Neisseria

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 mediated by the 10G33 epitope present on mCD14 . These results suggest that either the 10G33 epitope on sCD14 is involved in the binding of LPS/sCD14 complexes to the cells, or that 10G33 mAb inhibits the binding of FITC-LPS to sCD14 . Taken together, these data indicate that sCD14 which is present in normal serum, in addition to membrane CD14, enables LPS to bind monocytes through an as yet unidentified molecule and that sCD14 does not simply serve as a shuttle for transfer of LPS to membrane CD14.

J Antimicrob Chemother, 1997 Dec, 40(6), 817 - 22
Plasmid analysis of Neisseria gonorrhoeae isolates and dissemination of tetM genes in southern Africa 1993-1995; Chalkley LJ et al.; One group (145 isolates) of Neisseria gonorrhoeae was collected from municipal clinics in Bloemfontein in 1994 and a second group (65 isolates) in 1995 . Penicillin and tetracycline MICs were determined and plasmid analysis performed to monitor antimicrobial susceptibilities in conjunction with the occurrence of plasmids in these isolates . The prevalence of penicillin resistance caused by beta-lactamase plasmids remained constant at 9% during the study period . Three high-level tetracycline-resistant strains (MICs 16 mg/L), the first to be detected in South Africa, were isolated in 1994 . Although there was a reduction in the percentage of isolates harbouring 24.5 MDa conjugative plasmids (from 79% in 1994 to 46% in 1995), this was partially counteracted by an increase in TetM-encoding conjugative plasmids (25.2 MDa) from 2% to 18.5% . The tetM genes of 13 isolates shown to exhibit high-level tetracycline resistance were characterized as the American type . The American-type tetracycline resistance plasmid was demonstrated in 11 isolates . Digestion with Bg/l showed that two isolates harboured tetM-encoding plasmids that differed from the American- and Dutch-type plasmids described previously: one isolate contained a plasmid that produced two fragments of different sizes from those of the American-type plasmid and the second isolate possessed an American/Dutch hybrid plasmid . Auxotyping/serotyping and random amplified polymorphic DNA analysis revealed a predominant tetracycline-resistant family (NR/IA-6, genomic group I) in Bloemfontein . As there is a high incidence of chlamydial infections in southern Africa requiring tetracycline therapy, selective pressures exist in the environment for the maintenance and rapid spread of high-level tetracycline-resistant N . gonorrhoeae . It is possible that tetM genes may have emanated from Botswana and/or Namibia to Bloemfontein . The establishment of high-level tetracycline-resistant N . gonorrhoeae in Bloemfontein was seen to be complex as a related group of strains was identified, plasmid dissemination was evident and two new TetM-encoding plasmids were demonstrated . The appearance of these TetM-encoding plasmids indicates either that the American- and Dutch-type plasmids are continuing to evolve or that tetM genes are being introduced into different families of 24.5 MDa conjugative plasmids.

J Clin Pathol, 1997 Nov, 50(11), 947 - 50
Evaluation of 2-SP transport medium for detection of Chlamydia trachomatis and Neisseria gonorrhoeae by two automated amplification systems and culture for chlamydia; Dubuis O et al.; AIMS: To assess the performance of 2-sucrose-phosphate based transport medium (2-SP) for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by an automated commercial polymerase chain reaction (PCR) and ligase chain reaction (LCR) compared to centrifugation culture on McCoy cells for C trachomatis . Second, to compare both amplification systems for initial diagnostic testing of a low prevalence population for sexually transmitted diseases . METHODS: Four hundred and eighty one consecutive urogenital and conjunctival specimens were examined . All tests were performed on the same specimen collected with a dacron swab and transported in 2-SP medium . Samples that were positive by culture or by both PCR and LCR were considered to be true positives . RESULTS: The prevalences of C trachomatis and of N gonorrhoeae were 2.7% and 0.4%, respectively . PCR had a resolved sensitivity and specificity of 100% and 99.8%, respectively, for C trachomatis, and 100% and 98.9%, respectively, for N gonorrhoeae . LCR was 100% sensitive and specific for both pathogens . The resolved sensitivity of the shell vial assay was 85% . No culture positive sample would have been missed by PCR or LCR . The inhibition rate for PCR was 4.8% . CONCLUSIONS: 2-SP medium proved to be suitable for both PCR and LCR . It is not limited to any one test manufacturer and allows the performance of amplification techniques and viral and chlamydia culture from the same specimen . The LCR was more reliable than PCR on initial testing . However, hands on time is longer, and no amplification control is available for LCR.

Diagn Microbiol Infect Dis, 1997 Dec, 29(4), 281 - 3
Characterization of DFA-negative, probe-positive Neisseria gonorrhoeae by pulsed field gel electrophoresis; Billings SD et al.; Eight Neisseria gonorrhoeae isolates, negative by direct fluorescent antibody (DFA) but positive by a DNA probe, were characterized by pulsed field gel electrophoresis and compared to eight DFA-positive, probe-positive isolates . Results indicate that DFA-negative, probe-positive Neisseria gonorrhoeae isolates may be clonal.

J Clin Microbiol, 1998 Jan, 36(1), 191 - 7
An internal control for routine diagnostic PCR: design, properties, and effect on clinical performance; Rosenstraus M et al.; We constructed internal controls (ICs) to provide assurance that clinical specimens are successfully amplified and detected . The IC nucleic acids contain primer binding regions identical to those of the target sequence and contain a unique probe binding region that differentiates the IC from amplified target nucleic acid . Because only 20 copies of the IC are introduced into each test sample, a positive IC signal indicates that amplification was sufficient to generate a positive signal from targets present at the limit of test sensitivity . The COBAS AMPLICOR Chlamydia trachomatis, Neisseria gonorrhoeae, Mycobacterium tuberculosis, and human hepatitis C virus tests exhibited inhibition rates ranging from 5 to 9% . Approximately 64% of these inhibitory specimens were not inhibitory when a second aliquot was tested . Because repeatedly inhibitory specimens were not reported as false negative and because additional infected specimens were detected during retesting, test sensitivities were 1 to 6% greater than they would have been if the IC had not been used.

Zh Mikrobiol Epidemiol Immunobiol, 1995 Jul-Aug, (4), 24 - 9
{The ultrastructural characteristics of Neisseria meningitidis strains isolated in different forms of infection}; Vysotskii VV et al.; The electron microscopic study of cell populations of the first generation of several N.meningitidis strains, isolated from humans and grown on artificial culture medium (Hottinger's serum agar) has made it possible to establish that these organisms are morphologically heterogeneous . It was manifested by the presence of 7 morphological variants of cells in their cultures (different strains have different proportions of these variants): (a) "normal" and similar to normal diplococcal variants; (b) atypically dividing cells; (c) spheroplastic forms; (d) protoplastic forms (typical and atypical); (e) variably gemmating forms; (f) microforms and (g) gigantic forms . These data are of interest for the microbiological diagnosis of meningitis.

Arch Pediatr Adolesc Med, 1998 Jan, 152(1), 52 - 6
Urine-based screening of adolescents in detention to guide treatment for gonococcal and chlamydial infections . Translating research into intervention; Oh MK et al.; OBJECTIVES: To determine the utility of urine-based ligase chain reaction assays for Neisseria gonorrhoeae and Chlamydia trachomatis in (1) the acceptability of such testing to adolescent detainees, (2) the potential use of these tests for identifying asymptomatic infections, and (3) the effectiveness of this approach for ensuring treatment of infected adolescents . DESIGN: Cross-sectional screening and verification of treatment for infected cases . SUBJECTS: Adolescents admitted to a short-term juvenile detention center . MAIN OUTCOME MEASURES: Neisseria gonorrhoeae and C trachomatis infection rates, and timing and location of treatment for infected patients . RESULTS: Refusal rate was 1.5% . Of 263 participants, 46 (17.5%) were female subjects . Chlamydia trachomatis infections were identified in 28.3% of the female and 8.8% of the male subjects . Neisseria gonorrhoeae infections were present in 13.1% of the female and 2.8% of the male subjects . Overall, 37 participants (14%) were positive for N gonorrhoeae, C trachomatis, or both, only one of whom had symptoms . Almost 70% (25/36) of asymptomatic infected subjects were treated within 28 days of screening . A treatment was documented in 36 of the 37 infected youth, including 20 who were followed up and treated after release from the detention center, by 6 months after testing . CONCLUSION: Urine ligase chain reaction tests were effective for identifying and guiding treatment of unsuspected N gonorrhoeae and C trachomatis infections in teenagers admitted to a short-term detention center where traditional swab specimens may be difficult to obtain.

Infect Immun, 1998 Feb, 66(2), 505 - 13
Dynamics of the murine humoral immune response to Neisseria meningitis group B capsular polysaccharide; Colino J et al.; Immunization with Neisseria meningitidis group B capsular polysaccharide (CpsB) elicited responses in adult mice that showed the typical dynamic characteristics of the response to a thymus-independent antigen, in contrast to the thymus-dependent behavior of antibody responses to CpsC . The former had a short latent period and showed a rapid increase in serum antibodies that peaked at day 5, and immunoglobulin M (IgM) was the major isotype even though IgG (mainly IgG2a and IgG2b) was also detectable . This response was of short duration, and the specific antibodies were rapidly cleared from the circulation . The secondary responses were similar in magnitude, kinetics, IgM predominance, and IgG distribution . Nevertheless, a threefold IgG increase, a correlation between IgM and IgG levels, and dose-dependent secondary responses were observed . Hyperimmunization considerably reinforced these responses: 10-fold for IgM and 300-fold for IgG . This favored isotype switch was accompanied by a progressive change in the subclass distribution to IgG3 (62%) and IgG1 (28%), along with the possible generation of B-cell memory . The results indicate that CpsB is being strictly thymus independent and suggest that unresponsiveness to purified CpsB is due to tolerance.

Infect Immun, 1998 Feb, 66(2), 469 - 73
Transcellular passage of Neisseria gonorrhoeae involves pilus phase variation; Ilver D et al.; Piliated and nonpiliated Neisseria gonorrhoeae organisms were added on top of confluent layers of HEC-1-B cells, each maintained on a microporous Transwell-COL membrane . The bacteria released into the lower chamber were characterized with respect to the following virulence determinants: pili, which mediate adherence to target host cells; PilE, the major pilus subunit protein; and PilC, which is involved in pilus biogenesis and adherence . Even if >99% of the added bacteria of N . gonorrhoeae MS11 were piliated, bacteria recovered on the other side of the cell layer were predominantly nonpiliated . The recovered clones still expressed unassembled PilE protein, but 50% had lost PilC production . Nonpiliated gonococci, in which the 5' end of pilE had been deleted, were released in reduced numbers, and piliated recA bacteria added to the cell layer were not released at all, at time points when piliated recA+ clones were found at high numbers in the lower chamber . Our data indicate that bacteria producing unassembled PilE protein are selected for during passage through an epithelial cell layer . The finding that the pilE gene sequence was altered in the transmigrants suggests that pilin sequence variation is involved in the transcellular passage of N . gonorrhoeae.

Cent Afr J Med, 1997 Jul, 43(7), 192 - 5
Prevalence of gonorrhoea and knowledge of sexually transmitted infections in a farming community in Zimbabwe; Gomo E et al.; OBJECTIVE: To determine the prevalence of gonorrhoea and knowledge about sexually transmitted infections in a farming community . DESIGN: Cross sectional study . SETTING: Commercial farming area, Zimbabwe . SUBJECTS: 1,005 individuals aged between 15 and 60 years . MAIN OUTCOME MEASURES: Prevalence rates, knowledge of gonorrhoea and other sexually transmitted infections (STIs) by age and sex . RESULTS: Knowledge of gonorrhoea and other STIs was higher while knowledge of AIDS was lower among males than females . Frequency of past history of STIs in five years was significantly higher among males than females (OR 3.22; 95% CI 2.45 to 4.25) . Physical examination revealed that 14% of males and 22% of females had other STIs besides gonorrhoea . The prevalence of gonorrhea was 18.4% (95% CI 16.0 to 20.8) and was higher among females than males (OR 2.77; 95% CI 1.97 to 3.90) . Thirty one percent of the gonorrhoea infections were due to penicillinase producing Neisseria gonorrhoeae . CONCLUSIONS: Our findings emphasize the disparity between knowledge and practices with regards to STIs, and the importance of STIs as potential targets for and indices in the fight against human immunodeficiency virus (HIV) infectionPIP: A cross-sectional study of 1005 males and females 15-60 years of age from a commercial farming community (Hippo Valley Sugar Estates) in Zimbabwe investigated knowledge and prevalence rates of gonorrhea and other sexually transmitted infections (STIs) . Although over 70% of respondents were aware of gonorrhea, only 45.5% were knowledgeable about AIDS . Males had higher knowledge of gonorrhea and other STIs than females, but lower knowledge of HIV/AIDS . 47% of subjects had a history of an STI in the 5 years preceding the study . The frequency of such a history was significantly higher among males compared to females (odds ratio (OR), 3.22; 95% confidence interval (CI), 2.45-4.25) . Physical examination indicated that 20.5% of males and 37.9% of females had signs of STIs . The majority of STI cases affected men in the 20-29 year age group and women 30-39 years of age . Gonorrhea prevalence was 18.4% and significantly higher among females than males (OR, 2.77; 95% CI, 1.97-3.90) . 31% of gonorrhea infections were associated with penicillinase-producing Neisseria gonorrhoeae . Since STIs have been associated with increased transmission of HIV, these infections should be targeted for campaigns aimed at reducing the disparity between knowledge and practices . Moreover, examination of the prevalence of STIs such as gonorrhea is a quick way of assessing the impact of HIV control measures in a community .

Mol Microbiol, 1997 Oct, 26(1), 25 - 35
Energy-dependent changes in the gonococcal transferrin receptor; Cornelissen CN et al.; The pathogenic Neisseria spp . are capable of iron utilization from host iron-binding proteins including transferrin and lactoferrin . Transferrin iron utilization is an energy-dependent, receptor-mediated event in which two identified transferrin-binding proteins participate . One of these proteins, TbpA, is homologous to the TonB-dependent family of outer membrane receptors that are required for high-affinity uptake of vitamin B12 and ferric siderophores . The 'TonB box' is a conserved domain near the amino-terminus of these proteins that has been implicated in interaction with TonB . Interaction between a periplasmic domain of TonB and the TonB box allows energy transduction to occur from the cytoplasmic membrane to the energy-dependent receptor in the outer membrane . We created a TonB box mutant of gonococcal TbpA and demonstrated that its binding and protease accessibility characteristics were indistinguishable from those of gonococcal Ton system mutants . The protease exposure of the second transferrin-binding protein, TbpB, was affected by the energization of TbpA, consistent with an interaction between these proteins . TbpB expressed by the de-energized mutants was readily accessible to protease, similar to TbpB expressed in the absence of TbpA . The de-energized mutants exhibited a marked decrease in transferrin diffusion rate, suggesting that receptor energization was necessary for ligand release . We propose a model to explain the observed Ton-dependent changes in the binding parameters and exposures of TbpA and TbpB.

Commun Dis Intell, 1997 Dec 25, 21(23), 349 - 53
Antimicrobial resistance in gonococci, WHO Western Pacific Region, 1996 . The WHO Western Pacific Region Gonococcal Antimicrobial Surveillance Programme; Exploring hydrophobic sites in proteins with xenon or krypton; LURE, Universite Paris-Sud, Orsay, France . prange@lure.u-psud.fr

X-ray diffraction is used to study the binding of xenon and krypton to a variety of crystallised proteins: porcine pancreatic elastase; subtilisin Carlsberg from Bacillus licheniformis; cutinase from Fusarium solani; collagenase from Hypoderma lineatum; hen egg lysozyme, the lipoamide dehydrogenase domain from the outer membrane protein P64k from Neisseria meningitidis; urate-oxidase from Aspergillus flavus, mosquitocidal delta-endotoxin CytB from Bacillus thuringiensis and the ligand-binding domain of the human nuclear retinoid-X receptor RXR-alpha . Under gas pressures ranging from 8 to 20 bar, xenon is able to bind to discrete sites in hydrophobic cavities, ligand and substrate binding pockets, and into the pore of channel-like structures . These xenon complexes can be used to map hydrophobic sites in proteins, or as heavy-atom derivatives in the isomorphous replacement method of structure determination.

Medicina (B Aires), 1997, 57(1), 1 - 6
{The impact of HIV infection on the incidence of syphilis and gonorrhea at a university hospital (1985-1994)}; Griemberg G et al.; Syphilis and gonorrhea are two sexually transmitted diseases (STD) which still persist in spite of the existence of effective treatment . In this paper the incidence of these two diseases was evaluated between 1985-1994, a decade during which the problem of HIV infection arose . Every patient who attended the STD Program at the Hospital and was suspected of having syphilis and/or gonorrhea was clinically evaluated and was subjected to the following tests: a) search for Neisseria gonorrhoeae (Ng) and penicinilase producers (PPNG) identification, b) search for Treponema pallidum (Tp) by darkfield microscopy, c) serological studies for syphilis . From 1985 to 1991 the average of positive cases for Ng was 14.8% while from 1992 to 1994 it was 4.0% (p < 0.0001) showing a dramatic decrease between 1991 and 1992 . PPNG increased from 8.7% in 1985 to 44.6% in 1988 and then slowly decreased down to 26.9% in 1994 . From 1985 to 1990 the average of positive darkfield examinations was 20.7% while from 1991 to 1994 it was 12.2% (p < 0.0001) . Between 1985 to 1988 syphilis serological studies showed 45.7% positive cases dropping to 34.3% between 1989 and 1994 (p < 0.0001) . HIV infection appearance had a wide repercussion throughout the world generating behavioral and sexual habit changes which have probably helped to prevent these two STD . Gonorrhea incidence was also influenced by the appearance of new effective treatments, even against PPNG, such as fluorated quinolones and third generation cephalosporines . The population studied belongs to the most crowded area in Argentina, so that these observations should not be extrapolated to the rest of the country.

Arch Biochem Biophys, 1997 Dec 15, 348(2), 357 - 62
Isolation and biochemical characterization of the PilA protein of Neisseria meningitidis; Arvidson CG et al.; PilA is the response regulator of a two-component regulatory system that controls a number of genes in the pathogenic Neisseria . Previous work has shown that Neisseria gonorrhoeae (GC) PilA binds DNA and also hydrolyzes GTP . Here, we report the cloning, sequencing, purification, and biochemical characterization of PilA from N . meningitidis (MC) strain 8013 . MC pilA is 94% identical to GC pilA at the nucleotide level . Of the 78 nucleotide changes, 52 are silent, while 26 result in a total of 20 amino acid changes . Additionally, the MC homolog has a 4-amino acid insertion between the putative DNA-binding and GTP-binding domains . Purified MC PilA binds to the same DNA fragment we have previously shown to be bound by GC PilA specifically and also hydrolyzes GTP . The K(m) of MC PilA for GTP is 8.6 microM, similar to that determined for the GC protein . However, the maximum velocity (Vmax) is approximately 35-fold greater than the GC PilA activity . Additionally, the nucleotide specificity of MC PilA differs from that of GC PilA . While GC PilA hydrolyzes only GTP, MC PilA hydrolyzes GTP and ATP equally well, and CTP and UTP also compete for this activity.

Hum Reprod, 1997 Nov, 12(11 Suppl), 121 - 6
Clinical aspects of pelvic inflammatory disease; Munday PE; Pelvic inflammatory disease (PID) is a common and poorly managed condition . Untreated or inadequately treated, it leads to tubal infertility, ectopic pregnancy and chronic pelvic pain . Diagnostic difficulties are compounded by the wide variety of clinical presentations and the insensitivity and poor specificity of laboratory tests . Better recognition of mild and atypical disease needs a high index of suspicion whenever young, sexually active women present with gynaecological symptoms . Laparoscopy supplemented by microbiological tests and fimbrial minibiopsy should be regarded as the diagnostic 'gold standard' for research studies; new studies are required to identify techniques which might reduce under- and over-diagnosis . Early treatment reduces the risk of an adverse effect on fertility . Any therapeutic regimen selected should be effective against the common aetiological agents Chlamydia trachomatis, Neisseria gonorrhoeae, genital mycoplasmas and aerobic and anaerobic bacteria . Since at least 60% of cases of PID can be attributed to infection with a sexually transmitted organism, partner notification forms an essential part of management.

Hum Reprod, 1997 Nov, 12(11 Suppl), 107 - 12
Preventing infective sequelae of abortion; Penney GC; Pelvic infection complicates up to 12% of induced abortions and has an adverse effect on future reproductive outcome . The presence in the lower genital tract of Neisseria gonorrhoeae, Chlamydia trachomatis or the anaerobic organisms characterizing bacterial vaginosis is associated with an increased risk of post-abortion infective morbidity . Meta-analysis of randomized trials has shown that prophylaxis with antibiotics effective against either C . trachomatis or bacterial vaginosis reduces the risk of post-abortion infective morbidity by around a half . Other strategies which have been advocated for minimizing the risk of infective morbidity are screening for lower genital tract infections, with treatment of positive cases only, and a combined strategy where women are screened for sexually transmitted infections as well as receiving prophylaxis . These strategies provide the opportunity for appropriate follow-up and partner notification of those women found to have sexually transmitted infections . A multicentre study designed to determine the prevalence of genital tract infections among Scottish women seeking induced abortion, and to compare strategies of 'universal prophylaxis' and 'screen and treat' for minimizing infective morbidity in such women has been undertaken . A total of 1672 women were recruited . Prevalence rates of lower genital tract gonorrhoea, chlamydia and bacterial vaginosis were found to be similar to those reported in other UK studies . Women managed by the 'screen and treat' strategy (particularly those whose genital tract swabs were reported negative) had slightly higher rates of infective morbidity in the 8 weeks after abortion than those managed by 'prophylaxis' . Using currently available screening tests and genitourinary medicine services, 'prophylaxis' appears to be the more cost effective of the two strategies studiedPIP: Reported rates of post-abortion pelvic inflammatory disease (PID) range from 5-29% . The risk of infection has been associated with the presence of Neisseria gonorrhoeae, Chlamydia trachomatis, and anaerobic organisms in the lower genital tract . The present study analyzed the prevalence of genital tract infections in 1672 women undergoing induced abortion at 3 centers in Scotland and evaluated the efficacy of two preventive interventions . Prevalence rates of lower genital tract gonorrhea, chlamydia, and bacterial vaginosis before abortion were similar to those identified in other UK studies . Women were randomly allocated to receive either prophylactic metronidazole (immediately before abortion) and doxycycline (for 7 days after abortion) or received antibiotics only if pre-abortion genital tract swabs were positive for any of the 3 infections . During the 8-week post-abortion follow-up period, women managed by the screen-and-treat protocol had slightly less favorable outcomes in terms of hospital readmissions, general practitioner consultations, antibiotic prescriptions, time off work, and limitations on domestic activities than women who received prophylactic treatment . Differences were statistically significant, however, only for women whose swabs were negative for all 3 infections . The rate of post-abortion PID/endometritis in this groups was 3% among women who received prophylactic antibiotics and 6% in those who were screened and not treated . These findings suggest that universal antibiotic prophylaxis may represent the most cost-effective approach to minimizing the risk of infective morbidity . Advocated for consideration is a third strategy involving prophylaxis at the time of abortion followed by screening for gonorrhea and chlamydia to ensure adequate follow-up of treatment results and partner notification .

Arch Med Res, 1997 Winter, 28(4), 591 - 5
Evaluation of the antimeningococcal immunoglobin activity against five strains of Neisseria meningitidis group B from Latin America using mice as model; Perez-Ramirez PJ et al.; To know the activity of antimeningococcal immunoglobulin, Balb/c mice of 18-22 g of body weight were challenged with 5 serotype B strains of Neisseria meningitidis (Nm) isolated from patients of different Latin American countries . The specific antimeningococcal Ig was extracted from the serum of volunteers previously vaccinated with the antimeningococcal BC vaccine VA-MENGOC-BC (Finlay Institute, Havana, Cuba) . The Ig was intraperitoneally (IP) administered in a unique dose of 10 mg/mouse . The strains A, B, C, CH and D were inoculated IP in the following charges: strain A, 20 LD50; B, 25 LD50; C, 44.5 LD50; CH, 36 LD50, and D, 200, 20 and 2 LD50 . For each strain, a control group received living bacteria and virulent stimulating factor (VSF) . The Ig was injected 30 min before or 30 min after the challenge dose had been given, except for strain D, which only received the Ig 30 min after the challenge . As VSF, 0.5 mg of iron in the form of iron dextran was used . The experiment was analyzed considering the survival time after the challenge for each strain compared to the corresponding control group (C) . When the Ig was used 30 min before the challenge, the protection period for the A strain was (C:18.1h) more than 72h (P<0.001) and 100% survival; for the B strain, (C:29.5h) 42h (P<0.05) and almost 20% survival; for the C strain (C:16.5h) 35h (P<0.01) with a 40% survival, and the CH strain (C:18.1h) 26.5h (P<0.02), with a 20% survival . When the Ig was injected 30 min after the challenge, the average survival time and the survival for the A strain was 28h (P<0.05) with 62.5%; for the B strain it was 42 h (P<0.005) and 0.0%; for the C strain 27.3 h (P<0.05) and 30%; for the CH strain 25.8h (P<0.05) and 0.0%, and for the D strain 19.1h, 26h, and more than 72h with a 0.0%, 60% and 100%, depending on the challenging dose . In general, the specific Ig used showed a protective effect in mice against the different Latin American strains tested . Additionally, the experimental model proved to be useful for the study of the antimeningococcal human Ig.

Mol Microbiol, 1997 Dec, 26(5), 971 - 80
Differential Opa specificities for CD66 receptors influence tissue interactions and cellular response to Neisseria gonorrhoeae; Gray-Owen SD et al.; The ability of all 11 variable opacity (Opa) proteins encoded by Neisseria gonorrhoeae MS11 to interact directly with the five CD66 antigens was determined . Transfected HeLa cell lines expressing individual CD66 antigens were infected with recombinant N . gonorrhoeae and Escherichia coli strains expressing defined Opas . Based upon the ability of these bacteria to bind and invade and to isolate specifically CD66 antigens from detergent-soluble extracts of the corresponding cell lines, distinct specificity groups of Opa interaction with CD66 were seen . Defining these specificity groups allowed us to assign a specific function for CD66a in the Opa-mediated interaction of gonococci with two different target cell types, which are both known to co-express multiple CD66 antigens . The competence of individual Opas to interact with CD66a was strictly correlated with their ability to induce an oxidative response by polymorphonuclear neutrophils . The same Opa specificity was observed for the level of gonococcal binding to primary endothelial cells after stimulation with TNFalpha, which was shown to increase the expression of CD66a rather than CD66e . As CD66e alone is expressed on other target tissues of gonococcal pathogenicity, Opa variation probably contributes to the cell tropism displayed by gonococci.

J Bacteriol, 1998 Jan, 180(1), 119 - 27
Substrate ambiguity of 3-deoxy-D-manno-octulosonate 8-phosphate synthase from Neisseria gonorrhoeae in the context of its membership in a protein family containing a subset of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthases; Subramaniam PS et al.; 3-Deoxy-D-manno-octulosonate 8-phosphate (KDOP) synthase and 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase catalyze similar phosphoenolpyruvate-utilizing reactions . The genome of Neisseria gonorrhoeae contains one gene encoding KDOP synthase and one gene encoding DAHP synthase . Of the two nonhomologous DAHP synthase families known, the N . gonorrhoeae protein belongs to the family I assemblage . KDOP synthase exhibited an ability to replace arabinose-5-P with either erythrose-4-P or ribose-5-P as alternative substrates . The results of periodate oxidation studies suggested that the product formed by KDOP synthase with erythrose-4-P as the substrate was 3-deoxy-D-ribo-heptulosonate 7-P, an isomer of DAHP . As expected, this product was not utilized as a substrate by dehydroquinate synthase . The significance of the ability of KDOP synthase to substitute erythrose-4-P for arabinose-5-P is (i) recognition of the possibility that the KDOP synthase might otherwise be mistaken for a species of DAHP synthase and (ii) the possibility that the broad-specificity type of KDOP synthase might be a relatively vulnerable target for antimicrobial agents which mimic the normal substrates . An analysis of sequences in the database indicates that the family I group of DAHP synthase has a previously unrecognized membership which includes the KDOP synthases . The KDOP synthases fall into a subfamily grouping which includes a small group of DAHP synthases . Thus, family I DAHP synthases separate into two subfamilies, one of which includes the KDOP synthases . The two subfamilies appear to have diverged prior to the acquisition of allosteric-control mechanisms for DAHP synthases . These allosteric control specificities are highly diverse and correlate with the presence of N-terminal extensions which lack homology with one another.

Antimicrob Agents Chemother, 1997 Dec, 41(12), 2776 - 80
Sulfated carbohydrate compounds prevent microbial adherence by sexually transmitted disease pathogens; Herold BC et al.; Heparan sulfate (HS) serves as a receptor for adherence of herpes simplex viruses, Chlamydia trachomatis, Neisseria gonorrhoeae, and, indirectly, human immunodeficiency virus . Using primary human culture systems, we identified sulfated carbohydrate compounds that resemble HS and competitively inhibit infection by these pathogens . These compounds are candidates for intravaginal formulations for the prevention of sexually transmitted diseases.

Sex Transm Dis, 1997 Nov, 24(10), 576 - 81
Antimicrobial resistance in Neisseria gonorrhoeae in Finland, 1976 to 1995 . The Finnish Study Group For Antimicrobial Resistance; Nissinen A et al.; BACKGROUND AND OBJECTIVES: The worldwide increase in antimicrobial resistance in Neisseria gonorrhoeae prompted the authors to evaluate the status and course of resistance in gonococci in Finland . GOALS: The minimal inhibitory concentrations (MIC) of penicillin, tetracycline, spectinomycin, ciprofloxacin, ceftriaxone, and cefixime were tested for 337 consecutive clinical N . gonorrhoeae isolates collected in 19 Finnish microbiology laboratories in 1993 . STUDY DESIGN: The results were compared with data obtained in three Finnish laboratories in 1986 and contrasted with the development of the incidence of gonorrhea and the prevalence of penicillinase-producing N . gonorrhoeae (PPNG) in Finland, 1976 to 1995 . The number of strains with an elevated MIC to ciprofloxacin was assessed by questionnaire . RESULTS: A decrease, from more than 50% in 1986 to 20% in 1993, of strains susceptible to penicillin and tetracycline was observed . The prevalence of PPNG increased from 0% (1976) to 5.7% (1995) . In 1995, two strains with a ciprofloxacin MIC of > or = 32 micrograms/ml were reported . No resistance to ceftriaxone or spectinomycin was detected . CONCLUSIONS: In spite of the rarity of gonorrhea and the availability of efficient antimicrobials in Finland, monitoring of the antimicrobial resistance of N . gonorrhoeae remains important.

Sex Transm Dis, 1997 Nov, 24(10), 561 - 6
Genomic homogeneity of the AHU/IA-1,2 phenotype of Neisseria gonorrhoeae during its disappearance from an urban population; Xia M et al.; BACKGROUND: Isolates of Neisseria gonorrhoeae requiring arginine, hypoxanthine, and uracil (AHU) appeared in Denmark in 1946, were preponderant in Seattle during the 1970s, were associated with disseminated gonococcal infections (DGI), and were primarily of the IA-1,2 serovar . GOALS: To investigate the disappearance of the AHU/IA-1,2 phenotype and to examine by pulsed-field gel electrophoresis (PFGE) the genomic homogeneity of this unique phenotype isolated from Seattle-King County during the past decade . STUDY DESIGN: This retrospective study used data extracted from previous publications for the period 1971 through 1984, and from existing records at the Neisseria Reference Laboratory, University of Washington for the period 1985 through 1996 . Samples (n = 68) of AHU/IA-1,2 isolates from 1984 to 1985 and 1988 to 1993 were analyzed using endonucleases NheI and XbaI . For comparison, 10 AHU/IB isolates were included in the study . RESULTS: AHU isolates, predominantly IA-1,2 strains accounted for 52% of the gonococcal isolates for the period 1971 through 1974, 40% for 1974 through 1976, 16% for 1984, and then declined from 7% in 1986 to 0% for each of the last 3 years, 1994 through 1996 . Isolates with < or = 1 band difference after digestion with either NheI or XbaI were considered to belong to a single closely related pattern . Pulsed-field gel electrophoresis (PFGE) type designation was made from the combination of NheI and XbaI patterns . These criteria yielded 5 NheI and 8 XbaI PFGE patterns, and 11 PFGE types based on the combination of NheI and XbaI pattern . The most frequently occurring NheI and XbaI patterns accounted for 74% and 57% of isolates, respectively . One type persisted throughout the decade and accounted for 54% of the 68 isolates . Analysis of the 10 AHU/IB isolates yielded 7 NheI and 8 XbaI patterns that gave 9 types that were distinct from the types found in the AHU/IA-1,2 strains . CONCLUSION: The AHU/IA-1,2 phenotype first documented 50 years ago in Denmark still shows a high degree of genomic homogeneity during the past decade in Seattle . The implementation of screening, decreased rates of sexual exposure in the acquired immune deficiency syndrome era, or other factors may explain its apparent elimination in Seattle-King County.

J Antimicrob Chemother, 1997 Oct, 40(4), 561 - 5
Molecular evolution of the tet(M) gene in Gardnerella vaginalis; Huang R et al.; Five clinical isolates of Gardnerella vaginalis known to carry the tetracycline resistance determinant Tet M were examined by hybridization and nucleotide sequencing . Four of the strains carried tet(M) genes with identical sequences . The two versions of the tet(M) gene found in G . vaginalis did not show complete identity with other published tet(M) sequences, but showed mosaic structures with regions of homology to tet(M) gene sequences from Tn916, Tn1545 and the American type plasmid found in Neisseria gonorrhoeae . Hybridization studies showed that all isolates carried the tet(M) gene on a single HindII restriction fragment of variable length . No evidence was found for the presence of sequences homologous to the transposition functions of Tn916.

Immunol Invest, 1997 Aug-Dec, 26(5-7), 661 - 79
Immunoassay of human Neisseria meningitidis serogroup A antibody; Diakun KR et al.; An enzyme immunoassay is described which quantitates antibodies to Neisseria meningitidis serogroup A capsular polysaccharide in human sera . Modifications of a previously developed two-day assay by Carlone et al . were made to permit analysis in one day and to be compatible with automation . The allowable variations in assay conditions and the areas in which stringent control must be maintained for consistent assay performance are described . Antigen-coating parameters, the kinetics of primary and secondary antibody incubation steps, the buffer compositions, including detergents, serum requirements, and the need for blocking steps were examined . Our modified one-day assay showed excellent agreement with the standardized method of Carlone, with a correlation coefficient between the two methods of 0.989 . This assay is adaptable within a permissible range of parameters thus facilitating the implementation of the standardized assay . This will maximize the consistency of results from serum analysis for conjugate vaccine trials related to serotype A Neisseria meningitidis.

Infect Immun, 1997 Dec, 65(12), 5028 - 34
Natural proteoglycan receptor analogs determine the dynamics of Opa adhesin-mediated gonococcal infection of Chang epithelial cells; van Putten JP et al.; Many bacterial pathogens possess a complex machinery for the induction and/or secretion of factors that promote their uptake by mammalian cells . We searched for the molecular basis of the 60- to 90-min lag time in the interaction of Neisseria gonorrhoeae carrying the heparin-binding Opa adhesin with Chang epithelial cells . Infection assays in the presence of chloramphenicol demonstrated that the Opa-mediated gonococcal infection of Chang cells required bacterial protein synthesis when the microorganisms were derived from GC agar but not when grown in liquid media . Further analysis indicated that contact with agar ingredients rather than the growth state of the microorganisms determined the infection dynamics . DEAE chromatography of GC agar extracts and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses and testing of collected fractions in infection assays identified negatively charged high-molecular-weight polysaccharides in the agar as inhibitors of the cellular infection . Electron microscopy showed that agar-grown gonococci were surrounded by a coat of alcian blue-positive material, probably representing accreted polysaccharides . Similar antiphagocytic material was isolated from bovine serum, indicating that in biological fluids gonococci producing the heparin-binding Opa adhesin may become covered with externally derived polysaccharides as well . Binding assays with gonococci and epithelial proteoglycan receptors revealed that polysaccharides derived from agar or serum compete with the proteoglycans for binding of the heparin-binding Opa adhesin and thus act as receptor analogs . Growth of gonococci in a polysaccharide-free environment resulted in optimal proteoglycan receptor binding and rapid bacterial entry into Chang cells . The recognition that gonococci with certain phenotypes can recruit surface polysaccharides that determine in vitro infection dynamics adds a different dimension to the well-recognized biological significance of genetic variation for this pathogen.

Gene, 1997 Oct 1, 198(1-2), 89 - 97
The normally silent sigma54 promoters upstream of the pilE genes of both Neisseria gonorrhoeae and Neisseria meningitidis are functional when transferred to Pseudomonas aeruginosa; Carrick CS et al.; The pilE gene encodes the pilin subunit in Neisseria gonorrhoeae and Neisseria meningitidis . Transcriptional analysis of promoters upstream of pilE in N . gonorrhoeae has been described previously (Fyfe et al . (1995) J . Bacteriol . 177, 3781-3787) . Transcription from the sigma54-dependent promoter P3 was detected in Pseudomonas aeruginosa . Here we show that this transcription is dependent on the P . aeruginosa transcriptional activator PilR, and a specific upstream sequence with a high degree of similarity to the PilR-binding site found upstream of the P . aeruginosa pilin gene . This implies there is an upstream activator site (UAS) present 5' of pilE . Sequencing upstream of the N . meningitidis MC58 c2 pilE gene shows this region to be very similar to that in N . gonorrhoeae . P3 and the UAS are conserved, although insertions were noted on either side of the UAS . Transcriptional analysis has shown that the N . meningitidis P3 promoter is used in P . aeruginosa, provided PilR and an upstream region that includes sequence similar to the UAS are present . Transcription from the N . meningitidis PpilE is stronger than from the N . gonorrhoeae equivalent . N . meningitidis uses the sigma70 promoter P1 to transcribe pilE.

Mol Microbiol, 1997 Sep, 25(5), 893 - 901
A peptidoglycan hydrolase similar to bacteriophage endolysins acts as an autolysin in Neisseria gonorrhoeae; Dillard JP et al.; We have identified a gene encoding an autolysin (atlA) from Neisseria gonorrhoeae . The deduced amino acid sequence of AtlA shows significant similarity to the peptidoglycan degrading transglycosylases (endolysins) of bacteriophages lambda and P2, suggesting that the encoded protein also functions in peptidoglycan hydrolysis . An atlA mutant was identical to the wild-type strain in exponential growth rate, but demonstrated reduced lysis and peptidoglycan turnover in the stationary phase of growth . When transferred into a buffer solution, at a pH non-permissive for other gonococcal autolysins, an autolytic activity was detectable in the wild-type strain that was not present in the mutant . The most dramatic phenotype of the mutant occurred after extended time in stationary phase . After approximately 16h in stationary phase, both strains underwent an apparent replication event, after which the wild-type strain died rapidly whereas the atlA mutant survived considerably longer . Even after both the wild-type and mutant cells were dead, many of the mutant cells maintained intact morphology, whereas the wild-type cells were lysed . These results suggest that AtlA is a peptidoglycan transglycosylase related to bacteriophage endolysins and acts as an autolysin in the stationary phase.

Eur J Biochem, 1997 Oct 1, 249(1), 187 - 94
Characterization of a recombinant Neisseria meningitidis alpha-2,3-sialyltransferase and its acceptor specificity; Gilbert M et al.; The structure and specificity of the recombinant alpha-2,3-sialyltransferase from Neisseria meninigitidis are reported . This enzyme showed an unusual acceptor specificity in that it could use alpha-terminal and beta-terminal Gal residues as acceptors . In addition (beta1-->4)-linked and (beta1-->3)-linked terminal Gal served as acceptors . These properties distinguish the bacterial enzyme from the more widely investigated mammalian equivalents . The protein was expressed as a membrane-associated protein in Escherichia coli at a level of 750 U/l (approximately 250 mg/l) . The protein could be extracted with buffers containing 0.2% Triton X-100 and purified to homogeneity using immobilized-metal-affinity chromatography . Electrospray-ionization mass spectrometry of peptides obtained by cleavage with cyanogen bromide and trypsin confirmed over 95% of the deduced amino acid sequence . When used for enzymatic synthesis in coupled reactions with recombinant CMP-Neu5Ac synthetase, the alpha-2,3-sialyltransferase could sialylate fluorescent derivatives of N-acetyllactosamine with N-acetylneuraminic acid, N-propionylneuraminic acid and N-glycoloylneuraminic acid.

J Infect Dis, 1997 Nov, 176(5), 1269 - 76
High-level tetracycline-resistant Neisseria gonorrhoeae in Ontario, Canada--investigation of a cluster of isolates, showing chromosomally mediated resistance to penicillin combined with plasmid-mediated resistance to tetracycline; Harnett N et al.; Between 1991 and 1994, plasmid-mediated, tetracycline-resistant Neisseria gonorrhoeae (TRNG) increased from 61.8% to 85.96% of all resistant isolates in Ontario, Canada . Ninety-nine isolates with tetracycline MICs >32 mg/L were characterized by auxotype/serovar (A/S) class, plasmid profile, hybridization with eight tetracycline-resistant probes, and pulsed-field gel electrophoresis (PFGE) of genomic DNA after digestion with NheI and SpeI restriction endonucleases . A cluster of 82 isolates with penicillin MICs of 2-4 mg/L and tetracycline MICs of 128 mg/L (chromosomally mediated resistance) belonged to A/S class NR/IB-1 and had identical or closely related PFGE profiles . Seventeen isolates, TRNG (10) and penicillinase-producing TRNG (7), with tetracycline MICs of 64-256 mg/L, belonged to eight A/S classes and displayed 12 different PFGE profiles . The 99 isolates hybridized only with the TetM probe . Phenotypic and molecular characterization indicated a diverse population throughout the Province of Ontario.

Infect Immun, 1997 Nov, 65(11), 4770 - 7
Display of a PorA peptide from Neisseria meningitidis on the bacteriophage T4 capsid surface; Jiang J et al.; The exterior of bacteriophage T4 capsid is coated with two outer capsid proteins, Hoc (highly antigenic outer capsid protein; molecular mass, 40 kDa) and Soc (small outer capsid protein; molecular mass, 9 kDa), at symmetrical positions on the icosahedron (160 copies of Hoc and 960 copies of Soc per capsid particle) . Both these proteins are nonessential for phage infectivity and viability and assemble onto the capsid surface after completion of capsid assembly . We developed a phage display system which allowed in-frame fusions of foreign DNA at a unique cloning site in the 5' end of hoc or soc . A DNA fragment corresponding to the 36-amino-acid PorA peptide from Neisseria meningitidis was cloned into the display vectors to generate fusions at the N terminus of Hoc or Soc . The PorA-Hoc and PorA-Soc fusion proteins retained the ability to bind to the capsid surface, and the bound peptide was displayed in an accessible form as shown by its reactivity with specific monoclonal antibodies in an enzyme-linked immunosorbent assay . By employing T4 genetic strategies, we show that more than one subtype-specific PorA peptide can be displayed on the capsid surface and that the peptide can also be displayed on a DNA-free empty capsid . Both the PorA-Hoc and PorA-Soc recombinant phages are highly immunogenic in mice and elicit strong antipeptide antibody titers even with a weak adjuvant such as Alhydrogel or no adjuvant at all . The data suggest that the phage T4 hoc-soc system is an attractive system for display of peptides on an icosahedral capsid surface and may emerge as a powerful system for construction of the next generation multicomponent vaccines.

J Clin Microbiol, 1997 Nov, 35(11), 2985 - 8
Development of an antimicrobial susceptibility surveillance system for Neisseria gonorrhoeae in Malawi: comparison of methods; Daly CC et al.; Susceptibility of Neisseria gonorrhoeae to gentamicin, the primary treatment for gonorrhea in Malawi since 1993, was determined by using agar dilution MICs, E-test MICs, disc diffusion, and clinical cure rate . Agar dilution MICs were slightly higher in 1996 than in 1993 isolates, with a concomitant drop in the clinical cure rate . E-test MICs were substantially lower than agar dilution determinations, with only 77.4% within 1 log2 concentrationPIP: Gentamicin (240 mg single intramuscular dose) was adopted in 1993 as the treatment of choice for gonorrhea in Malawi, following findings of high-level gonococcal resistance to penicillin, tetracycline, and streptomycin . This study sought to assess the clinical efficacy of gentamicin in the treatment of Neisseria gonorrhoeae 2 years after the onset of its widespread use and to evaluate the potential roles of the disc diffusion and E test in the development of a surveillance system for monitoring antimicrobial susceptibility . 206 consecutive men presenting to a sexually transmitted disease clinic in Lilongwe in 1996 and 198 urethritis patients treated at a hospital in Blantyre during 1992-93 were enrolled . The clinical cure rate for gentamicin treatment of gonorrhea was 91.8% in 1996 compared to 95.0% in 1993 . The proportion of highly susceptible isolates declined from 17.2% in 1993 to 9.5% in 1996, but the majority of isolates remained in the moderately susceptible category . The E test yielded substantially lower minimum inhibitory concentrations (MICs) than the standard agar dilution assay, with only 77.4% within 1 log2 concentration . Disc diffusion was not helpful in determining the efficacy of gentamicin, since there was little difference in the range of disc diameters at different MICs as determined by agar dilution . Establishment of an MIC that clearly shows resistance to gentamicin was not possible . These findings suggest that the agar dilution assay remains the method of choice for MIC determination, although the less expensive E test may be capable of monitoring antibiotic efficacy under field conditions .

Clin Rheumatol, 1997 Sep, 16(5), 450 - 3
Osteoarticular and muscle infectious lesions in patients with the human immunodeficiency virus; Belzunegui J et al.; Between 1988 and 1995, 1832 HIV positive patients were evaluated in our institution . We studied the epidemiologic, immunologic and bacteriologic data, laboratory tests, and X-Ray films in those with musculoskeletal infection . We reviewed twenty-one cases of musculoskeletal infection in twenty patients aged 23-35 years (mean 28,6 years, M:F= 15:5) . In all of them risk factor for HIV was intravenous drug abuse . The number of CD4 positive lymphocytes ranged from 0,003 to 0,5 10(9)/l . Staphylococcus aureus was the organism responsible of the infection in twelve cases, all active intravenous drug abusers at the time the diagnosis was done . The remaining causative agents were: Mycobacterium tuberculosis (3 cases), Candida albicans (2 cases), Salmonella subgroup 1 (1 case), Neisseria gonorrhoeae (1 case), Pseudomona aeruginosa (1 case) and Streptococcus agalactiae (1 case) . Fifteen infections were diagnosed between 1988 and 1991 and 6 between 1992 and 1995 . Musculoskeletal infectious lesions in HIV positive patients in our country are related in the majority of cases to intravenous drug abuse . In the last four years due to a National medical health care plan conducted to educate this group of people the number of musculoskeletal infections is decreasing.

Diagn Microbiol Infect Dis, 1997 Aug, 28(4), 193 - 200
Antimicrobial activity of trovafloxacin tested against ciprofloxacin-susceptible and -resistant Neisseria gonorrhoeae . Interpretive criteria and comparisons with Etest results; Jones RN et al.; Trovafloxacin, a new fluorinated naphthyridine, has enhanced activity against Gram-positive cocci, while retaining an excellent spectrum against Gram-negative pathogens . It has been used successfully in clinical trials for therapy of gonorrhea, and this investigation proposes in vitro susceptibility testing criteria for trovafloxacin . A total of 150 Neisseria gonorrhoeae clinical isolates (50 resistant to ciprofloxacin; MICs > or = 0.12 microgram/mL) were tested by methods recommended by the National Committee for Clinical Laboratory Standards (NCCLS) and the Etest (AB BIODISK, Solna, Sweden) . Trovafloxacin was very active against gonococci (MIC90, 0.008 to 0.015 microgram/mL), but was generally eightfold less potent versus ciprofloxacin-resistant strains . Etest results correlated well (r = 0.96; 98% of MICs +/- one log2 dilution) compared to the reference agar dilution test . Reference agar dilution and Etest MICs were compared to disk-diffusion test zones (10-micrograms trovafloxacin disk), and excellent categorical agreement (89.4 to 99.3%) was achieved without significant false-susceptible or -resistant error (< or = 1.3%) . Tentative breakpoints were suggested initially to outline the ciprofloxacin-susceptible and trovafloxacin-susceptible as susceptible (MIC, < or = 0.015 microgram/mL; zones > or = 47 mm), and strains with various well-characterized mutations of the gyr A and par C genes as either intermediate or resistant to trovafloxacin . When the results of clinical studies treating ciprofloxacin-resistant N . gonorrhoeae with trovafloxacin become available, the alternative breakpoints could be utilized for resistant MIC breakpoints of > or = 0.06 microgram/mL or > or = 0.5 microgram/mL . Trovafloxacin was stable in supplemented GC agar for 21 days stored at refrigerated temperatures . These in vitro results indicate that trovafloxacin should be a very acceptable agent for therapy of gonorrhea and other common sexually transmitted pathogens.

Chirurg, 1997 Jun, 68(6), 591 - 5
{Current status of vaccination and antibiotic prophylaxis in splenectomy . II: Children}; Funk EM et al.; Children are exposed to a greater risk than adults for severe late postsplenectomy infection . Therefore, prophylaxis against bacterial infections need to be more extensive . This paper presents a protocol for preventive measures in children . Repeated education of the patient and his/her parents about the consequences of splenic loss is mandatory . Vaccinations against Streptococcus pneumoniae and Hemophilus influenzae are highly recommended and also against Neisseria meningitidis in certain situations . The importance of long-term antibiotic prophylaxis in children is emphasized . Recent advances in vaccine development and the increasing problems with antibiotic resistance are discussed.

Chirurg, 1997 Jun, 68(6), 586 - 90
{The current status of vaccination and antibiotic prophylaxis in splenectomy . I: Adults}; Funk EM et al.; The prevention of severe late postsplenectomy infection by vaccination and antibiotic prophylaxis is controversial . This paper reviews the literature and presents a protocol for postsplenectomy adults . Prophylaxis should be adapted to the underlying disease . Every patient has to be informed about the consequences of splenectomy . Depending on the underlying disease, vaccination is recommended against pneumococci, Hemophilus influenzae and Neisseria meningitidis . Long-term antibiotic prophylaxis is not advised in adults.

Biotechniques, 1997 Oct, 23(4), 714 - 6, 718-20
Specific inhibition of PCR by non-extendable oligonucleotides using a 5' to 3' exonuclease-deficient DNA polymerase; Yu D et al.; The Stoffel fragment of Taq DNA polymerase lacks the 5' to 3' exonuclease activity that hydrolyzes potentially blocking DNA strands during primer extension . We there-fore asked whether by using this fragment in the PCR, non-extendable, base-paired oligonucleotides could inhibit amplification in a sequence-dependent manner . Model targets were chosen from the partially conserved ribosomal 16S rDNA of three bacterial species: E . coli, Bacillus subtilis and Neisseria gonorrhoea . A single pair of primers was capable of amplifying a homologous 240-bp region from all three . Two non-extendable "blocking" oligonucleotides were synthesized with sequences complementary to the inter-primer regions of E . coli and B . subtilis, respectively . Both blockers were shown specifically to prevent amplification of their complementary targets, but not of the reciprocal control targets or of the reciprocal control targets or of the non-complementary N . gonorrhea . Specificity was further confirmed by an internal positive control . Similar inhibition was seen with mixtures of targets in a single reaction . With intact Taq DNA polymerase, no blocking was observed . Primers and blockers targeting specific regions of N . gonorrhoea rDNA were used to confirm the requirement that blockers be directed to the inter-primer region . Sequence-dependent amplification inhibition, such as that demonstrated here, would be applicable to PCR-related strategies using primers capable of using multiple targets, where such selective inhibition could be useful.

AIDS, 1997 Oct, 11(12), F85 - 93
The associations between cervicovaginal HIV shedding, sexually transmitted diseases and immunosuppression in female sex workers in Abidjan, Côte d'Ivoire; Ghys PD et al.; OBJECTIVE: To measure the frequency and associated factors of cervicovaginal HIV shedding and to determine the impact of sexually transmitted disease (STD) treatment on HIV shedding . DESIGN: Cross-sectional study with 1-week follow-up . SETTING: Confidential clinic for female sex workers in Abidjan, Cote d'Ivoire . PARTICIPANTS: A total of 1201 female sex workers . INTERVENTIONS: STD treatment based on clinical signs . MAIN OUTCOME MEASURES: HIV serostatus; cervicovaginal HIV shedding at enrollment and at 1-week follow-up; STD status at enrollment and at 1-week follow-up . RESULTS: Cervicovaginal shedding of HIV-1 in HIV-1-seropositive women was more frequent (96 out of 404, 24%) than shedding of HIV-2 in HIV-2-seropositive women {one out of 21, 5%; odds ratio (OR), 6.2; 95% confidence interval (CI), 1.0-261} . Among 609 HIV-1-seropositive or dually seroreactive women, HIV-1 shedding was significantly more frequent in immunosuppressed women {adjusted OR (AOR), 6.3; 95% CI, 3.4-11.9; and AOR, 2.9; 95% CI, 1.6-5.0 for CD4 < 14% and CD4 14-28%, respectively, versus CD4 > 28%}, and in women with Neisseria gonorrhoeae (AOR, 1.9; 95% CI, 1.2-3.0), those with Chlamydia trachomatis (AOR, 2.5; 95% CI, 1.1-5.8), and with a cervical or vaginal ulcer (AOR, 3.9; 95% CI, 2.1-7.4) . HIV-1 shedding decreased from 42 to 21% (P < 0.005) in women whose STD were cured . CONCLUSIONS: These data help to explain the difference in transmissibility between HIV-1 and HIV-2 and the increased infectiousness of HIV in the presence of immunosuppression and STD . In addition, they lend biological plausibility to arguments for making STD control an integral part of HIV prevention strategies in Africa.

Zh Mikrobiol Epidemiol Immunobiol, 1997 Jul-Aug, (4), 96 - 8
{The antilysozyme activity of the causative agent in the diagnosis and prognosis of the course of gonococcal infection}; Voronina LG; Differences in the frequency and level of antilysozyme activity in Neisseria gonorrhoeae cultures isolated from patients with different forms and stages of gonococcal infection were revealed . The possibility of using the antilysozyme sign of N . gonorrhoeae for the differential diagnostics of the clinical forms of this infection and for prognosticating the chronization of the inflammatory process in newly infected patients and in cases of relapses in chronic gonorrhea patients.

Zh Mikrobiol Epidemiol Immunobiol, 1997 Jul-Aug, (4), 73 - 7
{The characteristics of the hydrophobic properties of bacteria when interacting with blood serum}; Brudastov IuA et al.; Considerable variability of the hydrophobic properties of bacterial surface was demonstrated on 132 representatives of Escherichia coli, Neisseria, staphylococci and enterococci, belonging to museum strains and isolated from clinical material . Taxonomic and pathovariant differences in their susceptibility to hydrophobic modification induced by the thermostable and thermolabile components of blood serum were experimentally established . Extracellular bacterial products were found to be capable of preventive and restorative counteraction to such hydrophobic modifications . The antilysozyme and anticomplement activities of bacteria were shown to take part in the expression of hydrophobic/hydrophilic properties in the interaction of bacteria with blood serum.

Zh Mikrobiol Epidemiol Immunobiol, 1997 Jul-Aug, (4), 63 - 7
{The anticomplement activity of Neisseria gonorrhoeae}; Akhunova NR et al.; Bacteria of the species N . gonorrhoeae have anticomplementary activity whose absolute values exceed the level of this activity both in other representatives of the genus Neisseria and in microorganisms of other taxonomic groups found in the microbiocenosis of the reproductive tract . The specificity of the anticomplementary action of N . gonorrhoeae extracellular products with respect to individual components of the complement system, as well as their role in the formation of the state of seroresistance and in the determination of the effectiveness of the interaction of gonococci with neutrophilic phagocytes in the system of opsonic cooperation, have been characterized.

Zh Mikrobiol Epidemiol Immunobiol, 1997 Jul-Aug, (4), 102 - 5
{The suppressive action of a magnetic-laser ray and of an electrolytic solution of sodium hypochlorite on the factors of causative agent persistence}; Usviatsov BIa et al.; The suppressive action of a magnet-laser ray and electrolyzed sodium hypochlorite solution on the persistence factors (antilysozyme, "anti-interferon") of Staphylococcus aureus and Neisseria gonorrhoeae is shown . The optimum conditions (time, dose, concentration) for the regulation of the persistence properties of the pathogens has been determined . The use of physicochemical factors in the proposed parameters has been shown to be effective for the therapy of purulent inflammatory diseases and the sanitation of S.aureus carriers.

Sex Transm Dis, 1997 Oct, 24(9), 495 - 502
Chlamydial and gonococcal cervicitis in HIV-seropositive and HIV-seronegative pregnant women in Bangkok: prevalence, risk factors, and relation to perinatal HIV transmission; Chaisilwattana P et al.; OBJECTIVES: To determine the prevalence and risk factors associated with cervicitis caused by Chlamydia trachomatis and Neisseria gonorrhoeae in human immunodeficiency virus (HIV) type 1-seropositive and HIV-seronegative pregnant women in Bangkok, and the relation to perinatal HIV transmission . METHODS: As part of a multicenter perinatal HIV transmission study in an antenatal population with 2% HIV seroprevalence, endocervical swabs obtained at mid-pregnancy from a consecutive sample of 222 HIV-seropositive and 219 HIV-seronegative pregnant women at two large hospitals in Bangkok were tested for the presence of C . trachomatis and N . gonorrhoeae by DNA hybridization probe (Gen-Probe) . Clinical risk factors and DNA probe results were analyzed in relation to the women's and newborns' HIV infection status . RESULTS: The prevalence of C . trachomatis was 16.2% in HIV-seropositive pregnant women and 9.1% in HIV-seronegative pregnant women (P = 0.03) . The prevalence of N . gonorrhoeae was 2.7% in HIV-seropositive pregnant women and 1.4% in HIV-seronegative pregnant women (P = 0.5) . The overall population prevalence estimate was 9.2% for C . trachomatis and 1.4% for N . gonorrhoeae . Women with gonococcal infection were more likely to be positive for C . trachomatis (RR(MH) = 5.2, P < 0.01) . Young age (<21 years) and primigravid status were associated with C . trachomatis infection among HIV-seropositive women; history of multiple sex partners (>1) were associated with C . trachomatis infection among HIV-seronegative women . For HIV-seropositive women, primigravida status also was associated with C . trachomatis infection . The perinatal HIV transmission rates were similar for those with and without C . trachomatis (24.1% and 23.2%, P = 0.9) and among those with and without N . gonorrhoeae (20% and 23.5%, P = 1.0) . CONCLUSIONS: Among pregnant women in Bangkok, C . trachomatis infection was considerably more common than N . gonorrhoeae infection and was associated with HIV infection, young age and first pregnancy (HIV-seropositive women), and multiple partners (HIV-seronegative women) . Our data do not suggest an association between perinatal HIV transmission and maternal C . trachomatis or N . gonorrhoeae infection identified and treated during pregnancy . The high prevalence of C . trachomatis found using a test not readily available in Thailand emphasizes the need for improved, inexpensive ways to screen for and diagnose these sexually transmitted infections in developing countriesPIP: Numerous studies have suggested that Chlamydia trachomatis and Neisseria gonorrhoeae facilitate heterosexual HIV transmission; the impact of these sexually transmitted diseases (STDs) on perinatal HIV transmission is unknown, however, due to the expense of routine screening for STDs during pregnancy in developing countries . As part of a multicenter perinatal HIV transmission study, 222 HIV-positive and 219 HIV-negative women presenting for prenatal care at 2 hospitals in Bangkok, Thailand, during 1993-94 were enrolled . At mid-pregnancy, endocervical swabs were obtained and tested for the presence of C . trachomatis and N . gonorrhoeae by DNA hybridization probe . There were 36 cases (16.2%) of C . trachomatis infection among HIV-positive women and 20 cases (9.1%) among HIV-negative women . There were 6 cases (2.7%) of N . gonorrhoeae among HIV-positive women and 3 cases (1.4%) among HIV-negative women . Based on an estimated antenatal HIV seroprevalence of 2%, these findings imply a general antenatal prevalence of 9.2% for C . trachomatis and 1.4% for N . gonorrhoeae . Women with gonococcal infection were more likely (relative risk, 5.2) to be positive for C . trachomatis as well . C . trachomatis infection among HIV-infected pregnant women was associated with age under 21 years and primigravidity . The overall perinatal HIV transmission rate was 24.2%, with no significant difference according to STD infection status . However, since all women diagnosed with STDs received treatment by the mid-third trimester of pregnancy, it remains possible that untreated STDs facilitate perinatal HIV transmission . The high prevalence of C . trachomatis detected in this study through use of a test not readily available in Thailand emphasizes the need for inexpensive, reliable methods to screen for STDs among pregnant women in developing countries .

J Antimicrob Chemother, 1997 Sep, 40(3), 437 - 9
In-vitro antimicrobial activity of HSR-903, a new fluoroquinolone, against clinical isolates of Neisseria gonorrhoeae with quinolone resistance-associated alterations in GyrA and ParC; Deguchi T et al.; The in-vitro antimicrobial activity of HSR-903, a new fluoroquinolone, was tested against 51 clinical Neisseria gonorrhoeae isolates in comparison with ciprofloxacin, levofloxacin and sparfloxacin . The MICs of HSR-903 for 11 isolates with alterations in both GyrA and ParC, for 19 isolates with alterations only in GyrA and for 21 isolates without alterations in either GyrA or ParC ranged from 0.03 mg/L to 1.0 mg/L (MIC90 = 0.25 mg/L), from 0.03 mg/L to 0.5 mg/L (MIC90 = 0.125 mg/L) and from < or = 0.001 mg/L to 0.008 mg/L (MIC90 = 0.004 mg/L), respectively . Levofloxacin and ciprofloxacin were the least active of the four quinolones tested, particularly against the mutant strains . Sparfloxacin was more active, but HSR-903 exhibited the most potent in-vitro activity against the clinical N . gonorrhoeae isolates, including those harbouring quinolone-resistance-associated alterations in GyrA and ParC.

Acad Emerg Med, 1997 Oct, 4(10), 962 - 7
Underrecognition of cervical Neisseria gonorrhoeae and Chlamydia trachomatis infections in the emergency department; Yealy DM et al.; OBJECTIVES: 1) To quantify the frequency of underrecognized Neisseria gonorrhoeae and Chlamydia trachomatis cervical infections in women tested in the ED, 2) to describe and compare the characteristics of those treated and not treated during the initial visit, and 3) to quantify the delay interval until treatment was provided . METHODS: A 2-year, retrospective consecutive case series was performed from June 1, 1992, to May 31, 1994 . There were 148 women with > or = 1 discrete occurrence of culture-proven cervical N . gonorrhoeae or C . trachomatis infection studied . All the patients were evaluated in a university-affiliated, tertiary care hospital-based ED with a large rural referral area . The main outcome measures were the proportions of patients with positive cultures both treated and not treated in the ED, the clinical characteristics of each group, and the proportion remaining untreated or experiencing treatment delays of > 2 weeks after attempted phone, mail, and public health follow-up . RESULTS: Of 157 occurrences of positive cultures for N . gonorrhoeae or C . trachomatis, 86 (53%) were treated with a regimen suggested by the CDC prior to ED release . The proportion of women with isolated C . trachomatis infections that were underrecognized and untreated initially was larger than the proportions with isolated N . gonorrhoeae or combined infections (79% vs 27% and 53%, respectively, p < 0.0001) . Women with findings suggestive of advanced disease (history of fever or chills, or examination evidence of temperature > 38 degrees C, purulent vaginal discharge, or any uterine/salpinx/ovarian tenderness) were more often recognized and treated with appropriate antibiotics initially (p = 0.02 to < 0.00001 for all) . After phone, mail, and public health follow-up, treatment could not be documented for 25% of the occurrences, in all cases due to an inability to locate the patient . An additional 20% of the women did not receive treatment for 14-60 days . CONCLUSIONS: In this population, both N . gonorrhoeae and C . trachomatis cervical infections are frequently underrecognized in the ED, with isolated C . trachomatis infections associated with significantly higher proportions of underrecognition . Many affected women remain untreated for extended intervals, creating public and individual health risks . Improved point of contact detection, follow-up, and treatment policies are needed to limit these risks.

Biol Pharm Bull, 1997 Sep, 20(9), 1010 - 2
Evaluation of the standardized disk diffusion and agar dilution antibiotic susceptibility test methods by using strains of Neisseria gonorrhoeae from Tucumán, Argentina; de Castillo MC et al.; At present, most Neisseria gonorrhoeae testing is done with beta-lactamase and agar dilution tests using common therapeutic agents . Generally, in bacteriological diagnosis laboratories in Argentina, study of antibiotic susceptibility of N . gonorrhoeae is based on beta-lactamase determination and agar dilution method using common therapeutic agents . The National Committee for Clinical Laboratory Standards (NCCLS) recently described a disk diffusion test that produces results similar to the reference agar dilution method for antibiotic susceptibility of N . gonorrhoeae . We obtained 57 gonococcal isolates from patients attending a clinic for sexually transmitted diseases in Tucuman, Argentina . Antibiotic susceptibility tests using agar dilution and disk diffusion techniques were compared . The established NCCLS interpretive criteria for both susceptibility methods appeared to be applicable to domestic gonococcal strains . The correlation between the minimum inhibitory concentration (MIC's) and the zones of inhibition was studied for penicillin, ampicillin, cefoxitin, spectinomycin, cefotaxime, cephaloridine, cephalexin, tetracycline, norfloxacin and kanamycin . Dispersion diagrams showed a high correlation between both methods, with a sensitivity of 89% and specificity of 91%.

Genitourin Med, 1997 Jun, 73(3), 194 - 7
Serological and clinical correlates of gonorrhoea and syphilis in fertile and infertile Nigerian women; Okonofua FE et al.; OBJECTIVE: To test the hypothesis that infertile Nigerian women have higher serum levels of antibodies against Neisseria gonorrhoea and Treponema pallidum compared with fertile controls . DESIGN: The prevalence of serum antibodies against N gonorrhoea and T pallidum was compared in fertile and infertile Nigerian women . SETTING: Population based case-control study in Ile-Ife, southwestern Nigeria . SUBJECTS: 60 women with infertility identified from a community based questionnaire survey of 1075 women were compared with 53 age matched fertile controls . METHODS: Sera of fertile and infertile women were tested for the presence of gonococcal antibodies with indirect agglutination test and syphilis antibodies using rapid reagin method . MAIN OUTCOME MEASURES: Prevalence of anti-gonococcal and anti-treponemal antibodies in cases and controls . Frequency of self reports of sexually transmitted infections (STIs) in cases and controls . RESULTS: In comparison with fertile women, infertile women were more likely to report having had repeated lower abdominal pains (p < 0.01), yellow vaginal discharge (p < 0.004), and whitish vaginal discharge (p < 0.02) . There was no significant difference between cases and controls in the proportions reporting previous STI diagnoses . However, two infertile women reported previous gonococcal infection compared with none in the fertile group . Sixteen of the infertile women (26.7%) demonstrated anti-gonococcal antibodies in their sera compared with only four of the 53 fertile controls (7.5%) (p < 0.02; OR 4.5) . There was no significant difference between fertile and infertile women in the proportion showing serological reactivity to T pallidum . CONCLUSION: Infertile women have a higher prevalence of anti-gonococcal antibodies compared with fertile controls . Infertile women are also more likely to report previous lower abdominal pains and vaginal discharge . These results provide credible evidence implicating STIs and N gonorrhoea in particular as important factors contributing to female infertility in this population . Public health measures are warranted to address the high rate of STIs and N gonorrhoea in Nigeria.

Am J Obstet Gynecol, 1997 Sep, 177(3), 680 - 4
Antimicrobial prophylaxis in pregnancy: a randomized, placebo-controlled trial with cefetamet-pivoxil in pregnant women with a poor obstetric history; Gichangi PB et al.; OBJECTIVE: This study was undertaken to measure the impact of a single oral dose of cefetamet-pivoxil on pregnancy outcome in a population with substantial rates of low birth weight and high prevalence rates of maternal infections . STUDY DESIGN: A total of 320 pregnant women with a poor obstetric history, defined as a history of low birth weight or stillbirth, were randomized to receive a single oral dose of 2 gm of cefetamet-pivoxil or a placebo at a gestational age between 28 and 32 weeks . Patients were assessed at delivery and 1 week post partum for pregnancy outcome, postpartum endometritis, human immunodeficiency virus-1 and gonococcal infections . RESULTS: A total of 253 (79%) women gave birth at the maternity hospital, of whom 210 (83%) attended the follow-up clinic . Overall, 18.1% of these pregnant women were human immunodeficiency virus-1 seropositive, whereas 9.5% had antibodies against Treponema pallidum . There was a significant difference between cefetamet-pivoxil- and placebo-treated women in infant birth weight (2927 gm vs 2772 gm, p = 0.03) and low birth weight (< 2500 gm) rates (18.7% vs 32.8%, p = 0.01, odds ratio 2.1, 95% confidence interval 1.2 to 3.8) . The stillbirth rate was 2.2% in the cefetamet-pivoxil group and 4.2% in the placebo group (not significant) . Postpartum endometritis was found in 17.3% in the intervention arm versus 31.6% in the placebo group (p = 0.03, odds ratio 2.2, 95% confidence interval 1.1 to 7.6) . Neisseria gonorrhoeae was isolated from the cervix in 5 of 103 (4.9%) women in the intervention and in 14 of 101 (13.9%) in the placebo group (p = 0.04, odds ratio 3.2, 95% confidence interval 1.1 to 10.5) . CONCLUSION: A single oral dose of cefetamet-pivoxil administered to pregnant women with a poor obstetric history seemed to improve pregnancy outcome in this population with high rates of maternal infections . Larger studies should be carried out to examine the public health impact, the feasibility, and the overall cost/benefit ratio of this intervention.

Infect Immun, 1997 Oct, 65(10), 4122 - 9
Differential response of human monocytes to Neisseria gonorrhoeae variants expressing pili and opacity proteins; Knepper B et al.; Experiments in vitro suggest that Neisseria gonorrhoeae surface variation plays a key role in gonococcal pathogenesis by providing the appropriate bacterial phenotypes to go through different stages of the infection . Here we report on the effects of phase and antigen variation of two major gonococcal adhesins, pili and opacity (Opa) outer membrane proteins, on the interaction of the gonococci with human monocytes . Using a set of recombinants of gonococcus strain MS11 that each express 1 of 11 genetically defined Opa proteins or a defined type of pilus, we found that both Opa proteins and pili promote bacterial phagocytosis by monocytes in the absence of serum and that this feature largely depends on the type of protein that is expressed . One of the Opa proteins (Opa{50}) strongly promoted uptake by monocytes but had little effect on the interaction with polymorphonuclear leukocytes under the conditions employed . Similarly, the phagocytosis-promoting effect of the pili was much more pronounced in monocytes than in neutrophils (4-fold versus 22-fold stimulation of uptake, respectively) . Only a subpopulation of both types of phagocytes actively ingested bacteria, as has been observed during natural infections . Measurements of luminol-enhanced chemiluminescence demonstrated that phagocytosis of opaque but not piliated gonococci was accompanied by an increase in oxygen-reactive metabolites . These findings demonstrate that the monocyte response towards gonococci is highly dependent on the bacterial phenotype and differs from the neutrophil response . This diversity in bacterial behavior towards various types of human phagocytic cells underlines the biological impact of gonococcal surface variation and may explain previous contradictory results on this subject.

J Clin Microbiol, 1997 Oct, 35(10), 2556 - 60
Multiplex AMPLICOR PCR screening for Chlamydia trachomatis and Neisseria gonorrhoeae in women attenting non-sexually transmitted disease clinics . The European Chlamydia Epidemiology Group; Bassiri M et al.; A new PCR kit (AMPLICOR CT/NG; Roche Diagnostic Systems, Inc., Branchburg, N.J.) was used as a screening tool for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in first-void urine (FVU) specimens from 3,340 asymptomatic women attending European health care units for contraceptive advice or pregnancy termination . All samples were kept frozen (-20 degrees C) prior to testing . Chlamydia-positive samples were retested once by the plasmid-based PCR kit and also by a major outer membrane protein (MOMP) primer-based PCR . Discrepancies were resolved by using the direct immunofluorescence test (DIF) with the centrifuged sediment of the FVU specimens . Samples positive for N . gonorrhoeae were retested by chromosomal primer-based PCR and verified by a 16S RNA PCR . Of the samples tested, 1.8% were considered inhibitory by using the internal amplification control . Of 81 samples positive for C . trachomatis, 74 samples were positive by both plasmid- and MOMP-based PCRs, 6 samples were positive by plasmid-based PCR and DIF, and one sample was positive by both MOMP-based PCR and DIF . Nine samples (0.3%) were positive for N . gonorrhoeae by the chromosomal primer-based PCR; however, none of the results could be confirmed . The test offers the unique ability to identify inhibition of amplification with the optional internal control.

Mol Microbiol, 1997 Aug, 25(4), 639 - 47
Membrane cofactor protein (MCP or CD46) is a cellular pilus receptor for pathogenic Neisseria; Kallstrom H et al.; Pili of Neisseria gonorrhoeae and Neisseria meningitidis mediate binding of the bacteria to human cell-surface receptors . We found that purified pili bound to a 55- to 60-kDa doublet band on SDS-PAGE of separated human epithelial cell extracts . This is a migration pattern typical of membrane cofactor protein (MCP or CD46) . MCP is a widely distributed human complement regulatory protein . Attachment of the bacteria to epithelial cells was blocked by polyclonal and monoclonal antibodies directed against MCP, suggesting that this complement regulator is a receptor for piliated Neisseria . We proved this hypothesis by demonstrating that piliated, but not non-piliated, gonococci bound to CHO cells transfected with human MCP-cDNA . We also demonstrated a direct interaction between purified recombinant MCP and piliated Neisseria . Finally, recombinant MCP protein produced in E . coli inhibited attachment of the bacteria to target cells . Taken together, our data show that MCP is a human cell-surface receptor for piliated pathogenic Neisseria.

Vaccine, 1997 Aug-Sep, 15(12-13), 1344 - 8
Neisseria gonorrhoeae: vaginal clearance and its correlation with resistance to infection in subcutaneous chambers in orally immunized estradiol-primed mice; Arko RJ et al.; The rate of clearance of Neisseria gonorrhoeae in vaginal specimens harvested from inbred Balb/c and outbred (Institute of Cancer Research {ICR}) white mice, previously primed with saline or beta-estradiol and then challenged with live gonococci, was determined . Survival of gonococci was significantly (P < 0.05) better in estradiol-primed mice from both inbred and outbred lines at 24 h after challenge . Vaginal clearance of gonococci from ICR mice parenterally primed with a gonococcal PI-B synthetic peptide and then orally immunized with gamma irradiated, protein-III deficient gonococcal cells was significantly (P < 0.001) faster than in corresponding nonimmunized, estradiol-primed controls . The more rapid vaginal clearance in the orally immunized mice was correlated with an elevated subcutaneous chamber ID50% (> 100,000 colony forming units) determined for the same groups of mice . Peptide-primed mice orally immunized with irradiated Escherichia coli cells demonstrated a chamber ID50% of < 100 CFU and were significantly (P < 0.01) slower to clear gonococci following vaginal challenge . High levels of specific IgA and IgG antibodies to gonococci were detected by an enzyme-linked immunosorbent assay in vaginal wash specimens from orally immunized mice . However, high antibody titers were not always predictive of an increased resistance to vaginal challenge . These models may be helpful in providing more economical and accessible in vivo methods for screening certain gonococcal vaccine candidates before more expensive testing in chimpanzees or humans.

Clin Diagn Lab Immunol, 1997 Sep, 4(5), 522 - 9
Heterogeneity of tbpB, the transferrin-binding protein B gene, among serogroup B Neisseria meningitidis strains of the ET-5 complex; Rokbi B et al.; ET-5 complex strains of Neisseria meningitidis were traced intercontinentally and have been causing hyperendemic meningitis on a worldwide scale . In an attempt to develop a fully broad cross-reactive transferrin-binding protein B (TbpB)-based vaccine, we undertook to assess the extent of variability of TbpB proteins among strains of this epidemiological complex . For this purpose, a PCR-based method was developed to study the heterogeneity of the tbpB genes from 31 serogroup B N . meningitidis strains belonging to the ET-5 complex . To define adequate primers, the tbpB gene from an ET-5 complex strain, 8680 (B:15:P1.3; isolated in Chile in 1987), was cloned and the nucleotide sequence was determined and compared to two other previously published tbpB sequences . A tbpB fragment was amplified from genomic DNA from each of the 31 strains . By this method, heterogeneity in size was observed and further characterized by restriction pattern analysis with four restriction enzymes and by sequencing tbpB genes from three other ET-5 complex strains . Four distinct tbpB gene types were identified . Fifty-five percent of the strains studied (17/31) harbored tbpB genes similar to that of strain BZ83 (B:15:-) isolated in The Netherlands in 1984 . Ten of the 31 strains (32.2%) had tbpB genes close to that of strain M982 . Only 3 of the 31 (9.6%) were found to harbor tbpB genes close to that of strain 8680, and finally one strain, 8710 (B:15:P1.3; isolated in Chile in 1987), was found to harbor a tbpB gene different from all the others . These results demonstrated a pronounced variability among tbpB alleles within a limited number of ET-5 complex strains collected over a 19-year period . Despite the genetic heterogeneity observed, specific antisera raised to purified Tbps from ET-5 complex strains showed broad cross-reactivity between different TbpBs both by Western blot analysis and bactericidal assay, confirming that a limited number of TbpB molecules included in a vaccine are likely to induce broadly cross-reactive antibodies against the different strains.

Pathol Biol (Paris), 1997 May, 45(5), 371 - 5
High-level tetracycline resistant Neisseria gonorrhoeae isolated in Portugal; Ferreira E et al.; The first high-level tetracycline resistance (MIC > or = 16 mg/l) isolates of Neisseria gonorrhoeae (TRNG) were reported in 1990 from patients attending a Sexual Transmitted Disease (STD) Center in Lisbon . The TRNG prevalence was 4% in 1991, 5.3% in 1992 and 10,8% in 1994, exploding to 52.2% in 1995 . The tet M determinant was evaluated by PCR . The digests of PCRP using HpaII produced the restriction pattern 2 for all the strains, except one (pattern 3) . 78.3% of the TRNG strains were beta-lactamase producers and the 4.5 MDa penicillinase plasmid was the dominant (83%), 90% and 93.3% of the TRNG strains belonged to the auxotype NR and to the serogroup IA, respectively . The IA-8/NR class represented 58.3% of the TRNG isolates, suggesting a clonal spreading.

Sex Transm Dis, 1997 Sep, 24(8), 449 - 55
Risk factors for urethritis in heterosexual men . The role of fellatio and other sexual practices; Schwartz MA et al.; BACKGROUND: Nonchlamydial nongonococcal urethritis (NGU) is a common sexually transmitted disease (STD) in heterosexual men . Prior studies have suggested that NGU may be acquired by insertive oral sex . GOAL: To assess the association of oral sex and other sexual practices with nonchlamydial NGU in heterosexual men in order to better understand this syndrome and to guide its prevention and treatment . Risk factors for urethral gonorrhea and chlamydial infection were explored to contrast with NGU . STUDY DESIGN: A retrospective case-control study was conducted among heterosexual men attending as STD clinic during 1993 and 1994 . The study included 4,848 men who were sexually active within the prior 2 months and had urethral specimens obtained for Gram's stain, culture for Neisseria gonorrhoeae, and culture for Chlamydia trachomatis . RESULTS: Insertive oral sex was not shown to be an independent risk factor for NGU . Independent predictors of nonchlamydial NGU by multivariate analysis included African-American race (odds ratio {OR} 3.71, 95% confidence interval {95% CI} 3.06 to 4.50) and having > or = two sex partners in the prior 2 months (OR 1.45, 95% CI 1.20 to 1.75) . History of using condoms "always" was negatively associated with NGU (OR 0.59, 95% CI 0.43 to 0.79), gonorrhea (OR 0.31, 95% CI 0.17 to 0.56), and chlamydial infection (OR 0.67, 95% CI 0.44 to 1.03) . CONCLUSIONS: This study supports the sexually transmitted nature of nonchlamydial NGU but did not confirm an association with oral sex . However, the analysis was compromised by the rarity of insertive oral sex as patients' only sexual exposure . Consistent condom use protects against all causes of sexually acquired urethritis.

Am J Obstet Gynecol, 1997 Aug, 177(2), 375 - 80
Use of antibiotics to prevent preterm birth; Gibbs RS et al.; Our purpose is to review recent data and provide a clinical opinion on the use of antibiotics to prevent preterm birth or related maternal-neonatal complications . A literature review and a synthesis of opinion are provided . During prenatal care, standard practices should be applied regarding Neisseria gonorrhoeae, Chlamydia trachomatis, and bacteriuria . In addition, screen for and treat bacterial vaginosis in patients at high risk for preterm birth but do not treat Ureaplasma urealyticum or group B streptococci genital colonization . With preterm labor and intact membranes, standard practices should be applied regarding group B streptococci prophylaxis . Do not give antibiotics routinely to prolong pregnancy, but in patients with bacterial vaginosis and Trichomonas vaginalis specific treatment should be given . With preterm premature rupture of membranes, standard practices should be applied regarding group B streptococci prophylaxis, but additional antibiotics should also be given to prolong pregnancies at 24 to 32 weeks' gestation . Reported adverse effects have been few to date . However, increased diligence is needed for resistant organisms . In selected clinical settings antibiotic therapy is now indicated to prolong pregnancy and prevent maternal-neonatal complications associated with preterm birth.

Infect Immun, 1997 Sep, 65(9), 3857 - 66
Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invasion of HeLa cells; Gomez-Duarte OG et al.; Neisseria gonorrhoeae induces local infections in the human genitourinary tract and can disseminate to other organs to cause severe disease . Blood-derived factors present in the genital mucosa have been suggested to facilitate the spread of N . gonorrhoeae in disseminated gonococcal infections . Using gentamicin invasion assays and confocal microscopy, we observed a strong stimulatory effect of fetal calf serum (FCS) on the gonococcal invasion of HeLa cells . FCS-mediated invasion was dependent on the expression of the epithelial cell invasion-associated Opa protein (plasmid-encoded Opa50 or its chromosomal homolog Opa30), while N . gonorrhoeae expressing noninvasive Opa proteins (Opa(51-60)) or no Opa protein (Opa-) was not invasive even in the presence of FCS . Incubation of N . gonorrhoeae MS11 with biotinylated FCS revealed a 78-kDa protein as the prominent protein binding to Opa50- or Opa30-expressing gonococci . This protein was recognized by antibodies against vitronectin (VN) in Western blots . Purified human or bovine VN efficiently bound to Opa50-expressing gonococci, while binding to noninvasive Opa- or Opa52-expressing gonococci was significantly lower . Binding of VN was inhibited by heparin in a concentration-dependent manner, indicating that the heparin binding sites present in VN or Opa50 may play an essential role in this interaction . Based on gentamicin invasion assays and confocal microscopy studies, VN binding was associated with an increased invasion of Opa50- and Opa30-expressing gonococci into HeLa cells . The ability of VN to mediate entry into epithelial cells may constitute an important event in the pathogenesis of local as well as disseminated gonococcal infections.

Infect Immun, 1997 Sep, 65(9), 3736 - 42
A proposed role for the lutropin receptor in contact-inducible gonococcal invasion of Hec1B cells; Spence JM et al.; We previously reported the existence of a contact-inducible, enhanced invasion phenotype in the obligate human pathogen Neisseria gonorrhoeae . Our present studies showed that the ability of glutaraldehyde-fixed eucaryotic cells to convert gonococci (GC) to this invasive phenotype (Inv+) is limited to cells derived from reproductive tissues . We present evidence that GC recognize the lutropin receptor (LHr), which recognizes both luteinizing hormone and human chorionic gonadotropin (hCG), as the tissue-specific environmental signal that induces the conversion of GC to the Inv+ phenotype . By competitive binding studies, we showed that Inv+ GC bind to Hec1B cells, a human endometrial cell line, by a unique adhesin not present on noninduced GC and that this Inv+ GC-specific binding is completely blocked by the addition of hCG . We demonstrated that limiting the access of GC to LHr decreases the ability of the host cell to both convert GC to the Inv+ phenotype and serve as a target for Inv+ GC invasion . We propose a model of GC invasion of Hec1B cells in which the LHr plays a dual role both as an induction signal and as part of the internalization mechanism . This utilization of LHr could account for both the preponderance of complicated GC disease in women and the observed correlation of the disease with the onset of menses.

J Clin Microbiol, 1997 Sep, 35(9), 2408 - 9
Tampon sampling for diagnosis of bacterial vaginosis: a potentially useful way to detect genital infections?
Wilkinson D, Ndovela N, Kharsany A, Connolly C, Sturm AW.
Genital tract infections are important causes of ill health in developing countries, but diagnosis is difficult . Bacterial vaginosis (BV) was correctly diagnosed by using a vaginal specimen obtained by tampon sampling in 22 of 24 women (91.6%) for whom BV was diagnosed by Gram staining . The yield for other vaginal infections was higher (28% for Trichomonas vaginalis and 32.7% for Candida albicans) than it was for cervical infections (0% for Neisseria gonorrhoeae and 30% for Chlamydia trachomatis) . Tampon sampling was acceptable to patients and may facilitate diagnosis of genital infections in developing countries.

Wien Klin Wochenschr, 1997 Aug 8, 109(14-15), 584 - 9
Mycoplasma hominis and Ureaplasma urealyticum in patients with sexually transmitted diseases; Koch A et al.; Mycoplasma hominis and Ureaplasma urealyticum can be isolated with considerable frequency from the human urogenital tract and are thought to cause various syndromes such as nongonococcal urethritis, pelvic inflammatory disease, pyelonephritis or infertility . The aim of this study was the evaluation of the presence of different genital pathogens in patients with sexually transmitted diseases (STD) and, in particular, the detection of mycoplasmas in individuals infected with genital microbes and an assessment of the presence of genital microorganisms in patients harbouring Mycoplasma hominis or Ureaplasma urealyticum . Furthermore, the occurrence of mycoplasmas in women with bacterial vaginosis was established . Specimens were collected from a total of 41,980 persons attending the Outpatients' Centre for Infectious Venero-Dermatological Diseases in Vienna from 1994 to 1996 . Of all genital pathogens, Ureaplasma urealyticum was cultured most frequently in men and women . Mycoplasma hominis and Ureaplasma urealyticum were detected more often in the vaginal fluid than in the male urethra . By contrast, infection rates with Neisseria gonorrhoeae and Chlamydia trachomatis were higher in men than in women . In both men and women, trichomoniasis increased colonisation with Mycoplasma hominis, while mycoplasmas occurred less frequently together with genital candidiasis . Mycoplasma hominis was cultivated significantly more often in women with bacterial vaginosis than in those without . In contrast to urethral infections in men, cervical infections with Neisseria gonorrhoeae or Chlamydia trachomatis raised the incidence of Mycoplasma hominis in the vaginal fluid.

J Biol Chem, 1997 Aug 8, 272(32), 19688 - 96
Shortened hydroxyacyl chains on lipid A of Escherichia coli cells expressing a foreign UDP-N-acetylglucosamine O-acyltransferase; Odegaard TJ et al.; The first reaction of lipid A biosynthesis in Gram-negative bacteria is catalyzed by UDP-N-acetylglucosamine (UDP-GlcNAc) O-acyltransferase, the product of the lpxA gene . The reaction involves the transfer of an acyl chain from hydroxyacyl-acyl carrier protein (ACP) to the glucosamine 3-OH position of UDP-GlcNAc . The lipid A isolated from Escherichia coli contains (R)-3-hydroxymyristate at the 3 and 3' positions . Accordingly, LpxA of E . coli is highly selective for (R)-3-hydroxymyristoyl-ACP over ACP thioesters of longer or shorter acyl chains . We now demonstrate that the lpxA gene from Neisseria meningitidis encodes a similar acyltransferase that selectively utilizes 3-hydroxylauroyl-ACP . Strains of E . coli harboring the temperature-sensitive lpxA2 mutation make very little lipid A and lose viability rapidly at 42 degrees C . We have created an E . coli strain in which the chromosomal lpxA2 mutation is complemented by the N . meningitidis lpxA gene introduced on a plasmid . This strain, RO138/pTO6, grows similarly to wild type cells at 42 degrees C and produces wild type levels of lipid A . However, the lipid A isolated from RO138/pTO6 contains mostly hydroxylaurate and hydroxydecanoate in the 3 and 3' positions . The strain RO138/pTO6 is more susceptible than wild type to certain antibiotics at 42 degrees C . This is the first report of an E . coli strain growing with shortened hydroxyacyl chains on its lipid A . The lpxA gene product appears to be a critical determinant of the length of the ester-linked hydroxyacyl chains found on lipid A in living cells.

Vaccine, 1997 Aug, 15(11), 1225 - 34
Outer membrane vesicle vaccines made from short-chain lipopolysaccharide mutants of serogroup B Neisseria meningitidis: effect of the carbohydrate chain length on the immune response; Andersen SR et al.; Outer membrane vesicle (OMV) vaccines were made from Neisseria meningitidis strain 44/76 and its two short-chain lipopolysaccharide (LPS) mutants, Mu-1 and Mu-4 . Only the 44/76 vaccine contained LPS with the host antigen lacto-N-neotetraose . The protein composition of the vaccines was similar . The LPS carbohydrate chain length proved to influence drastic changes in the LPS immunogenicity as well as the outer membrane proteins (OMPs) ability to elicit functional antibodies in mice . Only LPS in the Mu-1 and Mu-4 vaccines were immunogenic, and the 44/76 vaccine differed also by not inducing antibodies to the class 4 OMP . The Mu-1 vaccine, with a LPS carbohydrate chain comprising only two residues of 2-keto-3-deoxy-octonic acid, induced lower bactericidal activity and less antibodies to the class 1 OMP, compared to the two other vaccines . This indicates that LPS of a certain carbohydrate chain length is required for adequate exposure of the class 1 OMP epitopes essential for inducing bactericidal antibodies.

Sex Transm Dis, 1997 Aug, 24(7), 422 - 8
Comparisons of cost and accuracy of DNA probe test and culture for the detection of Neisseria gonorrhoeae in patients attending public sexually transmitted disease clinics in Los Angeles County; Ciemins EL et al.; BACKGROUND: Strict handling and transport requirements for the successful use of culture in the detection of Neisseria gonorrhoeae warrant investigation of accurate and cost-effective test alternatives such as the Gen-Probe PACE 2 DNA probe assay (Gen-Probe, Inc., San Diego, CA) . STUDY DESIGN: The Gen-Probe PACE 2 DNA probe assay for N . gonorrhoeae was compared with conventional culture methods in the principal Los Angeles County (LAC) Department of Health Services (DHS) Public Health Laboratory and three of its branch laboratories . Urethral and endocervical samples were collected from 1,566 patients (921 males; 645 females) attending six LAC DHS sexually transmitted disease clinics . Cost analysis was performed comparing material and labor costs of the two test methods . RESULTS: The overall prevalence based on culture was 11.8% (15.7% for males; 6.4% for females) . Nine samples were culture positive, Gen-Probe negative and four samples were culture negative, Gen-Probe positive and remained discordant after discrepant analysis . The sensitivity and specificity were 94.6% and 99.7%, respectively, for the PACE 2 assay compared with culture . The positive and negative predictive values were 97.8% and 99.3%, respectively . No statistically significant difference was found between the two tests . A cost analysis found an average cost of $3.11/test for culture and $3.85/test for PACE 2, given the approximate 12% disease prevalence in this population . CONCLUSIONS: Gen-Probe's PACE 2 assay may provide an acceptable, cost-effective alternative to culture, especially among high-risk males.

Sex Transm Dis, 1997 Aug, 24(7), 409 - 17
Molecular epidemiology of infections with Neisseria gonorrhoeae among visitors to a sexually transmitted diseases clinic; van Duynhoven YT et al.; OBJECTIVES: To identify determinants for plasmid-mediated resistance to penicillin (penicillinase-producing Neisseria gonorrhoeae {PPNG}) and tetracycline (tetracycline-resistant N . gonorrhoeae {TRNG}) among gonococci, to determine the distribution of bacterial characteristics, and to correlate these with antibiograms and patient characteristics . STUDY DESIGN: Gonococcal isolates from 131 patients attending a sexually transmitted diseases clinic in The Netherlands in 1994 were auxotyped and serotyped and antimicrobial susceptibility was tested . Information on patient characteristics was collected at the initial visit . RESULTS: The most prevalent serotype, IB-1 (26%), proved to be related to sexual contact with casual partners, especially commercial sex partners . In addition, IB-1 strains were associated with PPNG and displayed higher minimum inhibitory concentrations (MICs) for ceftriaxone, cefuroxime, and ciprofloxacin . Homosexual men were more often infected with nonrequiring, IB-2, and IB-6 strains than heterosexuals . These strains were very sensitive to ceftriaxone and ciprofloxacin . Overall, one strain showed decreased susceptibility to ciprofloxacin (MIC 0.5 microgram/ml), but no resistance to ceftriaxone, ciprofloxacin, or cefuroxime was observed . However, 31% of the isolates were TRNG, PPNG, or both . Determinants for these resistant strains among men were the use of antibiotics (odds ratio {OR} = 4.8, 90% confidence interval {CI} 1.3-19.1), Surinam or Morrocan origin (OR = 3.3, 90% CI 1.3-8.4), and homosexual contacts (OR = 0.1, 90% CI 0.03-0.4) . CONCLUSIONS: Different types, with variable susceptibility, were associated with homosexual and commercial sexual behavior . PPNG and TRNG were more commonly isolated from antibiotic users, heterosexual individuals, and ethnic minorities . Continuous surveillance of susceptibility is needed to follow the spread of PPNG and TRNG and to detect resistance to the currently recommended agents in a timely fashion.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1843 - 5
Comparison of single-dose oral grepafloxacin with cefixime for treatment of uncomplicated gonorrhea in men . The STD Study Group; Hook EW 3rd et al.; In a randomized open study, 351 male patients with uncomplicated gonorrhea were given single oral doses of grepafloxacin (400 mg) or cefixime (400 mg) . In the 299 microbiologically evaluable patients, urethral infections were cured in 99% (147 of 149) of those receiving grepafloxacin and 97% (145 of 150) of those given cefixime . Eradication rates for both regimens were 100% in the 16% (47 of 299) of participants who were infected with penicillin-resistant Neisseria gonorrhoeae and 97% in the 21% (62 of 299) of participants infected with tetracycline-resistant strains . Grepafloxacin is a well-tolerated alternative to cefixime for treatment of uncomplicated gonorrhea in males.

Infect Immun, 1997 Aug, 65(8), 3391 - 8
Experimental infection of native human ureteral tissue with Neisseria gonorrhoeae: adhesion, invasion, intracellular fate, exocytosis, and passage through a stratified epithelium; Mosleh IM et al.; The exact mechanisms by which Neisseria gonorrhoeae invades the mucosal lining to cause local and disseminated infections are still not fully understood . The ability of gonococci to infect the human ureter and the mechanism of gonococcal infection in a stratified epithelium were investigated by using distal ureters excised from healthy adult kidney donors . In morphological terms, this tissue closely resembles parts of the urethral proximal epithelium, a site of natural gonococcal infection . Using piliated and nonpiliated variants of N . gonorrhoeae MS11, we demonstrated the importance of pili in the attachment of gonococci to native epithelial cells as well as their association with epithelial damage . By electron microscopy we elucidated the different mechanisms of colonization and invasion of a stratified epithelium, including adherence to surface cells, invasion and eventual release from infected cells, disintegration of intercellular connections followed by paracellular tissue infiltration, invasion of deeper cells, and initiation of cellular destruction and exfoliation resulting in thinning of the mucosa.

Infect Immun, 1997 Aug, 65(8), 3203 - 8
Neisserial porins may provide critical second signals to polysaccharide-activated murine B cells for induction of immunoglobulin secretion; Snapper CM et al.; Resting B cells stimulated with dextran-conjugated anti-immunoglobulin D (anti-IgD) antibodies (anti-Ig-dex), a model for B-cell activation in response to polysaccharide antigens, proliferate but secrete little if any Ig, unless additional stimuli are present . In order to elucidate the parameters which costimulate T-cell-independent antipolysaccharide antibody responses during bacterial infections, we tested the capacities of highly purified porin proteins from Neisseria meningitidis and Neisseria gonorrhoeae to augment in vitro proliferation and induce Ig secretion by anti-Ig-dex-activated B cells . Resting B cells, from lipopolysaccharide (LPS)-nonresponsive C3H/HeJ mice, proliferated and secreted IgM in response to each of three distinct porins acting alone . Further, porins, even at concentrations that were minimally inductive when acting alone, were strongly synergistic with anti-Ig-dex for proliferation and Ig secretion . Similar synergistic effects of porins with CD40-ligand were also observed . These effects of porins were shown to occur directly at the level of the B cell . The predominant Ig isotype elicited in response to porins plus anti-Ig-dex or CD40-ligand was IgM (>97%), with the remainder comprising IgG . Surprisingly, picogram-per-milliliter amounts of neisserial LPS were also found to be highly synergistic with anti-Ig-dex for induction of IgM secretion by LPS-responsive C3H/HeN, but not C3H/HeJ, B cells . Thus, these data suggest that porins, as well as LPS, may provide critical second signals for T-cell-independent induction of polysaccharide-specific Ig in response to neisserial and other gram-negative porin-expressing bacterial pathogens, without a requirement for the participation of non-B cell types . These data may also help to explain the potent immunopotentiating effects of porins for polysaccharide-specific, as well as protein-specific, humoral responses in vivo.

J Clin Microbiol, 1997 Aug, 35(8), 2129 - 32
Diagnosis of Neisseria gonorrhoeae infections in women by using the ligase chain reaction on patient-obtained vaginal swabs; Hook EW 3rd et al.; The increased sensitivities of nucleic acid amplification tests such as ligase chain reaction (LCR) have the potential to simplify specimen collection for gonorrhea diagnosis . In this study patients took their own vaginal swab specimens for gonorrhea culture and LCR testing . Immediately following specimen collection by patients, a trained clinician obtained endocervical swab specimens for the same tests . By using LCR to diagnose gonorrhea, 54 (17.5%) of 309 patients had positive tests . Forty-five patients with positive cervical LCR tests also had positive vaginal LCR tests; for one patient, only a cervical LCR specimen was positive, and for eight patients, only vaginal specimens were positive . For specimens from patients whose gonorrhea cultures were positive, all vaginal swab specimens were positive by LCR and 42 (91%) of 46 cervical swab specimens were positive by LCR . LCR-positive specimens from eight patients with negative cultures (four with positive vaginal specimens only, one with a positive cervical specimen only, and three with positive vaginal and cervical specimens) were further evaluated with unrelated probe sets for gonococcal pilin B . Following resolution of the discrepancies between culture-negative and LCR-positive specimens, a diagnosis of gonorrhea could be confirmed for 52 of 54 patients with positive LCR tests . LCR testing with vaginal swabs was 100% sensitive and 99.6% specific and had a positive predictive value of 98.1% and a negative predictive value of 100% . In this study LCR testing of vaginal swab specimens obtained by patients themselves was significantly more sensitive for gonorrhea diagnosis of women than cervical LCR or culture (100% versus 84.6% for cervical LCR or culture; Mantel-Haenszel chi-square test result, 8.58; P = 0.003).

J Exp Med, 1997 Jul 21, 186(2), 247 - 58
Neisseria gonorrhoeae epithelial cell interaction leads to the activation of the transcription factors nuclear factor kappaB and activator protein 1 and the induction of inflammatory cytokines; Naumann M et al.; We have studied the effect of human bacterial pathogen Neisseria gonorrhoeae (Ngo) on the activation of nuclear factor (NF)-kappaB and the transcriptional activation of inflammatory cytokine genes upon infection of epithelial cells . During the course of infection, Ngo, the etiologic agent of gonorrhea, adheres to and penetrates mucosal epithelial cells . In vivo, localized gonococcal infections are often associated with a massive inflammatory response . We observed upregulation of several inflammatory cytokine messenger RNAs (mRNAs) and the release of the proteins in Ngo-infected epithelial cells . Moreover, infection with Ngo induced the formation of a NF-kappaB DNA-protein complex and, with a delay in time, the activation of activator protein 1, whereas basic leucine zipper transcription factors binding to the cAMP-responsive element or CAAT/enhancer-binding protein DNA-binding sites were not activated . In supershift assays using NF-kappaB-specific antibodies, we identified a NF-kappaB p50/p65 heterodimer . The NF-kappaB complex was formed within 10 min after infection and decreased 90 min after infection . Synthesis of tumor necrosis factor alpha and interluekin (IL)-1beta occurred at later times and therefore did not account for NF-kappaB activation . An analysis of transiently transfected IL-6 promoter deletion constructs suggests that NF-kappaB plays a crucial role for the transcriptional activation of the IL-6 promoter upon Ngo infection . Inactivation of NF-kappaB conferred by the protease inhibitor N-tosyl--phenylalanine chloromethyl ketone inhibited mRNA upregulation of most, but not all, studied cyctokine genes . Activation of NF-kappaB and cytokine mRNA upregulation also occur in Ngo-infected epithelial cells that were treated with cytochalasin D, indicating an extracellular signaling induced before invasion.

Commun Dis Intell, 1997 Jul 10, 21(14), 189 - 92
Annual report of the Australian Gonococcal Surveillance Programme 1996; Tapsall J; In 1996 the Australian Gonococcal Surveillance Programme (AGSP) examined 2,753 isolates of Neisseria gonorrhoeae . The source of isolates, site of infection and antibiotic susceptibility patterns showed considerable regional variation . Strains examined in Adelaide, Sydney and Melbourne were predominantly from male patients where rectal and pharyngeal isolates were common . Cases in other centres had a much lower male:female ratio and most were genital tract isolates . Resistance to the penicillin and quinolone groups of antibiotics was highest in Sydney and Melbourne . Gonococcal resistance to the penicillins was similar to previous years . Quinolone-resistant Neisseria gonorrhoeae (QRNG) were isolated mostly from overseas travellers . However, some local transmission of QRNG was documented in Sydney . All isolates were sensitive to spectinomycin and ceftriaxone.

J Laryngol Otol, 1997 Jul, 111(7), 614 - 5
Bacterial colonization of pacifiers of infants with acute otitis media; Brook I et al.; The presence of aerobic and facultative anaerobic bacteria on the surface of pacifiers used by children with acute otitis media was investigated . The surface of 40 recently used pacifiers was swabbed after they were allowed to dry for five to six minutes . The swabs were processed quantitatively for the presence of aerobic bacteria . The antibacterial activity of the pacifier material was tested in vitro . Microorganisms were isolated from 21 (52.5 per cent) pacifiers . The number of colonies per pacifier varied between one and 35 (average six) . The isolates included eight alpha-haemolytic streptococci, six Staphylococcus epidermis, five Candida albicans, five alpha-haemolytic streptococci, three Neisseria spp . and two Staphylococcus aureus . The pacifier material was shown to be inhibitory against S . aureus . This study illustrated that pacifiers do not contain high numbers of organisms and therefore are not likely to serve as a source of persistence of transfer of organisms.

Eur J Clin Microbiol Infect Dis, 1997 Jul, 16(7), 541 - 4
Pulsed-field gel electrophoresis analysis of clonal relationships among Neisseria meningitidis A strains from different outbreaks; Nicolas P et al.; Pulsed-field gel electrophoresis was used to analyze the DNA of 67 Neisseria meningitidis A strains from five African outbreaks occurring between 1988 and 1996 . Endonuclease Bg/II was used to cut chromosomal DNA, generating 19 analyzable fragments . This technique allowed comparison of clonal relationships between outbreak strains and revealed that the same pulsotype was responsible for the outbreaks . This pulsotype is closely related to strain B54, subgroup III reference strain, Finland, 1975, showing only two fragment differences.

Mol Gen Genet, 1997 Jul, 255(3), 285 - 93
Transcriptional regulation of pilC2 in Neisseria gonorrhoeae: response to oxygen availability and evidence for growth-phase regulation in Escherichia coli; Mellies J et al.; The type-4 pilus of Neisseria gonorrhoeae is a dominant surface antigen which facilitates adhesion to host target cells, an essential event in gonococcal infection . pilC2 encodes a 110-kDa protein involved in pilus assembly, pilus-mediated adherence to human epithelial cells in culture and natural competence for DNA transformation . Luciferase activity directed from a chromosomal pilC2::luxAB transcriptional fusion was reduced approximately 4-fold when cells were grown anaerobically . We observed a concomitant reduction in gonococcal piliation by electron microscopy and a reduction in the ability to adhere to ME-180 human epithelial cells when bacteria were grown in the absence of oxygen . Furthermore, we present evidence for growth-phase regulation of the gonococcal pilC2 gene in Escherichia coli, and show that all sequences necessary for growth-phase regulation are contained on a 121-bp pilC2 fragment . Expression from the minimal pilC2 fragment fused to lacZ in single-copy in E . coli was induced 2-fold when cells entered stationary phase . Surprisingly, induction does not require rpoS, the gene, which encodes the starvation-induced sigma factor RpoS . In summary, we have demonstrated that pilC2 is both positively and negatively regulated at the level of transcription . This regulation is most probably relevant to physiological conditions within the human host which influence gonococcal infections.

Microbiology, 1997 Jul, 143 ( Pt 7), 2127 - 33
Involvement of the gonococcal MtrE protein in the resistance of Neisseria gonorrhoeae to toxic hydrophobic agents; Delahay RM et al.; Low-level resistance of Neisseria gonorrhoeae to toxic hydrophobic agents (HAs), including some antibiotics, is chromosomally mediated via the multiple transferable resistance (mtr) efflux system . The gene encoding the 48:3 kDa outer-membrane protein MtrE, which is associated with the mtr phenotype, was identified and is homologous to export-associated outer-membrane proteins, including the OprM (formerly OprK) lipoprotein of Pseudomonas aeruginosa . Insertional inactivation of the mtrE gene in N . gonorrhoeae strain FA19 resulted in the loss o the outer-membrane protein, with concomitant hypersusceptibility of the mutant strain to a range of HAs . The properties of this mutant confirmed the role of MtrE in multidrug resistance mediated by an active efflux mechanism . Secondary structure predictions for MtrE indicated a largely hydrophilic protein with a single alpha-helical transmembrane region . A transposon-like element, similar to that found downstream of the region containing the promoters for mtrR and mtrC in Neisseria meningitidis, was identified 63 bp downstream of the mtrE gene.

Microbiology, 1997 Jul, 143 ( Pt 7), 2117 - 25
The MtrD protein of Neisseria gonorrhoeae is a member of the resistance/nodulation/division protein family constituting part of an efflux system; Hagman KE et al.; The mtr (multiple transferable resistance) system of Neisseria gonorrhoeae mediates resistance of gonococci to structurally diverse hydrophobic agents (HAs) through an energy-dependent efflux process . Recently, complete or partial ORFs that encode membrane proteins (MtrC, MtrD, MtrE) forming an efflux pump responsible for removal of HAs from gonococci were identified and appeared to constitute a single transcriptional unit . In this study, the complete nucleotide sequence of the mtrD gene was determined, permitting the characterization of the MtrD protein . The full-length MtrD protein has a predicted molecular mass of nearly 114 kDa, putatively containing a 56 amino acid signal peptide . MtrD displays significant amino acid sequence similarity to a family of cytoplasmic membrane proteins, termed resistance/nodulation/division (RND) proteins, which function as energy-dependent transporters of antibacterial agents and secrete bacterial products to the extracellular fluid . The predicted topology of the MtrD transporter protein revealed 12 potential membrane-spanning domains, which were clustered within the central and C-terminal regions of the primary sequence . Loss of MtrD due to insertional inactivation of the mtrD gene rendered gonococci hypersusceptible to several structurally diverse HAs, including two fatty acids (capric acid and palmitic acid) and a bile salt (cholic acid), but not hydrophilic antibiotics such as ciprofloxacin and streptomycin . Since gonococci often infect mucosal sites rich in toxic fatty acids and bile salts, the expression of the mtr efflux system may promote growth of gonococci under hostile conditions encountered in vivo.

Sex Transm Dis, 1997 Jul, 24(6), 378 - 85
Antimicrobial susceptibility and types of Neisseria gonorrhoeae in Greece . Data for the period 1990 to 1993; Tzelepi E et al.; BACKGROUND: Surveillance of the rapidly changing patterns of antimicrobial resistance of Neisseria gonorrhoeae is imperative for monitoring gonococcal infection . GOAL: To describe the types and the antimicrobial susceptibility profile of a representative samples of gonococci isolated in Greece between 1990 and 1993 . STUDY DESIGN: The antimicrobial susceptibilities, serovar/auxotypes classes, and plasmid contents of 263 consecutive isolates of N . gonorrhoeae, recovered from cases of male gonococcal urethritis, were determined . RESULTS: Penicillinase-producing N . gonorrhoeae (PPNG) were isolated at a rate of 6.1% and were mostly from imported cases of infection . Six (2.3%) of the isolates (one PPNG and five non-PPNG) were highly resistant to tetracycline, and one PPNG strain was resistant to norfloxacin and ciprofloxacin . Strains with chromosomal resistance to penicillin, tetracycline, erythromycin, and chloramphenicol accounted for 18.5%, 12.5%, 19%, and 16% of the isolates, respectively; much higher proportions of strains were intermediately susceptible to these antibiotics . Spectinomycin and cefotaxime were active against all gonococci studied . A shift to IB serovars and to sporadic types of strains was noted from previous years among the non-PPNG isolates . This is compatible with the marked increase in the rate of imported cases of infection caused by n on-PPNG strains . CONCLUSIONS: The emergence of high-level resistance to tetracycline and resistance to fluoroquinolones was ascertained . At the present, however, the main problem with gonococcal resistance in Greece seems to ensue from the increasing rates of chromosomally resistant strains . Moreover, the increasing frequency of imported gonococci underlines the necessity for continuous epidemiologic surveillance.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1547 - 51
Differential intracellular efficacies of ciprofloxacin and cefixime against Neisseria gonorrhoeae in human fallopian tube organ culture; Phanucharas JP et al.; This study compared the abilities of ciprofloxacin and cefixime to kill intracellular Neisseria gonorrhoeae in a human fallopian tube organ culture assay . When invasion was inhibited by cytochalasin D, 0.996% of the tissue-associated gonococci survived ciprofloxacin exposure compared to 1.70% of gonococci exposed to cefixime (95% confidence interval for the ratio of the means, 0.267 to 1.30), indicating that the two antibiotics did not significantly differ in the ability to kill extracellular attached organisms . In the absence of cytochalasin D, 1.63% survived ciprofloxacin exposure while 9.76% survived cefixime treatment (95% confidence interval for the ratio of the means, 0.067 to 0.418) . These results suggest that ciprofloxacin penetrated epithelial cells and killed intracellular gonococci better than did cefixime . Thus, at concentrations achievable in serum, ciprofloxacin was more effective in total gonococcal killing than cefixime in this human fallopian tube organ culture model.

Chemotherapy, 1997 Jul-Aug, 43(4), 239 - 44
Comparison of in vitro antimicrobial activity of AM-1155 with those of tosufloxacin and fleroxacin against clinical isolates of Neisseria gonorrhoeae harboring quinolone resistance alterations in GyrA and ParC; Deguchi T et al.; The in vitro antimicrobial activities of AM-1155, a new fluoroquinolone, tosufloxacin and fleroxacin were tested against 55 clinical isolates of Neisseria gonorrhoeae using the agar dilution method . In our previous study, all the strains had been examined for mutations in the region corresponding to the quinolone-resistance determining region of the Escherichia coli gyrA gene and the analogous region of the parC gene, and tested for susceptibility to ciprofloxacin . In this study, the 55 isolates of N . gonorrhoeae were assigned to one of three categories based on the presence or absence of alterations in GyrA and ParC . In each category, the antimicrobial activity of AM-1155 against the isolates was compared with those of tosufloxacin and fleroxacin . The MICs of AM-1155 for 11 highly fluoroquinolone-resistant isolates with alterations in both GyrA and ParC ranged from 0.06 to 1.0 microgram/ml . The MICs inhibiting 50% (MIC50) and 90% (MIC90) of these isolates were 0.125 and 1.0 microgram/ml, respectively . The MICs of AM-1155 for 20 moderately fluoroquinolone-resistant isolates with alterations only in GyrA ranged from 0.03 to 0.25 microgram/ml (MIC50, 0.06 microgram/ml; MIC90m, 0.125 microgram/ml) . The MICs of AM-1155 for 24 of the quinolone-susceptible isolates without alterations in either GyrA or ParC ranged from 0.004 to 0.03 microgram/ml (MIC50, 0.008 microgram/ml . MIC90, 0.015 microgram/ml) . There were significant differences between the MIC distribution of AM-1155 and each corresponding MIC distribution of tosufloxacin and fleroxacin in these three categories to which the 55 isolates were assigned (p < 0.05) . Based on the MIC90S of the tested fluoroquinolones, AM-1155 was two- and eightfold more active against the highly fluoroquinolone-resistant isolates than tosufloxacin and fleroxacin, respectively . Against the moderately fluoroquinolone-resistant isolates, AM-1155 was four- and sixteenfold more active than tosufloxacin and fleroxacin, respectively . Against the quinolone-susceptible strains, AM-1155 was also two- to fourfold more active than the other fluoroquinolones . Overall, AM-1155 exhibited more potent in vitro activity against both quinolone-resistant and quinolone-susceptible isolates of N . gonorrhoeae than tosufloxacin and fleroxacin . In ciprofloxacin treatment failures of gonorrhea at single doses of 500 mg . MICs for the causative organisms have ranged from 1.0 to 16.0 micrograms/ml . The MICs of AM-1155 for the isolates harboring quinolone resistance-associated genetic alterations, including strains exhibiting ciprofloxacin MICs of 2.0 and 8.0 micrograms/ml, still ranged from 0.03 to 1.0 microgram/mL A single-dose study in humans has demonstrated higher peak serum concentrations and longer half-lives of AM-1155, resulting in the AUC0-00 values of AM-1155, which are threefold greater than those of ciprofloxacin at the single doses of 400 and 600 mg . Because of its potent in vitro antimicrobial activity and advantageous pharmacokinetic behavior, AM-1155 may be a clinically useful agent for treating gonorrhea including that caused by quinolone-resistant strains.

J Bacteriol, 1997 Jul, 179(13), 4123 - 8
The MtrR repressor binds the DNA sequence between the mtrR and mtrC genes of Neisseria gonorrhoeae; Lucas CE et al.; Gonococcal resistance to antimicrobial hydrophobic agents (HAs) is due to energy-dependent removal of HAs from the bacterial cell by the MtrCDE membrane-associated efflux pump . The mtrR (multiple transferrable resistance Regulator) gene encodes a putative transcriptional repressor protein (MtrR) believed to be responsible for regulation of mtrCDE gene expression . Gel mobility shift and DNase I footprint assays that used a maltose-binding protein (MBP)-MtrR fusion protein demonstrated that the MtrR repressor is capable of specifically binding the DNA sequence between the mtrR and mtrC genes . This binding site was localized to a 26-nucleotide stretch that includes the promoter utilized for mtrCDE transcription and, on the complementary strand, a 22-nucleotide stretch that contains the -35 region of the mtrR promoter . A single transition mutation (A-->G) within the MtrR-binding site decreased the affinity of the target DNA for MtrR and enhanced gonococcal resistance to HAs when introduced into HA-susceptible strain FA19 by transformation . Since this mutation enhanced expression of the mtrCDE gene complex but decreased expression of the mtrR gene, the data are consistent with the notion that MtrR acts as a transcriptional repressor of the mtrCDE efflux pump protein genes.

Blood, 1997 Jul 1, 90(1), 148 - 55
Downregulation of c-kit expression in human endothelial cells by inflammatory stimuli; Konig A et al.; In recent studies we have shown that the expression of stem cell factor (SCF) in human endothelial cells is regulated by inflammatory processes . Gram-negative bacteria, interleukin-1 (IL-1), and lipopolysaccharide were able to upregulate the expression of SCF in human umbilical vein endothelial cells (HUVEC) (Blood 83:2836, 1994) . Interestingly enough c-kit, the receptor of SCF, is coexpressed on HUVEC, suggesting an autoregulatory mechanism . To investigate the relation of c-kit and inflammatory processes we stimulated HUVEC with IL-1alpha and we established an in vitro model of inflammation . Binding experiments with 125I-SCF were performed to study the c-kit receptor expression on HUVEC . Scatchard analysis revealed both high-affinity receptors (K(d) approximately 0.36 nmol/L) and low-affinity receptors (K(d) approximately 2.9 nmol/L) . Exposure to IL-1alpha led to a significant 50% reduction of c-kit high-affinity receptors, whereas the number of low-affinity receptors was not affected, in comparison to a control group of untreated HUVEC . Furthermore, using Northern blot analysis we studied the regulation c-kit mRNA expression in HUVEC after stimulation with IL-1alpha . Kinetic experiments showed a time-dependent downregulation of c-kit specific transcripts . In addition, we cocultured HUVEC with diverse bacterial strains . Experiments were performed over time with 1 x 10(6) bacteria/mL . Our data showed that, in contrary to the previously reported upregulation of SCF mRNA expression, stimulation with Yersinia enterocolitica or with Neisseria meningitidis led to a significant time-dependent downregulation of c-kit mRNA within 3 hours . These data indicate that inflammatory stimuli such as IL-1 or living bacteria activate a mechanism that downregulates c-kit receptor expression in human endothelial cells during the state of inflammation.

J Infect Dis, 1997 Jul, 176(1), 289 - 92
A self-administered technique for the detection of sexually transmitted diseases in remote communities; Tabrizi SN et al.; The control of sexually transmitted diseases (STDs) in remote rural communities would be enhanced by a sensitive self-administered method for the detection of asymptomatic infection . Results of conventional methods for the detection of STDs were compared with results of tampon-collected specimens analyzed by polymerase chain reaction (PCR) for 480 women . Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis were detected by routine methods in 4 (1%), 14 (3%), and 41 (9%) samples, respectively, while PCR detected these organisms from 52 (11%), 26 (5%), and 75 (16%) tampons, respectively . The detection of each organism was significantly greater by PCR in tampon-collected samples than by routine conventional methods (P < .01) . Discrepant results were confirmed by separate primers in 40 of 48 specimens for N . gonorrhoeae, in 11 of 12 specimens for C . trachomatis, and in 31 of 32 specimens for T . vaginalis . Tampons tested by PCR provide an acceptable and sensitive method for detection of STDs in women living in remote areas.

Infect Immun, 1997 Jul, 65(7), 2970 - 4
A monoclonal antibody directed against the 97-kilodalton gonococcal hemin-binding protein inhibits hemin utilization by Neisseria gonorrhoeae; Lee BC et al.; Neisseria gonorrhoeae expresses two hemin-binding proteins (HmBPs) of 97,000 and 44,000 in molecular weight . A murine monoclonal antibody (MAb) produced against the 97-kDa HmBP from N . gonorrhoeae PID543 specifically inhibited in a concentration-dependent manner the ability of hemin to promote growth . The anti-97-kDa HmBP MAb competitively inhibited binding of the 97-kDa HmBP to a hemin-agarose affinity column . In Western immunoblots, the MAb recognized the 97-kDa homologs from a limited survey of clinical gonococcal isolates . These results support the contention that the 97-kDa HmBP is involved in the gonococcal hemin acquisition pathway.

FEMS Microbiol Lett, 1997 Jun 15, 151(2), 225 - 30
Identification and characterization of thymidylate synthase from Neisseria gonorrhoeae; Carlson JH et al.; Thymidylate synthase converts deoxyuridylate to deoxythymidylate . The thyA gene has been cloned and sequenced from Neisseria gonorrhoeae MS11 . This gene has an open reading frame of 795-801 bp and encodes a product which shares 71% identity with its Escherichia coli homolog . Unlike its E . coli counterpart, gonococcal thyA has a large, upstream transcribed region (300+ bp) that lacks a translatable reading frame . Gonococcal thyA is capable of complementing an E . coli thyA null mutant and shares similar levels of sensitivity with trimethoprim.

Gene, 1997 Jun 11, 192(1), 135 - 40
Microevolution and epidemic spread of serogroup A Neisseria meningitidis--a review; Achtman M; An extensive and representative strain collection of serogroup A Neisseria meningitidis was established . These bacteria were obtained from different endemic and epidemic/pandemic sources and include strains from diseased patients and healthy carriers . The genetic relationships of the bacteria were defined by multi-locus enzyme electrophoresis and sequence polymorphisms of genetically variable antigens have been analyzed in closely-related groupings . The results are interpreted as reflecting a balance of recombination events, which disrupt clonal relationships, and sequential bottlenecks, which purify the bacterial population of genetic variants during epidemic spread.

Gene, 1997 Jun 11, 192(1), 125 - 34
Transformation competence and type-4 pilus biogenesis in Neisseria gonorrhoeae--a review; Fussenegger M et al.; In Neisseria gonorrhoea (Ngo), the processes of type-4 pilus biogenesis and DNA transformation are functionally linked and play a pivotal role in the life style of this strictly human pathogen . The assembly of pili from its main subunit pilin (PilE) is a prerequisite for gonococcal infection since it allows the first contact to epithelial cells in conjunction with the pilus tip-associated PilC protein . While the components of the pilus and its assembly machinery are either directly or indirectly involved in the transport of DNA across the outer membrane, other factors unrelated to pilus biogenesis appear to facilitate further DNA transfer across the murein layer (ComL, Tpc) and the inner membrane (ComA) before the transforming DNA is rescued in the recipient bacterial chromosome in a RecA-dependent manner . Interestingly, PilE is essential for the first step of transformation, i.e., DNA uptake, and is itself also subject to transformation-mediated phase and antigenic variation . This short-term adaptive mechanism allows Ngo to cope with changing micro-environments in the host as well as to escape the immune response during the course of infection . Given the fact that Ngo has no ecological niche other than man, horizontal genetic exchange is essential for a successful co-evolution with the host . Horizontal exchange gives rise to heterogeneous populations harboring clones which better withstand selective forces within the host . Such extended horizontal exchange is reflected by a high genome plasticity, the existence of mosaic genes and a low linkage disequilibrium of genetic loci within the neisserial population . This led to the concept that rather than regarding individual Neisseria species as independent traits, they comprise a collective of species interconnected via horizontal exchange and relying on a common gene pool.

J Antibiot (Tokyo), 1997 Jun, 50(6), 509 - 13
A new type of chemical modification of glycopeptides antibiotics: aminomethylated derivatives of eremomycin and their antibacterial activity; Pavlov AY et al.; A series of derivatives of eremomycin aminomethylated at the 7d position of the resorcinol ring of the amino acid No . 7 was prepared by interaction of eremomycin with formaldehyde and various primary and secondary amines and ammonia . The most active compound obtained was 7d-decylaminomethyl derivative, whose minimal inhibitory concentrations for clinical isolates of staphylococci are 2 approximately 8 times lower than those of the parent antibiotic . 7d-Decylaminomethyl derivative was also active against vancomycin-resistant VanA enterococci (8 microg/ml) and Neisseria gonorrhoeae (16 microg/ml).

Am J Public Health, 1997 Jun, 87(6), 1012 - 5
Low prevalences of HIV infection and sexually transmitted disease among female commercial sex workers in Mexico City; Uribe-Salas F et al.; OBJECTIVES: This study tried to determine human immunodeficiency virus (HIV)/sexually transmitted disease (STD) prevalences among female commercial sex workers in Mexico City . METHODS: A sampling frame was constructed that included bars, massage parlors, and street corners . RESULTS: Prevalences for Treponema pallidum, herpes simplex virus type 2, HIV, Neisseria gonorrhoeae, and Chlamydia trachomatis were 6.4%, 65%, 0.6%, 3.7%, and 11.1%, respectively . A significant association was found between higher STD frequencies and working at street sites . CONCLUSIONS: Most STD frequencies were lower in comparison with rates found for female sex workers in other countries . However, preventive programs against STD/ HIV are needed in this population.

J Antimicrob Chemother, 1997 Jun, 39(6), 747 - 55
Rifampicin resistance in Neisseria meningitidis: evidence from a study of sibling strains, description of new mutations and notes on population genetics; Nolte O; To provide direct evidence for the mechanism leading to resistance to rifampicin, two Neisseria meningitidis strains from one clonal lineage (so-called sibling strains) were studied; one of these strains was resistant, the other sensitive to rifampicin . The polymerase chain reaction (PCR) was used to amplify fragments of the known rifampicin resistance region on the rpoB gene coding for the beta subunit of DNA-directed RNA polymerase and the amplimers were then sequenced . In addition to the DNA from the sibling strains, DNA from further strains was analysed, including two Spanish, rifampicin-resistant strains, eight further N . meningitidis strains, strains of four further Neisseria spp . and one reference strain . The results demonstrated how quickly and easily N . meningitidis can acquire resistance to rifampicin, and also suggest a clonal population structure within the collection of strains studied . This finding is discussed with respect to recent studies that indicate a more panmictic population structure within particular serogroups of N . meningitidis.

Mol Microbiol, 1997 Jun, 24(5), 1083 - 94
The Neisseria type 2 IgA1 protease cleaves LAMP1 and promotes survival of bacteria within epithelial cells; Lin L et al.; Infection of human epithelial cells by Neisseria meningitidis (MC) and Neisseria gonorrhoeae (GC) increases the rate of degradation of LAMP1, a major integral membrane glycoprotein of late endosomes and lysosomes . Several lines of evidence indicate that the neisserial IgA1 protease is directly responsible for this LAMP1 degradation . LAMP1 contains an IgA1-like hinge region with potential cleavage sites for the neisserial type 1 and type 2 IgA1 proteases . Neisserial type 2 IgA1 protease cleaves purified LAMP1 in vitro . Unlike its wild-type isogenic parent, an iga mutant of N . gonorrhoeae cannot affect LAMP1 turnover and its growth in epithelial cells is dramatically reduced . Thus, IgA1 protease cleavage of LAMP1 promotes intracellular survival of pathogenic Neisseria spp.

EMBO J, 1997 Jun, 16(12), 3435 - 45
CD66 carcinoembryonic antigens mediate interactions between Opa-expressing Neisseria gonorrhoeae and human polymorphonuclear phagocytes; Gray-Owen SD et al.; Colonization of urogenital tissues by the human pathogen Neisseria gonorrhoeae is characteristically associated with purulent exudates of polymorphonuclear phagocytes (PMNs) containing apparently viable bacteria . Distinct variant forms of the phase-variable opacity-associated (Opa) outer membrane proteins mediate the non-opsonized binding and internalization of N . gonorrhoeae by human PMNs . Using overlay assays and an affinity isolation technique, we demonstrate the direct interaction between Opa52-expressing gonococci and members of the human carcinoembryonic antigen (CEA) family which express the CD66 epitope . Gonococci and recombinant Escherichia coli strains synthesizing Opa52 showed specific binding and internalization by transfected HeLa cell lines expressing the CD66 family members BGP (CD66a), NCA (CD66c), CGM1 (CD66d) and CEA (CD66e), but not that expressing CGM6 (CD66b) . Bacterial strains expressing either no opacity protein or the epithelial cell invasion-associated Opa50 do not bind these CEA family members . Consistent with their different receptor specificities, Opa52-mediated interactions could be inhibited by polyclonal anti-CEA sera, while Opa50 binding was instead inhibited by heparin . Using confocal laser scanning microscopy, we observed a marked recruitment of CD66 antigen by Opa52-expressing gonococci on both the transfected cell lines and infected PMNs . These data indicate that members of the CEA family constitute the cellular receptors for the interaction with, and internalization of, N . gonorrhoeae.

Infect Dis Clin North Am, 1997 Jun, 11(2), 313 - 29
Frontiers of occupational health . New vaccines, new prophylactic regimens, and management of the HIV-infected worker; Beekmann SE et al.; New prophylactic or treatment options are available for a number of infectious diseases that may be transmitted in the health care setting . Infectious diseases that can now be prevented by vaccination of the employee include hepatitis A, pertussis, hepatitis B, and primary varicella . New prophylactic or treatment regimens are available for Neisseria meningitidis, Streptococcus pyogenes, and Bordetella pertussis; treatment of multidrug-resistant tuberculosis is also discussed . Finally, management of the HIV-infected health care worker is reviewed.

J Infect Dis, 1997 Jun, 175(6), 1396 - 403
Antimicrobial resistance in Neisseria gonorrhoeae in the United States, 1988-1994: the emergence of decreased susceptibility to the fluoroquinolones; Fox KK et al.; Antimicrobial susceptibilities of Neisseria gonorrhoeae have been prospectively determined in the Gonococcal Isolate Surveillance Project of the Centers for Disease Control and Prevention . From 1988 through 1994, susceptibilities were determined for 35,263 isolates from 27 clinics . Patients were demographically similar to those in nationally reported gonorrhea cases . In 1994, 30.5% of isolates had chromosomally or plasmid-mediated resistance to penicillin or tetracycline . Penicillin resistance increased from 1988 (8.4%) to 1991 (19.5%) and then decreased in 1994 (15.6%) . Tetracycline resistance decreased from 1988 (23.4%) to 1989 (17.3%) and then increased in 1994 (21.7%) . Most isolates (99.9%) were highly susceptible to broad-spectrum cephalosporins . Isolates with decreased susceptibility to ciprofloxacin increased from 1991 (0.4%) to 1994 (1.3%); 4 isolates were ciprofloxacin-resistant . Ciprofloxacin-resistant strains may not respond to therapy with recommended doses of fluoroquinolones, and the clinical importance of strains with decreased susceptibility is unknown . The emergence of fluoroquinolone resistance in N . gonorrhoeae in the United States threatens the future utility of this class of antimicrobials for gonorrhea therapy.

J Bacteriol, 1997 Jun, 179(11), 3541 - 8
Porin polypeptide contributes to surface charge of gonococci; Swanson J et al.; Each strain of Neisseria gonorrhoeae elaborates a single porin polypeptide, with the porins expressed by different strains comprising two general classes, Por1A and Por1B . In the outer membrane, each porin molecule folds into 16 membrane-spanning beta-strands joined by top- and bottom-loop domains . Por1A and Por1B have similar membrane-spanning regions, but the eight surface-exposed top loops (I to VIII) differ in length and sequence . To determine whether porins, and especially their top loop domains, contribute to bacterial cell surface charge, strain MS11 gonococci that were identical except for expressing a recombinant Por1A, Por1B, or mosaic Por1A-1B polypeptide were compared by whole-cell electrophoresis . These porin variants displayed different electrophoretic mobilities that correlated with the net numbers of charged amino acids within surface-exposed loops of their respective porin polypeptides . The susceptibilities of porin variants to polyanionic sulfated polymers correlated roughly with gonococcal surface charge; those porin variants with diminished surface negativity showed increased sensitivity to the polyanionic sulfated compounds . These observations indicate that porin polypeptides in situ contribute to the surface charge of gonococci, and they suggest that the bacterium's interactions with large sulfated compounds are thereby affected.

Infect Immun, 1997 Jun, 65(6), 2420 - 7
Ultrastructural analysis of primary human urethral epithelial cell cultures infected with Neisseria gonorrhoeae; Harvey HA et al.; In men with gonococcal urethritis, the urethral epithelial cell is a site of infection . To study the pathogenesis of gonorrhea in this cell type, we have developed a method to culture primary human urethral epithelial cells obtained at the time of urologic surgery . Fluorescent analysis demonstrated that 100% of the cells stained for keratin . Microscopic analyses indicated that these epithelial cells arrayed in a pattern similar to that seen in urethral epithelium . Using immunoelectron and confocal microscopy, we compared the infection process seen in primary cells with events occurring during natural infection of the same cell type in men with gonococcal urethritis . Immunoelectron microscopy studies of cells infected with Neisseria gonorrhoeae 1291 Opa+ P+ showed adherence of organisms to the epithelial cell membrane, pedestal formation with evidence of intimate association between the gonococcal and the epithelial cell membranes, and intracellular gonococci present in vacuoles . Confocal studies of primary urethral epithelial cells showed actin polymerization upon infection . Polyclonal antibodies to the asialoglycoprotein receptor (ASGP-R) demonstrated the presence of this receptor on infected cells in the primary urethral cell culture . In situ hybridization using a fluorescent-labeled probe specific to the ASGP-R mRNA demonstrated this message in uninfected and infected cells . These features were identical to those seen in urethral epithelial cells in exudates from males with gonorrhea . Infection of primary urethral cells in culture mimics events seen in natural infection and will allow detailed molecular analysis of gonococcal pathogenesis in a human epithelial cell which is commonly infected.

Infect Immun, 1997 Jun, 65(6), 2353 - 61
Differential recognition of members of the carcinoembryonic antigen family by Opa variants of Neisseria gonorrhoeae; Bos MP et al.; Opacity (Opa) protein variation in Neisseria gonorrhoeae is implicated in the pathogenesis of gonorrhea, possibly by mediating adherence and entry of the bacteria into human tissues . One particular Opa protein mediates adherence to epithelial cells through cell surface proteoglycans . Recently, two other eukaryotic cell receptors for Opa proteins have been reported . These receptors are members of a subgroup of the carcinoembryonic (CEA) gene family that express CD66 antigens . CEA family members vary in their distribution in human tissues . In order to understand whether interactions between Opa and CEA-like molecules play any role in pathogenesis, we must investigate which CEA family members are able to serve as Opa receptors and which Opa proteins recognize CEA-like molecules . We therefore studied HeLa cells that were stably transfected with five different members of the CEA family, i.e., CEA, CEA gene family member 1a (CGM1a), CGM6, nonspecific cross-reacting antigen (NCA), and biliary glycoprotein a (BGPa) . We infected these transfectants with all possible 11 Opa variants of gonococcal strain MS11 and determined the numbers of bacteria that were bound and internalized . To account for proteoglycan-mediated adherence, infection assays were also performed in the presence of heparin . Our results show that of the 11 Opa variants of MS11, the same 4 recognized CGM1a and NCA . CGM6, however, was not recognized by any Opa variant of MS11 . CEA was recognized by at least 9 of 11 Opa variants, and the BGP transfectants specifically bound and internalized 10 of 11 Opa variants and also bound Opa-negative gonococci . Immunofluorescence experiments showed that clustering of CEA-like molecules occurred upon infection of HeLa transfectants with those Opa variants that interacted specifically with the CEA family member . Together these data show that CEA family members are differentially recognized by gonococcal Opa variants, suggesting that this phenomenon may contribute to cell tropism displayed by gonococci.

Infect Immun, 1997 Jun, 65(6), 2094 - 9
Pelvic inflammatory disease isolates of Neisseria gonorrhoeae are distinguished by C1q-dependent virulence for newborn rats and by the sac-4 region; Nowicki S et al.; The virulence mechanism of Neisseria gonorrhoeae in pelvic inflammatory disease (PID) is not well understood, and an objective diagnostic method to identify patients with PID is lacking . We investigated the hypothesis that development of PID was associated with a C1q-dependent virulence property of gonococcal strains . Recent development of a C1q-dependent experimental model of gonococcal infection (S . Nowicki, M . Martens, and B . Nowicki, Infect . Immun . 63:4790-4794, 1995) created an opportunity to evaluate this hypothesis in vivo . Therefore, the virulence of 32 clinical isolates (18 PID isolates and 14 local infection {LI} isolates) was evaluated in experimental rat pups . A serum bactericidal assay was used to characterize a gonococcal serum-resistant (ser(r)) phenotype . PCR primers designed to amplify a suitable-size gonococcal sac-4 DNA fragment (unique for serum-resistant donor JC1) were used to evaluate the association of serum-resistant genotype sac-4 with two phenotypes: C1q-dependent virulence expressed in vivo and resistance to bactericidal activity of human serum expressed in vitro . Strains were also characterized by auxotyping and serotyping . Of 32 gonococcal strains, 15 (46.7%) caused C1q-dependent bacteremia in rat pups and were sac-4 positive and ser(r) . However, of the 15 isolates, 13 (87%) represented strains associated with human PID and 2 (13%) were associated with LI . None of the strains that were completely serum-sensitive (ser(s)) and sac-4 negative produced C1q-dependent bacteremia in rat pups, suggesting that both ser(r) and sac-4 were required for infection . The serum-resistant recombinant recipient of sac-4 produced C1q-dependent bacteremia in the rat model similarly to the serum-resistant donor of sac-4; the serum-sensitive parent strain did not produce bacteremia . These data suggest that sac-4-mediated serum resistance conferred C1q-dependent virulence and is a unique characteristic associated with PID . These newly identified features may contribute to the understanding of the pathogenic mechanism of PID-associated strains and open perspectives for establishing novel diagnostic methods.

J Clin Microbiol, 1997 Jun, 35(6), 1536 - 40
Detection of Neisseria gonorrhoeae and Chlamydia trachomatis in genitourinary specimens from men and women by a coamplification PCR assay; Crotchfelt KA et al.; A coamplification PCR test for the direct detection of Neisseria gonorrhoeae and Chlamydia trachomatis in urethral and endocervical swabs and urine samples from men and women was compared to standard culture techniques . Processed specimens were amplified in single reaction tubes containing primers for both organisms, and PCR products were detected by a colorimetric microwell plate hybridization assay specific for each pathogen . Of 344 specimens from men, 45 (13.1%) urine specimens were PCR positive for C . trachomatis, 51 (14.8%) urethral swab specimens were PCR positive, and 29 urethral swab specimens (8.4%) were culture positive . After analysis of discrepancies, the resolved sensitivity and specificity of PCR for C . trachomatis were 96.2 and 99.3%, respectively, in urethral swab specimens, compared to 88.2 and 98.6% for urine specimens . Of the 192 specimens from women, 28 (14.6%) urine specimens were PCR positive for C . trachomatis, 32 (16.7%) endocervical specimens were PCR positive, and 19 (9.9%) endocervical specimens were culture positive . After analysis of discrepancies, the resolved sensitivity and specificity of PCR for C . trachomatis for endocervical specimens were both 100% compared to 100 and 99.4%, respectively, for urine specimens from women . In men, 68 (19.8%) urine specimens were PCR positive for N . gonorrhoeae, 73 (21.2%) urethral swabs were PCR positive, and 59 (17.2%) urethral swabs were culture positive . After analysis of discrepancies, the resolved sensitivity and specificity of PCR for N . gonorrhoeae were 97.3 and 97.0%, respectively, for urethral specimens compared to 94.4 and 98.5% for urine specimens . In women, 18 (9.4%) urine specimens were PCR positive for N . gonorrhoeae, 23 (12.0%) were endocervical swab PCR positive, and 15 (7.8%) endocervical specimens were culture positive . After analysis of discrepancies, the resolved sensitivity and specificity of PCR for N . gonorrhoeae were 100 and 99.4%, respectively, for endocervical specimens compared to 90.0 and 95.9% for female urine specimens . These results indicate that a multiplex PCR is highly sensitive for detecting both C . trachomatis and N . gonorrhoeae from a single urine or genital swab, providing a more cost-effective way of screening multiple pathogens.

J Mol Biol, 1997 May 30, 269(1), 129 - 41
Molecular structure of the lipoamide dehydrogenase domain of a surface antigen from Neisseria meningitidis; Li de la Sierra I et al.; The protein p64k from the surface of the Neisseria meningitidis bacteria has been characterized as a two-domain protein . It contains a dihydrolipoamide dehydrogenase domain of 482 residues, involving a FAD prosthetic group as a cofactor, and a smaller lipoic acid binding domain of 86 residues . The two domains are joined by a flexible segment rich in alanine and proline residues . The structure of the dihydrolipoamide dehydrogenase domain was determined by X-ray diffraction . It was solved by a combination of molecular replacement and multiple isomorphous replacement techniques and refined to 2.7 A resolution . In the crystal, the recombinant p64k mimics the functional homo-dimer by using one of the crystallographic 2-fold axes . The reactive disulphide bridge Cys161-Cys166 is in the oxidised state and the FAD is bound in an extended conformation . This main domain contains the major antigenic determinant of the protein, an extended loop of 32 residues at the surface of the protein . A mis-attribution at residue 553 in the sequence has been detected by inspection of electron density maps and the geometry . However, when compared to the other dihydrolipoamide dehydrogenases, there are some significant differences: (1) an unusual number of cis-proline residues and (2) a new motif built around a 2-fold axis by the sulphur atoms of residues Met558, Cys560 and their symmetry related equivalents.

Mol Gen Genet, 1997 May 20, 254(5), 479 - 85
Cloning, nucleotide sequence and transcriptional analysis of the uvrA gene from Neisseria gonorrhoeae; Black CG et al.; A recombinant plasmid capable of restoring UV resistance to an Escherichia coli uvrA mutant was isolated from a genomic library of Neisseria gonorrhoeae . Sequence analysis revealed an open reading frame whose deduced amino acid sequence displayed significant similarity to those of the UvrA proteins of other bacterial species . A second open reading frame (ORF259) was identified upstream from, and in the opposite orientation to the gonococcal uvrA gene . Transcriptional fusions between portions of the gonococcal uvrA upstream region and a reporter gene were used to localise promoter activity in both E . coli and N . gonorrhoeae . The transcriptional starting points of uvrA and ORF259 were mapped in E . coli by primer extension analysis, and corresponding sigma70 promoters were identified . The arrangement of the uvrA-ORF259 intergenic region is similar to that of the gonococcal recA-aroD intergenic region . Both contain inverted copies of the 10 bp neisserial DNA uptake sequence situated between divergently transcribed genes . However, there is no evidence that either the uptake sequence or the proximity of the promoters influences expression of these genes.

J Immunol, 1997 May 15, 158(10), 5043 - 9
The human complement C9 gene: identification of two mutations causing deficiency and revision of the gene structure; Witzel-Schlomp K et al.; The ninth component of human complement (C9) is the last of the terminal complement components creating the membrane attack complex . C9 is a single-chain serum protein that is encoded by a gene located on chromosome 5p . Deficiency of terminal complement components is generally associated with recurrent neisseria infections . We studied a previously described Swiss family with inherited C9 deficiency . To identify the genetic basis of C9 deficiency, we developed an approach using exon-specific PCR and direct DNA sequencing . As a cause of C9 deficiency, we found two different point mutations, both generating TGA stop codons in the coding sequence . One mutation, a C to A exchange, was detected in exon 2 at cDNA position 166, the other, a C to T exchange, was located in exon 4 (cDNA position 464) . In family studies of three first-degree relatives with heterozygous C9 deficiency, we demonstrated that the two mutations are segregating independently . Therefore, these mutations are sufficient to explain the complete deficiency of both the probands studied . DNA sequencing of the exon-intron junctions revealed a number of revisions regarding the boundaries between exons 4, 5, and 6 as well as between exons 10 and 11 . No additional introns were detected in exons 6 and 10 . Furthermore, DNA marker studies were conducted using known polymorphisms of the C6, C7, and C9 genes, confirming the linkage of the observed C9 mutations with defined haplotypes.

J Exp Med, 1997 May 5, 185(9), 1557 - 64
Several carcinoembryonic antigens (CD66) serve as receptors for gonococcal opacity proteins; Chen T et al.; Neisseria gonorrhoeae (GC) is a human pathogen that adheres to and invades genital surfaces . Although pili are required for the initial adherence, the interaction of GC with epithelial cells is also promoted by a family of outer membrane proteins, the opacity (Opa) proteins such as OpaA protein from strain MS11 . Studies have demonstrated that the interaction of the OpaA GC with epithelial cells involves binding to heparan sulfate attached to syndecan receptors . However, other Opa proteins interact with CEA gene family member 1 (CGM1) or biliary glycoprotein (BGP), members of the CD66 antigen family . In this study, we demonstrate that, in addition, the 180-kD carcinoembryonic antigen (CEA) is a receptor for Opa proteins . This conclusion was based on the following observations . First, transfected HeLa cells expressing CEA (HeLa-CEA) and the CEA-expressing colon cancer cell line (LS 174T) bound and subsequently engulfed the Opa+ bacteria . These interactions were inhibited by anti-CEA antibody, but could not be inhibited by addition of heparin . Furthermore, OpaI E . coli directly bound purified CEA . We also compared the adherence and invasion by Opa+ bacteria of CD66 transfected HeLa cells: HeLa-BGPa, HeLa-CGM6, HeLa-NCA, HeLa-CGM1a, HeLa-CEA, and HeLa-Neo serving as negative control . Using OpaI as the prototype, the relative ability of the transfected HeLa cell lines to support adherence was (CEA = BGPa >CGM1a >NCA >>CGM6 = Neo) . The ability to mediate invasion of the transfectant cells was (CGM1a >CEA >BGPa >NCA >CGM6 = Neo) . Among the Opa proteins tested, OpaC proved to be bifunctional, able to mediate adherence to both syndecan receptors and to CD66 antigens.

Ophthalmologe, 1997 May, 94(5), 317 - 20
{Ophthalmia neonatorum caused by Chlamydia trachomatis . Rapid diagnosis and therapy}; Schaller U et al.; BACKGROUND: In up to 73% of ophthalmia neonatorum, Chlamydia trachomatis is the causative agents . Untreated sequelae to the eyes and organs may be the result . The aim of this study was to determine the bacterial spectrum of ophthalmia neonatorum with special regard to chlamydia and their diagnostic tests . MATERIALS AND METHODS: We compared the results of 15 newborn with ophthalmia neonatorum . For the diagnosis we used a rapid diagnostic test, Immunofluorescent Antibody Staining and Culture on McCoy cells . Bacteria that were cultured on culture media were also identified . RESULTS: In 66% the newborn showed a positive rapid diagnostic test result that was confirmed by Immunofluorescent Antibody Staining . In 5 patients all tests were negative . CONCLUSIONS: In this study C . trachomatis was the most frequent pathogen . In the culture media we isolated mostly gram-positive cocci but not Neisseria gonorrhoeae . We point out the value of an exact rapid diagnosis and specific treatment to avoid sequelae to the eye and organs.

Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi, 1997 May-Jun, 38(3), 229 - 31
Endocarditis due to Neisseria sicca: report of one case; Chao HC et al.; Many species of the Neisseria, which are respiratory commensals in humans, have been regarded as being nonpathogenic or as causing disease in only immunocompromised hosts . We report a case in which Neisseria sicca was the cause of infective endocarditis in a child with a ventricular septal defect and review the literature on endocarditis due to N . sicca infection . Most of these patients had an underlying heart disease . Dental caries and poor oral hygiene may be two factors that predispose patients to the infection . N . sicca endocarditis usually results in a subacute onset of symptoms and, if not diagnosed early and treated, is associated with a high rate of embolic complications.

Trop Med Int Health, 1997 May, 2(5), 428 - 32
Increasing prevalence of penicillinase-producing Neisseria gonorrhoeae and the emergence of high-level, plasmid-mediated tetracycline resistance among gonococcal isolates in The Gambia; Adegbola RA et al.; One hundred and three strains of Neisseria gonorrhoeae isolated from a periurban STD clinic in The Gambia were studied for antimicrobial susceptibility, plasmid profile, and serogroup using standard procedures . Seventy-nine (77%) were penicillinase producers (PPNG) and fully resistant to penicillin (MIC > or = 8 mg/l) . One isolate showed chromosomally induced resistance to penicillin (MIC 2 mg/l) . None of the isolates was sensitive to tetracycline; 16 (16%) showed intermediate resistance (MICs 1-8 mg/l) and 87 (84%) showed high-level plasmid-mediated resistance (TRNG) (MICs > 10 mg/l) . This is the first report of TRNG in The Gambia . Only 6 (6%) strains were fully sensitive to trimethoprim-sulphamethoxazole (MIC < 8 mg/l); 78 (76%) showed intermediate level resistance (MICs 8-16 mg/l) and 19 (18%) were fully resistant (MIC > 32 mg/l) . Indications of an increase in MIC to ciprofloxacin and ceftriaxone were found in 6 (6%) and 1 (1%) strains, respectively, although all remained fully sensitive (MICs 0.004-0.03 mg/l and 0.001-0.015 mg/l) . All PPNG and TRNG strains carried the 3.2 MDa and 25.2 MDa plasmids, respectively . All isolates carried the 2.6 MDa cryptic plasmid and 9 (3 PPNG and 6 non-PPNG) carried the 24.5 MDa conjugative plasmid . Forty-four (43%) strains were typed group W1, 58 (56%) W11/111 and 1 had cross-reacting antigens . Because PPNG are frequently encountered and high-level TRNG is now prevalent, the newer cephalosporins and quinolones must now be considered as first-line drugs for the treatment of gonorrhoea in The Gambia.

Pathology, 1997 May, 29(2), 206 - 8
Selection of optimum laboratory tests for the identification of Moraxella catarrhalis; Singh S et al.; We evaluated a variety of conventional and rapid tests and examined the erythromycin susceptibility of a collection of Moraxella catarrhalis and commensal neisseria strains in order to determine the optimum method for routine identification . One hundred and fifty three strains were tested by Gram stain, catalase, oxidase, carbohydrate degradation by two methods and the presence of esterases using indoxyl acetate, 4-methylumbelliferyl butyrate (MUB) . Tween 80 and tributyrin as substrates . Erythromycin MICs and zone diameters around 1, 5 and 15 micrograms discs were determined by the NCCLS method for 151 of the strains . A combination of Gram stain, oxidase and either indoxyl acetate, spot MUB or tributyrin hydrolysis test proved to be reliable and potentially the most convenient for routine testing . MICs and zone diameters easily distinguished between the erythromycin-sensitive M . catarrhalis and the erythromycin-resistant commensal neisserias and would provide confirmation of identification if used for susceptibility testing.

Acta Obstet Gynecol Scand, 1997 May, 76(5), 438 - 41
Reduced prevalence of cervical Chlamydia infection among women requesting termination; Jonsdottir K et al.; BACKGROUND: The prevalence and pattern of Chlamydia trachomatis infection among women requesting induced abortion in the three year period 1992-95 was evaluated and compared to the results of a previous study in 1982-84, where the prevalence of chlamydial infection had been 13.5% . METHODS: A total of 1995 women requested termination, 1855 (93%) of whom were tested for Chlamydia and were included in the study . Two types of tests for chlamydial infection, ELISA and PCR, were used in two consecutive periods . In addition cultures for gonorrhea were done in each case . Information on age, marital status, parity, gestational age and the results of chlamydia and gonorrhea tests of the women and sexual partners were recorded . RESULTS: Chlamydia trachomatis positive women were 149 (8.0%), a significant reduction from the previous 1982-84 study (p<0.001) . Women with positive tests were significantly younger (80% < or = 25 years of age; p<0.001) and more frequently single (86.6%; p<0.001), than those with negative tests, as in the previous period . Of the partners, 80.4% were contacted, and 52.1% presented for investigation . Of those tested 42.1% were Chlamydia positive . Four women (0.2%) had Neisseria gonorrhea but none of the partners . CONCLUSIONS: The prevalence of Chlamydia trachomatis is receding among women coming for termination of pregnancy . As treatment before or at operation has repeatedly been shown to be of benefit and since the prevalence is still considerable, continued screening of these women is justified.

Infection, 1997 May-Jun, 25(3), 154 - 8
Comparative effects of cefadroxil and phenoxymethylpenicillin on the normal oropharyngeal and intestinal microflora; Adamsson I et al.; The ecological effects on the commensal microflora in saliva and stool samples were studied during administration of two commonly used antibiotics: cefadroxil 500 mg b.i.d . for 10 days and phenoxymethylpenicillin 1 g b.i.d . for 10 days . Twenty healthy volunteers participated in the study . In the oropharyngeal microflora the aerobic microflora was significantly suppressed during administration of cefadroxil while no significant changes were noticed in the anaerobic microflora . Administration of phenoxymethylpenicillin caused a strong decrease in the number of viridans streptococci and an overgrowth of Neisseria cocci . The total numbers of anaerobic oropharyngeal microorganisms were suppressed during phenoxymethylpenicillin administration . In the intestinal microflora the variation in numbers of aerobic and anaerobic microorganisms was minor in both groups . The microflora became normalised 2 weeks after withdrawal of the drugs . It was concluded that peroral administration of cefadroxil to healthy volunteers resulted in minor ecological disturbances in the oropharyngeal and intestinal microflora, which were in the same range as for phenoxymethylpenicillin.

J Pediatr Adolesc Gynecol, 1997 May, 10(2), 93 - 4
Gonococcal infection in cerebrospinal fluid and the presence of a ventriculoperitoneal shunt; Morgan DC et al.; BACKGROUND: Neisseria gonorrhoeae is one of the most common organisms associated with pelvic disease in a woman of reproductive age . CASE: We present an unusual case of cerebrospinal fluid infection with N . gonorrhoeae in a woman with a ventriculoperitoneal shunt who complained of abdominal pain . Her shunt was removed and after adequate antibiotic therapy, it was re-inserted . CONCLUSION: Sexually active women, especially those with ventriculoperitoneal shunts, should be encouraged to use a barrier method of contraception, and should have a pelvic examination as part of their evaluation when they present with complaints of abdominal pain.

Int J STD AIDS, 1997 May, 8(5), 299 - 302
Azithromycin and erythromycin resistant Neisseria gonorrhoeae following treatment with azithromycin; Young H et al.; A pre-treatment and a 3-week post-treatment isolate of Neisseria gonorrhoeae from a 13-year-old boy treated with azithromycin in a single 1 g oral dose were characterized microbiologically . Both isolates were of the same serovar/auxotype (1B6/non-requiring) and had similar antibiograms apart from erythromycin and azithromycin: the pre- and post-treatment MICs (minimum inhibitory concentrations) were: 1 mg/L and 32 mg/L to erythromycin and 0.125 mg/L and 3 mg/L to azithromycin . The finding that both isolates were 1B6/NR, had similar antibiograms (other than azithromycin and erythromycin), and no other 1B6/NR isolates were resistant to erythromycin supports the view that macrolide resistance developed following treatment . A high overall level of azithromycin susceptibility was confirmed by testing 67 clinical isolates: MIC90 0.5 mg/L (range 0.023-0.75 mg/L) . We conclude that the long half-life of azithromycin which is beneficial in treating chlamydial infection may result in increased selective pressure for resistance in gonococci . This report also highlights the importance of antibiotic susceptibility surveillance of gonococci and stresses the need for appropriate treatment of gonococcal infection, particularly when it is prescribed outwith departments of genitourinary medicine.

Microbiology, 1997 May, 143 ( Pt 5), 1757 - 64
Control of Neisseria gonorrhoeae pilin gene expression by environmental factors: involvement of the pilA/pilB regulatory genes; Larribe M et al.; The control of the expression of the pilin gene (pilE) in Neisseria gonorrhoeae under a wide variety of growth conditions has been studied . The expression of pilE was measured using transcriptional fusions between pilE and the gene encoding chloramphenicol acetyltransferase (CAT), and the level of pilin production was measured by Western blot analysis . Many of the conditions tested affected both growth rate and pilin gene expression (e.g . isoleucine, high osmolarity, high temperature, anaerobic growth, pH 6, urea and iron depletion) . Changes in the level of many other proteins were also observed, depending on the conditions, indicating that gonococci undergo an adaptive response to environmental variations . Moreover, environment-induced changes in the level of many proteins, including pilin, seem to involve the pilA/pilB regulatory system, which has been previously proposed to modulate the expression of the gonococcal pilin gene.

Microb Pathog, 1997 May, 22(5), 265 - 74
Induction of human endothelial tissue factor expression by Neisseria meningitidis: the influence of bacterial killing and adherence to the endothelium; Heyderman RS et al.; Tissue factor (TF), a small membrane bound high affinity receptor for factor VII, has an important procoagulant role in the haemostatic dysfunction associated with severe sepsis . Using an in vitro model of human endothelial TF expression, defined strains of Neisseria meningitidis were found to upregulate endothelial cell procoagulant activity (PCA) in a dose dependent manner . This TF response was detected with as little as 10(4) cfu/ml and reached similar levels to those seen with high concentrations of purified endotoxin (> 1 ng/ml) . Treatment of N . meningitidis with either adult donor immune serum, penicillin or gentamicin failed to enhance this PCA . Limulus amoebocyte lysate assay of lipopolysaccharide in bacterial culture filtrates together with polymyxin B inhibition experiments suggest that endotoxin is largely responsible for endothelial TF induction by N . meningitidis . Incubation of endothelial cells with N . meningitidis B1940 and B1940 siaD- (an eight-fold more adherent unencapsulated isogenic strain), revealed a significantly greater TF response to B1940 siaD- (P < 0.01) . In conclusion, bacterial adhesion to the vessel wall and therefore local levels of endotoxin may be important determinants of the endothelial procoagulant response to N . meningitidis and the consequent coagulopathy commonly associated with the disease.

Sex Transm Dis, 1997 May, 24(5), 261 - 6
Validity of self-reported sexual behaviors in adolescent women using biomarker outcomes; Orr DP et al.; OBJECTIVES: To determine the criterion-related validity of alternative approaches to the measurement of sexual intercourse using sexually transmitted diseases (STD) as a biomarker (the criterion) . STUDY DESIGN: Analyses are based on an urban sample of 255 adolescent women, 15 to 19 years of age, treated for genitourinary infections with Neisseria gonorrhoeae, Chlamydia trachomatis or Trichomonas vaginalis who returned 3 months later for reexamination . Subjects completed self-administered questionnaires at enrollment and at 3 months . Genitourinary cultures were obtained at enrollment, 2 to 4 weeks later at a test-of-treatment visit (TOT), and at 3 months . RESULTS: Two hundred fourteen of the 255 adolescents also returned for a TOT culture; 186 of these 214 (73%) were free of infection 2 to 4 weeks after enrollment and 30% (56/186) acquired a subsequent STD by 3 months . The validity of questions about sexual behavior differed . No adolescent who denied interim intercourse by reporting "0" sexual partners or "0" coitions acquired an interval STD . Adolescents who denied regular intercourse (vaginal sex) or failed to indicate the number of interim coitions were at high risk for new STD-23% and 21%, respectively . A new measure of sexual intercourse using both the number of sexual partners and the number of coitions contained no missing data; adolescents classified as not having had interim sexual intercourse were free of infection at 3 months, whereas 32% of those who reported intercourse acquired an interim infection . CONCLUSIONS: These data suggest that high-risk urban adolescent women can accurately report whether they have engaged in vaginal intercourse . The validity of the report appears sensitive to the wording and content of the questions.

Sex Transm Dis, 1997 May, 24(5), 257 - 60
Comparison of E-test with agar dilution methods in testing susceptibility of N . gonorrhoeae to azithromycin; Yasin RM et al.; BACKGROUND AND OBJECTIVES: The antimicrobial susceptibility pattern of Neisseria gonorrhoeae varies from one country to another and may also change with time . To monitor these variations and changes, it is desirable to have a method that is simple and reproducible . This study was undertaken to determine the in vitro susceptibility of N . gonorrhoeae to azithromycin and to assess the reliability of results obtained using E-test methodology for determination of the minimum inhibitory concentration (MIC) of azithromycin . STUDY DESIGN: The MICs for 135 clinical isolates of N . gonorrhoeae were determined by a modified Kirby-Bauer method recommended by the National Committee for Clinical Laboratory Standards against penicillin, cefuroxime, ceftriaxone, norfloxacin, tetracycline, kanamycin, spectinomycin, and azithromycin . The MIC of azithromycin was determined by both the E-test and agar dilution method . All tests were done simultaneously . RESULTS: The MIC of azithromycin to all 135 isolates ranged from 0.078 to 0.25 microgram/ml with the agar dilution method and from 0.016 to 0.50 microgram/ml with the E-test . The MIC50 and MIC90 of azithromycin were 0.064 microgram/ml and 0.125 microgram/ml, respectively, by the agar dilution method, whereas they are slightly higher by the E-test method . Seventy-six of the isolates were beta-lactamase producers and 69 were high-level tetracycline-resistant N . gonorrhoeae . There was no difference in the MIC50 and MIC90 of azithromycin in these groups of isolates . The percentage agreement within the acceptable +/-1 log2 dilution difference between MICs obtained by E-test and those obtained by the agar dilution method was 97.8% . CONCLUSIONS: Azithromycin has a very good in vitro antigonococcal activity, and the E-test is a reliable method to determine the MIC of azithromycin against N . gonorrhoeaePIP: A single dose of a new antibiotic, azithromycin, has been shown to be effective in the treatment of uncomplicated Neisseria gonorrhoeae . A clinical study was conducted to assess the in vitro susceptibility of N gonorrhoeae to azithromycin and compare the reliability of results obtained using the new E-test methodology for determination of the minimum inhibitory concentration (MIC) of antibiotic with those obtained through the standard agar dilution method . 135 clinical isolates of N gonorrhoeae were obtained from patients attending hospital-based sexually transmitted disease clinics in five geographic locations in Malaysia . 76 of the isolates were penicillinase-producing N gonorrhoeae and 69 were high-level tetracycline-resistant N gonorrhoeae . All isolates were susceptible to azithromycin based on the susceptible MIC breakpoint of 2.0 mcg/ml . The MICs ranged from 0.0078-0.25 mcg/ml by agar dilution method and from 0.016-0.50 mcg/ml by E-test . Agreement between these two methods was 97.8% . The single-dose regime and good antigonococcal and antichlamydial activity of azithromycin make this antibiotic a suitable treatment choice . Moreover, the findings of this study suggest that the simpler, faster E-test is as reliable as the agar dilution method . Given the tendency of the antimicrobial susceptibility pattern of N gonorrhoeae to change rapidly, it is important to monitor MICs to detect the emergence of resistance .

Sex Transm Dis, 1997 May, 24(5), 247 - 50
Fluoroquinolone treatment failure in gonorrhea . Emergence of a Neisseria gonorrhoeae strain with enhanced resistance to fluoroquinolones; Deguchi T et al.; BACKGROUND AND OBJECTIVES: Although emergence of clinical isolates of Neisseria gonorrhoeae with decreased susceptibilities to fluoroquinolones and treatment failures in gonorrhea have been reported, there have been no clinical reports that fluoroquinolone treatments actually select quinolone-resistant strains, nor have isolates that exhibited clinically significant resistance been analyzed for resistance mechanisms . GOALS: To report a case of fluoroquinolone treatment failure in gonorrhea and emergence of a posttreatment isolate with enhanced resistance to fluoroquinolones; and to study mechanisms of quinolone resistance in the isolates from this patient . STUDY DESIGN: A patient with gonococcal urethritis treated with ofloxacin, 200 mg, three times daily for 5 days is described . Pretreatment and posttreatment isolates were tested for minimum inhibitory concentrations (MICs) of antimicrobial agents and analyzed for alterations in DNA gyrase and topoisomerase IV . They were also examined for ofloxacin uptake . RESULTS: Treatment failure with multiple doses of ofloxacin was observed in this case of gonorrhea . The pretreatment isolate showed decreased susceptibilities to fluoroquinolones (MIC of ofloxacin, 1.0 mg/l; MIC of ciprofloxacin, 0.25 mg/l), and had amino acid changes of Ser-91-->Phe in GyrA and Ser-87-->Ile in ParC . The posttreatment isolate exhibited an increase in resistance to fluoroquinolones (MIC of ofloxacin, 8.0 mg/l; MIC of ciprofloxacin, 1.0 mg/l) . This isolate had identical alterations in GyrA and ParC, but exhibited significantly reduced uptake of ofloxacin . This isolate also showed a small decrease in susceptibilities to cephalosporins . CONCLUSIONS: Alterations in DNA gyrase and topoisomerase IV confer clinically significant resistance to fluoroquinolones in N . gonorrhoeae strains . Treatment with multiple doses of fluoroquinolones is likely to bring about selection of more fluoroquinolone-resistant strains of N . gonorrhoeae and to influence susceptibilities to cephalosporins.

Sex Transm Dis, 1997 May, 24(5), 241 - 6
Sexual behavior risk factors associated with bacterial vaginosis and Chlamydia trachomatis infection; Nilsson U et al.; BACKGROUND AND OBJECTIVES: Few studies have demonstrated that bacterial vaginosis (BV) is associated with sexual behavior risk factors similar to those for other sexually transmitted diseases . The objective of this study was to determine the prevalence of these factors in a multivariate analysis of data from women infected with BV and Chlamydia trachomatis, and noninfected control subjects . GOALS: To study detailed sexual behavior risk factors reported by women with BV versus genital C . trachomatis infection (CT) and by non-BV-infected controls . STUDY DESIGN: A cross-sectional study was conducted with 1,011 women (Swedish Women's Health Study) recruited from family planning and youth clinics in Eskilstuna and Stockholm, Sweden (November, 1989-January, 1991) . Participants were evaluated for the presence of BV, CT, Neisseria gonorrhoeae, Treponema pallidum, Trichomonas vaginalis, and human immunodeficiency virus, and interviewed in detail with respect to sexual behaviors . Statistical comparisons were made using chi-square test (Pearson and likelihood ratio), t test, and logistic regression multivariate analysis . RESULTS: Of 956 women eligible for analysis, the prevalence of BV and CT was 13.7% and 8.9%, respectively . The comparison group consisted of the remaining 825 women without BV . After excluding those with concomitant CT infection, there were 118 women with BV who were compared with 72 women with CT infection only . Sexual factors associated with BV versus the comparison group were a short-term relationship before and after sexual debut, high number of lifetime sexual partners, multiple partners during the last month, high orgasm ability, and more frequent history of group sex, sexual abuse, and rape . When the BV group was compared with the CT group, there were no significant differences in sexual activity risk factors, except for a higher frequency of experience of casual sex in the CT group . CONCLUSIONS: Bacterial vaginosis is associated with sexual behavior risk factors similar to those associated with genital CT infection.

J Med Microbiol, 1997 May, 46(5), 383 - 90
Analysis of Neisseria gonorrhoeae in Ontario, Canada, with decreased susceptibility to quinolones by pulsed-field gel electrophoresis, auxotyping, serotyping and plasmid content; Harnett N et al.; The incidence of Neisseria gonorrhoeae with reduced susceptibility to quinolones increased from 0.18% (63 of 3285) in 1992 to 0.56% (15 of 2663) in 1993 and 0.62% (46 of 2846) in 1994 . In all, 65 of the 67 isolates of Neisseria gonorrhoeae with decreased susceptibility to quinolones were characterised by pulsed-field gel electrophoresis (PFGE), auxotyping, serotyping and plasmid content . The strains were distributed among 14 auxotype/serovar (A/S) classes . Thirty isolates (46.2%) which were penicillin-susceptible with ciprofloxacin MIC90 of 0.12 mg/L and norfloxacin MIC90 of 1.0 mg/L belonged to a single A/S class, OUHL/IA-2 . All but two of the 30 isolates had identical PFGE restriction profiles with NheI restriction endonuclease . Fifteen isolates (23.1%) with MICs in the intermediate (or resistant) categories for penicillin and with ciprofloxacin and norfloxacin MIC90 of 0.25 and 4.0 mg/L and (0.5 and 4.0 mg/L) respectively, belonged to A/S class P/IB-1 . The 15 isolates showed nine different patterns with NheI and eight patterns with SpeI restriction endonucleases . Two of three beta-lactamase-producing (PPNG) isolates belonged to A/S class P/IB-5 and had a dissimilar PFGE restriction profile with NheI endonuclease; the other isolate belonged to A/S class P/IB-8 . The remaining 17 isolates were distributed among 11 A/S classes . Three isolates within the common A/S class NR/IB-1 were subdivided into two types by PFGE as were three isolates belonging to A/S class NR/IB-2 . Overall the 65 isolates of N . gonorrhoeae were distributed into 30 NheI and 26 SpeI macrorestriction profiles . All but one isolate harboured the 2.6-MDa cryptic plasmid and 18 isolates carried the 24.5-MDa transferable plasmid . The three PPNG isolates carried the 4.5-MDa Asian beta-lactamase-producing plasmid and a 25.2-MDa conjugative plasmid was found in the two TRNG isolates.

J Bacteriol, 1997 May, 179(10), 3324 - 30
Cloning and characterization of the gene for amylosucrase from Neisseria polysaccharea: production of a linear alpha-1,4-glucan; Buttcher V et al.; The gene for the amylosucrase from Neisseria polysaccharea (ATCC 43768) was cloned by use of a functional expression system in Escherichia coli XL1-Blue . The deduced amino acid sequence of the protein has homology to the sequences of the alpha-amylase class of enzymes, with the highest similarities being found to the sequences of the trehalose synthase from Pimelobacter sp . strain R48 (17) and amylomaltase from Thermotoga maritima (11) . However, the regions of highest homology within the alpha-amylase class of enzymes, which are essential for the catalytic activity, are only scarcely found in the sequence of amylosucrase . By using the enzyme isolated from culture supernatants of transformed E . coli cells, it is possible to synthesize linear alpha-1,4-glucans from sucrose, indicating that the enzyme is not capable of producing alpha-1,6-glycosidic linkages on its own.

J Bacteriol, 1997 May, 179(9), 3047 - 52
Promoter mapping and transcriptional regulation of the iron-regulated Neisseria gonorrhoeae fbpA gene; Forng RY et al.; In this study, we have mapped the promoter region of the Neisseria gonorrhoeae ferric iron binding protein-encoding gene fbpA, determined the start point of transcription, and examined the accumulation of fbpA mRNA Primer extension analysis of the fbpA promoter region indicated a single transcriptional start site located 51 bp upstream of the ATG translational start site . Northern blot analysis with a 200-bp fbpA structural gene probe detected one transcript of 1.0 kb in RNAs extracted from gonococcal cultures grown under iron-restricted conditions; the 1.0-kb transcript was observed to accumulate at a steady rate throughout the growth cycle . In comparison, in cultures grown under iron-sufficient conditions, the intensity of the 1.0-kb transcript was reduced considerably . Isolation of total RNA from rifampin-treated cells indicated that the half-life of the 1.0-kb fbpA transcript in cells grown under iron-restricted conditions was 1.2 +/- 0.2 min, while that of the 1.0-kb fbpA transcript obtained from cultures grown under iron-sufficient conditions was 0.5 +/- 0.1 min . Taken together, our results indicate that the fbpA promoter is regulated by iron and that transcription and translation of FbpA are closely linked.

J Infect Dis, 1997 May, 175(5), 1153 - 8
Antibody to chlamydial hsp60 predicts an increased risk for chlamydial pelvic inflammatory disease; Peeling RW et al.; To determine whether serum antibody to Chlamydia trachomatis antigens alters the risk of C . trachomatis pelvic inflammatory disease (PID), 280 female sex workers were prospectively evaluated over a 33-month period for incident C . trachomatis and Neisseria gonorrhoeae cervical infection and for clinical PID . At enrollment, women were tested for antibody to C . trachomatis elementary bodies by an indirect microimmunofluorescence assay and to recombinant chlamydial hsp60 (Chsp60) by an ELISA format . At each follow-up visit, women were tested for cervical chlamydial and gonococcal infection and were identified as having clinical PID if they complained of lower abdominal pain and were found to have uterine and adnexal tenderness on pelvic examination . The data demonstrate that antibody to Chsp60 predicts a 2- to 3-fold increased risk for C . trachomatis PID.

Infect Immun, 1997 May, 65(5), 1863 - 9
An in vitro-differentiated human cell line as a model system to study the interaction of Neisseria gonorrhoeae with phagocytic cells; Hauck CR et al.; The extreme host specificity of pathogenic neisseriae limits investigations aimed at the analysis of bacterial-host interactions almost completely to the use of in vitro models . Although permanent epithelial and endothelial cell lines are already indispensable tools with respect to initial infection processes, studies concerning the interaction of neisseriae with phagocytic cells have been confined to primary human blood cells . We investigated the use of human leukemia-derived monocytic and myelomonocytic cell lines that can be differentiated in vitro towards phagocytic cells by a panel of chemical and biological reagents including cytokines, vitamin analogs, and antileukemia drugs . Whereas tumor necrosis factor alpha, gamma interferon, bufalin, or granulocyte-macrophage colony-stimulating factor only marginally increased the ability of monocytic MonoMac-6 and myelomonocytic JOSK-M cells to interact with the bacteria, retinoic acid and vitamin D3 treatment for 2 to 4 days led to highly phagocytic cells that internalized gonococci in an Opa protein-specific manner . This is comparable to the phagocytosis by primary monocytes from human blood, where more than 80% of cells are infected with intracellular bacteria . The increased phagocytic activity of JOSK-M cells following in vitro differentiation was paralleled by enhanced oxidative burst capacity . Whereas undifferentiated cells responded to neither phorbol 12-myristate 13-acetate nor other known soluble and particulate stimuli, cells incubated with retinoic acid and bufalin showed the same pattern and the same intensity of oxidative burst activity in response to Neisseria gonorrhoeae as primary cells: Opa-expressing gonococci elicited an oxidative burst, whereas Opa- gonococci did not . The surface expression of major histocompatibility complex (MHC) class II molecules was only slightly changed after retinoic acid treatment . Also, phagocytosis of gonococci had no influence on MHC class II surface expression . Taken together, our results demonstrate that in vitro-differentiated human myelomonocytic JOSK-M cells provide a suitable model for the study of a variety of aspects of the gonococcal interaction with phagocytes.

Gene, 1997 Apr 11, 189(1), 107 - 12
Transcriptional analysis of the groESL operon of Neisseria gonorrhoeae; Tauschek M et al.; The nucleotide sequence upstream of the groEL gene of Neisseria gonorrhoeae has been determined . Upstream of groEL is a homolog of groES and the divergently transcribed frpB gene . The promoter region of groES lacks the inverted repeat sequences (IR) that act as a regulatory element controlling the expression of similar operons in many other bacterial species . This region contains overlapping consensus sequences for sigma 32-dependent and sigma 70-dependent promoters, and an appropriately placed transcription start point was mapped downstream of these promoters . Northern hybridization demonstrated that synthesis of a full-length groES-groEL transcript was induced in heat-stressed cells . These experiments also revealed the presence of a shorter groES-specific transcript, apparently the result of the premature termination of transcription at an IR situated between the groES and groEL genes.

J Clin Microbiol, 1997 Apr, 35(4), 948 - 50
Rapid screening of point mutations of the Neisseria gonorrhoeae parC gene associated with resistance to quinolones; Deguchi T et al.; To detect quinolone resistance-associated mutations within the Asp-86, Ser-87, Ser-88, and Glu-91 codons of the Neisseria gonorrhoeae parC gene, we developed a rapid and simple assay based on amplification of the regions of the parC gene containing the mutations sites by PCR and digestion of the PCR products with restriction enzymes . By using the method of primer-specified restriction site modification, artificial SalI, PstI, EcoRI, and HinfI restriction sites were created in the regions containing the Asp-86, Ser-87, Ser-88, and Glu-91 codons, respectively . The mutations generating alterations at Asp-86, Ser-87, Ser-88, and Glu-91 were detected as failures of SalI, PstI, EcoRI, and HinfI to digest the respective PCR products . Fifty-five clinical strains of N . gonorrhoeae were examined for mutations in the parC gene by this assay . Appropriate mutations at either the Asp-86, Ser-87, Ser-88, or Glu-91 codon were detected in each of 11 strains in which a mutation had previously been observed by DNA sequencing . This rapid and simple assay could be a useful device for screening genetic alterations in the parC gene associated with resistance to quinolones in N . gonorrhoeae.

Microbiology, 1997 Apr, 143 ( Pt 4), 1415 - 22
Variation within serovars of Neisseria gonorrhoeae detected by structural analysis of outer-membrane protein PIB and by pulsed-field gel electrophoresis; Cooke SJ et al.; Outer-membrane protein PI is the antigen responsible for serovar specificity of Neisseria gonorrhoeae and is a potential vaccine target . In order to investigate possible hidden variation within a serovar, the sequence of the por genes encoding protein PIB have been obtained from a series of strains, including isolates known to be epidemiologically linked . The inferred amino acid sequences of the PIB molecules of isolates from known sexual contacts were identical, but non-related isolates showed significant heterogeneity in PIB sequence . These differences were not confined to the two variable regions (Var1 and Var2) which have previously been identified, but were largely, although not exclusively, located in regions predicted to form one of eight surface-exposed loops . The isolates were subjected to pulsed-field gel electrophoresis of restriction digests of chromosomal DNA, which also demonstrated identity between linked strains but revealed diversity within a serovar . The deduced amino acid sequences of PIB were also used to synthesize peptides for epitope-mapping experiments . These revealed that some mAbs, used to define serovar specificity, recognized linear epitopes located in loops 5 and 6, while others appeared to recognize conformational epitopes elsewhere in the molecule . The occurrence of the sequence differences within a serovar, which are not detected by the serotyping reagents, reveals that PIB represents a potential source of information which should permit considerably more detailed epidemiological studies than are currently possible and focuses attention on more conserved regions of the protein as potential targets for vaccination.

Mol Microbiol, 1997 Apr, 24(1), 169 - 79
Cloning and functional characterization of Neisseria gonorrhoeae tonB, exbB and exbD genes; Biswas GD et al.; Neisseria gonorrhoeae is able to utilize iron (Fe) from a variety of sources including transferrin (TF) and lactoferrin (LF) . To gain insight into the molecular mechanisms used by gonococci to scavenge Fe from TF and LF, we cloned a 3.5 kb segment of wild-type DNA that repaired the defect in tlu mutants, which are unable to take up Fe from either TF or LF despite exhibiting apparently normal ligand binding to the receptor . Nucleotide sequence determination identified three open reading frames (ORFs), designated ORF1, ORF2, and ORF3, which were arranged in tandem . The deduced amino acid sequence of the 852 bp ORF1 encoded a 28 kDa protein that exhibited 26-32% identity with TonB proteins of nine other bacteria . The 663 bp ORF2 predicted a 24 kDa protein and the 435 bp long ORF3 predicted a 15 kDa protein . These predicted protein sequences exhibited 32-38% and 24-36% identity, respectively, with ExbB and ExbD proteins of three other bacteria . Thus, the sequence comparison identified the ORF1, ORF2 and ORF3 as gonococcal homologues of the E . coli tonB, exbB and exbD genes . An insertional mutation in the tonB homologue resulted in the failure of gonococci to grow with TF, LF or human haemoglobin (HB) as sole Fe sources and in the inability to take up 55Fe from TF and LF . The tonB mutation did not prevent the utilization of Fe from citrate (CT) or haemin (HM) . Binding of TF, LF and HB to whole cells in a solid-phase binding assay was largely unaffected by the tonB mutation . We conclude that the pathways for utilization of Fe bound to TF, LF and HB but not to HM or CT were dependent on the TonB system.

Gene, 1997 Apr 1, 188(2), 215 - 20
Insertionally inactivated and inducible recA alleles for use in Neisseria; Seifert HS; Two classes of recA mutations have been constructed for use in Neisseria gonorrhoeae: three insertionally inactivated ('knockout') mutations and three LacI-regulatable constructs that can be shifted between Rec- and Rec+ by the removal or addition of IPTG . The effects of regulating recA expression on the processes of DNA transformation, DNA repair and pilin-phase variation are described . These regulatable cassettes can also be used to control the expression of any chromosomal gene.

Immunol Cell Biol, 1997 Apr, 75(2), 190 - 2
Plant immunomodulators for termination of unwanted pregnancy and for contraception and reproductive health; Talwar GP et al.; Neem (Azadirachta indica) seed and leaf extracts have spermicidal, anti-microbial, anti-fungal and anti-viral properties . They are also immunomodulators that induce primarily a TH1 type response . These properties are being exploited to develop two different useful methods of fertility control . Neem extracts given orally at early post-implantation stage terminate pregnancy in rodents and primates . Treatment has no residual permanent effect and fertility is regained in subsequent cycles . The mechanism by which the action occurs is not fully clear . A transient increase in CD4 and more significantly in CD8 cells is noticed in mesenteric lymph nodes and spleen . A rise in immunoreactive and bioactive TNF-alpha and IFN-gamma in draining lymph nodes, serum and foetal-placental tissue is observed . A polyherbal cream and pessary have been developed containing three active ingredients of plant origin . These have synergistic spermicidal properties on human sperm as determined by the Sander Cramer test . Their use before mating has high contraceptive efficacy in rabbits and baboons . Another interesting property is their inhibitory action on a wide spectrum of micro-organisms, including Candida albicans, C . tropicalis, Neisseria gonorrhoeae, the multidrug-resistant Staphylococcus aureus and urinary tract Escherichia coli, Herpes simplex-2 and HIV-1 . Phase I clinical trials have been completed in India, Egypt and the Dominican Republic, and indicate the safety of the formulation, its acceptability and beneficial action invaginosis due to infections.

J Bacteriol, 1997 Apr, 179(8), 2783 - 7
Cloning and characterization of the ponA gene encoding penicillin-binding protein 1 from Neisseria gonorrhoeae and Neisseria meningitidis; Ropp PA et al.; The ponA gene encoding penicillin-binding protein 1 (PBP 1) from Neisseria gonorrhoeae was cloned by a reverse genetic approach . PBP 1 was purified from solubilized membranes of penicillin-susceptible strain FA19 by covalent ampicillin affinity chromatography and used to obtain an NH2-terminal amino acid sequence . A degenerate oligonucleotide based on this protein sequence and a highly degenerate oligonucleotide based on a conserved amino acid motif found in all class A high-molecular-mass PBPs were used to isolate the PBP 1 gene (ponA) . The ponA gene encodes a protein containing all of the conserved sequence motifs found in class A PBPs, and expression of the gene in Escherichia coli resulted in the appearance of a new PBP that comigrated with PBP 1 purified from N . gonorrhoeae . A comparison of the gonococcal ponA gene to its homolog isolated from Neisseria meningitidis revealed a high degree of identity between the two gene products, with the greatest variability found at the carboxy terminus of the two deduced PBP 1 protein sequences.

J Infect Dis, 1997 Apr, 175(4), 833 - 8
Urethral infection in a workplace population of East African men: evaluation of strategies for screening and management; Jackson DJ et al.; Transport workers (n = 504) in Mombasa, Kenya, were screened for urethral infection by history, clinical examination, and laboratory testing of urethral swabs and first-catch urine specimens . The prevalence of Neisseria gonorrhoeae was 3.4%, Chlamydia trachomatis, 3.6%, and Trichomonas vaginalis, 6.0%; more than two-thirds of infections were asymptomatic . A complaint of urethral discharge, dysuria, or both was twice as sensitive as the sign of discharge on physical examination (34.5% vs . 15.5%) in identifying infection . A positive leukocyte esterase dipstick (LED) test on urine predicted infection with a sensitivity of 95.0% and a specificity of 59.3% in symptomatic men and with a sensitivity of 55.3% and a specificity of 82.8% in asymptomatic men . Demographic and behavioral factors were not independent predictors of infection . In resource-poor settings with high prevalences of urethral infection, an effective screening and management strategy would be to treat symptomatic men, as well as asymptomatic men with a positive LED test, for all three infections.

J Bacteriol, 1997 Apr, 179(7), 2382 - 8
Inversion of Moraxella lacunata type 4 pilin gene sequences by a Neisseria gonorrhoeae site-specific recombinase; Rozsa FW et al.; A plasmid library of Neisseria gonorrhoeae sequences was screened for the ability to mediate recombinations on a sequence containing the Moraxella lacunata type 4 pilin gene invertible region in Escherichia coli . A plasmid containing the N . gonorrhoeae sequence encoding the putative recombinase (gcr) was identified and sequenced . Plasmids containing gcr were able to mediate site-specific recombinations despite a weak amino acid homology to Piv, the native M . lacunata pilin gene invertase . The gcr gene is present only in pathogenic strains of Neisseria tested; however, in our assays gene knockouts of gcr did not alter the variation of surface features that play a role in the pathogenesis of N . gonorrhoeae.

Arch Med Res, 1997 Spring, 28(1), 41 - 5
The hyperferremic mouse model for the evaluation of the effectiveness of VA-MENGOC-BC against Neisseria meningitidis B clinical isolates; Sifontes S et al.; VA-MENGOC-BC is a vaccine against B and C serogroups of Neisseria meningitidis . Its effectiveness at population level has been shown after the application of the vaccine in Cuba, Brazil, Argentina and Colombia . In vitro assays are not always able to reproduce the microorganism-host relationships and this makes it necessary to compile and standardize results obtained in animal models to extrapolate them with a greater degree of safety for humans . We evaluated the effectiveness of VA-MENGOC-BC against Neisseria meningitidis group B isolates from clinically ill patients in Latin America (Argentina, B not typeable: P1; Chile, not typed; Colombia, B4:P1.15 and Cuba B4:P1.15) using Balb/cJ mice treated with iron to make them susceptible to Neisseria meningitidis . The lethal median dose of each strain and of two others that were not included in challenge assays (Brazil: P1.15 and Argentina, B2b:P1.10) were determined . Results were 2.68 x 10(6), 3.16 x 10(7), 1.98 x 10(8), 1.28 x 10(9), 6.42 x 10(6) and 3.88 x 10(7) colony forming units (CFU), respectively . Non-immunized animals and mice treated with one and two doses of VA-MENGOC-BC were challenged with 10(3)-(10) CFU . Protection ranged from 30 to 100% with one dose and was equal to or higher than 70% with the two-dose immunization schedule . A significant protection could not be observed against the Colombian isolate from the lethality point of view, but the mean time of survival lengthened in immunized animals in relation to the controls . The applied inoculum of this strain was much higher (505 x LD50) than the remaining ones . The protection conferred was evident; nevertheless, more data are needed to determine how relevant the results are to humans.

Nucleic Acids Res, 1997 Apr 1, 25(7), 1362 - 8
Analysis of protein binding to the Sma/Cla DNA repeat in pathogenic Neisseriae; Wainwright LA et al.; Antigenic variation of the pilus is an essential component of Neisseria gonorrhoeae pathogenesis . Unidirectional recombination of silent pilin DNA into an expressed pilin gene allows for substantial sequence variation of this highly immunogenic surface structure . While the RecA protein is required for pilin gene recombination, the factors which maintain the silent reservoir of pilin sequences and/or allow unidirectional recombination from silent to expression loci remain undefined . We have previously shown that a conserved sequence at the 3'end of all pilin loci (the Sma/Cla repeat) is required to be present at the expression locus for efficient recombination from the silent loci . In this study, the binding of gonococcal proteins to this DNA sequence was investigated . Gel mobility shift assays and competition experiments using deletion derivatives of the repeat, show that multiple activities bind to different regions of the Sma/Cla repeat and define the boundaries of the binding sequences . Moreover, only the pathogenic Neisseria harbor proteins which specifically bind to this repeat, suggesting a correlation between the expression of these DNA binding proteins and the potential to cause disease.

FEBS Lett, 1997 Mar 17, 405(1), 86 - 90
The lysosomal/phagosomal membrane protein h-lamp-1 is a target of the IgA1 protease of Neisseria gonorrhoeae; Hauck CR et al.; Lysosomal/phagosomal membranes of mammalian cells are coated by highly conserved glycoproteins (lamps) that are thought to protect the membranes from degradation . Interestingly, we identified an amino acid sequence in human lamp-1 characteristic of a cleavage site for the Neisseria gonorrhoeae IgA1 protease . Furthermore, gonococci are detected in h-lamp-1-positive vacuoles after their uptake by professional phagocytes and epithelial cells . Here we demonstrate cleavage of glycosylated h-lamp-1 by the secreted gonococcal IgA1 protease . The cleavage was observed with h-lamp-1 purified from epithelial cells but not from professional phagocytes . The biological role of lamp-1 cleavage by the gonococcal protease is discussed.

Eur J Biochem, 1997 Mar 15, 244(3), 755 - 60
The complete sequence, expression in Escherichia coli, purification and some properties of carbonic anhydrase from Neisseria gonorrhoeae; Chirica LC et al.; The complete nucleotide sequence of the carbonic anhydrase gene from Neisseria gonorrhoeae has been determined . The gene encodes a 252-residue polypeptide with a molecular mass of 28085 Da . The gene has been cloned and overexpressed in Escherichia coli, and the enzyme has been purified . A 26-residue signal peptide is cleaved off by the E . coli processing machinery . Thus, the isolated enzyme contains 226 amino acid residues with a molecular mass of 25314 Da . Most of the enzyme seems to be produced as a soluble protein located in the periplasm of E . coli . The enzyme is homologous to carbonic anhydrases from the animal kingdom; it is an alpha-carbonic anhydrase . A comparison with the amino acid sequences of human carbonic anhydrases I and II suggests that the secondary structures are essentially intact in the bacterial enzyme but that several loops are much shorter than in the human forms . Most of the active-site residues are identical to those found in the high-activity human isozyme II . The bacterial enzyme has a high CO2 hydration activity with a k(cat) of 1.1 x 10(6) s(-1) and Km of 20 mM at pH 9 and 25 degrees C . The enzyme also catalyzes the hydrolysis of 4-nitrophenyl acetate . The pH/rate profile can be described as a titration curve with pKa of 6.7 and a maximal value of the catalytic second-order rate constant, k(enz), of 130 M(-1) x s(-1).

Enferm Infecc Microbiol Clin, 1997 Mar, 15(3), 134 - 9
{Extent of endemic Chlamydia trachomatis in the metropolitan area of Buenos Aires (Argentina)}; de Cristofano MA et al.; BACKGROUND: In this report we inform laboratory results accumulated over ten years (1986-1995) . The number of cases, and the geographic distribution, allow us to present a very reliable data about the dimension of Chlamydia trachomatis urogenital infections in Buenos Aires city and we also compare this profile with the prevalence of Neisseria gonorrhoeae in the same area and period of time . METHODS: Patients were females and males (aged from 15 to 49 years old) attending clinics not specialize in Sexually Transmitted Diseases (STD) . Intent to isolation of C . trachomatis was done in McCoy cells culture . Patients for Neisseria gonorrhoeae investigation were a population assisted at the Clinic for STD of an University Hospital (aged from 15 to 75 years old) . Study for detection of N . gonorrhoeae was developed by direct and conventional culture technics . RESULTS: 4128 endocervical samples from women with lower genital tract pathology were studied and C . trachomatis infection was detected in 25.6 +/- 4.8% . Over 1206 male urethral samples 29.5 +/- 4.47% shows positive cultures . Except for years 1989 and 1990 in which annual percentage of infected women showed slightly higher percentage over the global average, the results shows a very stable annual values, as it was also found in male patients . Infection in males shows a discrete tendency to be higher compare with values obtained in women . Global results of the evolution of prevalence of N . gonorrhoeae infection shows a very different pattern . Since 1992 we demonstrate a very significant decrease in the number of confirmed cases . CONCLUSIONS: We concluded that patients not attending a STD clinic, reveal a high and very stable endemic level of C . trachomatis lower tract urogenital infections . N . gonorrhoeae in this population is a very sporadic or null finding . Prevalence of Chlamydial infection in STD centers is even higher and also shows an stable profile . In people attending STD clinics N . gonorrhoeae shows a very different kinetics, with an important decrease in prevalence in the last five years.

Burns, 1997 Mar, 23(2), 174 - 5
A thermal burn to a prolapsed uterus; Sagi A et al.; Burns to the concealed area of the perineum, are relatively rare and usually associated with massive burns and a high mortality rate . A rare case of a thermal burn to a prolapsed uterus is described . The victim was a 72-year-old Bedouin woman, with a 70 per cent total body surface area deep burn from an open fire . In addition to the conventional treatment dictated by such a burn, two unique problems must be considered: (1) the common pathogens of the uterus, Neisseria gonorrhoeae, Chlamydia trachomatis and mycoplasma, are different from those of the skin; (2) the lymphatics of the uterus drain directly into the abdominal cavity and the risk of peritonitis and generalized infection is potentially higher . Intravenous, prophylactic, broad-spectrum antibiotics were therefore initiated immediately following admission . These included: cefoxitin, gentamicin and metronidazole a combination that covers both the potential pathogens of the uterus and the common pathogens of the skin . In addition, and for the same reason, Betadine substituted Flamazine for the local treatment of the exposed uterus . Our patient did not survive the burn, but in a similar, unusual case, the local and systematic remedies must protect against uterine pathogens that are not commonly seen in a burn victim.

Rev Panam Salud Publica, 1997 Mar, 1(3), 208 - 12
{Detection of Neisseria meningitidis group B antigens by MB-Dot-Elisa test in patients with meningitis}; Alkmin MG et al.; Infection with Neisseria meningitidis group B has been difficult to detect, partly because this bacterial group's polysaccharide is a weak immunogen . This article describes work carried out to test a new procedure (MB-Dot-ELISA) employing a high-titered horse antiserum for detection of N . meningitidis group B antigens . The study assayed cerebrospinal fluid samples from 585 subjects, 574 with suspected meningitis cases and 11 with neurologic disorders . The results of the assay indicated a sensitivity of 0.991 and a specificity of 0.826 . These results were superior to those obtained with latex agglutination and in substantial agreement with the results of counterimmunoelectrophoresis and bacteriologic methods . Overall, the MB-Dot-ELISA was found to be sensitive, inexpensive, and suitable for public health laboratory investigations.

Sex Transm Dis, 1997 Mar, 24(3), 142 - 8
Antimicrobial susceptibilities of strains of Neisseria gonorrhoeae in Bangkok, Thailand: 1994-1995; Knapp JS et al.; BACKGROUND AND OBJECTIVES: Failure of uncomplicated gonococcal infections acquired in the Far East to respond to doses of ciprofloxacin and ofloxacin recommended by the Centers for Disease Control and Prevention have been identified in Australia, the United Kingdom, and the United States . In the Republic of the Philippines, 54.3% of strains exhibited decreased susceptibility to fluoroquinolones; 12% of strains were resistant to ciprofloxacin . This study was undertaken to compare the antimicrobial susceptibilities of gonococcal isolates in Bangkok, Thailand, with those in the Republic of the Philippines . GOAL: To determine the frequency and diversity of antimicrobial resistance, particularly to fluoroquinolones, in gonococcal strains in Bangkok, Thailand . STUDY DESIGN: Strains of Neisseria gonorrhoeae isolated from 101 patients with uncomplicated gonorrhea in Bangkok, Thailand, in July, 1994 (46 strains) and November, 1994 to July, 1995 (55 strains), were characterized by auxotype/serovar class, antimicrobial susceptibilities, and plasmid profile . Susceptibilities were determined to penicillin G, tetracycline, ceftriaxone, cefixime, cefoxitin, ciprofloxacin, ofloxacin, norfloxacin, erythromycin, kanamycin, and thiamphenicol . RESULTS: Of 101 strains, 89.1% (90/101) were resistant to penicillin or tetracycline . Plasmid-mediated resistance to penicillin or tetracycline was identified in 33.7% (34/101) of the isolates: penicillinase-producing Neisseria gonorrhoeae (17.8%; 18/101), tetracycline-resistant Neisseria gonorrhoeae (7.9%; 8/101), and penicillinase-producing/tetracycline-resistant Neisseria gonorrhoeae (7.9%; 8/101) . Most penicillinase-producing strains (96.2%; 25/26) possessed the 4.4-megadalton (Md) beta-lactamase plasmid; one strain possessed the 3.2-Md beta-lactamase plasmid . Chromosomally mediated resistance to penicillin and tetracycline was exhibited by 51.5% (52/101) of strains, and 4.0% (4/101) were tetracycline resistant . All strains were susceptible to spectinomycin . Of 21.8% (22/101) strains exhibiting decreased susceptibility to ciprofloxacin (minimal inhibitory concentration {MIC} > or = 0.125 microgram/ml), one strain (ciprofloxacin MIC, 0.5 microgram/ml; ciprofloxacin inhibition zone diameter of 23 mm) had MICs of 2.0 and 8.0 micrograms/ml for ofloxacin and norfloxacin, respectively, indicating resistance to these agents . Decreased susceptibility to ciprofloxacin was identified in strains with chromosomally mediated resistance to penicillin or tetracycline and in penicillinase-producing strains . CONCLUSIONS: In Bangkok, Thailand, gonococcal isolates exhibit resistance to penicillin, tetracycline, kanamycin, and thiamphenicol . Decreased susceptibility to fluoroquinolones is emerging in a variety of strains of N . gonorrhoeae . Thus, all gonococcal infections should be treated with antimicrobial therapies known to be active against all gonococcal strains to reduce the spread of strains exhibiting decreased susceptibilities to fluoroquinolonesPIP: To assess the frequency and diversity of antimicrobial resistant strains of Neisseria gonorrhoeae in Thailand, endocervical or urethral strains were isolated from 101 patients at a Bangkok sexually transmitted disease clinic . Of the 101 strains, 90 (89.1%) were resistant to penicillin or tetracycline . Plasmid-mediated resistance to penicillin or tetracycline was found in 33.7% of isolates; this included penicillinase-producing N gonorrhoeae (17.8%), tetracycline-resistant N gonorrhoeae (7.9%), and penicillinase-producing/tetracycline-resistant N gonorrhoeae (7.9%) . Moreover, 51.5% of all strains exhibited chromosomally mediated resistance to penicillin and tetracycline . All strains were susceptible to spectinomycin . 22 strains (21.8%) showed decreased susceptibility to ciprofloxacin, while another fifth demonstrated resistance to both fluoroquinolones and norfloxacin . Finally, more than 75% of strains exhibited decreased susceptibility to kanamycin and thiamphenicol . The penicillin/tetracycline resistance phenotypes identified in Bangkok were more similar to those of isolates in the US than in the Philippines, where decreased susceptibility to fluoroquinolones predominates . Given evidence of antimicrobial resistance to penicillin, tetracycline, kanamycin, thiamphenicol, and fluoroquinolones in Thailand, the choice of agents for the treatment of uncomplicated gonococcal infection should be considered carefully and periodic surveillance of antimicrobial resistance is recommended to permit timely revision of treatment protocols .

Mol Microbiol, 1997 Mar, 23(6), 1121 - 31
Random shuttle mutagenesis: gonococcal mutants deficient in pilin antigenic variation; Mehr IJ et al.; Shuttle mutagenesis has been adapted to randomly mutate the genome of Neisseria gonorrhoeae (gonococcus; Gc) . A size-restricted plasmid library of Gc strain FA1090 was mutated with the mini-transposon mTnEGNS . Radomness was tested by checking for transposon insertion bias between vector and insert DNA, Gc transformation efficiency of individual mutated clones, and representation of unique clones before and after Gc transformation with a mutated pool of DNA . Mutants created by random shuttle mutagenesis were screened, using a colony-based polymerase chain reaction assay, for the ability to undergo pilin antigenic variation . Out of 8,064 mutants screened, 22 unique transposon insertion mutants were found to be antigenic variation deficient (Avd) . The Avd mutants were separated into five types according to recombination defect-associated phenotypes, including colony growth, natural DNA transformation competence, and repair of DNA damage caused by ultraviolet radiation.

Mol Biol Evol, 1997 Mar, 14(3), 239 - 47
A likelihood method for the detection of selection and recombination using nucleotide sequences; Grassly NC et al.; Different regions along nucleotide sequences are often subject to different evolutionary forces . Recombination will result in regions having different evolutionary histories, while selection can cause regions to evolve at different rates . This paper presents a statistical method based on likelihood for detecting such processes by identifying the regions which do not fit with a single phylogenetic topology and nucleotide substitution process along the entire sequence . Subsequent reanalysis of these anomalous regions may then be possible . The method is tested using simulations, and its application is demonstrated using the primate psi eta-globin pseudogene, the V3 region of the envelope gene of HIV-1, and argF sequences from Neisseria bacteria . Reanalysis of anomalous regions is shown to reveal possible immune selection in HIV-1 and recombination in Neisseria . A computer program which implements the method is available.

Infect Immun, 1997 Mar, 65(3), 964 - 70
Vitronectin mediates internalization of Neisseria gonorrhoeae by Chinese hamster ovary cells; Duensing TD et al.; Gonococci producing a distinct opacity protein (OpaA in strain MS11) adhere to and are efficiently internalized by cultured epithelial cells such as the Chang conjunctiva cell line . Both adherence and uptake require interactions between OpaA and heparan sulfate proteoglycans on the mammalian cell surface . Chinese hamster ovary (CHO) cells also support adherence of gonococci through interactions of OpaA with cell surface heparan sulfate proteoglycans . However, despite this similarity in the requirements for adherence, CHO cells are not capable of internalizing gonococci . In this report, we characterized this apparent deficiency and identified a factor in fetal calf serum (FCS) which is capable of mediating uptake of gonococci by CHO cells . In the absence of FCS, OpaA+ gonococci adhered to but were not internalized by CHO cells, whereas in the presence of up to 15% FCS, the bacteria were efficiently internalized by the cells . Preincubation of bacteria, but not cells, with FCS also stimulated internalization, suggesting that a factor present in FCS was binding to the surface of gonococci and subsequently stimulating entry . Using a combination of chromatographic purification procedures, we identified the adhesive glycoprotein vitronectin as the serum factor which mediates the internalization of gonococci by CHO cells . Vitronectin-depleted serum did not support gonococcal entry, and this deficiency was restored by the addition of purified vitronectin . Further experiments using a set of gonococcal recombinants, each expressing a single member of the family of Opa outer membrane proteins, demonstrated that vitronectin bound to the surface of OpaA-producing gonococci only and that the vitronectin-mediated uptake by the CHO cells was limited to this bacterial phenotype . To our knowledge, our data are the first example that vitronectin can serve as a molecule that drives bacterial entry into epithelial cells.

Mol Microbiol, 1997 Feb, 23(4), 657 - 68
PilP, a pilus biogenesis lipoprotein in Neisseria gonorrhoeae, affects expression of PilQ as a high-molecular-mass multimer; Drake SL et al.; Studies of gonococcal pilus biogenesis are fundamental to understanding organelle structure/function relationships and identifying new approaches to controlling disease . This area of research is also relevant to elucidating the basic mechanisms of outer membrane translocation of macromolecules, which requires components highly related to those involved in type IV pilus expression . Previous studies have shown that products of several ancillary pil genes are required for organelle biogenesis but of these only PilQ, a member of the GspD protein family, is a component of the outer membrane . DNA sequencing of the region upstream of pilQ revealed the presence of two open reading frames (ORFs) whose deduced polypeptides shared significant identities with proteins required for pilus expression in Pseudomonas aeruginosa and Pseudomonas syringae, the genes for which are arrayed upstream of a gene encoding a PilQ homologue . Gonococcal mutants bearing transposon insertions in these ORFs were non-piliated and failed to express pilus-associated phenotypes, and the corresponding genes were designated PilO and pilP . The piliation defects in the mutants could not be ascribed to polarity on distal pilQ expression as shown by direct measurement of PilQ antigen in those backgrounds and the use of a novel technique to create tandem duplications in the gonococcus (Gc) genome . As predicted by the presence of a consensus lipoprotein signal sequence, PilP expressed in both Escherichia coli and Gc could be labelled with {3H}-palmitic acid . PilP- as well as PilQ- mutants shed PilC, a protein which facilitates pilus assembly and is implicated in epithelial cell adherence, in a soluble form . Combined with the finding that levels of multimerized PiIQ were greatly reduced in PilP- mutants, the results suggest that PilP is required for PilQ function and that PilQ and PilC may interact during the terminal stages of pilus biogenesis . The findings also support the hypothesis that the Gc PilQ multimer corresponds to a physiologically relevant form of the protein required for pilus biogenesis.

Mol Microbiol, 1997 Feb, 23(4), 649 - 56
Integration host factor is a transcriptional cofactor of pilE in Neisseria gonorrhoeae; Hill SA et al.; Integration host factor (IHF) is a small, heterodimeric DNA-binding protein with pleiotropic function . IHF was purified to apparent homogeneity from Neisseria gonorrhoeae . Gel-retardation assays demonstrated binding of IHF to the pilE promoter region . The IHF-binding site was identified by DNase I protection assays and mapped proximal to three previously defined pilE promoters . Removal of the putative IHF-binding domain from pilE promoter DNA negated retardation of the DNA fragment when assessed by gel-shift analysis . Kleinschmidt electron microscopy showed pronounced kinking of pilE promoter DNA following incubation with IHF . Isogenic N . gonorrhoeae strains were constructed that contained either a wild-type pilE locus or a deleted pilE locus where the IHF-binding domain was removed . Primer-extension analysis and Northern blotting of total gonococcal RNA showed that in the absence of IHF binding at the pilE promoter, transcription was reduced 10-fold . Together, these data indicate that IHF is a transcriptional co-activator of pilE.

Genitourin Med, 1997 Feb, 73(1), 33 - 8
STD rapid assessment in Rwandan refugee camps in Tanzania; Mayaud P et al.; OBJECTIVE: To obtain baseline information on sexually transmitted diseases (STDs) in the Rwandan refugees camps in Tanzania, prior to establishment of STD services . SETTING: The largest camps of Rwandan refugees in the Ngara District of Tanzania (estimated population 300,000) . The study was carried out in 8 days in August 1994 . SUBJECTS AND METHODS: A rapid assessment technique was used to measure STD prevalences among: (i) 100 antenatal clinic attenders (ANC); (ii) 239 men from outpatient clinics (OPD); and (iii) 289 men from the community . Interviews (by questionnaire) and genital examination were performed for all participants; sampling for females included genital swabs for the the diagnosis of Neisseria gonorrhoeae (NG), Candida albicans (CA), Trichomonas vaginalis (TV), bacterial vaginosis (BV) and a blood sample for syphilis serology . Men provided urine samples which were screened for leucocytes using the leucocyte esterase (LE) dipstick; urethral swabs for Gram stain were taken from men with a reactive LE test and from those with symptoms or signs of urethritis . OPD males provided a blood sample for syphilis serology . RESULTS: All groups reported frequent experience with STDs and engaging in risky sexual behaviour prior to the survey . During the establishment of the camps, sexual activity was reportedly low . Over 50% of ANC attenders were infected with agents causing vaginitis (TV/BV/CA) and 3% were infected with NG . The prevalence of active syphilis was 4% . In the male outpatients, the prevalence of urethritis was 2.6% and of serological syphilis was 6.1% . Among males in the community, the prevalence of urethritis was 2.9% (the majority being asymptomatic infections) . We noted frequent over-reporting of STD symptoms, unconfirmed clinically or biologically . CONCLUSIONS: STD case detection and management should be improved by training health workers in using the WHO syndromic approach, and through IEC campaigns encouraging attendance at clinics . Rapid epidemiological methods provide quick and useful information at low cost in refugee camps.

Arch Oral Biol, 1997 Feb, 42(2), 101 - 11
Variations in the predominant cultivable microflora of dental plaque at defined subsites on approximal tooth surfaces in children; Babaahmady KG et al.; The distribution and composition of the resident microflora were determined in approximal gingival margin plaque from 21 premolars extracted from schoolchildren (mean age 12.0 +/- 1.8 yr) . Indigo carmine (5% w v) was used to visualize plaque to facilitate sampling . About 1 mm2 of plaque was removed from sites away from (A), to the side of (S), and below (B) the contact area . Plaque samples were dispersed, serially diluted, and cultured on selective and non-selective agar media . An average of seven to nine species was found at each subsite . Streptococcus and Actinomyces were subdivided on the basis of a range of biochemical tests . The predominant Actinomyces and streptococcal species at most subsites were A . naeslundii and Strep . mitis biovar I . A . naeslundii and A . odontolyticus were isolated more often at subsite B (90.5 and 57.1%, respectively), and A . israelii at subsite S (66.7%) Strep . mitis 1 and Strep . sanguis were found more frequently at subsite S (76.2 and 66.7% respectively), whereas Strep mutans, Strep . sobrinus, Strep . gordonii and Veillonella spp . were recovered most commonly from subsite B (85.7, 33.3, 38.1 and 76.2%, respectively) . The isolation frequencies of Strep . mutans and Strep . sobrinus were significantly higher at subsite B (A B p < 0.01 and p < 0.05, respectively) . Veillonella spp . were significantly higher at subsites B and S (A < B, p > 0.01; B > S, p < 0.05), while Neisseria spp . were most common at subsite A (A > B . p > 0.03) . IgAl protease-producing species were found at each subsite, but they formed only a small proportion of the total Streptococcus population . This study has shown that local variations were evident at different subsites, both with respect to species prevalence and to proportions of each species within each subsite . The population shifts in gingival margin plaque appear to relate to the location of plaque in relation to the most caries-prone site below the contact area B.

Br J Rheumatol, 1997 Feb, 36(2), 270 - 2
Gonococcal endocarditis in a patient with systemic lupus erythematosus; Tikly M et al.; There is mounting evidence that patients with systemic lupus erythematosus (SLE) are prone to disseminated neisserial infections . We describe the first proven case of gonococcal endocarditis affecting the pulmonary valve in a patient known to have SLE . The clinical clues and pitfalls in diagnosis are discussed, and the role of echocardiography is highlighted . Possible reasons for the association of gonococcal endocarditis with SLE include pre-existing Libman . Sacks endocarditis, complement deficiency and abnormalities of the reticuloendothelial system.

Clin Infect Dis, 1997 Feb, 24(2), 170 - 8
Oral clindamycin and ciprofloxacin versus intramuscular ceftriaxone and oral doxycycline in the treatment of mild-to-moderate pelvic inflammatory disease in outpatients; Arredondo JL et al.; This multicenter, prospective, double-blind study compared the safety and efficacy of clindamycin and ciprofloxacin versus ceftriaxone and doxycycline in the treatment of outpatients with mild to moderate pelvic inflammatory disease (PID) diagnosed by laparoscopy . Samples taken from the endocervix, endometrium, and abdominal cavity before treatment and from the endocervix after treatment were cultured for aerobes, anaerobes, Neisseria gonorrhoeae, and Chlamydia trachomatis . Of the 138 patients enrolled, 131 were evaluable for efficacy . The most prevalent bacteria were streptococci, staphylococci, and Escherichia coli (among aerobes) and Bacteroides species and peptostreptococci (among anaerobes) . N . gonorrhoeae was present in 2% (3) of the 131 evaluable patients, and C . trachomatis was in 11% (15) . The clinical cure rate was 97% (65 of 67) in the clindamycin and ciprofloxacin group and 95% (61 of 64) in the ceftriaxone and doxycycline group . Side effects were similar in both groups . In conclusion, the two regimens for the outpatient treatment of mild to moderate PID were similarly effective and safe.

Sex Transm Dis, 1997 Feb, 24(2), 109 - 15
Evaluation of a screening algorithm for the diagnosis of genital infections with Neisseria gonorrhoeae and Chlamydia trachomatis among female sexworkers in Bénin; Germain M et al.; BACKGROUND AND OBJECTIVES: In developing countries, simple and cheap procedures for the diagnosis of sexually transmitted diseases (STDs) are urgently needed, especially for screening purposes in high risk groups . GOALS: To evaluate the sensitivity and specificity of a screening algorithm for STDs among 364 female sex workers in Benin, in comparison with reference laboratory tests . STUDY DESIGN: The algorithm relied on the following criteria, which were evaluated in sequence: the presence of endocervical mucopus on visual inspection of the cervix, a positive swab test, or a microscopic examination of vaginal fluid showing more than 10 polymorphonuclear cells per field . The algorithm diagnosed an infection if any one of these criteria was fulfilled . True infectious status was determined by the combined results of culture for Neisseria gonorrhoeae, enzyme immunoassay for Chlamydia trachomatis, and polymerase chain reaction assays for both infections . RESULTS: Gonococcal or chlamydial infection was diagnosed in 39.8% of the study population according to the reference tests . The algorithm had a sensitivity of 57.9% and a specificity of 61.2% . In the presence of Candida sp or Trichomonas vaginalis, specificity decreased to 39.1%, but sensitivity increased to 67.5% . CONCLUSIONS: These results underscore the limitations of simple, nonlaboratory diagnostic tools for screening STDs in high-risk groups in developing countries . Further research is needed to increase the validity--especially the sensitivity--of these algorithms.

J Antimicrob Chemother, 1997 Feb, 39(2), 247 - 9
Antimicrobial activity of a new fluoroquinolone, DU-6859a, against quinolone-resistant clinical isolates of Neisseria gonorrhoeae with genetic alterations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV; Deguchi T et al.; The in-vitro antimicrobial activity of DU-6859a, a new fluoroquinolone, was tested against 55 clinical isolates of Neisseria gonorrhoeae . The MIC of DU-6859a inhibiting 90% (MIC90) of the isolates with genetic alterations of both the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV was 0.125 mg/L . The MIC90 for isolates with alterations of GyrA alone or without alterations of GyrA or ParC was 0.03 mg/L and 0.004 mg/L, respectively . The potency of DU-6859a against clinical isolates bearing genetic alterations associated with quinolone resistance was significantly greater than that of currently available fluoroquinolones.

Antonie Van Leeuwenhoek, 1997 Feb, 71(1-2), 129 - 36
Response of Neisseria meningitidis to iron limitation; Pettersson A et al.; In the human body, the concentration of free iron is limiting for bacterial growth, since iron is bound to transport and storage proteins such as transferrin and lactoferrin . When grown under iron starvation, Neisseria meningitidis produces receptors for these proteins in the outer membrane . These receptors are presently being characterized at the molecular level . Here, we summarize our current knowledge of these receptors, with special emphasis on the LbpA and FrpB proteins, which are studied in our laboratories . Furthermore, the genetic and antigenic variability of these proteins and their vaccine potential are discussed.

Microbiology, 1997 Feb, 143 ( Pt 2), 633 - 40
Arginine-, hypoxanthine-, uracil-requiring isolates of Neisseria gonorrhoeae are a clonal lineage with a non-clonal population; Gutjahr TS et al.; Multilocus enzyme electrophoresis has shown that a collection of 101 arginine-, hypoxanthine-, uracil-requiring (AHU-) isolates of Neisseria gonorrhoeae, recovered over a 39 year period from the UK and Denmark, were of a single electrophoretic type (91% of strains), or differed from the predominant electrophoretic type at only a single locus . The striking uniformity of the AHU-isolates, and the correlation between auxotype, serovar and overall genetic background, contrasts with previous studies of gonococcal populations (that included very few AHU-strains), and a small sample of non-AHU-isolates studied here, which demonstrated a non-clonal population structure and a lack of association between auxotype, serovar and genetic background . There was no marked difference in the ability of AHU-isolates to be transformed with their own DNA, or with DNA from gonococci of other auxotypes, and the relative genetic stability of AHU-isolates does not appear to be due to a defect in their ability to be transformed . An alternative possibility is that AHU-gonococci recombine with other lineages, but that the resulting recombinants are not maintained in the population . This would occur, for example, if AHU-gonococci competed poorly in mixed infections, within which effective recombination between lineages occurs, and are usually only transmitted from individuals who are singly infected with an AHU-strain . The association between AHU-gonococci and asymptomatic infections may lead to an increased rate of transmission of these strains which under this scenario would be needed to prevent them from being lost from the population.

Obstet Gynecol, 1997 Feb, 89(2), 184 - 92
Acute pelvic inflammatory disease: associations of clinical and laboratory findings with laparoscopic findings; Eschenbach DA et al.; OBJECTIVE: To assess the relation of clinical variables and laboratory data to pelvic laparoscopic observations of tubal occlusion, adnexal adhesions, and peritoneal exudate in women with acute salpingitis . METHODS: Clinical and laboratory evaluations were performed systematically before laparoscopy in 155 women with suspected acute pelvic inflammatory disease (PID), 82% of whom proved to have acute salpingitis confirmed with laparoscopy . Laparoscopic findings were scored in three categories (tubal patency, adhesions, and exudate.) RESULTS: Two general categories of laparoscopic findings were present: 1) tubal occlusion and moderate to severe adhesions in 30 women, and 2) pelvic-abdominal exudate in 27 women . In the remaining 16 women, these laparoscopic findings occurred alone or in other combinations . Among women with acute salpingitis, tubal occlusion was associated positively with older age, palpable adnexal mass, and moderate to severe pelvic adhesions; negative associations were found with abdominal rebound tenderness, mean abdominal-pelvic tenderness score, pelvic-abdominal exudate, and isolation of either Neisseria gonorrhoeae or Chlamydia trachomatis . Moderate or severe pelvic adhesions were associated positively with increased duration of abdominal pain (5 versus 3 days) compared with limited or no pelvic adhesions, but they were associated negatively with mean abdominal-pelvic tenderness score and with pelvic-abdominal exudate (47% versus 73%) . Free exudate in the pelvis or abdomen as compared with limited or no exudate was associated positively with abdominal rebound tenderness (86% versus 65%), abdominal-pelvic tenderness score, elevated white blood cell count (83% versus 52%), and recovery of N gonorrhoeae (79% versus 57%) . Free exudate was associated negatively with the median duration of pain (3 versus 6 days), oral contraceptive use (4% versus 26%), and palpable adnexal mass (7% versus 25%) . Analyses limited to women without a history of PID gave similar results . CONCLUSIONS: Although clinical and laboratory criteria traditionally used to judge the clinical severity of acute PID partially predict the degree of tubal or other pelvic abnormalities among women with acute salpingitis and tend to distinguish those with tubal occlusion or moderate to severe adhesions from those with peritonitis, these criteria have low predictive value and are not reliable in the individual patient.

Infect Immun, 1997 Feb, 65(2), 645 - 50
Selection of Opa+ Neisseria gonorrhoeae by limited availability of normal human serum; Bos MP et al.; Experimental infections of human male volunteers with Neisseria gonorrhoeae have provided valuable insights into the early stages of gonorrheal disease . Bacterial variants expressing outer membrane opacity (Opa) proteins appear to be selected from the inoculum during a period in which total recoverable numbers of bacteria decrease rapidly . This apparent survival advantage occurs simultaneously with the onset of an inflammatory response, characterized by local production of interleukin 6 (IL-6) and IL-8 and the appearance of leukocytes in urine . Since the inflammatory response may also result in the presence of serum factors on the mucosal surface, we investigated the possibility that killing in normal human serum (NHS) leads to the selection of Opa+ variants . We therefore studied killing of separate populations and mixtures of Opa- and Opa+ N . gonorrhoeae MS11mk in NHS . Expression of an Opa protein conferred a survival advantage upon the organism; i.e., the Opa+ variants were more serum resistant than their isogenic Opa- counterparts, resulting in a selection for Opa+ phenotypes when a mixture of Opa+ and Opa- gonococci (GC) was exposed to submaximal doses of NHS . This selection was observed in three different lipooligosaccharide (LOS) backgrounds, indicating that it was not due to a difference in LOS expression between Opa- and Opa+ phenotypes . Incubation in NHS of sialylated GC resulted in a similar selection for Opa+ variants . The presence of normal human urine during the serum killing assay had no effect on the selection phenomenon but drastically depleted NHS of bactericidal activity, which was found to be at least partly due to complement inhibition . The results suggest that serum killing may contribute to the transition from Opa- to Opa+ phenotypes during the early stages of infection of the male urethra.

Infect Immun, 1997 Feb, 65(2), 636 - 9
Protegrin structure and activity against Neisseria gonorrhoeae; Qu XD et al.; Protegrin 1 (PG-1) is a broad-spectrum antimicrobial peptide that contains 18 amino acid residues (RG GRLCYCRRRFCVCVGR) and has two intramolecular cystine disulfide bonds . To determine the minimal structure responsible for protegrin-mediated activity against Neisseria gonorrhoeae, we synthesized 15 protegrin variants and tested them against two well-characterized gonococcal strains . The MICs of PG-1 were 0.61 microM (1.31 microg/ml) for the serum-sensitive strain F 62 and 0.98 microM (2.11 microg/ml) for the serum-resistant strain FA 19 . Six amino acid residues (Arg1, Gly2, Gly3, Arg4, Gly17, and Arg18) and either disulfide bond could be deleted from PG-1 without impairing its potency against strain F 62 . In contrast, only Gly17 and Arg18 could be removed without decreasing its activity against FA 19 . Protegrin congener 64a (PC-64a; LTYCRRRFCVTV), a variant of PG-1 with 12 amino acid residues and one disulfide bond, displayed MICs of 0.45 microM (0.68 microg/ml) for strain F 62 and 1.37 microM (2.07 microg/ml) for strain FA 19, which approximated those of intact PG-1 . Serum-sensitive sac-1+ and sac-3+ transformants of N . gonorrhoeae FA 19 and two FA 19 derivatives with truncated lipooligosaccharide structures were more susceptible to PG-1 and variants with altered disulfide structures . These data suggest that structurally simpler protegrin variants, such as PC-64a, could be used as topical microbicides for N . gonorrhoeae . They also suggest that the cystine-stabilized antiparallel beta-sheet formed by PG-1 residues 5 to 16 is principally responsible for its activity against gonococci.

J Bacteriol, 1997 Feb, 179(3), 982 - 6
Antigenic variation in Neisseria gonorrhoeae: production of multiple lipooligosaccharides; Burch CL et al.; Individual cells of Neisseria gonorrhoeae may express a single lipooligosaccharide (LOS) component on their cell surfaces, or they may simultaneously express multiple LOS structures . Strain FA19 expresses LOS components that react with monoclonal antibodies (MAbs) 2-1-L8 and 1B2 . The genetic locus responsible for this phenotype in FA19 was identified by isolating a clone that is able to impart the ability to simultaneously express both LOS molecules to strain 1291, a strain expressing only the MAb 1B2-reactive LOS . This clone, pCLB1, was characterized, and the gene responsible for the expression of both LOS components was determined to be lsi2 . DNA sequence analysis of lsi2(Fa19) indicates that there are several differences between the DNA sequences of lsi2(FA19) and lsi2(1291) . The region responsible for the LOS-specific phenotype change in lsi2(FA19) was identified by deletion and transformation analysis, mapping to a polyguanine tract within lsi2 where lsi2(FA19) possesses a +2 frameshift relative to lsi2(1291) . The polyguanine tract in lsi2(FA19) was modified by site-directed mutagenesis to change the sequence to GGGAGGTGGCGGA to prevent frameshifting during DNA replication, transcription, and/or translation . Transformants of strain 1291 containing this DNA sequence express a single MAb 2-1-L8-reactive LOS component, the same phenotype exhibited by lsi2-defective strains . These data indicate that FA19 is able to generate a small amount of functional Lsi2 protein via transcriptional and/or translational frameshifting, and this limited amount of protein allows for the expression of MAb 1B2-reactive LOS molecules.

Am J Med, 1997 Jan 27, 102(1A), 30S - 34S
Monarthritis: differential diagnosis; Sack K; Acute monarthritis should be regarded as infectious until proved otherwise . Early evaluation is crucial because of the capacity of some infectious agents to destroy cartilage rapidly . The history and physical examination can provide highly suggestive clues, but a definitive diagnosis may depend on arthrocentesis and analysis of synovial fluid . The diagnosis of acute monarthritis is rarely established by radiography . The most common cause of bacterial arthritis is Neisseria gonorrhoeae . Staphylococcus aureus and streptococci are the organisms most frequently implicated in nongonococcal bacterial arthritis, although the possibility of Gram-negative bacteria or anaerobes should not be overlooked in intravenous drug users or immunocompromised patients . Inflammation in a large joint, particularly the knee, might arouse suspicion of Lyme disease . Other, less frequently encountered infectious causes of acute monarthritis include tuberculosis and other mycobacteria, fungi, and viruses . Arthroscopic examination and synovial tissue biopsy may be necessary to diagnose such processes . Microscopic examination of the synovial fluid may reveal a crystalline etiology for monarthritis . Monosodium urate crystals induce gout, usually in the toe, ankle, or midfoot, while calcium pyrophosphate crystals cause pseudogout, most often in the knee or wrist . Acute monarthritis is sometimes a manifestation of osteoarthritis or an early sign of a systemic arthritis such as rheumatoid or reactive arthritis . Processes underlying acute monarthritis can also evolve into a more chronic clinical picture as exemplified by the spondyloarthropathies.

Bull Soc Pathol Exot, 1997, 90(2), 81 - 2
{The microbial etiology of genital discharges in Nouakchott, Mauritania}; Lo BB et al.; The survey carried on 579 patients of both sexes, consulting for genital discharges (spontaneous or referred), showed that in 61.1% men cases, STD agents were isolated (Neisseria gonorrhoeae 51.5%) and 64.4% in women cases (Gardnerella vaginalis 24%, Candida albicans 20% . Trichomonas vaginalis 14%) . 17% of Neisseria gonorrhoeae strains were penicillinase-producing (PPNG).

Am J Ophthalmol, 1997 Jan, 123(1), 135 - 6
Exogenous Neisseria meningitidis endophthalmitis; Saperstein DA et al.; PURPOSE: To report a case of Neisseria meningitidis endophthalmitis in association with a leaking filtering bleb and to consider antibiotic prophylaxis of those people with whom the patient had contact . METHOD: We treated an 81-year-old man who had a chronic, leaking filtering bleb and who developed exogenous N meningitidis endophthalmitis . RESULT: N meningitidis endophthalmitis was controlled with antibiotic therapy . Antibiotic prophylaxis for those with whom the patient had contact was not recommended . CONCLUSION: In this case, the N meningitidis strain was not considered invasive because the bacteria apparently entered the eye through a leaky filtering bleb and not through the bloodstream . Recovery of noninvasive N meningitidis does not require prophylaxis for patient contacts . In cases of endogenous or idiopathic N meningitidis endophthalmitis, antibiotic prophylaxis of close patient contacts may be warranted.

Mikrobiol Z, 1997 Jan-Feb, 59(1), 82 - 6
{A comparison of nutrient media for the cultivation of Neisseria gonorrhoeae}; Skliar TV et al.; It is established that the highest growth peculiarities are characteristic of the nutrient medium for gonococcus isolation of the following composition: MPA of fresh bovine hearts enriched with casein hydrolysate, yeast autolysate, cattle serum (CS) and 0.01% addition of sodium humate . The given nutrient medium has the highest diagnostic potentialities . A possibility to change bovine hearts MPA for swine heart MPA, CS serum and plasmol . The nutrient medium on the basis of fresh swine heart, enriched with plasmol, casein hydrolysate and yeast autolysate may be used to isolate and cultivate gonococci under the laboratory conditions.

Kansenshogaku Zasshi, 1997 Jan, 71(1), 34 - 8
{Analysis of prognosis of bronchopulmonary infectious disease with lung cancer}; Mori K et al.; Bronchopulmonary infection affects the prognosis of lung cancer patients . Thus, we investigated the relationship between the prognosis of bronchopulmonary infectious diseases and their causative bacteria isolated by transtracheal aspiration (TTA) in lung cancer patients . In the present study, we determined which factor is more predisposing for the outcome of bronchopulmonary infections, the type of causative bacteria or the host nutritional status . A total of 107 lung cancer patients, which consisted of 105 males and 5 females (mean age 67.3 +/- 8.0), were included in this study . The study was conducted from 1981 to 1994 . They were classified into the survival group and the deceased group . Causative agents of infection were compared between these 2 groups . S . pneumoniae, alpha-Streptococcus sp., M . catarrhalis, and Neisseria sp . were predominant in organisms isolated from TTA-specimens of lung cancer patients with bronchopulmonary infections, regardless of prognosis . Nutritional status, as determined by serum levels of cholinesterase, albumin, and cholesterol, was poor in the deceased group than in the survival group . These results indicate that the outcome of bronchopulmonary infections in lung cancer patients are affected mainly by the nutritional status of the host.

Emerg Infect Dis, 1997 Jan-Mar, 3(1), 33 - 9
Fluoroquinolone resistance in Neisseria gonorrhoeae; Knapp JS et al.; Fluoroquinolones and broad-spectrum cephalosporins are the most effective antimicrobial agents for the treatment of gonorrhea . However, clinically significant resistance to fluoroquinolones has emerged in Neisseria gonorrhoeae . Fluoroquinolone-resistant strains account for approximately 10% of all gonococcal strains in Hong Kong and the Republic of the Philippines . As many as 50% of strains from some Far Eastern countries exhibit decreased susceptibility (intermediate resistance) to fluoroquinolones . Strains with intermediate resistance and clinically significant resistance are being isolated sporadically in North America, where resistant strains have been associated with an outbreak and with failure of infections to respond to treatment with doses of ciprofloxacin and ofloxacin recommended by the Centers for Disease Control and Prevention; strains exhibiting decreased susceptibility to these agents are endemic in at least one metropolitan area . Monitoring for fluoroquinolone resistance is now critical for ensuring adequate treatment of infections with resistant strains and for maximizing the time during which fluoroquinolones may be used to treat gonorrhea.

Soz Praventivmed, 1997, 42(1), 30 - 6
Surveillance of sexually transmitted diseases in Switzerland, 1973-1994: evidence of declining trends in gonorrhoea and syphilis; Paget WJ et al.; The HIV/AIDS epidemic has led to growing interest in the epidemiology of sexually transmitted diseases (STDs) in Switzerland . STD surveillance data from three sources are presented: reports from six policlinics of dermatovenereology since 1973, laboratory reports of Neisseria gonorrhoeae, Treponema pallidum and Chlamydia trachomatis since 1988, and reports by the Swiss Sentinel Network between June 1991 and December 1993 . The data indicate that there has been a decline in the number of cases of syphilis and, in particular, gonorrhoea since the early 1980s in Switzerland . Since many factors could explain the declines (e.g . more effective treatments, more widespread use of antibiotics, marked changes in behaviour such as increased levels of condom use) it is difficult to identify their exact causes . Evidence for two of the before-mentioned factors exists and these probably played important roles in the declines . Firstly, as a result of the emergence of penicillinase producing strains of N . gonorrhoeae, the introduction of new treatments for gonorrhoea in the early 1980s which were simpler to administer and more effective (in particular the use of spectinomycin) . Secondly, the national AIDS prevention campaign which began in 1987 and has been associated with major increases in condom use in the Swiss population.

Mol Microbiol, 1997 Jan, 23(2), 281 - 93
Two-dimensional structure of the Opc invasin from Neisseria meningitidis; Merker P et al.; A two-dimensional structural model was devised for the Opc outer membrane protein invasin which contains 10 transmembrane strands and five surface-exposed loops . One continuous epitope recognized by three monoclonal antibodies was localized to the tip of loop 2 by synthetic peptides and site-directed mutagenesis while a second, discontinuous epitope recognized by a fourth antibody was localized to loops 4 and 5 by insertion mutagenesis . These monoclonal antibodies are bactericidal and inhibit adhesion and invasion . Most of the T-cell epitopes defined by Wiertz et al . (1996) were localized to the transmembrane strands . Oligonucleotides encoding a foreign epitope (nabla) from Semliki Forest virus were inserted into BglII restriction sites created by site-directed mutagenesis . The nabla epitopes inserted in all five predicted loops were recognized on the cell surface of live Escherichia coli bacteria by a monoclonal antibody and are exposed while nabla epitopes in the N-terminus or three predicted turns were not . The results thus confirm important predictions of the model and define five permissive sites within surface-exposed loops which can be used to insert foreign epitopes.

Int J STD AIDS, 1997 Jan, 8(1), 32 - 5
Heterosexual gonorrhoea at St Thomas'--I: Patient characteristics and implications for targeted STD and HIV prevention strategies; Daker-White G et al.; This paper compares the socio-demographic characteristics of patients with gonorrhoea with a control group of other attendees to an inner-London genitourinary medicine (GUM) clinic . Between 16 May and 21 December 1994 inclusive there were 312 culture-confirmed heterosexually-acquired cases of Neisseria gonorrheae treated in our clinic: 192 (61.54%) men and 120 (38.46%) women . There were significant differences between the population of cases and controls . Both male (z = -5.36, P < 0.001) and female (z = -6.6, P < 0.001) cases were younger than controls . Cases were more likely to be black African-Caribbean than were controls and these differences were more marked in men (chi 2 = 47.85, P < 0.001) . Cases were also more likely to reside in south London postal districts than were controls (chi 2 = 24.98, P < 0.001) . The implications of these findings for targeted health interventions are discussed and we suggest avenues for further work.

Sex Transm Dis, 1997 Jan, 24(1), 32 - 7
Increasing resistance of Neisseria gonorrhoeae in west and central Africa . Consequence on therapy of gonococcal infection; Van Dyck E et al.; BACKGROUND AND OBJECTIVES: Antimicrobial resistant strains of Neisseria gonorrhoeae have spread with remarkable rapidity in many African countries . Chromosomal resistance to penicillin, tetracycline, and thiamphenicol is frequent now, and reported prevalences of penicillinase-producing N . gonorrhoeae isolates vary between 15% and 80% . Plasmid-mediated tetracycline-resistant N . gonorrhoeae isolates have been observed in several African countries . GOALS: To characterize gonococcal isolates from three sites in West and Central Africa, to determine antimicrobial susceptibility patterns, to document the spread of plasmid-mediated resistance to penicillin and tetracycline in these three sites, and to discuss the consequences of rising antimicrobial resistance on the management of gonococcal infection in Africa . STUDY DESIGN: Over time, a total of 2,288 gonococcal isolates were obtained from Abidjan, Ivory Coast (1992-1993, n = 251), from Kigali, Rwanda (1988-1993, n = 952), and from Kinshasa, Zaire (1988-1990, n = 1,085) . The isolates were characterized by auxotyping and serotyping . Plasmid-mediated resistance to penicillin and to tetracycline was determined . Antimicrobial susceptibility testing to ceftriaxone, ciprofloxacin, penicillin, spectinomycin, tetracycline, and thiamphenicol was performed with an agar dilution method . RESULTS: The prevalence of penicillinase-producing N . gonorrhoeae increased significantly over time from 44% to 57% in Kigali and remained stable at a high level in Abidjan (73%) and in Kinshasa (67%) . The frequency of tetracycline-resistant N . gonorrhoeae increased significantly during the observation periods in all three sites: from 20% to 65% in Abidjan, from 0% to 64% in Kigali, and from 14% to 41% in Kinshasa . Chromosomal resistance to penicillin was common in Kigali and Kinshasa, and chromosomal resistance to tetracycline and thiamphenicol was frequent in all three sites . All gonococcal isolates were susceptible to ceftriaxone, ciprofloxacin, and spectinomycin . Prototrophic and proline requiring strains were predominant, and IA-6 was the most common serovar in the three sites . IB-specific serovars were more common among penicillinase-producing N . gonorrhoeae and IA-specific serovars were more frequent among tetracycline-resistant N . gonorrhoeae, but there was no evidence for a clonal spread of resistant strains . CONCLUSIONS: This study illustrates the high frequency of resistant gonococci in Africa and shows that tetracycline-resistant N . gonorrhoeae have become highly endemic in different geographic areas of the continent . The use of effective drugs is essential to reduce gonorrhea transmission . Surveillance of temporal changes in antimicrobial resistance in gonococcal strain populations should be part of sexually transmitted diseases control programs.

Sex Transm Dis, 1997 Jan, 24(1), 18 - 22
Antibiotic susceptibility patterns and plasmid profiles of penicillinase-producing Neisseria gonorrhoeae strains in Durban, South Africa, 1990-1993; Chenia HY et al.; BACKGROUND AND OBJECTIVES: The appearance of strains of Neisseria gonorrhoeae resistant, both chromosomally and plasmid-mediated, to penicillin and other antibiotics makes this versatile pathogen difficult to treat . There is, therefore, a need for surveillance of N . gonorrhoeae strains to determine the efficacy of current therapeutic measures . GOALS: To survey the antibiotic susceptibilities and plasmid profiles of penicillinase-producing N . gonorrhoeae strains isolated over a 4-year period . STUDY DESIGN: Penicillinase-producing N . gonorrhoeae strains were detected by the chromogenic cephalosporin test . Minimum inhibitory concentrations to penicillin G, tetracycline, ceftriaxone, and ciprofloxacin were determined using the E-test . Plasmid DNA was obtained by the alkaline lysis method and profiles generated . RESULTS: Penicillinase-producing N . gonorrhoeae strains increased from 16.4% to 19.0% in the period from 1990 through 1993 . Although all strains were resistant to penicillin, strains were susceptible to varying levels of ciprofloxacin, ceftriaxone, and even tetracycline . All penicillinase-producing N . gonorrhoeae strains possessed the 2.6-megadalton cryptic plasmid, and in addition 87.7% contained the 24.5-megadalton conjugative plasmid . Of the six known gonococcal beta-lactamase plasmids, the 4.4-megadalton Asian and 3.2-megadalton African plasmids were predominant . The most prevalent plasmid profile contained the 2.6-megadalton cryptic, 24.5-megadalton conjugative, and 4.4-megadalton Asian plasmids . CONCLUSIONS: To ensure effective treatment of gonorrhea, continued surveillance of the antimicrobial susceptibilities and plasmid profiles of penicillinase-producing N . gonorrhoeae strains is necessaryPIP: In South Africa, 321 gonococcal strains were isolated from urethral exudates of adult men presenting with acute urethritis at the Sexually Transmitted Diseases Clinic at King Edward VIII Hospital in Durban during 1990-1993 . The purpose was to examine the in vitro susceptibility patterns and plasmid profiles of penicillinase-producing Neisseria gonorrhoeae (PPNG) strains . 17.8% of the strains were PPNG strains . During 1990-1993, the PPNG prevalence increased from 16.4% to 19% . 96.5% of the PPNG strains were sensitive to ceftriaxone with minimum inhibitory concentrations (MICs) of no more than 0.25 g/ml . 94.7% were sensitive to ciprofloxacin with MICs of no more than 0.06 g/ml . 82.5% were sensitive to tetracycline with MICs of no more than 0.25 g/ml . The fact that all PPNG strains had the 2.6 megadalton (MDa) cryptic plasmid suggests that this plasmid confers a selective advantage to the gonococcus . 87.7% also had the 24.5 MDa conjugative plasmid . High prevalence of this plasmid in the PPNG strains may account for the increased prevalence of these strains . The predominant beta-lactamase plasmid type was the 4.4 MDa Asian plasmid (77.2%), not the 3.2 MDa African plasmid . The high prevalence of the Asian plasmid may be because Durban is an important port city and tourist destination for people from Asia . The most common plasmid profile of the PPNG strains contained the 2.6 MDa cryptic, 24.5 MDa conjugative, and 4.4 MDa Asian plasmids (68.4%) . Since N gonorrhoeae has several antibiotic resistance mechanisms, epidemiologists should monitor the effectiveness of single-dose treatment and conduct surveillance of trends in resistance in order to achieve effective control of gonorrhea .

Sex Transm Dis, 1997 Jan, 24(1), 2 - 7
Molecular epidemiology, in 1994, of Neisseria gonorrhoeae in Manila and Cebu City, Republic of the Philippines; Knapp JS et al.; BACKGROUND AND OBJECTIVES: Failure of gonococcal infections to respond to 500 mg of ciprofloxacin or 400 mg of ofloxacin has been reported from Australia, the United Kingdom, and the United States . Recently, high rates of decreased susceptibility to the fluoroquinolones have been detected in penicillinase-producing Neisseria gonorrhoeae in the Republic of the Philippines . GOALS: To assess the diversity of antimicrobial-resistant gonococcal strains isolated from female sex workers in Manila and Cebu City in the Republic of the Philippines in 1994 . STUDY DESIGN: Isolates of N . gonorrhoeae isolated from 92 female sex workers in Manila (n = 28) and Cebu City (n = 64), respectively, were characterized by plasmid profile, auxotype/serovar class, and antimicrobial susceptibility profile . RESULTS: Plasmid-mediated resistance to penicillin or tetracycline was identified in 79.3% (73/92) of the isolates: penicillinase-producing N . gonorrhoeae (65/92; 70.7%), tetracycline-resistant N . gonorrhoeae (6/92; 6.5%), and penicillinase-producing/tetracycline-resistant N . gonorrhoeae (1/92; 1.1%) . A beta-lactamase plasmid of 3.9 megadaltons was discovered . Of 54.3% (50/92) of strains resistant to nalidixic acid, 84% (42/50) of strains had minimum inhibitory concentrations of > or = 0.125 microgram/ml ciprofloxacin; penicillinase-producing N . gonorrhoeae (possessing the 3.05-, 3.2-, 3.9-, and 4.4-megadalton beta-lactamase plasmids, respectively) accounted for 68% (34/50) of these strains . CONCLUSIONS: In the Republic of the Philippines, gonococcal isolates resistant to penicillin or tetracycline accounted for 85.9% (79/92) of the isolates examined and included strains exhibiting resistance to fluoroquinolones . All gonococcal infections should be treated with antimicrobial therapies known to be active against all gonococcal strains to reduce the spread of strains exhibiting decreased susceptibilities to fluoroquinolonesPIP: During July-October 1994 in the Philippines, Neisseria gonorrhoeae were isolated from 92 female sex workers in Manila and Cebu City . The purpose was to characterize the gonococcal strains by plasmid content, auxotype, serovar, and antimicrobial susceptibilities in order to examine the diversity of antimicrobial-resistant N gonorrhoeae strains in these sex workers . Penicillinase-producing N gonorrhoeae (PPNG) comprised 70.7% of the isolates . Strains with the 3.2-megadalton (Mda) beta-lactamase plasmid were more common in Cebu City than in Manila (57.8% vs . 28.6%; p = 0.02) . They had significantly lower minimum inhibitory concentrations (MICs) to penicillin, tetracycline, ceftriaxone, ciprofloxacin, and erythromycin than did strains with 3.05-, 3.9-, or 4.4-Mda plasmid (p 0.01) . One PPNG strain had a previously undescribed 3.9-Mda beta-lactamase plasmid . It also had a 24.5-Mda conjugative plasmid . Only 15.2% of all 92 isolates were susceptible to both penicillin and tetracycline . Tetracycline-resistant N gonorrhoeae (TRNG) comprised 6.5% of the isolates . 1.1% of isolates were resistant to both penicillin and tetracycline . 54.3% of all strains were resistant to nalidixic acid (fluoroquinolone), 84% of which had MICs of at least 0.125 mcg/ml ciprofloxacin . 68% of the nalidixic acid-resistant strains had 4.4 Mda beta-lactamase plasmids . 85.9% of all gonococcal isolates exhibited resistance to penicillin or tetracycline . They also included strains exhibiting resistance to fluoroquninolones . These findings reveal the need for periodic surveillance for resistance in N gonorrhoeae to the antimicrobial agents used for primary gonorrhea therapy in order to improve the treatments of choice .

Mol Microbiol, 1997 Jan, 23(1), 123 - 35
Promoter strength influences phase variation of neisserial opa genes; Belland RJ et al.; The opa multigene family of Neisseria gonorrhoeae encodes 11 related outer-membrane proteins which phase vary in vitro and in vivo . Illegitimate recombination within direct pentameric DNA repeats, encoding the signal-peptide region of pre-Opas, leads to switches in expression states . Despite the conserved nature of the variation mechanism, specific genes are expressed at high frequencies in the transition from Opa- to Opa+ . The genes which are expressed at elevated frequencies differ from the rest of the family with respect to promoter structure, based on sequence comparisons between the opa genes of strain MS11mk . We have analysed transcription of the opa gene family of N . gonorrhoeae MS11mk, focussing on the different promoters found among the 11 genes to determine whether increased levels of expression are associated with increased phase-variation rates . Primer extension and Northern blotting was used to assess the levels of transcription of three representative opa genes (opaA, B and C) in 'on' and 'off' states . Full-length opa mRNA was detected primarily in strains expressing the homologous gene . Truncated opa mRNA was constitutively expressed from all opa genes regardless of their expression state . Quantitative comparisons in N . gonorrhoeae were complicated by the simultaneous expression of all 11 genes and the cross-reactivity of mRNA probes . Expression levels from the individual promoters were therefore assessed by creating transcriptional and translational lacZ fusions to each of the representative opa promoters which lacked the DNA repeats responsible for variation . The expression levels were compared to the phase-variation rates of translational opa::phoA fusions containing the same promoters in addition to the corresponding coding repeat regions . A strong correlation was found between expression levels from the different promoters and the variation rates at which 'on' variants appeared from an 'off' population (i.e . opaA > opaB > opaC) . These results provide an explanation for the favoured expression of specific Opa proteins and indicate that expression of opa genes may be regulated at the level of transcription.

Clin Infect Dis, 1997 Jan, 24 Suppl 1, S93 - 7
Antimicrobial resistance in Neisseria gonorrhoeae; Lind I; The changing patterns of antimicrobial resistance in Neisseria gonorrhoeae have been reviewed regularly since the introduction of antimicrobial therapy in the 1930s . At present, ceftriaxone, fluoroquinolones, and spectinomycin have remained efficient as single-dose treatment of gonorrhea worldwide . To ensure that limited resources can be used in the best possible way, continuous surveillance of gonococcal resistance to antimicrobials is needed.

Kyobu Geka, 1997 Jan, 50(1), 78 - 81
{A case of acute mediastinitis with pyothorax secondary to peritonsillar abscess}; Iriyama T et al.; A 41-year-old man was admitted to a hospital elsewhere because of tonsillitis with high grade fever . On the 9th day of hospitalization, the patient complained of dysphagia and dyspnea . A chest X-ray film and a CT scan showed right pleural effusion and pericardial effusion, and he was referred to our hospital . Immediately after admission, he underwent pericardiotomy to relieve cardiac tamponade, and a right thoracic tube was inserted for pyothorax . Next day, mediastinal drainage was accomplished through a cervical incision and a right thoracotomy . Eight drainage tubes were left in place . Cultures revealed alpha-Streptococcus, Neisseria and group F Streptococci . Continuous closed irrigation with diluted Isodine (povidone iodine) solution was performed . The last extubation of the drainage tube was done on the 140th day after operation . He was cured and discharged on the 162nd day after operation . In patients with extensive acute mediastinitis secondary to deep cervical infection, early complete mediastinal drainage via a cervical and a transthoracic incision is essential.

J Clin Microbiol, 1997 Jan, 35(1), 239 - 42
Comparison of ligase chain reaction and culture for detection of Neisseria gonorrhoeae in genital and extragenital specimens; Stary A et al.; In addition to the urogenital tract, Neisseria gonorrhoeae infects extragenital sites such as the pharynx and anorectal canal . Culture and a ligase chain reaction (LCR)-based assay were compared for their performance for the diagnosis of N . gonorrhoeae infection with specimens from various urogenital and extragenital sites of 200 men and 125 women . The sensitivity and specificity of the LCR assay with male urethral swabs were both 100%, compared to values of 95.9 and 100%, respectively, for culture of urethral swabs or 98.0 and 100%, respectively, for LCR with first-void urine (FVU) . For women, LCR with FVU showed the highest sensitivity (94.7%), and culture of urethral samples showed the lowest sensitivity (63.2%) (P < 0.05) . In a selected subgroup of 47 men and 22 women at increased risk, the rates of pharyngeal infection were 15 and 18%, respectively, and those of anorectal infection were 13 and 45%, respectively . The sensitivity of LCR was greater than that of culture for both pharyngeal and anorectal specimens . Thus, the overall performance of LCR testing with swabs or FVU was better than that of culture for the diagnosis of genital or extragenital gonorrhea.

FEMS Microbiol Lett, 1996 Dec 15, 145(3), 371 - 6
Pseudo-transposition of a Tn5 derivative in Neisseria gonorrhoeae; Thomas CE et al.; We constructed a Tn5 derivative for potential use in transposon mutagenesis of Neisseria gonorrhoeae . It was incorporated into the chromosome apparently at random following transformation, but the insertion events were dependent on a functional RecA and independent of a functional transposase . Furthermore, in most cases there was an incomplete transposon inserted with little or no IS50 insertion sequence . These observations suggest that Tn5 transposition may not be possible in N . gonorrhoeae and that this organism may have an unexplored illegitimate recombination system.

Gene, 1996 Dec 12, 183(1-2), 215 - 8
Molecular cloning and expression of NlaIII restriction-modification system in E . coli; Morgan RD et al.; The NlaIII restriction enzyme isolated from Neisseria lactamica recognizes the sequence 5'-CATG-3', cleaving after the G to generate a four base 3' overhang . The NlaIII methylase and a portion of the NlaIII endonuclease gene were cloned into E . coli by the methylase selection method, and the remaining portion of the NlaIII endonuclease gene was cloned by inverse PCR . The nucleotide sequence of the endonuclease gene and the methylase gene were determined . The NlaIII endonuclease gene is 693 bp, encoding a protein with predicted molecular weight of 26487 . The NlaIII methylase gene was identical with that previously reported {Labbe, D., Joltke, H.J . and Lau, P.C . (1990) Cloning and characterization of two tandemly arranged DNA methyltransferse genes of Neisseria lactamica: an adenine-specific M.NlaIII and a cytosine-type methylase . Mol . Gen . Genet . 224, 101-110} . The endonuclease and methylase genes overlap by four bases and are transcribed in the same orientation . The endonuclease gene was cloned into an improved T7 vector, and a high level of NlaIII endonuclease expression was achieved in E . coli.

Gene, 1996 Dec 12, 183(1-2), 115 - 21
Cloning, sequencing and expression of an Eikenella corrodens gene encoding a component protein of the lectin-like adhesin complex; Yumoto H et al.; A lectin-like substance (LS), that was isolated from Eikenella corrodens (Ec) 1073, migrated as proteins of about 300 and 45 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions . In this study, we cloned the gene encoding the 45-kDa protein and predicted its structure and function . Based on the N-terminal 23-amino acid (aa) sequence of this protein, we cloned the region for its N-terminus . We cloned the entire gene by means of gene walking using polymerase chain reaction and Southern hybridization . The nucleotide sequences of cloned fragments revealed an open reading frame encoding a polypeptide of 330 aa (M(r), 35748) . This ORF displayed high homology to those of porins of Neisseria species . Using the T7-expression system, the 45-kDa protein was produced in E . coli . Our results suggested that the 45-kDa protein of Ec 1073 is a component of the EcLS complex, and that it is the major outer membrane protein.

Proc Natl Acad Sci U S A, 1996 Dec 10, 93(25), 14851 - 6
CGM1a antigen of neutrophils, a receptor of gonococcal opacity proteins; Chen T et al.; Neisseria gonorrhoeae (GC) or Escherichia coli expressing phase-variable opacity (Opa) protein (Opa+ GC or Opa+ E . coli) adhere to human neutrophils and stimulate phagocytosis, whereas their counterparts not expressing Opa protein (Opa- GC or Opa- E . coli) do not . Opa+ GC or E . coli do not adhere to human lymphocytes and promyelocytic cell lines such as HL-60 cells . The adherence of Opa+ GC to the neutrophils can be enhanced dramatically if the neutrophils are preactivated . These data suggest that the components binding the Opa+ bacteria might exist in the granules . CGM1a antigen, a transmembrane protein of the carcinoembryonic antigen family, is exclusively expressed in the granulocytic lineage . The predicted molecular weight of CGM1a is approximately 30 kDa . We observed specific binding of OpaI+ E . coli to a 30-kDa band of polymorphonuclear leukocytes lysates . To prove the hypothesis that the 30-kDa CGM1a antigen from neutrophils was the receptor of Opa+ bacteria, we showed that a HeLa cell line expressing human CGM1a antigen (HeLa-CGM1a) bound Opa+ E . coli and subsequently engulfed the bacteria . Monoclonal antibodies (COL-1) against CGM1 blocked the interaction between Opa+ E . coli and HeLa-CGM1a . These results demonstrate that HeLa cells when expressing the CGM1a antigens bind and internalize OpaI+ bacteria.

Arch Mal Coeur Vaiss, 1996 Dec, 89(12), 1689 - 93
{Neisseria Sicca endocarditis disclosed by multiple organ failure syndrome}; Rozenberg V et al.; The authors report a case of Neisseria sicca endocarditis presenting with multiple organ failure in a 33 year old intravenous drug user . The diagnosis was confirmed by transthoracic and transoesophageal echocardiography showing vegetations on the aortic valve and three blood cultures positive for Neisseria Sicca . Massive aortic regurgitation occurred on the 4th day . The patient died of complications of intracerebral haematomas before surgical intervention . Contrary to pathogenic Neisseria gonorrhoeae and meningitides, saprophytic Neisseria, including Neisseria sicca, are commensal organisms of the upper respiratory tract . They are exceptionally rare causes of endocarditis . A review of the literature from the era of antibiotic therapy, found about thirty cases of saprophytic Neisseria endocarditis of which only five were due to Neisseria sicca . The clinical characteristics were the young age, the mainly left heart valve disease and the high incidence of cerebrovascular accidents . The originality of this case was the exceptionally rare involvement of this organism and the multiplicity of the extracardiac manifestations, especially renal and neurological.

Antimicrob Agents Chemother, 1996 Dec, 40(12), 2775 - 80
Comparison of single-dose cefuroxime axetil with ciprofloxacin in treatment of uncomplicated gonorrhea caused by penicillinase-producing and non-penicillinase-producing Neisseria gonorrhoeae strains; Thorpe EM et al.; A randomized, multicenter, investigator-blind trial was conducted to compare the efficacies of cefuroxime axetil and ciprofloxacin for treatment of patients with uncomplicated gonorrhea caused by penicillinase-producing Neisseria gonorrhoeae (PPNG) . A total of 832 patients (434 females and 398 males) received a single oral dose of cefuroxime axetil (1,000 mg {417 patients}) or ciprofloxacin (500 mg {415 patients}) . N . gonorrhoeae was eradicated from the cervix in 114 of 118 (97%) and 118 of 119 (99%) bacteriologically evaluable females treated with cefuroxime axetil and ciprofloxacin, respectively (P = 0.213; difference, -2%; 95% confidence interval, -6 to 1%), and from the urethra in 154 of 166 (93%) and 171 of 171 (100%) bacteriologically evaluable male patients treated with cefuroxime axetil and ciprofloxacin, respectively (P < 0.001; difference, -7%; 95% confidence interval, -11 to -3%) . Both treatments were effective in eradicating N . gonorrhoeae in females with rectal infections (cefuroxime axetil, 29 of 30 {97%}; ciprofloxacin, 25 of 25 {100%}; P = 1.00) . In small numbers of patients, cefuroxime axetil was less effective than ciprofloxacin in treating males with pharyngeal infections (eradication in 4 of 10 and in 8 of 8 patients, respectively; P = 0.013) . PPNG was eradicated from the cervix in 22 of 23 (96%) and 32 of 32 (100%) bacteriologically evaluable female patients treated with cefuroxime axetil and ciprofloxacin, respectively (P = 0.418; difference, -4%; 95% confidence interval, -13 to 4%), and from the urethra in 35 of 36 (97%) and 34 of 34 (100%) bacteriologically evaluable male patients treated with cefuroxime axetil and ciprofloxacin, respectively (P = 1.00; difference, -3%; 95% confidence interval, -8 to 3%) . The incidences of drug-related adverse events were similar for the two study drugs . In summary, treatment with a single oral dose of cefuroxime axetil is as effective as treatment with a single oral dose of ciprofloxacin in eradicating PPNG from males and females with uncomplicated gonorrhea (urethral and endocervical), and both regimens are well-tolerated . However, in the present study, cefuroxime axetil was less effective than ciprofloxacin in treating urethral gonococcal infections in male patients, although both study drugs were highly effective in treating cervical gonococcal infections in female patients.

Mol Biotechnol, 1996 Dec, 6(3), 231 - 40
Generation of antiserum to specific epitopes; Marchion DC et al.; The ability to prevent disease by immunization with subunit vaccines that incorporate specific epitopes was demonstrated by DiMarchi et al . (1), who used a synthetic peptide to protect cattle against foot-and-mouth disease . However, generation of antibody to peptide antigens is often difficult owing to the small molecular mass and limited chemical complexity . We tested the hypothesis that recombinant DNA and synthetic peptide techniques would make it possible to stimulate vigorous immune responses to specific epitopes of an outer membrane protein of Neisseria gonorrhoeae . The MtrC AP1 sequence from the invariant MtrC gonococcal lipoprotein was genetically fused to maltose binding protein . The resultant fusion protein was used as the primary immunogen to stimulate MtrC AP1-specific antiserum . To enhance antibody production specific to MtrC AP1, boosting immunizations were performed with synthetic MtrC AP1 sequence contained in a multiple antigenic peptide system immunogen . The MtrC AP1-specific antiserum strongly recognized the MtrC protein on Western blots and appeared to bind native MtrC protein in situ . The generation of antibody in this fashion provides the technology to produce antibody to defined epitopes of any protein, including those found in the gonococcal outer membrane . The ability of those antibodies to inhibit bacterial growth or to activate complement protein can then be tested.

J Antibiot (Tokyo), 1996 Dec, 49(12), 1236 - 48
Synthesis and antibacterial activity of derivatives of the glycopeptide antibiotic A-40926 and its aglycone; Hermann R et al.; Starting from the antibiotic A-40926 and the aglycone of A-40926 a series of compounds were prepared by modifying the free functionalities . Their antimicrobial activity was determined, particularly against Neisseria gonorrhoeae, against which A-40926, unlike other natural glycopeptides, is active . Improved in vivo activity was displayed by the monomethyl ester of A-40926 esterified at the carboxyl group of the N-acylamino-glucuronyl moiety.

Lab Anim Sci, 1996 Dec, 46(6), 656 - 62
Microbiologic, radiographic, and anatomic study of the nasolacrimal duct apparatus in the rabbit (Oryctolagus cuniculus); Marini RP et al.; This study was motivated by the sporadic observation of epiphora in two male rabbits . The epiphora was unilateral and not associated with conjunctivitis or Pasteurella infection . To characterize the cause of epiphora, we studied 15 specific-pathogen-free New Zealand White rabbits . This study group was composed of the two affected males, four unaffected males, and nine unaffected females . Clinical evaluation consisted of bacterial culture of conjunctival specimens, examination of conjunctival scrapings for chlamydial inclusions, culture and cytologic examination of specimens from the nasolacrimal duct, plain and contrast radiography, latex casting, histologic examination, and the Schirmer tear test . Important differences found in the rabbits with epiphora included an opalescent, gritty, nasolacrimal duct flush fluid and marked unilateral dilatation of the duct proximal to a dorsal flexure at the caudal limit of the incisor tooth root . The flush solution from one affected rabbit cleared with ether, suggesting the presence of triglycerides or cholesterol . The organisms most commonly isolated from the conjunctiva were Moraxella sp., Oligella urethralis, Staphylococcus aureus, coagulase-negative Staphylococcus sp., and Streptococcus viridans . The organisms most commonly isolated from the nasolacrimal duct flush fluid were Moraxella sp., S . viridans, and Neisseria sp . Culture of the nasolacrimal duct flush fluid yielded microorganisms more consistently than did culture of the conjunctival specimens . All microorganisms isolated from affected rabbits also were isolated from unaffected rabbits . There was no apparent contribution of microorganisms to the development of epiphora, and Schirmer tear test results for affected animals were within the range seen in unaffected animals . Occlusion of the nasolacrimal duct was presumed to be attributable to fat droplets . This study augments the existing literature and represents the first report of anomalous nasolacrimal duct anatomic features in the rabbit.

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