea due to high prevalence of quinolone-resistant isolates with the GyrA mutation . The Pazufloxacin STD Group; Tanaka M et al.; Forty-two men with gonococcal urethritis were treated with an oral dosage of 200 mg of pazufloxacin, a new fluoroquinolone, three times daily for 3 days . Only 28 of the 42 men (66.7%) had negative culture results for Neisseria gonorrhoeae during follow-up . Of the 42 isolates, 41 could be recultured for antibiotic susceptibility testing and DNA sequencing . In 26 of the 41 isolates (63.4%), GyrA mutations with or without ParC mutations were identified . Among the 26 isolates, 23 contained a single GyrA mutation, 1 contained two GyrA mutations, and 2 contained three mutations including double GyrA and single ParC mutations . A single Ser-91-to-Phe mutation, which was detected in 14 of the 26 isolates, was the most common GyrA mutation, followed by an Ala-75 to Ser mutation and an Asp-95 to Asn or Gly mutation in GyrA . All three isolates with two or three mutations contained the Ser-91-to-Phe GyrA mutation . Eleven of the 14 isolates with the single Ser-91-to-Phe mutation within GyrA and all 3 isolates with two or three mutations persisted after pazufloxacin treatment . On the other hand, all 15 wild-type and 9 mutant isolates with a substitution at codon Ala-75 or Asp-95 were eradicated . The mean MIC of pazufloxacin for mutants with the single Ser-91-to-Phe mutation in GyrA was 66-fold higher than that for the wild type . The results obtained in this study suggest that a high prevalence of fluoroquinolone-resistant gonococcal isolates with the Ser-91-to-Phe mutation in GyrA reduced the efficacy of pazufloxacin as treatment for gonococcal urethritis. J Exp Med, 1998 Mar 2, 187(5), 743 - 52 A novel sialic acid binding site on factor H mediates serum resistance of sialylated Neisseria gonorrhoeae; Ram S et al.; Factor H (fH), a key alternative complement pathway regulator, is a cofactor for factor I-mediated cleavage of C3b . fH consists of 20 short consensus repeat (SCR) domains . Sialic acid binding domains have previously been localized to fH SCRs 6-10 and 13 . To examine fH binding on a sialylated microbial surface, we grew Neisseria gonorrhoeae in the presence of 5'-cytidinemonophospho-N-acetylneuraminic acid, which sialylates lipooligosaccharide and converts to serum resistance gonococci previously sensitive to nonimmune serum killing . fH domains necessary for binding sialylated gonococci were determined by incubating organisms with recombinant human fH (rH) and nine mutant rH molecules (deletions spanning the entire fH molecule) . rH and all mutant rH molecules that contained SCRs 16-20 bound to the sialylated strain; no mutant molecule bound to serum-sensitive nonsialylated organisms . Sialic acid was demonstrated to be the fH target by flow cytometry that showed a fourfold increase in fH binding that was reversed by neuraminidase-mediated cleavage of sialic acid off gonococci . Functional specificity of fH was confirmed by decreased total C3 binding and almost complete conversion to iC3b on sialylated gonococci . Sialic acid can therefore bind fH uniquely through SCRs 16-20 . This blocks complement pathway activation for N . gonorrhoeae at the level of C3. Gene, 1998 Feb 16, 208(1), 95 - 102 The pathogenic neisseriae contain an inactive rpoN gene and do not utilize the pilE sigma54 promoter; Laskos L et al.; The sigma54 promoter (P3) upstream of the pilE gene in Neisseria gonorrhoeae was shown to be non-functional by transcriptional analysis of a PpilE::lacZ fusion containing only P3 . A region on the chromosome of N . gonorrhoeae strain MS11-A was identified that potentially encodes a protein with a significant similarity to the Escherichia coli RpoN protein . However, this region (designated RLS for rpoN-like sequence) does not contain a single open reading frame (ORF) capable of encoding a functional RpoN protein . It appears that RLS may have arisen from an ancestral rpoN homologue that underwent a deletion removing the sequence encoding the essential helix-turn-helix (HTH) motif, and changing the subsequent reading frame . An RLS has been identified in several strains of N . gonorrhoeae and N . meningitidis . A 90-kDa gonococcal protein has previously been shown to react with a monoclonal antibody raised against the RpoN from Salmonella typhimurium . However, mutagenesis and Western blot analysis confirmed that the gene encoding this protein is not contained within RLS. Gene, 1998 Feb 16, 208(1), 61 - 6 Absence of an SOS-like system in Neisseria gonorrhoeae; Black CG et al.; The DNA repair capacities of Neisseria gonorrhoeae have not been well characterised, however, it is known that the gonococcus possesses an excision repair system . The fact that genes in this system are part of the SOS regulon in Escherichia coli prompted this investigation into the transcriptional regulation of genes involved in DNA repair in N . gonorrhoeae . Northern (RNA-DNA) dot blot hybridisation was used to investigate potential DNA damage-mediated induction of the gonococcal recA, uvrA and uvrB genes . In contrast to the situation in E . coli, transcription of these genes in N . gonorrhoeae was not induced in response to treatment with methyl methanesulfonate (MMS) and UV light . These data indicated that the gonococcus does not possess an SOS-like system that is induced in response to DNA damage. Gene, 1998 Feb 16, 208(1), 51 - 9 Characterization of genetic exchanges between various highly divergent tbpBs, having occurred in Neisseria meningitidis; Legrain M et al.; Transferrin-binding protein B (TbpB) from Neisseria is an outer membrane-associated extracellular protein involved in iron capture during bacterial infection . The tbpB genes display extensive divergences throughout the open reading frame (ORF) that have presumably been selected under the pressure of the immune system . Early studies suggested that they could possibly constitute two distantly related groups of genes (sharing less than 57% identical nt) . However, the analysis of one tbpB suggested the existence of a greater genetic diversity, and the occurrence of horizontal genetic exchanges leading to rearrangements of highly divergent ORFs . This study has confirmed this and revealed the occurrence of genetic exchanges having involved at least three types of very distantly related tbpBs . These rearrangements resulted from recombination events having occurred at very similar positions within an ORF region encoding a highly structured protein domain, probably due to constraints imposed by protein function and mode(s) of folding . These new data also provide valuable tools for epidemiological studies and evaluation of TbpBs as candidate vaccines. Mol Microbiol, 1998 Feb, 27(4), 763 - 75 A second prepilin peptidase gene in Escherichia coli K-12; Francetic O et al.; Escherichia coli K-12 strains grown at 37 degrees C or 42 degrees C, but not at 30 degrees C, process the precursors of the Neisseria gonorrhoeae type IV pilin PilE and the Klebsiella oxytoca type IV pseudopilin PulG in a manner reminiscent of the prepilin peptidase-dependent processing of these proteins that occurs in these bacteria . Processing of prePulG in Escherichia coli requires a glycine at position -1, as does processing by the cognate prepilin peptidase (PulO), and is unaffected by mutations that inactivate several non-specific proteases . These data suggested that E . coli K-12 has a functional prepilin peptidase, despite the fact that it does not itself appear to express either type IV pilin or pseudopilin genes under the conditions that allow prePilE and prePulG processing . The E . coli K-12 genome contains two genes encoding proteins with significant sequence similarity to prepilin peptidases: gspO at minute 74.5 and pppA (f310c) at minute 67 on the genetic map . We have previously obtained evidence that gspO encodes an active enzyme but is not transcribed . pppA was cloned and shown to code for a functional prepilin peptidase capable of processing typical prepilin peptidase substrates . Inactivation of pppA eliminated the endogenous, thermoinducible prepilin peptidase activity . PppA was able to replace PulO prepilin peptidase in a pullulanase secretion system reconstituted in E . coli when expressed from high-copy-number plasmids but not when present in a single chromosomal copy . The analysis of pppA-lacZ fusions indicated that pppA expression was very low and regulated by the growth temperature at the level of translation, in agreement with the observed temperature dependence of PppA activity . Polymerase chain reaction and Southern hybridization analyses revealed the presence of the pppA gene in 12 out of 15 E . coli isolates. Gac Med Mex, 1997, 133 Suppl 1, 133 - 7 Molecular diagnostics . The polymerase chain reaction and its use in the diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae; Uhrin M; The diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae was performed using classical microbiological techniques of chlamydial cell culture and agar isolation respectively in patients studied in Pittsburgh . The polymerase chain reaction (PCR) was compared to standard procedures used for diagnosing these sexually transmitted organisms . Statistically significant differences were observed in molecular diagnostics versus classical isolation techniques . Numerous specimen handling problems were identified in the handling of Neisseria . The enhanced ability to diagnose these sexually transmitted organisms is discussed in relation to the Human Immunodeficiency Virus (HIV). Proc Natl Acad Sci U S A, 1998 Feb 17, 95(4), 1829 - 33 Modulation of Neisseria gonorrhoeae susceptibility to vertebrate antibacterial peptides due to a member of the resistance/nodulation/division efflux pump family; Shafer WM et al.; We have previously described the antibacterial capacity of protegrin-1 (PG-1), a cysteine-rich, cationic peptide from porcine leukocytes, against Neisseria gonorrhoeae . We now report genetic and biochemical evidence that gonococcal susceptibility to the lethal action of PG-1 and other structurally unrelated antibacterial peptides, including a peptide (LL-37) that is expressed constitutively by human granulocytes and testis and inducibly by keratinocytes, is modulated by an energy-dependent efflux system termed mtr . These results indicate that such efflux systems may enable mucosal pathogens like gonococci to resist endogenous antimicrobial peptides that are thought to act during infection. J Bacteriol, 1998 Mar, 180(5), 1323 - 30 Sequence diversity, predicted two-dimensional protein structure, and epitope mapping of neisserial Opa proteins; Malorny B et al.; The sequence diversity of 45 Opa outer membrane proteins from Neisseria meningitidis, Neisseria gonorrhoeae, Neisseria sicca, and Neisseria flava indicates that horizontal genetic exchange of opa alleles has been rare between these species . A two-dimensional structural model containing four surface-exposed loops was constructed based on rules derived from porin crystal structure and on conservation of sequence homology within transmembrane beta-strands . The minimal continuous epitopes recognized by 23 monoclonal antibodies were mapped to loops 2 and 3 . Some of these epitopes are localized on the bacterial cell surface, in support of the model. West Indian Med J, 1997 Dec, 46(4), 107 - 10 Antibiotic susceptibility of Neisseria gonorrhoeae in Trinidad and Tobago; Swanston WH et al.; Treatment failures with standard doses of penicillin have been observed in the Sexually Transmitted Diseases (STD) clinics in Trinidad and Tobago . In the absence of an ongoing surveillance system, the antimicrobial susceptibility of 518 Neisseria gonorrhoeae strains was determined in order to guide treatment . 39 (7.6%) strains were resistant to penicillin, including 27 (5.2%) positive for beta-lactamase; that is penicillinase-producing Neisseria gonorrhoeae (PPNG) . 51 (10%) strains were resistant to tetracycline, with 26 (5.0%) of these exhibiting high levels of resistance compatible with tetracycline resistant Neisseria gonorrhoeae (TRNG) . Six strains showed evidence of having both PPNG and TRNG plasmids, and five strains showed chromosomally-mediated resistance to both penicillin and tetracycline . The overall resistance rate to penicillin and tetracycline was 17.7% . There was no resistance to spectinomycin, cefuroxime, ceftriaxone and norfloxacin . The resistance rates demonstrated in this study are sufficiently significant to preclude the use of penicillin and tetracycline in the STD clinics and to justify the use of newer antimicrobials . It is essential that resistance patterns be monitored by continued surveillancePIP: Neisseria gonorrhoeae from many areas of the world have developed resistance to a wide variety of antibiotics over the past 2 decades . The incidence of treatment failures with standard doses of penicillin observed in Trinidad and Tobago's sexually transmitted disease (STD) clinics suggests that such resistance also exists in Trinidad . 518 Neisseria gonorrhoeae strains were isolated from 1502 male and female patients attending the 7 STD clinics throughout Trinidad during May-October 1992, and tested to identify their antimicrobial susceptibility . 39 strains were resistant to penicillin, including 27 which were positive for beta-lactamase, indicating infection with penicillinase-producing Neisseria gonorrhoeae (PPNG) . 51 strains were resistant to tetracycline, of which 26 exhibited high levels of resistance compatible with tetracycline-resistant Neisseria gonorrhoeae (TRNG) . 6 strains showed evidence of having both PPNG and TRNG plasmids, and 5 strains showed chromosomally-mediated resistance to both penicillin and tetracycline . The overall resistance rate to penicillin and tetracycline was 17.7%, and there was resistance to neither spectinomycin, cefuroxime, ceftriaxone, nor norfloxacin . The resistance rates observed in this study are significant enough to warrant the cessation of penicillin and tetracycline use in STD clinics, and the use of newer antimicrobials . Neisseria gonorrhoeae resistance patterns demand ongoing monitoring . East Afr Med J, 1997 Jul, 74(7), 450 - 5 Bacterial pathogens and their antimicrobial susceptibility in Kumasi, Ghana; Ohene A; Between January, 1994 and June, 1996 a survey of bacterial isolates from clinical specimens and their antimicrobial susceptibility was performed at the Komfo Anokye Teaching Hospital, Microbiology Department, Kumasi, Ghana . A total of 11,380 bacterial isolates were cultured from eight different specimens . The sites of origin were wounds 32.2%, urine 28.1%, ear, nose and throat 3.6%, sputum 2.5% and aspirates 2.5% . Gram-negative bacteria accounted for 7955 (69.9%) isolates, the main species were Escherichia coli 47.1%, Pseudomonas spp . 16.8%, Proteus spp 14.6%, Klebsiella spp 10.2%, Neisseria gonorrhoeae 4.2%, Gram-positive bacteria contributed 3425 ((30.1%) of isolates, with Staphylococcus aureus 54.6% being the most predominant followed by Coagulase negative Staphylococcus 18.1%, Streptococcus pneumoniae 13.7% and Beta-haemolytic streptococci 4.1% . Escherichia coli showed 88% and 82% resistance to ampicillin and cotrimoxazole respectively with 78% being susceptible to gentamicin . Cefuroxime resistance in Gram-negative bacilli was 5% . As much as 30.6% and 21.7% of Streptococcus pneumoniae isolates were resistant to Penicillin and chloramphenicol respectively . Ten per cent of Staphylococcus aureus strains were susceptible to penicillin and 18% were resistant to flucloxacillin. East Afr Med J, 1997 Jul, 74(7), 447 - 9 Drug susceptibility of Neisseria isolates from patients attending clinics for sexually transmitted diseases in Addis Ababa; Meless H et al.; Sixty eight Neisseria gonorrhoeae strains were isolated from endocervical and urethral discharge of 233 patients attending health centres for sexually transmitted diseases (STDs) in Addis Ababa, were identified following conventional procedures and tested for susceptibility to penicillin, ampicillin, trimethoprim-sulphamethoxazole (bactrim), chloramphenicol, erythromycin and kanamycin by the agar disc diffusion technique . Penicillinase producing N . gonorrhoeae (PPNG) were identified using the chromogenic cephalosporin method and comprised 70% of the isolates . Seventy seven per cent, 73%, 64% and 17% of the isolates were found to be resistant to penicillin, ampicillin, bactrim and kanamycin, respectively . However, no resistance to erythromycin and chloramphenicol was observed . Multiple drug resistance was found to be 67% . This is a cause for concern in the control and treatment of gonococci. Am Fam Physician, 1998 Feb 15, 57(4), 735 - 46 Conjunctivitis; Morrow GL et al.; Conjunctivitis refers to any inflammatory condition of the membrane that lines the eyelids and covers the exposed surface of the sclera . It is the most common cause of "red eye" . The etiology can usually be determined by a careful history and an ocular examination, but culture is occasionally necessary to establish the diagnosis or to guide therapy . Conjunctivitis is commonly caused by bacteria and viruses . Neisseria infection should be suspected when severe, bilateral, purulent conjunctivitis is present in a sexually active adult or in a neonate three to five days postpartum . Conjunctivitis caused by Chlamydia trachomatis or Neisseria gonorrhoeae requires aggressive antibiotic therapy, but conjunctivitis due to other bacteria is usually self-limited . Chronic conjunctivitis is usually associated with blepharitis, recurrent styes or meibomianitis . Treatment requires good eyelid hygiene and the application of topical antibiotics as determined by culture . Allergic conjunctivitis is distinguished by severe itching and allergen exposure . This condition is generally treated with topical antihistamines, mast-cell stabilizers or anti-inflammatory agents. Bioorg Khim, 1997 Aug, 23(8), 635 - 41 {Enzyme linked immunosorbent assay for characterization of affinity and epitope specificity of antibodies to a polysaccharide antigen}; Kozlov LV et al.; The use of the Langmuir equation for processing ELISA data (the sandwich variant) helped to ascribe a physical sense to the parameters of the optimization of the antigen-antibody titration curve: the maximum response that characterizes complete binding corresponds to the saturation of all epitopes of the antigen, and the concentration at which half of the maximum response is attained corresponds to the dissociation constant of the immune complex, i.e., to the average affinity of the antibodies . The algorithm was tested for systems in which antibodies against IgE and IgD were sorbed on a support, and the antigen bound was determined by the antibodies conjugated with peroxidase . A good fit of the experimental and theoretical curves and reasonable values for the affinity constants were found . In another system, the binding of specific IgG, IgM, and IgA antibodies with the polysaccharide from Neisseria meningitidis serotype A was studied during vaccine testing . The structural simplicity of the antigen molecule made it possible to suggest the presence of two main epitopes and to reveal the dynamics of formation of the antibodies to them. Mol Microbiol, 1998 Feb, 27(3), 611 - 6 The transferrin receptor expressed by gonococcal strain FA1090 is required for the experimental infection of human male volunteers; Cornelissen CN et al.; Iron, an essential nutrient for most microorganisms, is sequestered by the host to decrease the concentration of iron available to bacterial pathogens . Neisseria gonorrhoeae, the causative agent of gonorrhoea, can acquire iron by direct interaction with human iron-binding proteins, including the serum glycoprotein, transferrin . Iron internalization from host transferrin requires the expression of a bacterial receptor, which specifically recognizes the human form of transferrin . Two gonococcal transferrin-binding proteins have been implicated in transferrin receptor function, TbpA and TbpB . We constructed a gonococcal transferrin receptor mutant without the introduction of additional antibiotic resistance markers and tested its ability to cause experimental urethritis in human male volunteers . The transferrin receptor mutant was incapable of initiating urethritis, although the same inoculum size of the wild-type parent strain, FA1090, causes urethritis in >90% of inoculated volunteers . To our knowledge, this is the first experimental demonstration that a bacterial iron acquisition system is an essential virulence factor for human infection. Mol Microbiol, 1998 Feb, 27(3), 599 - 610 Molecular characterization of LbpB, the second lactoferrin-binding protein of Neisseria meningitidis; Pettersson A et al.; The lbpA gene of Neisseria meningitidis encodes an outer membrane lactoferrin-binding protein and shows homology to the transferrin-binding protein, TbpA . Previously, we have detected part of an open reading frame upstream of lbpA . The putative product of this open reading frame, tentatively designated lbpB, showed homology to the transferrin-binding protein TbpB, suggesting that the lactoferrrin receptor, like the transferrin receptor, consists of two proteins . The complete nucleotide sequence of lbpB was determined . The gene encodes a 77.5 kDa protein, probably a lipoprotein, with homology, 33% identity to the TbpB of N . meningitidis . A unique feature of LbpB is the presence of two stretches of negatively charged residues, which might be involved in lactoferrin binding . Antisera were raised against synthetic peptides corresponding to the C-terminal part of the putative protein and used to demonstrate that the gene is indeed expressed . Consistent with the presence of a putative Fur binding site upstream of the lbpB gene, expression of both LbpA and LbpB was proved to be iron regulated in Western blot experiments . The LbpB protein appeared to be less stable than TbpB in SDS-containing sample buffer . Isogenic mutants lacking either LbpA or LbpB exhibited a reduced ability to bind lactoferrin . In contrast to the lbpB mutant, the lbpA mutant was completely unable to use lactoferrin as a sole source of iron. Infect Immun, 1998 Mar, 66(3), 987 - 93 Phase variation of hemoglobin utilization in Neisseria gonorrhoeae; Chen CJ et al.; Most Neisseria gonorrhoeae isolates are unable to use human hemoglobin as the sole source of iron for growth (Hgb-), but a minor population is able to do so (Hgb+) . This minor population grows luxuriously on hemoglobin, expresses two outer membrane proteins of 42 kDa (HpuA) and 89 kDa (HpuB), and binds hemoglobin under iron-stressed conditions . In addition to the previously reported HpuB, we identified and characterized HpuA, which is encoded by the gene hpuA, located immediately upstream of hpuB . Expression of both proteins was found to be controlled at the translational level by frameshift mutations in a run of guanine residues within the hpuA sequence encoding the mature HpuA protein . The "on-phase" hemoglobin-utilizing variants contained 10 G's, while the "off-phase" variants contained 9 G's . Insertional hpuB mutants of FA19 Hgb+ and FA1090 Hgb+ no longer expressed HpuB but still produced HpuA . A polar insertional mutation of the upstream hpuA gene in FA1090 Hgb+ eliminated production of both HpuA and HpuB, whereas a nonpolar insertional mutant expressed HpuB only . Insertional mutagenesis of either hpuA or hpuB or both substantially decreased the hemoglobin binding ability of the FA1090 Hgb+ variant and prevented growth on hemoglobin plates . Therefore, both HpuA and HpuB were required for the utilization of hemoglobin for growth. EMBO J, 1998 Jan 15, 17(2), 443 - 54 CD66-mediated phagocytosis of Opa52 Neisseria gonorrhoeae requires a Src-like tyrosine kinase- and Rac1-dependent signalling pathway; Hauck CR et al.; The interaction of Neisseria gonorrhoeae with human phagocytes is a hallmark of gonococcal infections . Recently, CD66 molecules have been characterized as receptors for Opa52-expressing gonococci on human neutrophils . Here we show that Opa52-expressing gonococci or Escherichia coli or F(ab) fragments directed against CD66, respectively, activate a signalling cascade from CD66 via Src-like protein tyrosine kinases, Rac1 and PAK to Jun-N-terminal kinase . The induced signal is distinct from Fcgamma-receptor-mediated signalling and is specific for Opa52, since piliated Opa- gonococci, commensal Neisseria cinerea or E.coli do not stimulate this signalling pathway . Inhibition of Src-like kinases or Rac1 prevents the uptake of Opa52 bacteria, demonstrating the crucial role of this signalling cascade for the opsonin-independent, Opa52/CD66-mediated phagocytosis of pathogenic Neisseria. Immunopharmacology, 1997 Dec, 38(1-2), 215 - 21 Molecular, genetic, and functional analysis of homozygous C8 beta-chain deficiency in two siblings; Kotnik V et al.; C8 deficiency is associated with an increased susceptibility to neisserial infections . We present a case of an 11 year old boy who suffered from infection with Neisseria meningitidis . Medical history of the patient and his family (n = 5) did not indicate any previous immunodeficiency symptoms . Results from the analysis of phagocyte and lymphocyte functions were within the normal range . No hemolytic activities of the classical (CH50) and the alternative (APH50) pathways of complement were measurable, and SC5b-9 protein complexes could not be detected in the patient's plasma . Further analysis by highly sensitive ELISA and functional assays revealed a complete deficiency of C8 . Upon the reconstitution with purified C8 total hemolytic activity could be restored . SDS-PAGE and Western blot analysis established a deficiency of the C8 beta chain . Genetic analysis at the genomic DNA level demonstrated the common C-T mutation in exon 9 of the C8B gene . Family analysis presented the older sister with non-detectable function of C8 in serum, both parents with about half-normal C8 titres, and the younger sister with normal C8 function . The parents and both sisters were asymptomatic, although the older of the sisters presented with the same complete C8 beta-chain deficiency as the patient described . In conclusion: the common C-T mutation in the C8B genes is the genetic basis of C8 beta-chain deficiency in two members of this Bosnian family. CMAJ, 1998 Jan 13, 158(1), 41 - 6 Chlamydial cervicitis: testing the practice guidelines for presumptive diagnosis; Sellors J et al.; OBJECTIVE: To test the recommendation from the Canadian guidelines for sexually transmitted diseases (STDs) that mucopurulent endocervical discharge and 10 or more polymorphonuclear leukocytes (PMNs) per high-power field of a Gram-stained endocervical smear or, when Gram staining is not possible, the presence of endocervical discharge and one of edema, erythema or induced mucosal bleeding of the cervix can be considered diagnostic for chlamydial cervicitis . METHODS: A total of 596 consecutive women attending 2 family planning clinics for routine care underwent vaginal speculum examination and were tested for Chlamydia trachomatis and Neisseria gonorrhoeae . PMN counts from Gram-stained endocervical smears and the presence or absence of putative indicators of chlamydial infection were recorded . RESULTS: The prevalence of chlamydial cervicitis was 6.2% (37/596), and no women tested positive for N . gonorrhoeae . Presumptive diagnosis of chlamydial cervicitis based on the guidelines criteria of mucopurulent endocervical discharge and 10 or more PMN per high-power microscopic field had a sensitivity and specificity of 18.9% and 97.0% respectively, a positive predictive value of 29.2% and a positive likelihood ratio (LR) of 6.2 (p = 0.003) . Presumptive diagnosis based on endocervical discharge with edema, erythema or induced mucosal bleeding of the cervix had a sensitivity and specificity of 43.2% and 80.0% respectively, a positive predictive value of 12.5% and a positive LR of 2.2 (p = 0.002) . In the presence of bacterial vaginosis or vaginitis, the LR for the criteria of mucopurulent endocervical discharge and 10 or more PMN per high-power field was 5.4 (p = 0.04), whereas the LR was 4.3 (p = 0.10) if bacterial vaginosis and vaginitis were absent . CONCLUSIONS: In this setting, the practice of making a presumptive diagnosis of chlamydial cervicitis on the basis of the criteria given in the Canadian STD guidelines was not supported. Microbiology, 1998 Jan, 144 ( Pt 1), 149 - 56 The phase-variable pilus-associated protein PilC is commonly expressed in clinical isolates of Neisseria gonorrhoeae, and shows sequence variability among strains; Backman M et al.; PilC is a phase-variable protein associated with pilus-mediated adherence of pathogenic Neisseria to target cells . In this study, 24 strains of Neisseria gonorrhoeae with known epidemiological data were examined for expression of PilC . All strains produced PilC independently of serovar and site of isolation . To investigate whether the PilC protein is conserved or variable among gonococcal strains, the complete nucleotide sequence of pilC in four strains, isolated from either rectum, throat or blood, was determined . The deduced amino acid sequence in these strains differed from each other and from the two PilC proteins of N . gonorrhoeae MS11 . These data demonstrate that PilC is commonly expressed, but the PilC sequence may vary among gonococcal strains. J Clin Microbiol, 1998 Feb, 36(2), 563 - 5 Genetic heterogeneity of strains of Neisseria meningitidis belonging to serotype 22 isolated in the Czech Republic; Musilek M et al.; Strains of Neisseria meningitidis of serogroup B isolated in the Czech Republic frequently belong to serotype 22 . We analyzed the genetic relationships among strains of this serotype by using the multilocus enzyme electrophoresis technique and the polymorphism of the pilA gene . Our results indicate that these strains correspond to a highly heterogeneous population rather than to the expansion of a single clone. Mol Microbiol, 1998 Jan, 27(1), 171 - 86 Using the yeast two-hybrid system to identify human epithelial cell proteins that bind gonococcal Opa proteins: intracellular gonococci bind pyruvate kinase via their Opa proteins and require host pyruvate for growth; Williams JM et al.; Neisseria gonorrhoeae opacity-associated (Opa) proteins are a family of outer membrane proteins involved in gonococcal adherence to and invasion of human cells . We wanted to identify additional roles for Opa in the infectious process and used the yeast two-hybrid system to identify human epithelial cell proteins that interact with Opa proteins . Although this system has been used successfully to identify many types of interacting proteins, it has not been used to screen a human cell cDNA library for binding partners of a prokaryotic outer membrane protein . Therefore, we were also interested in exploring the versatility of the yeast two-hybrid system in identifying bacteria-host interactions . Using OpaP from strain F62SF as bait, we screened a HeLa cell cDNA library for Opa-interacting proteins (OIPs) . We identified five different OIPs, designated OIP1-OIP5, two of which are homologous to human proteins--thyroid hormone receptor interacting protein (TRIP6) and pyruvate kinase isoenzyme M2 (PK) . In the studies presented here, we investigated the interaction between Opa proteins and PK in more depth . Opa-PK interactions were confirmed by in vitro and in vivo assays independent of the yeast two-hybrid system . Escherichia coli expressing six different Opa proteins from gonococcal strain FA1090 all bound more PK than Opa-negative E . coli in in vitro binding assays . Using anti-PK antibody and fluorescence microscopy, we showed that human epithelial cell PK co-localizes with intracellular Opa+ gonococci and E . coli expressing Opa proteins . Using a mutant of N . gonorrhoeae unable to grow on pyruvate or lactate, it appears that intracellular pyruvate is essential for gonococcal growth and survival . These results suggest a novel mechanism in bacterial pathogenesis, i.e . the requirement for direct molecular interaction with a host metabolic enzyme (PK) for the acquisition of an essential intracellular carbon source and growth substrate (pyruvate) . These results demonstrate that the yeast two-hybrid system is a valuable tool for identifying biologically relevant interactions between bacteria and host proteins, providing valuable leads for further investigations into novel mechanisms of bacterial pathogenesis. Eur J Immunol, 1997 Dec, 27(12), 3303 - 9 Lipopolysaccharide complexed with soluble CD14 binds to normal human monocytes; Blondin C et al.; Using flow cytometry we have compared the binding of Neisseria meningitidis lipopolysaccharide labeled with fluorescein isothiocyanate (FITC-LPS) to normal human monocytes in whole blood with the binding to chinese hamster ovary (CHO) cells transfected with human CD14 gene (hCD14-CHO cells) . Binding of FITC-LPS to cells was dose dependent, saturable and enhanced in the presence of increasing concentrations of serum . Blockade of membrane CD14 with saturating concentrations of anti-CD14 monoclonal antibody (mAb) My4 inhibited 50% of the binding of FITC-LPS to monocytes and 100% to hCD14-CHO cells . Similarly, removal of membrane CD14 by phosphatidylinositol phospholipase C (PI-PLC) treatment of the cells partially decreased the binding of FITC-LPS to monocytes but totally inhibited the binding to hCD14-CHO-transfected cells . These results suggest that binding of FITC-LPS to monocytes is not only mediated by membrane CD14 . Using two-color flow cytometry, we observed that FITC-LPS binds to My4-saturated monocytes in association with soluble (s)CD14 present in serum as revealed by staining with rhodamine-labeled My4 mAb . The binding of FITC-LPS/sCD14 complexes to monocytes treated with saturating amounts of unlabeled My4 prior to addition of the complexes was completely inhibited by anti-CD14 mAb 10G33 . When cells were first saturated with a mixture of My4 and 10G33 mAb, washed and further incubated with FITC-LPS/sCD14, inhibition of the binding of LPS was similar to that observed with cells saturated with My4 alone, showing that the binding of FITC-LPS is not