Neisseria

J Mol Evol, 1996 Dec, 43(6), 631 - 40
A comparison of the nucleotide sequences of the adk and recA genes of pathogenic and commensal Neisseria species: evidence for extensive interspecies recombination within adk; Feil E et al.; The sequences of the adenylate kinase gene (adk) and the RecA gene (recA) were determined from the same isolates of Neisseria gonorrhoeae, N . meningitidis, N . lactamica, N . polysaccharea, N . cinerea, N . mucosa, N . pharyngis var . flava, N . flavescens, and N . animalis . The patterns of sequence divergence observed at adk and recA were very different . Dendrograms constructed from the recA data using two different algorithms were statistically robust and were congruent with each other and with the relationships between the species previously proposed using other data . In contrast, the dendrograms derived from the adk data were noncogruent with each other, and with those from the recA data, and were statistically poorly supported . These results, along with the uniform distribution of pairwise sequence divergences between the species at adk, suggest there has been a history of interspecies recombination within the adk gene of the human Neisseria species which has obscured the phylogenetic relationships between the species . This view was supported by Sawyer's runs test, and the Index of Association (IA) between codons, which provided significant evidence for interspecies recombination between the adk genes from the human Neisseria species, but no evidence of interspecies recombination between the recA sequences.

FEMS Immunol Med Microbiol, 1996 Dec 1, 16(2), 127 - 39
Fimbrial adhesins: similarities and variations in structure and biogenesis; Smyth CJ et al.; Fimbriae are wiry (2 to 4 nm diam.) or rod-shaped (6 to 8 nm diam.), fibre-like structures on the surfaces of bacteria which mediate attachment to host cells . Much has been learned in recent years about the biogenesis, structure and regulation of expression of these adhesive organelles in Gram-negative bacteria . Analyses of the genetic determinants encoding the biogenesis of fimbriae has revealed that the adhesive interaction of fimbriae can be mediated by major subunits (CFA/I and CS1 fimbriae) or minor subunits (P, S, and type 1 fimbriae), with the adhesin being located either at the tip of the fimbria or along the length of the fimbrial shaft . Minor subunits can also act as adapters, anchors, initiators or elongators . Post-translational glycosylation of the type 4 pilins of Neisseria gonorrhoeae, Neisseria meningitidis and Pseudomonas aeruginosa has been demonstrated . The structures of the PapD chaperone of Escherichia coli and of N . gonorrhoeae type 4 fimbrin have been resolved at 2.0-2.6 A . Rod-shaped fimbriae should not be thought of as being rigid inflexible structures but rather as dynamic structures which can undergo transition from a helicoidal to a fibrillar conformation to provide a degree of elasticity and plasticity to the fimbriae so that they can resist shear forces, rather like a bungee cord . At least four mechanisms have been identified in the assembly of fimbriae from fimbrin subunits, namely the chaperone-usher pathway (e.g., P-fimbriae of uropathogenic E . coli), the general secretion assembly pathway (e.g., type 4 fimbriae or N-methylphenylalanine fimbriae of P . aeruginosa, the extracellular nucleation-precipitation pathway (e.g., curli of E . coli) and the CFA/I, CS1 and CS2 fimbrial pathway.

Eur J Epidemiol, 1996 Dec, 12(6), 651 - 4
Neisseria gonorrhoeae RNA/DNA hybridization and culture for screening of gonococcal infections in a low-prevalence population; Sednaoui P et al.; Gonorrhea is still a major sexually transmitted disease (STD) worldwide . Its etiologic diagnosis is based on identification of the causative agent, Neisseria gonorrhoeae, by culture of genital secretions, which is often hampered by difficulties of sample collection and transport . Alternatively, nucleic acid hybridization techniques for routine diagnosis of N . gonorrhoeae appear to be useful by eliminating problems associated with bacterial viability, particularly for surveillance of low-prevalence populations . Our study among 1,508 outpatients undergoing routine examination for common STDs used RNA/DNA hybridization with a DNA probe specific for N . gonorrhoeae (Gen Probe Pace 2) and classical culture . Of the 1,750 specimens tested, 12 were positive by DNA probe and culture . In 8 cases, only DNA probe was positive while culture was negative . In 3 of these discrepant cases clinical and epidemiological data suggested true N . gonorrhoeae infection . Thus, DNA probe assay for N . gonorrhoeae may greatly improve screening of N . gonorrhoeae among low-prevalence populations . However, culture remains mandatory for testing antimicrobial resistance of these highly communicable infectious agents.

Epidemiol Infect, 1996 Dec, 117(3), 423 - 8
An 18 year clinical review of septic arthritis from tropical Australia; Morgan DS et al.; A retrospective study of 191 cases of septic arthritis was undertaken at Royal Darwin Hospital in the tropical north of Australia . Incidence was 9.2 per 100,000 overall and 29.1 per 100,000 in Aboriginal Australians (RR 6.6; 95% CI 5.0-8.9) . Males were affected more than females (RR 1.6; 95% CI 1.2-2.1) . There was no previous joint disease or medical illness in 54% . The commonest joints involved were the knee (54%) and hip (13%) . Significant age associations were infected hips in those under 15 years and infected knees in those over 45 years . Seventy two percent of infections were haematogenous . Causative organisms included Staphylococcus aureus (37%), Streptococcus pyogenes (16%) and Neisseria gonorrhoeae (12%) . Unusual infections included three melioidosis cases . Polyarthritis occurred in 17%, with N . gonorrhoeae (11/23) more likely to present as polyarthritis than other organisms (22/168) (OR 6.0; 95% CI 2.1-16.7) . Univariate and multivariate analysis showed the hip to be at greater risk for S . aureus than other joints . Open arthrotomy was a more successful treatment procedure than arthroscopic washout or needle aspiration.

Clin Chem, 1996 Dec, 42(12), 1915 - 23
COBAS AMPLICOR: fully automated RNA and DNA amplification and detection system for routine diagnostic PCR; DiDomenico N et al.; The COBAS AMPLICOR system automates amplification and detection of target nucleic acids, making diagnostic PCR routine for a variety of infectious diseases . The system contains a single thermal cycler with two independently regulated heating/cooling blocks, an incubator, a magnetic particle washer, a pipettor, and a photometer . Amplified products are captured on oligonucleotide-coated paramagnetic microparticles and detected with use of an avidin-horseradish peroxidase (HRP) conjugate . Concentrated solutions of amplicon or HRP were pipetted without detectable carryover . Amplified DNA was detected with an intraassay CV of < 4.5%; the combined intraassay CV for amplification and detection was < 15% . No cross-reactivity was observed when three different target nucleic acids were amplified in a single reaction and detected with three target-specific capture probes . The initial COBAS AMPLICOR menu includes qualitative tests for diagnosing infections with Chlamydia trachomatis, Neisseria gonorrhoeae, Mycobacterium tuberculosis, and hepatitis C virus . All tests include an optional Internal Control to provide assurance that specimens are successfully amplified and detected.

FEMS Microbiol Lett, 1996 Dec 1, 145(2), 201 - 7
The prevalence of gentamicin 2'-N-acetyltransferase in the Proteeae and its role in the O-acetylation of peptidoglycan; Clarke AJ et al.; The prevalence of aac(2')-Ia, a gene coding for gentamicin 2'-N-acetyltransferase in Providencia stuartii, among species of the Proteeae was investigated to determine if it is a common resistance factor and whether the correlation observed in P . stuartii between its expression and the levels of peptidoglycan O-acetylation represents a general feature of bacteria producing this form of modified peptidoglycan . An evaluation of the MICs of gentamicin for each of the species of the Proteeae did not reveal any apparent relationship between resistance and the degree of O-acetylation of peptidoglycan . The entire aac(2')-Ia gene was used as a probe in Southern hybridization experiments against genomic DNA from each species of the Proteeae . A sequence with strong homology to aac(2')-Ia was present only in Proteus penneri while weak hybridization was also observed to the restriction digested DNA from Providencia rettgeri . Other bacteria that O-acetylate peptidoglycan were also screened with this probe and a homologous DNA sequence was only found in Neisseria subflava . These data suggest that AAC(2')-Ia may contribute to the O-acetylation of peptidoglycan in P . stuartii, but a more specific enzyme must also be produced for this function.

FEMS Microbiol Lett, 1996 Dec 1, 145(2), 173 - 9
Genome plasticity in Neisseria gonorrhoeae; Gibbs CP et al.; The pathogenic Neisseria have exploited the processes of horizontal DNA transfer and genetic recombination as mechanisms for the generation of extensive protein variation and modulation of gene expression . Localized recombinations have been well documented in members of multigene families as have alterations in short repetitive sequences . Here we report an analysis of the chromosomal structure of a defined lineage of Neisseria gonorrhoeae strain MSl1 pilin variants . This study reveals the occurrence of large rearrangements, including the amplification of a 26 kb region and an inversion involving more than a third of the chromosome . Additionally, a restriction site polymorphism that correlates with pilin expression has been observed . These findings highlight the flexibility of the gonococcal genome.

Infect Dis Clin North Am, 1996 Dec, 10(4), 797 - 809
Acute bacterial meningitis; Segreti J et al.; Despite improvements in antibiotic therapy and the use of vaccines and chemoprophylaxis, acute bacterial meningitis remains a significant cause of morbidity and mortality in the United States . Early diagnosis and therapy are important once the condition has been considered and the appropriate available specimens collected . Changes in epidemiologic frequencies and antimicrobial susceptibilities suggest that therapy will become more uniform across all age groups . Rapid, specific diagnostic modalities for all etiologic agents and improved vaccines for Neisseria meningitidis type B and Streptococcus pneumoniae are urgently needed.

Infect Immun, 1996 Dec, 64(12), 5263 - 8
Outer membrane protein of Neisseria meningitidis as a mucosal adjuvant for lipopolysaccharide of Brucella melitensis in mouse and guinea pig intranasal immunization models; Van De Verg LL et al.; A mucosal vaccine against brucellosis consisting of the lipopolysaccharide (LPS) of Brucella melitensis complexed with the outer membrane protein (GBOMP) of group B Neisseria meningitidis was tested in small-animal models of intranasal immunization . Mice given two doses of the vaccine developed high levels of immunoglobulin G (IgG) and IgA antibodies specific for B . melitensis LPS in lung lavages and specific IgG and IgA antibody-secreting cells in the lungs and spleen . Similarly, in guinea pigs immunized twice intranasally, IgG and IgA LPS-specific antibodies were detected in lung lavages, and specific antibody-secreting cells were isolated from the spleen and cervical nodes . In mice immunized with LPS only, pulmonary responses consisted mostly of IgM antibodies, while guinea pigs given LPS alone developed local antibody of all three isotypes, but at lower levels compared to animals given the complex vaccine . Both mice and guinea pigs also developed high levels of serum IgG and moderate levels of IgA as a result of intranasal immunization with the complex vaccine . The serum antibodies in both cases were found to cross-react with the LPS of B . abortus, which shares an immunogenic epitope with B . melitensis LPS . In mice given the complex vaccine, there was a prominent serum IgG1 response that was absent in the mice given LPS alone . In conclusion, the N . meningitidis GBOMP was an effective mucosal adjuvant for secretory IgA and IgG responses in the lungs of both mice and guinea pigs . The IgG1 subclass response in mice suggests that GBOMP may have favored a Th2 type of response to the LPS . A vaccine capable of stimulating high levels of antibody at local sites has the potential to protect against brucellae, since these pathogens gain entry to the host via mucosal routes.

J Clin Microbiol, 1996 Dec, 34(12), 3214 - 7
Comparative assessment of Etest for testing susceptibilities of Neisseria gonorrhoeae to penicillin, tetracycline, ceftriaxone, cefotaxime, and ciprofloxacin: investigation using 510(k) review criteria, recommended by the Food and Drug Administration; Biedenbach DJ et al.; We evaluated the ability of the Etest (AB Biodisk, Solna, Sweden) method to accurately and reproducibly determine the antimicrobial susceptibility of Neisseria gonorrhoeae . One hundred gonococcal isolates were used to evaluate the diagnostic performance of the Etest compared with the reference agar dilution method for penicillin, tetracycline, ciprofloxacin, and ceftriaxone . Between 92 and 99% of Etest MIC results for all drugs were within +/- 1 log2 dilution of the reference MIC . According to recommended interpretive criteria, ceftriaxone, cefotaxime, and ciprofloxacin had 100% categorical agreement, while penicillin (86%) and tetracycline (85%) categorical agreement percentages were lower because of the large number of strains that were within 0.5 to 1 log2 dilution of the susceptible or resistant breakpoints . Reproducibility data also demonstrated that the Etest was precise (99.1%) when subjected to replicate testing . On the basis of these data, the Etest method provides an effective, simple alternative to the reference agar dilution method for the direct quantification of N . gonorrhoeae susceptibility.

J Infect Dis, 1996 Dec, 174(6), 1238 - 48
Experimental immunization with a monoclonal anti-idiotope antibody that mimics the Neisseria gonorrhoeae lipooligosaccharide epitope 2C7; Gulati S et al.; An anti-idiotope monoclonal antibody (MAb), called CA1 (Ab2), was produced in mice against MAb 2C7, which recognizes a widely in vivo-expressed gonococcal lipooligosaccharide (LOS) epitope . Mice immunized with MAb CA1 initially had a 2.5-fold increase in IgG (12-fold after a booster) but no increase in IgM anti-LOS (Ab1') antibody . Control mice immunized with LOS had a 4.5-fold rise in IgG and 4-fold rise in IgM anti-LOS antibody . In rabbits, MAb CA1 elicited a 9-fold rise in IgG and a 3.3-fold rise in IgM anti-LOS (Ab1') antibody . Ab1' antibody bactericidal activity was 1-2 logs greater than that produced by immunization with LOS . Ab1' mediated complete human polymorphonuclear leukocyte phagocytosis of 2C7 epitope-positive (but not 2C7 epitope-negative) gonococci . MAb CA1 acts as a molecular surrogate (Ab2beta) for the nominal LOS antigen and may form the basis for vaccine candidates for human immunization against Neisseria gonorrhoeae.

J Infect Dis, 1996 Dec, 174(6), 1223 - 37
Immunogenicity of Neisseria gonorrhoeae lipooligosaccharide epitope 2C7, widely expressed in vivo with no immunochemical similarity to human glycosphingolipids; Gulati S et al.; Natural infection with Neisseria gonorrhoeae may elicit a substantial antibody response directed against gonococcal lipooligosaccharide . Monoclonal antibody (MAb) 2C7 recognized a gonococcal lipooligosaccharide epitope, identified the epitope directly in 94% of 68 consecutive culture-positive genital secretions, and recognized 95% of 101 randomly chosen fresh (second-passage) gonococcal isolates . The epitope was stably maintained after multiple in vitro passages and did not compete with any of the known cross-reactive human glycosphingolipid structures . MAb 2C7 mediated in vitro killing and phagocytosis by human polymorphonuclear leukocytes of 1 serum-sensitive (sialylated or not) and 1 stably serum-resistant gonococcal isolate that expressed the epitope . Gonococcal endometritis and disseminated infection elicited increases (6.5-fold IgM, 4.4-fold IgG; 18-fold IgM, 17-fold IgG, respectively) in anti-2C7 epitope antibody . Immunization with a gonococcal outer membrane vaccine elicited a mean 44.5-fold increase in IgG anti-2C7 epitope antibody in 20 of 28 subjects . The epitope identified by MAb 2C7 may represent an excellent target for a potentially protective gonococcal vaccine candidate.

J Biol Chem, 1996 Nov 8, 271(45), 28271 - 6
Cloning of the lipooligosaccharide alpha-2,3-sialyltransferase from the bacterial pathogens Neisseria meningitidis and Neisseria gonorrhoeae; Gilbert M et al.; The genes encoding the alpha-2,3-sialyltransferases involved in lipooligosaccharide biosynthesis from Neisseria meningitidis and Neisseria gonorrhoeae have been cloned and expressed in Escherichia coli . A high sensitivity enzyme assay using a synthetic fluorescent glycosyltransferase acceptor and capillary electrophoresis was used to screen a genomic library of N . meningitidis MC58 L3 in a "divide and conquer" strategy . The gene, denoted lst, was found on a 2 . 0-kilobase fragment of DNA, and its sequence was determined and then used to design probes to amplify and subsequently clone the corresponding lst genes from N . meningitidis 406Y L3, N . meningitidis M982B L7, and N . gonorrhoeae F62 . Functional sialyltransferase was produced from the genes derived from both L3 N . meningitidis strains and the N . gonorrhoeae F62 . However, the N . meningitidis M982B L7 gene contained a frameshift mutation that renders it inactive . The expression of the lst gene was easily detected using the enzyme assay, and the protein expression could be detected when an immunodetection tag was added to the COOH-terminal end of the protein . Using the synthetic acceptor N-acetyllactosamine-aminophenyl-(6-(5-(fluorescein-carboxamido)-hexan oic acid amide), the alpha-2,3 specificity of the enzyme was confirmed by NMR examination of the reaction product . The enzyme could also use synthetic acceptors with lactose or galactose as the saccharide portion . This study is the first example of the cloning, expression, and examination of alpha-2,3-sialyltransferase activity from a bacterial source.

Gene, 1996 Nov 7, 179(1), 147 - 55
The molecular genetics of type-4 fimbriae in Pseudomonas aeruginosa--a review; Mattick JS et al.; Type-4 fimbriae (or pili) are filaments found at the poles of a wide range of bacterial pathogens, including Neisseria gonorrhoeae, Moraxella bovis, Dichelobacter nodosus and Pseudomonas aeruginosa . They are composed of a small subunit which is highly conserved among different species and appear to mediate adhesion and translocation across epithelial surfaces via a phenomenon termed "twitching motility' . These fimbriae are key host colonisation factors and important protective antigens . We have analysed the genetics and biosynthesis of type-4 fimbriae in P . aeruginosa, which is an opportunistic pathogen of compromised individuals, including those suffering cystic fibrosis, AIDS or burns . A library of P . aeruginosa transposon mutants was constructed which exhibited loss of twitching motility, as determined by altered colony morphology . Analysis of these mutants, and of similar collections by other groups, have revealed that there are at least 22 genes involved in type-4 fimbrial assembly and function . A large number (pilA, B, C, D, E, M, N, O, P, Q, T, U, V and Z) appear to be involved in the biogenesis of the fimbriae and to represent a subset of a supersystem involved in the assembly of surface-associated protein complexes . Homologs of at least some of these genes have subsequently been identified in other type-4 fimbriate bacteria . In P . aeruginosa, the system is also regulated via two signal transduction pathways-a classic sensor-regulator system (encoded by pilS, pilR and rpoN) which controls transcription of the fimbrial subunit, presumably in response to host cues, and a chemotactic system (encoded by pilG, H, I, J, K and L) which may be involved in the directional or rate control of twitching motility in response to local environmental variables.

Mem Inst Oswaldo Cruz, 1996 Nov-Dec, 91(6), 789 - 93
Agar dilution method for susceptibility testing of Neisseria gonorrhoeae; de Castillo MC et al.; The antibiotic susceptibilities of Neisseria gonorrhoeae isolates obtained from patients attending a clinic for sexually transmitted diseases in Tucuman, Argentina, were determined by the agar dilution method (MIC) . 3.5% of the isolates produced beta-lactamase . A total of 96.5% of beta-lactamase negative isolates tested were susceptible to penicillin (MIC < or = 2 micrograms/ml-1); 14.03% of the tested isolates were resistant to tetracycline (MIC < or = 2 micrograms/ml-1), and 98% of the tested isolates were susceptible to spectinomycin (MIC < or = 64 micrograms/ml-1) . The MICs for 95% of the isolates, tested for other drugs were: < or = 2 micrograms/ml-1 for cefoxitin, < or = 0.06 microgram/ml-1 for cefotaxime, < or = 0.25 microgram/ml-1 for norfloxacin, < or = 10 micrograms/ml-1 for cephaloridine, < or = 10 micrograms/ml-1 for cephalexin, and < or = 50 micrograms/ml-1 for kanamycin . Antibiotic resistance among N . gonorrhoeae isolates from Tucuman, Argentina, appeared to be primarily limited to penicillin and tetracycline, which has been a general use against gonorrhoeae in Tucuman since 1960 . Periodic monitoring of the underlying susceptibility profiles of the N . gonorrhoeae strains prevalent in areas of frequent transmission may provide clues regarding treatment options and emerging of drug resistance.

Rev Clin Esp, 1996 Nov, 196(11), 741 - 6
{Neisseria meningitidis: isolation from low respiratory tract secretions of adult patients}; Ferrer Marcelles A et al.; OBJECTIVE: A bibliographic research was made using the Medline system of the clinico-microbiological features of reported cases of Neisseria meningitidis (NM) bronchopulmonary infection, as well as a retrospective study of NM isolation from lower respiratory tract secretions from adult inpatients . MATERIALS AND METHODS: All specimens from respiratory secretions were Gram stained and cultured onto blood . MacConkey and chocolate (quantitative) agar plates; a BCYE-alpha agar plate was also used when pneumonia was diagnosed . Fifty-five clinical records were retrospectively reviewed of patients with positive cultures for NM, for a 12-year period (1983-1994) . RESULTS: A total of 67 samples were positive among the 55 patients studied; sputum and tracheobronchial samples predominated . NM was recovered in pure culture from 48 specimens (71.6%) and with counts higher than 10(6) colony forming units/ml (CFU) . Twenty-seven isolates (40.3%) corresponded to serogroup B and 21 isolates (31.3%) did not group with serogroups A, B and C . Twenty-two patients were diagnosed of pneumonia; eleven of these 22 patients had an underlying chronic lung disease . Ten patients had a respiratory overinfection, in eight cases an episode of acute bronchitis was recorded and, finally, in 15 patients there was NM colonization only . CONCLUSION: The pathogenic role of NM in lower respiratory tract infections is probably underestimated because its isolation is difficult, particularly when there is oropharyngeal flora present, since in our study, in which only conventional culture media were used, samples which had NM recovered had a high number of colonies, in pure culture in most cases.

Int J STD AIDS, 1996 Nov-Dec, 7(7), 513 - 7
Determinants of the gonococcal serovar pattern in Edinburgh, Scotland: a multivariate analysis; Ross JD et al.; The geographical and temporal variety of gonococcal serovar patterns are well described but it remains uncertain what characteristics possessed by the organism, or sexual behaviour pattern in the patients, determine the particular serovar pattern in a given area at a given time . This study was designed to assess the relative contribution of various demographic and clinical features of infection to the observed pattern of serovars in Edinburgh between 1990 and 1993 . Five hundred and eight isolates were included in a multivariate analysis model to control for potential interactions between variables . Associations were noted between certain serovars and an asymptomatic clinical presentation, method of acquisition and site of infection . Certain physical characteristics of Neisseria gonorrhoeae in conjunction with the sexual behaviour patterns of patients are partially responsible for observed serovar patterns but more detailed analysis requires further sub-classification of serovars using molecular techniques.

Hum Reprod Update, 1996 Nov-Dec, 2(6), 519 - 29
Chlamydial pelvic inflammatory disease; Paavonen J et al.; Pelvic inflammatory disease (PID) is the most important complication present in the female lower genital tract, causing major medical, social and economic problems . Although PID can be caused by multiple microorganisms, it results most frequently from the ascent of sexually transmitted Chlamydia.trachomatis or Neisseria gonorrhoeae infections from the cervix to the upper genital tract . The importance of cervical chlamydial infection in the pathogenesis of PID is well recognized . Recent data from many developed countries have shown a striking decrease in the incidence of gonococcal infections, while the rates of chlamydial infections remain high in most countries . Complications of PID are common and usually irreversible . Emerging evidence suggests that universal or selected screening of defined populations for cervical chlamydial infection leads to a dramatic reduction in the incidence of PID . Recent technological advances should further enhance efforts to prevent chlamydial infection and PID . Gene amplification-based diagnostic tests, screening by testing first-void urine, and single dose antimicrobial therapy greatly facilitate chlamydia control programmes . Thus, screening for chlamydia is the key approach in the secondary prevention of PID . The obvious challenge is to make screening for chlamydia the standard for health care for young, sexually active individuals . Since PID is the most important consequence of sexually transmitted bacterial infections, it is also imperative to develop better treatments to prevent the long-term sequelae of this disease . The development and implementation of new and effective intervention programmes for prevention and control of PID is one of the major challenges for the year 2000 and beyond.

Lett Appl Microbiol, 1996 Nov, 23(5), 355 - 8
A new species of Neisseria from the dental plaque of the domestic cow, Neisseria dentiae sp . nov; Sneath PH et al.; A new species of the genus Neisseria is proposed, Neisseria dentiae sp . nov . The organism is found in dental plaque of domestic cows . It resembles N . animalis, N . canis and N . iguanae phenotypically but is distinguished from the first two by being positive for acidification of gluconate, D-glucose and usually D-fructose, and from the third by lack of predominant tetrad arrangement, lack of distinct alpha-haemolysis and by growing on nutrient agar and usually acidifying D-fructose . It is suggested that it may have significance for dental microbiology because members of the genus rapidly utilize oxygen and this may contribute to the anaerobic microenvironment found in dental plaque.

Sex Transm Dis, 1996 Nov-Dec, 23(6), 481 - 8
A tale of two sexually transmitted diseases . Prevalences and predictors of chlamydia and gonorrhea in women attending Colorado family planning clinics; Gershman KA et al.; BACKGROUND: The comparative prevalences and predictors of chlamydia and gonorrhea have not been studied in the family planning clinic population . GOALS: To determine the comparative prevalences and predictors of chlamydia and gonorrhea among Colorado family planning clinic patients . STUDY DESIGN: Cross-sectional study of public and private family planning clinic patients in Colorado tested for both chlamydia and gonorrhea (n = 12,926) . RESULTS: Among women tested for both infections, the chlamydia prevalence rate was 4.5% and the gonorrhea prevalence rate was 0.5% . Multivariate analysis showed that independent predictors of chlamydia were age younger than 25 years, black or Hispanic race-ethnicity, cervical friability, mucopus, exposure to a sex partner with chlamydia, or multiple recent sex partners . Independent predictors of gonorrhea were age younger than 20 years, black or Hispanic race-ethnicity, or exposure to a sex partner with gonorrhea; adjusted odds ratios for exposure to gonorrhea and black race were the highest for either infection . CONCLUSIONS: The gonorrhea prevalence rate was very low compared to that of chlamydia in patients at Colorado family planning clinics . Cost-effective gonorrhea testing strategies are needed for this populationPIP: In Colorado, state health department laboratory personnel used the DNA probe assay, Gen-Probe, to test specimens from 12,926 women for both Chlamydia trachomatis and Neisseria gonorrhoeae infections and from another 9416 women for Chlamydia alone . All the women had attended public and private family planning clinics state-wide during July 1994 to May 1995 . Researchers conducted a comparative analysis to determine the prevalence and the predictors of each sexually transmitted disease (STD) among the family planning clinic population in Colorado . Women tested for both chlamydia and gonorrhea had a higher chlamydia prevalence rate than those tested for only chlamydia (4.5% vs . 3.5%; p 0.001) . 64 (0.5%) of the women who were tested for both chlamydia and gonorrhea tested positive for gonorrhea . The gonorrhea prevalence was 9 times lower than that of chlamydia . Among the 64 women with gonorrhea, 25 (39.1%) also tested positive for chlamydia . Among the 577 women with chlamydia, 4.3% also tested positive for gonorrhea . The multivariate analysis revealed that variables independently associated with chlamydia included age under 20 or 20-24 years (odds ratio {OR} = 3.84 and 2.44, respectively), African-American or Hispanic race-ethnicity (OR = 2.41 and 1.65, respectively), cervical friability (OR = 2.26), mucopus (OR = 2.64), exposure to a sex partner with chlamydia (OR = 3.79), and multiple recent sex partners (OR = 1.4) . Variables independently associated with gonorrhea included age under 20 years (OR = 3.42), African-American or Hispanic race-ethnicity (OR = 12.71 and 3.07, respectively), and exposure to a sex partner with gonorrhea (OR = 39.29) . Cost-effective analyses would help determine appropriate selective screening strategies for gonorrhea . The researchers found that the criteria of urban residency, African-American race, and exposure to a sex partner with gonorrhea would involve the testing of 82% of patients, which would identify 94% of gonorrhea infections .

Mol Microbiol, 1996 Nov, 22(3), 509 - 22
The size and position of heterologous insertions in a silent locus differentially affect pilin recombination in Neisseria gonorrhoeae; Howell-Adams B et al.; Gonococcal pilus antigenic and phase variation result from unidirectional, RecA-dependent recombination of DNA sequences from a silent pilin copy (pilS) into the expressed pilin gene (pilE) . To develop a quantitative assay for pilin gene recombination that is independent of phase variation, a promoterless cat gene was inserted into pilS, and recombination of "cat into pilE was detected by selection of chloramphenicol-resistant (CmR) variants expressing "cat from the pilin promoter . Although RecA-dependent CmR variants occurred, none were generated by the simple transfer of "cat into pilE . Instead, each CmR variant contained a new pilin locus that was a hybrid of sequences from the pilE and the pilS1::cat loci in addition to the two starting loci . Therefore, this system could not be used to quantify antigenic variation . However, combined studies of these hybrid loci and of recombination products generated during additional pilS mutational analyses demonstrated that both the size and position of an insertion in pilS differentially affect pilin recombination . Also, the hybrid loci appear to be intermediates of antigenic variation . This enabled the creation of molecular models for the recombination reactions that result in pilin antigenic variation.

Acad Emerg Med, 1996 Nov, 3(11), 1030 - 4
High prevalence of sexually transmitted diseases in women with urinary infections; Berg E et al.; OBJECTIVE: To determine the prevalence and factors associated with unrecognized sexually transmitted diseases (STDs) in women who had pelvic examinations and were subsequently released from the ED with a sole diagnosis of urinary tract infection (UTI) . METHODS: A 3-month retrospective chart review was performed in an urban teaching hospital ED (> 70,000 visits/year) . Women aged 12-45 years who had pelvic examinations and were released from the ED with a sole diagnosis of UTI were included . Patient complaints, physical findings, and laboratory results were reviewed . Laboratory evaluations included the complete blood count, urinalysis, urine pregnancy test, and cervical cultures for Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas . RESULTS: Of the 94 women who met study criteria, 53% had proven STDs (19% N . gonorrhoeae, 22% C . trachomatis, 33% Trichomonas) . There was no difference between the patients with positive and negative tests for STDs with regard to complaints, physical findings, and laboratory results (all p > 0.05) . CONCLUSIONS: Women undergoing pelvic examinations who are subsequently released from this urban ED with the diagnosis of UTI have a high (> 50%) prevalence of occult STDs . No complaint, physical finding, or laboratory result reviewed was associated with the risk of an STD . Consideration should be given to empirical antibiotic therapy in similar urban populations.

Gynecol Oncol, 1996 Nov, 63(2), 258 - 60
Prevalence of mycoplasma conserved DNA in malignant ovarian cancer detected using sensitive PCR-ELISA; Chan PJ et al.; Mycoplasmas are tiny polymorphic prokaryotic organisms (0.2-0.3 microm) that lack a cell wall and reside ubiquitously at the cell membrane or internalized into the cell . The organisms have been implicated in many diseases including functioning as cofactors catalyzing the HIV disease state . The oncogenic potential of mycoplasmas was only recently realized when they were shown to cause chromosomal changes and in vitro cell transformations through gradual progressive chromosomal loss and translocations . While a recent study linked mycoplasmas with gastric cancer, the association between mycoplasmas and ovarian cancer has not been established . Recently, a commercial assay which combined polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) methods was developed for the detection of mycoplasmas . The present objective was to determine the prevalence of mycoplasmas in archived paraffin-embedded malignant ovarian cancer tissue . The combined PCR-ELISA procedure was used with consensus primers targeting for 15 species of mycoplasmas and acholeplasmas . Archived human malignant ovarian cancer tissues (N = 27 cases) embedded in paraffin blocks were processed, and DNA was extracted and the presence of DNA verified . The extracted DNA specimens were randomly divided into three groups for analyses . PCR-ELISA assays were performed on extracted DNA together with appropriate negative and positive controls . The results showed mycoplasmas were present in 59.3% of the malignant ovarian cancer specimens . PCR-ELISA analysis of Neisseria gonorrhea and Chlamydia trachomatis controls did not produce cross-reacting false-positive results . The results suggest an association between mycoplasmas and malignant ovarian cancer . A 59.3% prevalence rate was demonstrated for mycoplasmas in paraffin-embedded ovarian cancer tissues . The mechanism involved in oncogenesis by mycoplasmas remains to be elucidated.

J Clin Microbiol, 1996 Nov, 34(11), 2778 - 83
Evaluation of automated COBAS AMPLICOR PCR system for detection of several infectious agents and its impact on laboratory management; Jungkind D et al.; We evaluated the COBAS AMPLICOR (CA) PCR system (Roche Diagnostic Systems) designed for automated PCR amplification and detection of nucleic acids from infectious agents in clinical samples . The Roche AMPLICOR microwell plate (MWP) PCR was the reference method . CA amplifies target nucleic acid, captures the biotinylated amplification products by using magnetic particles coated with specific oligonucleotide probes, and detects the bound products colorimetrically . For Mycobacterium tuberculosis, the correlation of the results of CA tests with those of MWP tests was 100% with 230 samples, including 20 culture-positive samples . For hepatitis C virus, the correlation was 100% with 214 samples, including 60 positive samples . MultiPlex CA analysis of 199 cervical specimens for Chlamydia trachomatis, Neisseria gonorrhoeae, and the internal control gave 100% concordance . These samples included 19 C . trachomatis and 3 N . gonorrhoeae culture-positive samples . Overall, the agreement between PCR methods for all 842 comparisons was 100% . Compared with culture, the sensitivities of the assays for C . trachomatis and M tuberculosis were > or = 95% . After spiking alternating amplification tubes in the CA system with 10(14) copies of the Chlamydia amplicon per ml, we were unable to demonstrate any carryover cross-contamination of negative samples . Using the criteria of the College of American Pathologists workload recording method, we found that the total hands-on time to produce CA PCR results was 4.4, 7.9, and 3.3 min for M . tuberculosis, hepatis C virus, and the MultiPlexed assay for chlamydia plus gonorrhea and an internal control, respectively . The CA system brings true PCR automation to laboratories . In addition to the accuracy of automated results, the CA system provides labor savings, provides containment of the amplification and detection components of PCR, and supports both MultiPlex amplification and sequential algorithm (ReFlex) detection of analytes.

J Immunol, 1996 Nov 1, 157(9), 4239 - 43
Genetic bases of human complement C7 deficiency; Nishizaka H et al.; Complement C7 deficiency (C7D) is associated frequently with recurrent bacterial infections, especially meningitis caused by Neisseria meningitidis . We report in this work the molecular bases of C7D in two unrelated Japanese males . We used exon-specific PCR/single-strand conformation polymorphism analysis as a screening step for mutations . Subsequent direct sequencing of the target exons identified homozygous mutations in exon 16 of case 1 and in exon 15 of case 2 . The mutation of case 1 was a homozygous T to A transversion at nucleotide 2250, the third nucleotide of the codon TGT for Cys728, leading to a stop codon TGA (C728X) . In case 2, a homozygous 2-bp deletion (2137delTG/2138delGT/2139delTG) caused a frameshift, generating a premature termination codon 4 to 6 nucleotides downstream . Family study in case 1 confirmed the genetic nature of the defect . Moreover, we detected a novel polymorphism in intron 11 that presumably is linked to the mutation responsible for C7D in case 1 . Our results indicate that the pathogenesis of C7D is heterogeneous like most of the other deficiencies of complement components.

Infect Immun, 1996 Nov, 64(11), 4630 - 7
Regulation of gonococcal sialyltransferase, lipooligosaccharide, and serum resistance by glucose, pyruvate, and lactate; McGee DJ et al.; Strain F62 of Neisseria gonorrhoeae gonococci (GC) is sensitive to normal human serum unless CMP-N-acetylneuraminic acid (CMP-NANA) is present . NANA is transferred primarily to a 4.5-kDa lipooligosaccharide (LOS) structure by a GC sialyltransferase (Stase) . We investigated LOS and Stase expression and serum resistance in strain F62 grown in different carbon sources and growth conditions . Pyruvate-grown GC expressed 1.9- to 5.6-fold more Stase activity than did glucose-grown GC, whereas lactate-grown GC generally expressed intermediate Stase activities . Broth-grown GC expressed two- to fourfold more Stase activity than did plate-grown GC in all carbon sources . Pyruvate- or lactate-grown GC expressed significantly more of the sialylateable 4.5-kDa LOS species than did glucose-grown GC . Anaerobically, the 4.5-kDa LOS species was expressed in greater quantity than the 4.9-kDa N-acetyl galactosamine-terminating species in all carbon sources . Pyruvate-grown GC also incorporated up to threefold more radiolabelled CMP-NANA onto the 4.5-kDa LOS species than did glucose-grown GC . In serum resistance studies, pyruvate-grown GC were 6.5- to 16.1-fold more serum resistant than glucose-grown GC at limiting CMP-NANA concentrations (1.56 to 12.50 microg/ml) . Taken together, these results indicate that gonococcal expression of Stase activity is up-regulated by growth in pyruvate or lactate, which correlates with enhanced expression of the sialylateable 4.5-kDa LOS and, for growth in pyruvate, correlates with enhanced sialylation of gonococcal LOS and greater serum resistance . In different in vivo niches, gonococcal LOS sialylation, serum resistance, and interaction with host cells can be highly regulated.

Infect Immun, 1996 Nov, 64(11), 4472 - 9
Temperature- and medium-dependent secretion of proteins by Shiga toxin-producing Escherichia coli; Ebel F et al.; Infections due to Shiga toxin-producing Escherichia coli (STEC) are responsible for severe diarrheal disease in humans and livestock, and these bacteria have recently emerged as a leading cause of renal failure in children . In this study, we have examined medium- and temperature-dependent production of secreted proteins from a STEC O26 serotype strain . Growth of bacteria in Luria broth led to the detection of secreted polypeptides of 104, 55, 54, and 37 kDa (p104, p55, p54, and p37, respectively) . When grown in serum-free tissue culture medium, only p104, p37 and two additional polypeptides of 25 and 22 kDa (p25 and p22) were present in supernatant fluids . Production of these polypeptides was growth temperature dependent and induced in cultures grown at 37 degrees C . N-terminal amino acid sequencing revealed that p104 was homologous to the secreted p110 of enteropathogenic Escherichia coli (EPEC), and both proteins belong to a family of secreted proteins in pathogenic bacteria of which the immunoglobulin A protease of Neisseria gonorrhoeae is the prototype . The N-terminal amino acid sequences of p55 and p54 were unique to the STEC strain, while p37 and p25 were found to be highly homologous to the similarly sized EspA and EspB proteins, previously detected in culture supernatants of EPEC . Molecular cloning and sequencing of STEC espB alleles from two different serotypes showed that the encoded polypeptides were about 80% homologous . A monoclonal antibody raised against STEC EspB also cross-reacted with its EPEC analog and allowed us to demonstrate medium- and temperature-dependent production of this important virulence factor in STEC and EPEC strains of differing serotypes.

Gene, 1996 Oct 31, 178(1-2), 107 - 10
Absence of periplasmic DsbA oxidoreductase facilitates export of cysteine-containing passenger proteins to the Escherichia coli cell surface via the Iga beta autotransporter pathway; Jose J et al.; The Iga beta autotransporter function of IgA1 protease from Neisseria gonorrhoeae was assessed in Escherichia coli using the Vibrio cholerae toxin B subunit (CtxB) as a heterologous passenger . N-terminal fusions with Iga beta of native CtxB or mutant CtxB protein containing no cysteines were constructed and analysed in isogenic E . coli mutants carrying defects in either or both the ompT (outer membrane protease T) and dsbA (periplasmic disulfide oxidoreductase) determinants . While export of the cystein-less CtxB passenger was independent of the dsbA genotype, the native CtxB passenger was properly translocated across the outer membrane only in the dsbA mutant background . This effect was consistent in the presence and in the absence of the OmpT protease which rather determined the release of surface-bound CtxB into the medium . Therefore, in agreement with previous observations Iga beta-dependent protein secretion requires an unfolded conformation of the passenger domain and can be blocked by disulfide loop formation in the presence of DsbA . Since DsbA acts in the periplasm, this provides evidence for a periplasmic intermediate in the Iga beta-mediated export pathway . E . coli (dsbA ompT) is highly suitable as a strain for the surface display of recombinant proteins via Iga beta, whether or not they contain cysteine residues.

Arch Oral Biol, 1996 Oct, 41(10), 965 - 78
Stoichiometry of oxygen consumption and sugar, organic acid and amino acid utilization in salivary sediment and pure cultures of oral bacteria; Traudt M et al.; In each of 23 numerically or metabolically significant oral micro-organisms, and in each of the salivary sediments of 10 humans, oxygen uptake was determined quantitatively with various sugar and organic and amino acid substrates . With relatively few exceptions, the salivary sediments rapidly consumed oxygen with the array of substrates (23) tested . On the other hand, the individual pure cultures oxidized fewer substrates and did so selectively from this menu . The observation that the Gram-positive bacteria readily used oxygen when sugar substrates were provided, but were unable to use oxygen with all but one of the organic and none of the amino acids was significant . The Gram-negative bacteria, in contrast, used oxygen poorly with the sugars but most readily with many of the organic and amino acids, was significant . Only two of the Gram-positive but most of the Gram-negative micro-organisms tested showed oxygen uptake with L(+)-lactate; the Gram-negative bacteria were also active with D(-)-lactate, formate and succinate . Propionate was also tested and showed oxygen uptake only with the Gram-negative micro-organism, Neisseria subflava; acetate showed none or almost none with all of the examined bacteria . Where oxygen consumption occurred with the various pure or mixed cultures and substrates tested, the quantities of oxygen consumed were less than theoretically possible . For example, they ranged on average in the sediment results from 1.78 mumol oxygen per mumol of L(+)-lactate catabolized to 5.17 mumol oxygen per mumol of lactose . This was consistent with substrate oxidation by the oral bacteria being less than complete as in aerobic glycolysis, and with compounds other than water and carbon dioxide (such as acetate) being prominent amongst the end-products produced . The pure-culture oxygen data and other reports from this laboratory have made it possible to propose a speculative scheme as to which bacterial species might be involved in the various metabolic pathways used when different substrates are catabolized and oxidized by the mixed bacteria in salivary sediment or dental plaque . Also, it made it possible to suggest which bacteria and substrates are likely to be involved in the oxygen depletion that enables plaque to achieve anaerobiosis.

East Afr Med J, 1996 Oct, 73(10), 675 - 8
High frequency of sexually transmitted diseases among pregnant women in Dar es Salaam, Tanzania: need for intervention; Mwakagile D et al.; In order to determine the prevalence and characteristics of sexually transmitted diseases (STDs) in pregnant women (PW) attending a primary health care antenatal clinic (ANC) in metropolitan Dar es Salaam, Tanzania, a randomly selected sample of PW in their second or third trimesters were invited to participate at their first visit . They were interviewed using a questionnaire and underwent genital examination . Genital swabs were obtained for microscopy and/or culture isolation of Candida albicans, Trichomonas vaginalis, and Neisseria gonorrhoeae . Blood specimens were also obtained for serological testing for syphilis and for antibodies to the human immunodeficiency virus (HIV) . A total of 777 PW aged 14 to 40 years were seen . Parities ranged from 0 to 10 . Prevalence of syphilis, trichomoniasis, gonorrhoea and HIV infection were 4.0%, 22.7%, 3.6% and 15.2%, respectively . At least one acute STD (excluding HIV infection) was found in 32.8% of the PW . The prevalence of multiple STDs (excluding HIV infection) was higher in teenagers (45.3%, 77/170) than in PW in other age groups (29.2%, 177/607) (p < 0.001) . The prevalence of HIV infection in teenage PW was 10.0% . Most STDs were least prevalent in PW who were married monogamously . Of the 732 PW who had one or more genital infections (including infection with Candida species), 669 (91.4%) had one or more genital complaints . However, most of the genital complaints were not disease specific . Since this study has shown that the prevalences of acute STDs were high in PW, especially in teenagers, it is recommended that all PW in Tanzania should be screened for STDs syndromically including the use of appropriate clinical and laboratory examination whenever possiblePIP: A survey of 777 randomly selected pregnant women attending an antenatal clinic in Dar es Salaam, Tanzania, in 1993 revealed a high prevalence of sexually transmitted diseases (STDs), particularly among teenagers . The median age of survey respondents was 23.6 years (range, 14-40 years); 170 women (22%) were teenagers and 439 (56.7%) were married . 320 women (41.2%) had 1 or more STDs (excluding human immunodeficiency virus (HIV) infection); in 32.7%, there was active infection . STD prevalence was 45.3% in teenagers compared with 29.2% in adults . In the overall sample, the prevalences of syphilis, trichomoniasis, gonorrhea, and HIV were 4.0%, 22.7%, 3.6%, and 15.2%, respectively . 80 women (10.3%) showed serologic evidence of past syphilis infection and 4% had active syphilis . Syphilis was most prevalent in pregnant women aged 35 years and above (13.8%), while trichomoniasis was most common in teenagers (34.3%) . Of the 732 pregnant women with genital infections, 63 (8.6%) were asymptomatic; when symptoms did exist, they were generally not disease-specific . The most significant risk factor for STDs, including HIV, was single marital status . These findings suggest a need for the introduction of essential clinical and laboratory facilities for STD detection to antenatal clinics in Tanzania .

Biosci Biotechnol Biochem, 1996 Oct, 60(10), 1617 - 22
Biodegradation of cellulose acetate by Neisseria sicca; Sakai K et al.; Bacteria capable of assimilating cellulose acetate, strains SB and SC, were isolated from soil on a medium containing cellulose acetate as a carbon source, and identified as Neisseria sicca . Both strains degraded cellulose acetate membrane filters (degree of substitution, DS, mixture of 2.8 and 2.0) and textiles (DS, 2.34) in a medium containing cellulose acetate (DS, 2.34) or its oligomer, but were not able to degrade these materials in a medium containing cellobiose octaacetate . Biodegradation of cellulose acetate (DS, 1.81 and 2.34) on the basis of biochemical oxygen demand reached 51 and 40% in the culture of N . sicca SB and 60 and 45% in the culture of N . sicca SC within 20 days . A decrease in the acetyl content of degraded cellulose acetate films and powder was confirmed by infrared and nuclear magnetic resonance analyses . After 10-day cultivation of N . sicca SB and SC, the number-average molecular weight of residual cellulose acetate decreased by 9 and 5%, respectively . Activities of enzymes that released acetic acid and produced reducing sugars from cellulose acetate were mainly present in the culture supernatant . Reactivity of enzymes for cellulose acetate (DS, 1.81) was higher than that for cellulose acetate (DS, 2.34).

Diagn Microbiol Infect Dis, 1996 Oct, 26(2), 91 - 3
Recurrent bacterial peritonitis caused by Neisseria cinerea in a chronic ambulatory peritoneal dialysis (CAPD) patient; George MJ et al.; We present an unusual case of recurrent (chronic ambulatory peritoneal dialysis) CAPD-associated peritonitis caused by Neisseria cinerea . Using DNA restriction fragment length polymorphism (RFLP) analysis, we determined that the recurrent infection was caused by reinfection with a different N . cinerea strain rather than relapse with the index strain and that the probable origin of the reinfecting organism was the patient's upper respiratory tract.

Genitourin Med, 1996 Oct, 72(5), 358 - 61
Trends in sexually transmitted diseases and condom use patterns among commercial sex workers in Fukuoka City, Japan 1990-93; Tanaka M et al.; OBJECTIVE: To investigate trends in sexually transmitted diseases (STDs) among female commercial sex workers and in their condom use patterns during the period from 1990 to 1993 in Fukuoka, Japan . METHODS: The study group consisted of a total of 824 commercial sex workers who attended an STD clinic to undergo screening for STDs including chlamydia, gonorrhoea, syphilis, hepatitis B and HIV-1 infection during the period from 1990 to 1993 . For detection of Chlamydia trachomatis and Neisseria gonorrhoeae, endocervical smear specimens were taken from the women . Blood samples were obtained for serological diagnosis of syphilis, hepatitis B and HIV-1 . Commercial sex workers who visited the clinic during the period from November to December of 1993 were interviewed concerning past (1990 and 1991) and recent (1992 and 1993) condom use patterns . RESULTS: The annual detection rates of C trachomatis and N gonorrhoeae declined significantly from 16.3% in 1990 to 12.2% in 1993 (P < 0.0001) and from 1.5% in 1990 to 0.8% in 1993 (P = 0.0096), respectively . There was a remarkable reduction in the annual syphilis infection rate, from 7.5% in 1990 to 0.5% in 1993 (P = 0.0011) . The positive rate for the hepatitis B surface antigen in the women ranged from only 0.6% to 1.9% and none were found to be positive for HIV-1 during the 4-year period . During the same period, there was a significant increase in the proportion of commercial sex workers always using condoms from 6.3% in 1990-91 to 25.3% in 1992-93 (P = 0.0023) . CONCLUSION: The prevalences of chlamydia, gonorrhoea, and syphilis infections decreased significantly among commercial sex workers in Fukuoka from 1990 through 1993, and no commercial sex workers were HIV-1 seropositive . The reductions in the prevalence of major STDs may be related to the increased use of condomsPIP: A survey of commercial sex workers in Fukuoka City, Japan, during 1990-93 revealed significant declines in the prevalence of chlamydia, gonorrhea, and syphilis during the study period, presumably as a result of increased condom use . The study group consisted of 824 commercial sex workers who attended a sexually transmitted disease (STD) clinic in Fukuoka from 1990 to 1993 for voluntary STD check-ups . The annual Chlamydia trachomatis detection rate declined from 16.3% in 1990 to 12.2% in 1993; the annual detection rate for Neisseria gonorrhoeae fell from 1.5% to 0.8%, while that for syphilis dropped from 7.5% to 0.5% . None of the 791 women screened for human immunodeficiency virus was seropositive . In addition, a subsample of 79 commercial sex workers were interviewed about their condom use . The proportion of sex workers always using condoms rose from 6.3% in 1990 to 25.3% in 1993; in this period, never use of condoms decreased from 45.6% to 5.1% .

Infect Immun, 1996 Oct, 64(10), 4129 - 36
Expression of sialyltransferase is not required for interaction of Neisseria gonorrhoeae with human epithelial cells and human neutrophils; McGee DJ et al.; Sialyltransferase (Stase) in Neisseria gonorrhoeae organisms (gonococci {GC}) transfers sialic acid (N-acetylneuraminic acid {NANA}) from cytidine 5'-monophospho-N-acetylneuraminic acid (CMP-NANA) mainly to the terminal galactose (Gal) residue in the Gal beta-1,4 N-acetylglucosamine (Gal-GlcNAc)-R lipooligosaccharide (LOS) structure . Sialylated GC resist killing by normal human serum, sometimes show reduced invasion of epithelial cells, and have reduced adhesion to and stimulation of human neutrophils . We questioned whether Stase itself modulates the interactions of GC with human epithelial cells and neutrophils in the absence of exogenous CMP-NANA . To that end, we treated strain F62 with ethyl methanesulfonate and grew approximately 175,000 colonies on CMP-NANA plates, and screened them with monoclonal antibody 1B2-1B7 (MAb 1B2) . MAb 1B2 is specific for Gal-GlcNAc and reacts only with asialylated GC . We isolated 13 MAb 1B2-reactive mutants, including five null mutants, that had Stase activities ranging from barely detectable to fivefold less than that of wild-type (WT) F62 . The LOS phenotype of Stase null mutants was identical to that of WT F62, yet the mutants could not sialylate their LOS when grown with CMP-NANA . The Stase null phenotype was rescuable to Stase+ by transformation with chromosomal DNA from WT F62 . Stase null mutants remained serum sensitive even when grown with CMP-NANA . One Stase null mutant, ST94A, adhered to and invaded the human cervical epithelial cell line ME-180 at levels indistinguishable from that of WT F62 in the absence of CMP-NANA . In human neutrophil studies, ST94A stimulated the oxidative burst in and adhered to human neutrophils at levels similar to those of WT F62 . ST94A and WT F62 were also phagocytically killed by neutrophils at similar levels . These results indicate that expression of Stase activity is not required for interaction of GC with human cells.

Mol Microbiol, 1996 Oct, 22(1), 161 - 73
Fimbrial biogenesis genes of Pseudomonas aeruginosa: pilW and pilX increase the similarity of type 4 fimbriae to the GSP protein-secretion systems and pilY1 encodes a gonococcal PilC homologue; Alm RA et al.; Type 4 fimbriae of Pseudomonas aeruginosa are surface filaments involved in host colonization . They mediate both attachment to host epithelial cells and flagelia-independent twitching motility . Four additional genes, pilW, pilX, pilY1 and pilY2, are located on Spel fragment E in the 5 kb intergenic region between the previously characterized genes pilV and pilE, which encode prepilin-like proteins involved in type 4 fimbrial biogenesis . The phenotypes of a transposon insertion and other mutations constructed by allelic exchange show that these genes are involved in the assembly of type 4 fimbriae . The PilW and PilX proteins are membrane located, possess the hydrophobic N-terminus characteristic of prepilin-like proteins, and appear to belong to the GspJ and GspK group of proteins that are required for protein secretion in a wide range of Gram-negative bacteria . These findings increase the similarities between the fimbrial biogenesis and the Gsp-based protein-secretion supersystems . PilY1 is a large protein with C-terminal homology to the PilC2 protein of Neisseria gonorrhoeae, thought to be a fimbrial tip-associated adhesin, and which, like PilY1, is involved in fimbrial assembly . PilY1 appears to be located in both the membrane and the external fimbrial fractions . PilY2 is a small protein that appears to play a subtle role in fimbrial biogenesis and represents a new class of protein.

Mol Microbiol, 1996 Oct, 22(1), 31 - 42
Processing of the AIDA-I precursor: removal of AIDAc and evidence for the outer membrane anchoring as a beta-barrel structure; Suhr M et al.; The AIDA-I adhesin known to be responsible for the diffuse adherence (DA) phenotype of the diarrhoeagenic Escherichia coli (DAEC) strain 2787 has been shown previously to be synthesized as a precursor protein and to undergo additional C-terminal processing . Here, the C-terminal processing of the AIDA-I precursor and the outer membrane topology of the cleaved C-terminal fragment, AIDAc, were investigated . By isolation of the cleaved AIDAc fragment and N-terminal sequencing, the C-terminal cleavage site was identified between Ser-846 and Ala-847 thereby indicating a molecular mass of 47.5 kDa for AIDAc . The correct processing to AIDA-I and AIDAc in OmpT, OmpP and DegP protease-deficient E . coli strains as well as in avirulent salmonellae and shigellae points to an autocatalytic cleavage mechanism . The cleaved AIDAc was localized in the outer membrane . A leader sequence-AIDAc fusion was efficiently routed to the outer membrane . Analysis by protease digestion, secondary-structure prediction and modelling, by comparison with structurally related bacterial proteins like the IgA1 protease from neisseria, the vacuolating toxin from Helicobacter pylori, and the VirG protein of Shigella flexneri, strongly indicates that AIDAc is present in the outer membrane as a beta-barrel structure.

Microbiology, 1996 Oct, 142 ( Pt 10), 2951 - 7
Proline iminopeptidase gene from Xanthomonas campestris pv . citri; Alonso J et al.; The pip gene coding for the proline iminopeptidase (Pip) of Xanthomonas campestris pv . citri was cloned in an Escherichia coli leuB strain using a selective medium containing the dipeptide D-Ala-L-Leu as the sole source of L-leucine . Nucleotide sequencing of this gene revealed a 939 bp open reading frame encoding a 312 amino acid protein (35 126 Da) . The deduced amino acid sequence showed 47% identity with the Pip from Neisseria gonorrhoeae . A lacZ-pip fusion gene was overexpressed in E . coli under the control of the Plac promoter . The Pip of X . campestris hydrolysed L-prolyl-p-nitroanilide with the highest efficiency, but was also able to hydrolyse L-alanyl-p-nitroanilide and D-alanyl-p-nitroanilide . The molecular mass of Pip was found to be 35 kDa by SDS-PAGE and 120 kDa by gel filtration, suggesting that the active enzyme is a multimer.

Microbiology, 1996 Oct, 142 ( Pt 10), 2839 - 45
Uptake-sequence-independent DNA transformation exists in Neisseria gonorrhoeae; Boyle-Vavra S et al.; A DNA transformation dose-response curve of piliated (P+) gonococci with the use of cloned DNA containing a pilE2-cat fusion showed saturation at high and low levels of transforming DNA . At low DNA concentrations, transformation of the P+ strain MS11-A was effectively inhibited by a 1000-fold molar excess of the gonococcal transformation uptake sequence (GCUS) . The same molar excess of the GCUS did not inhibit transformation of MS11-A at high DNA concentrations . In MS11-B2, a nonpiliated (P-), pilin-nonproducing, isogenic variant of MS11-A, the GCUS did not inhibit transformation at any level of transforming DNA . These data suggest that two mechanisms of transformation exist in P+ cells: one which utilizes the GCUS and one which does not . In MS11-B2 P- cells, no evidence was found for the presence of the GCUS-dependent mechanism, suggesting that transformation in this background occurs solely by the GCUS-independent mechanism.

J Clin Microbiol, 1996 Oct, 34(10), 2548 - 51
Detection of Neisseria gonorrhoeae from air-dried genital samples by single-tube nested PCR; Herrmann B et al.; A single-tube nested PCR method was developed for the detection of Neisseria gonorrhoeae . The optimized assay had a detection limit of less than 0.3 cell . Five different storage conditions for gonococcal specimens were compared with respect to the PCR detection of bacteria . For air-dried gonococcal slides containing three bacteria, DNA was detected after 8 weeks at ambient temperature, and for slides containing 300 bacteria, DNA could be detected after 24 weeks at ambient temperature . Air-dried storage combined with analysis by the single-tube nested PCR and a commercially available PCR (Amplicor) was used to test 350 cervical specimens from women in the West African island nation of Cape Verde . The in-house PCR detected 17 cases of N . gonorrhoeae infection, while the Amplicor system detected 14 cases of N . gonorrhoeae infection . No specimen was negative by the in-house PCR assay and positive by the Amplicor PCR . This sensitive nested PCR assay, combined with air-dried storage, allows for the detection of gonococci when specimen storage and transport times are extended and freezing conditions are not available.

J Clin Microbiol, 1996 Oct, 34(10), 2395 - 400
Detection of Chlamydia trachomatis and Neisseria gonorrhoeae by ligase chain reaction-based assays with clinical specimens from various sites: implications for diagnostic testing and screening; Buimer M et al.; Ligase chain reaction (LCR)-based tests for the diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae infections in men and women attending a sexually transmitted disease clinic were evaluated . LCR testing of urethral swab and urine specimens from men and cervical swab and urine specimens from women was compared with culture of male urethral swabs and female cervical and urethral swabs, respectively . An expanded "gold standard" was defined as a positive culture or at least one specimen confirmed to be positive by LCR testing . The prevalence of C . trachomatis infection as detected by cell culture was 7.0% among 614 men and 5.0% among 602 women . By LCR, these values increased to 11.4 and 9.9% with urethral swabs and urine, respectively, for men and 9.6 and 9.1% with cervical swabs and urine, respectively, for women . Relative to the expanded gold standard, the sensitivity of cell culture with male urethral swabs or female cervical swabs was 57.3 and 45.5%, respectively, compared with corresponding values of 93.3 and 87.9% for LCR . The sensitivity of LCR with urine specimens was 77.3 and 78.8% for men and women, respectively . The prevalence of N . gonorrhoeae infection as detected by culture was 5.9% among 220 men and 2.9% among 383 women . The corresponding values were 8.2 and 5.5%, respectively, by LCR testing of swabs . Prevalence values by LCR testing of urine were 7.3% for men and 2.9% for women . The sensitivity of culture was 72.2% for men and 50.0% for women . The sensitivities of LCR were 100% with male urethral swabs, 95.4% with female cervical swabs, 88.9% with male urine, and 50.0% with female urine . These results indicate that the LCR-based assays represent a major improvement in C . trachomatis and N . gonorrhoeae diagnostics . The sensitivity of testing of urethral or cervical swabs by LCR was markedly greater than that by culture . The sensitivity of testing female or male urine specimens was equal to or greater than that of culturing cervical or urethral specimens . LCR testing of urine specimens may prove useful for screening for C . trachomatis.

J Exp Med, 1996 Oct 1, 184(4), 1233 - 41
Conservation of the lipooligosaccharide synthesis locus lgt among strains of Neisseria gonorrhoeae: requirement for lgtE in synthesis of the 2C7 epitope and of the beta chain of strain 15253; Erwin AL et al.; The present study was undertaken to examine the extent to which the lgt locus varies among strains of gonococci . This locus encodes five glycosyl transferases involved in the synthesis of the lipooligosaccharide (LOS) of Neisseria gonorrhoeae . We examined seven gonococcal strains and found that the structure of the lgt locus is conserved among six of these strains . The locus is strikingly altered in strain 15253 . This is one of the few strains where extensive structural analysis of its LOS is available, and therefore, we defined the altered lgt locus and focused on the reactivity of mAB 2C7 . We found that strain 15253 contains only two lgt genes, lgtA and lgtE . As in F62, lgtA encodes a GlcNAc transferase and is subject to phase variation . In addition, by analysis of deletion mutants, we found that lgtE, which encodes a galactosyl transferase that is required for elongating the alpha-chain, is also necessary for completing the beta chain.

Biochem J, 1996 Oct 1, 319 ( Pt 1), 99 - 102
Isolation and characterization of the gene encoding an aminopeptidase involved in the selective toxicity of ascamycin toward Xanthomonas campestris pv . citri; Sudo T et al.; An aminopeptidase gene named XAP has been isolated from Xanthomonas campestris pv . citri, a plant pathogenic bacterium . The bacterium is one of the rare micro-organisms susceptible to ascamycin, an aminoacyl nucleoside antibiotic that inhibits protein synthesis . Sequence analysis reveals that the gene encodes a 311 amino acid protein with a calculated molecular mass of 35134 Da and approx . 50% identity for amino acids to the proline iminopeptidase from Neisseria gonorrhoeae . The XAP gene product, Xap, expressed in Escherichia coli has proline iminopeptidase activity as well as ascamycin dealanylating activity in vitro.

J Med Microbiol, 1996 Oct, 45(4), 252 - 7
Blocking of iron uptake by monoclonal antibodies specific for the Neisseria meningitidis transferrin-binding protein 2; Pintor M et al.; The existence of epitopes common to different strains in the Neisseria meningitidis transferrin (Tf)-binding protein 2 (TBP2), combined with the ability of polyclonal anti-TBP2 antibodies to inhibit Tf binding and block iron uptake in this species, led to this study on the effect of anti-TBP1+2 monoclonal antibodies (MAbs) to determine the presence of epitopes inside the Tf-binding region . All MAbs used reacted exclusively with the homologous strain when tested by dot-blots of outer membrane vesicles, with the reaction being specific for TBP2 after SDS-PAGE and electroblotting . In contrast, ELISA and iron-uptake blocking assays were also positive with heterologous strains belonging to Rokbi's group II (high mol.wt TBP2) . The results confirmed the two group classification proposed by Rokbi and, in contrast to other studies, indicated the existence of epitopes in the Tf-binding region that are common only to strains of Rokbi's group II . These epitopes may become denatured after drying for dot-blot assays or after SDS-PAGE and electroblotting.

J Infect Dis, 1996 Oct, 174 Suppl 2, S230 - 9
Impact of a gonorrhea control program, including selective mass treatment, in female sex workers; Holmes KK et al.; A gonorrhea control program initiated in 1967 in registered female sex workers (FSWs) in the Philippines involved weekly endocervical cultures for Neisseria gonorrhoeae, with treatment of FSWs found infected or named as contacts by US Navy servicemen . Gonorrhea prevalence in FSWs in Olongapo city fell from 11.9% to 4.0% within 4 months, and gonorrhea incidence in servicemen at nearby Subic Bay fell by half . Selective mass treatment (SMT) with oral ampicillin-probenecid or tetracycline was then given to registered FSWs in an attempt to further reduce gonorrhea rates . N . gonorrhoeae was isolated from 105 (4.0%) of 2640 FSWs before SMT and from 43 (1.6%) 1 week later (P < .001) . However, gonorrhea incidence among servicemen fell no lower, and gonorrhea prevalence in FSWs quickly returned to higher levels . Thus, after implementation of weekly screening and treatment of FSWs found infected or named as contacts, SMT of FSWs (without increasing condom use or treating regular partners) contributed nothing further to gonorrhea control.

Ann Intern Med, 1996 Sep 15, 125(6), 465 - 70
The emergence of Neisseria gonorrhoeae with decreased susceptibility to ciprofloxacin in Cleveland, Ohio: epidemiology and risk factors; Gordon SM et al.; BACKGROUND: Until 1992, almost all strains of Neisseria gonorrhoeae that had been tested in the United States were susceptible to fluoroquinolones, including ciprofloxacin . However, among men with urethral gonococcal infections who attended one sexually transmitted disease clinic in Cleveland, Ohio, the prevalence of gonococci with decreased susceptibility to ciprofloxacin increased from 2% in 1991 to 16% in 1994 . OBJECTIVE: To describe the emergence of and risk factors for gonococcal urethritis caused by gonococci with decreased susceptibility to ciprofloxacin . Resistance to ciprofloxacin was considered to be decreased if the mean inhibitory concentration was at least 0.12 microgram/mL, and was less than or equal to 0.25 microgram/mL; this definition did not equate with the definition of clinical resistance . DESIGN: Case-control study . SETTING: An urban sexually transmitted disease clinic . PARTICIPANTS: 51 case-patients and 106 controls . MEASUREMENTS: Pulsed-field gel electrophoresis was used to identify individual genotypes of ciprofloxacin-resistant and ciprofloxacin-susceptible isolates . RESULTS: 55 of the 746 isolates of N . gonorrhoeae that were tested (7.4%) had decreased susceptibility to ciprofloxacin, and the prevalence of N . gonorrhoeae with decreased susceptibility significantly increased during the study period . Case-patients were significantly less likely to have gram-negative diplococci seen on microscopic examination of urethral discharge (P < or = 0.01) and were less likely to be treated for gonococcal urethritis than were controls (P < or = 0.001) . Molecular typing suggested the spread of a single genotype of N . gonorrhoeae . CONCLUSIONS: Strains of gonococci with decreased susceptibility to ciprofloxacin appear to have become endemic in Cleveland, Ohio . The clinical significance of these isolates is not clear, but the potential for the emergence of clinically important resistance may preclude the use of fluoroquinolones as an alternative treatment for uncomplicated gonorrhea.

Bull Pan Am Health Organ, 1996 Sep, 30(3), 212 - 7
Detection of N . meningitidis group B antigens by MB-Dot-ELISA in patients with meningitis; Alkmin M das G 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, 1996 Sep-Oct, 23(5), 425 - 8
Quinolone-resistant Neisseria gonorrhoeae isolated in Sydney, Australia, 1991 to 1995; Tapsall JW et al.; BACKGROUND AND OBJECTIVES: Quinolone antibiotics are used widely for the treatment of gonorrhea, but resistant strains appeared in Sydney in 1984, treatment failure with high-dose regimens in 1991, and isolates with very high minimal inhibitory concentrations (MICs) (16 mg/l) in 1994 . GOALS: To examine the frequency, source, and characteristics of Quinolone-resistant Neisseria gonorrhoeae (QRNG) in Sydney from 1991 to 1995 and to compare these data with those obtained from 1984 to 1990 . STUDY DESIGN: The antibiotic sensitivity, auxotype-serovar class, and geographic source of QRNG isolated in Sydney from January 1, 1991 to June 30, 1995 were analyzed . RESULTS: One hundred seven QRNG were isolated from 97 patients from 1991 to 1995 . The number, proportion, and MICs of QRNG increased slowly in the first 4 years of the study and rapidly in the last 6 months . Most QRNG were isolated from travelers entering Sydney from Asia . Twenty-seven different auxotype-serovar classes were detected including 6 auxotype-serovar classes in 14 isolates with high-level quinolone resistance (MIC, 16 mg/l) . CONCLUSIONS: QRNG isolated in Sydney during the past decade originated in Asia as multiple gonococcal subtypes and increased substantially in numbers and levels of resistance in 1995.

Sex Transm Dis, 1996 Sep-Oct, 23(5), 384 - 91
Decreasing incidences of gonorrhea- and chlamydia-associated acute pelvic inflammatory disease . A 25-year study from an urban area of central Sweden; Kamwendo F et al.; BACKGROUND AND OBJECTIVES: Acute pelvic inflammatory disease (PID) affects women in their reproductive years and is often a complication of a sexually transmitted disease (STD), particularly Neisseria gonorrhoeae and Chlamydia trachomatis . Infertility, ectopic pregnancy, and chronic lower abdominal pain are common long-term sequelae to acute PID . Through different preventive measures, endemic N . gonorrhoeae is almost eliminated, and C . trachomatis has been reduced almost fourfold in Sweden . GOALS: To investigate variations in STD-associated acute PID and the extent to which this influenced the yearly incidences of patients hospitalized for this complication during a 25-year-period . STUDY DESIGN: Hospital records of 2499 patients admitted and treated for acute PID from January 1, 1970 to December 31, 1994 were analyzed for infection with N . gonorrhoeae . Routine laboratory diagnosis for C . trachomatis infection started June 1, 1980 . Detailed statistical analysis for chlamydial-associated PID in this study, therefore, covers the period January 1, 1981 to December 31, 1994 and includes 1030 patients . RESULTS: Gonorrhea occurred in 42% of patients with acute PID in 1970 and decreased continuously to zero in 1988 and beyond . Concomitant urogenital chlamydial infection reduced almost fourfold from 28.4% in 1985 to 7.7% in 1994 . Yearly admissions for acute PID fluctuated slightly (< or = 16%) in the early 1970s and early 1980s but increased greatly (> 60%) in the middle and late 1970s; the highest was 180 per year in 1976 . This coincided with high incidence rates of gonorrhea in the general population, and probably of genital C . trachomatis infection as well, coupled with an increased use of intrauterine contraceptive device in nulliparous women . The largest increase in admissions for acute PID was in the 15- to 29-year-old group . A steady decrease started in 1987 and reached the low figure of 26 admissions in 1994 . The greatest decrease occurred in the 15- to 19-year-old group, from the relative age distribution of 28.9% in the period 1970 to 1974 to 12.9% in 1990 to 1994 . Yearly admissions for the > or = 35-year-old group remained almost constant during the study period, but the relative age distribution shifted from second lowest (excluding those 14 years or younger, totaling 15 admissions for the entire study period), 9.1% at the beginning of the study period, to the second largest, 24.9% at the end of it . The study also showed that the total and relative rates of recurrence decreased . CONCLUSIONS: Measures aimed at reducing incidences of gonorrhea and genital chlamydial infection will reduce the incidences of one of the most serious complications of these STDs, acute PID, and, in turn, its long-term sequelae.

Sex Transm Dis, 1996 Sep-Oct, 23(5), 378 - 83
Tetracycline-resistant Neisseria gonorrhoeae . Characteristics of patients and isolates at a London Genitourinary Medicine Clinic; Lewis DA et al.; OBJECTIVES: To compare auxotypes, serovars, and antibiograms of tetracycline-resistant Neisseria gonorrhoeae (TRNG) and non-TRNG isolated from patients attending an East London Genitourinary Medicine (GUM) Clinic . To obtain plasmid profiles for penicillinase-producing gonococci (PPNG) as well as presumptive TRNG . To identify differences in patient characteristics for the TRNG and non-TRNG patient groups . STUDY DESIGN: Gonococcal isolates were collected from 400 patients attending the GUM clinic at the Royal London Hospital GUM Clinic over a 1-year period . Isolates (378) were tested for susceptibility to various antibiotics, auxotyped, and serotyped . Plasmid profiles were obtained for PPNG and isolates exhibiting high-level tetracycline resistance (TRNG) . The presence of the tet M determinant was confirmed using the polymerase chain reaction (PCR) . The PCR product was digested with the restriction endonuclease (RE) Hpa II and electrophoresed on a 2.5% agarose gel to determine an "RE pattern." Patient data were collected by retrospective case-note review . RESULTS: TRNG (n = 42) accounted for 11% of the 378 isolates tested, and the remaining 336 (89%) isolates were non-TRNG . Non-requiring auxotrophy and P1B-2 serovar expression occurred more frequently among TRNG . PPNG accounted for 31% of TRNG and 5% of non-TRNG . Chromosomal resistance to penicillin (CMRNG) was absent among TRNG but accounted for 11% of non-TRNG . One TRNG isolate showed decreased susceptibility to ciprofloxacin (MIC 0.25 mg/l) . All isolates were sensitive to cefotaxime, cefixime, spectinomycin, and azithromycin . All TRNG possessed the 25.2 MDa plasmid and produced a PCR product of appropriate size after tet M gene sequence amplification . RE digests of the PCR product gave a single pattern . None of the TRNG in contrast to 18% of the non-TRNG were acquired homosexually . Ethnic distribution differed between the patients with TRNG and patients without non-TRNG (Afro-Caribbean 81% versus 58%; white 19% versus 36%) . Most TRNG were acquired in the United Kingdom . CONCLUSIONS: TRNG differ from the non-TRNG in their auxotype and serovar distribution . PPNG are more common among the TRNG isolates, whereas CMRNG appear absent . TRNG are isolated more commonly from Afro-Caribbean patients and were not represented among homosexually acquired isolates.

Fam Med, 1996 Sep, 28(8), 580 - 3
Use of wet mount to predict Chlamydia trachomatis and Neisseria gonorrhea cervicitis in primary care; Majeroni BA et al.; BACKGROUND AND OBJECTIVES: Cervicitis is associated with salpingitis, infertility, and complications of pregnancy . Universal screening has been recommended for high-prevalence populations but may not be appropriate in the family practice setting . Leukocytes on an endocervical gram stain have been associated with infectious cervicitis due to Chlamydia trachomatis and Neisseria gonorrhea . This study sought to determine whether the finding of leukocytes in a vaginal wet mount could be used to screen for infectious cervicitis in an urban family practice . METHODS: A consecutive sample of 357 women had cultures for C trachomatis and N gonorrhea and a standardized wet mount . RESULTS: All women with infectious cervicitis were under age 35 . Thirty-six percent of infected women had more leukocytes than epithelial cells in the wet mount, compared with 23% of women without these organisms . CONCLUSIONS: Wet mount findings did not reliably predict infectious cervicitis . Study of a larger population is needed to confirm these findings.

J Clin Microbiol, 1996 Sep, 34(9), 2255 - 8
Rapid detection of point mutations of the Neisseria gonorrhoeae gyrA gene associated with decreased susceptibilities to quinolones; Deguchi T et al.; Mutations in the gyrA gene resulting in amino acid changes at Ser-91 and Asp-95 are significantly associated with decreased susceptibilities to quinolones in Neisseria gonorrhoeae . To detect these mutations, we developed a rapid and simple assay based on amplification of the region of the gyrA gene containing the mutation sites by PCR and digestion of the PCR product with a restriction enzyme . A naturally occurring HinfI restriction site was present in the region containing the Ser-91 codon, and an artificial HinfI restriction site was created in the region containing the Asp-95 codon by the method of primer-specified restriction site modification . The mutations generating alterations at Ser-91 and Asp-95 were detected as restriction fragment length polymorphisms of the PCR products digested with HinfI . Fifty-five clinical strains of N . gonorrhoeae were examined for mutations in the gyrA gene by this method . Mutations at Ser-91 and/or Asp-95 were detected in all the 31 strains in which the mutations had been confirmed by DNA sequencing . Our method allows simultaneous testing of a large number of strains and provides results within 8 h . This rapid and simple assay could be a useful screening device for genetic alterations associated with decreased susceptibilities to quinolones in N . gonorrhoeae and could facilitate epidemiological studies on clinical isolates of N . gonorrhoeae with decreased susceptibilities to quinolones.

Microbiology, 1996 Sep, 142 ( Pt 9), 2635 - 45
IdiA, a 34 kDa protein in the cyanobacteria Synechococcus sp . strains PCC 6301 and PCC 7942, is required for growth under iron and manganese limitations; Michel KP et al.; In the cyanobacteria Synechococcus PCC 6301 and PCC 7942 a protein with an apparent molecular mass of about 34 kDa (called IdiA for iron-deficiency-induced protein A) accumulates under iron and managanese limitation . IdiA from Synechococcus PCC 6301 was partially sequenced, showing that the N-terminal amino acid is an alanine . Moreover, the gene encoding this protein in Synechococcus PCC 6301 has been identified and completely sequenced . The idiA gene codes for a protein starting with valine and consisting of 330 amino acid residues . Thus, IdiA is apparently synthesized as a precursor protein of 36.17 kDa and cleaved to its mature form of 35.01 kDa between two alanine residues at positions 9 and 10 . IdiA is a highly basic protein having an isoelectric point of 10.55 (mature protein) . Comparison of the amino acid sequence of IdiA with protein sequences in the database revealed that IdiA has similarities to two basic bacterial iron-binding proteins, SfuA from Serratia marcescens and Fbp from Neisseria gonorrhoeae . Insertional inactivation of the idiA gene in Synechococcus PCC 7942 resulted in a mutant which was unable to grow under iron- or manganese-limiting conditions . Manganese limitation of the mutant strain led to a drastic reduction of photosystem II activity (O2 evolution) within less than 48 h, while wild-type cells required a prolonged cultivation in Mn-deficient medium before an effect on photosystem II was observed . Thus, IdiA is a protein involved in the process of providing photosystem II with manganese.

Microbiology, 1996 Sep, 142 ( Pt 9), 2481 - 9
Identification of an EF-Tu protein that is periplasm-associated and processed in Neisseria gonorrhoeae; Porcella SF et al.; A 44 kDa protein is a dominant component of periplasmic extracts of Neisseria gonorrhoeae . Peptide sequence generated from a cyanogen-bromide-cleaved fragment of this protein indicated sequence homology with elongation factor-Tu (EF-Tu) . Polyclonal antiserum was made against the 44 kDa protein purified from periplasm extracts of N . gonorrhoeae . The preabsorbed antiserum was immunoblotted against whole-cell lysates on two-dimensional gels . A 44 kDa protein and a smaller 37 kDa protein were recognized by this antiserum . A N . gonorrhoeae gamma phage DNA library was screened and a clone expressing a 44 kDa protein was identified . The DNA insert in this clone contained several genes homologous to genes contained in the str operon of Escherichia coli . One ORF product with a calculated molecular mass of 43 kDa was highly homologous to the EF-TuA of E . coli . A synthetic peptide antiserum specific for a portion of the C terminus of EF-Tu confirmed that the 37 kDa protein in whole-cell lysates of N . gonorrhoeae was a processed form of EF-Tu . Deletion of the tufA gene homologue in N . gonorrhoeae was attempted but was unsuccessful.

Infect Immun, 1996 Sep, 64(9), 3641 - 5
Tolerance to appetite suppression induced by peptidoglycan; Biberstine KJ et al.; Physiologically realistic peptidoglycan (PG) fragments, derived from Neisseria gonorrhoeae, were shown previously to dose-dependently suppress food consumption and body weight gain in rats following single intraperitoneal injections . The present study, examining the effects of repeated daily injection of PG, provides additional support to our underlying hypothesis, i.e., that soluble PG fragments contribute to the loss of appetite commonly associated with bacterial infections . An initial intraperitoneal injection of purified, soluble, macromolecular, extensively O-acetylated PG fragments (S-O-PG) (240 micrograms/kg of body weight) decreased overnight food consumption in male Lewis rats (150 g) by approximately 35% relative to animals receiving diluent alone (P < 0.05) . However, subsequent daily injections of S-O-PG resulted in progressively smaller effects on food consumption until, by the fourth day, rats were completely nonresponsive (tolerant) to S-O-PG-induced hypophagia . Rats that developed tolerance to the effects of S-O-PG on appetite were also tolerant to three other known hypophagic agents, lipopolysaccharide (LPS), muramyl dipeptide, and interleukin-1, when challenged one day after establishment of S-O-PG tolerance . Similarly, rats developed tolerance to repeated injections of muramyl dipeptide or LPS and were cross-tolerant to S-O-PG when challenged 1 day later . However, 30 days after establishment of S-O-PG tolerance, rats remained nonresponsive to S-O-PG but regained full responsiveness to LPS-mediated hypophagia . Thus, at least two mechanisms of tolerance to S-O-PG hypophagia exist: an early tolerance which is nonspecific and a late tolerance which is specific for S-O-PG . Late, but not early, tolerance to S-O-PG-mediated suppression of appetite was associated with an increase in specific anti-PG antibody activity as measured in an enzyme-linked immunosorbent assay.

Biochim Biophys Acta, 1996 Aug 13, 1290(3), 210 - 4
Cloning and functional analysis of the pmmA gene encoding phosphomannomutase from the photosynthetic prokaryote Prochlorothrix hollandica; Dwivedi K et al.; The pmmA gene encoding a bifunctional phosphomannomutase/phosphoglucomutase (PMM/PGM) from the photosynthetic prokaryote, Prochlorothrix hollandica has been cloned and sequenced . The gene encodes a 51827 Da polypeptide 48% identical to the PMM of Azospirillum brasilense, 37% identical to the PGMs of pathogenic Neisseria sp . and 37% identical to the bifunctional AlgC PGM/PMM of Pseudomonas aeruginosa . The pmmA gene encodes an enzyme having both PGM and PMM activities as judged by both enzyme assays and complementation analysis in which the cloned gene partially corrected the pgm-1 mutation of Escherichia coli.

J Natl Cancer Inst, 1996 Aug 7, 88(15), 1068 - 75
Difficulty in elucidating the male role in cervical cancer in Colombia, a high-risk area for the disease; Munoz N et al.; BACKGROUND: Epidemiologic evidence has been inconclusive in linking men's sexual behavior and genital human papillomaviruses (HPVs) with cervical cancer risk in their sexual partners in areas with a high incidence of cervical cancer . PURPOSE: This study assesses the role of men's sexual behavior and the presence of penile HPV DNA on the risk of their wives' developing cervical neoplasia in an area in Colombia with a high incidence of cervical cancer . METHODS: A total of 210 husbands of women with cervical intraepithelial neoplasia grade III (n = 118) or invasive squamous cell carcinoma of the cervix (n = 92) and a total of 262 husbands of women recruited as control subjects (173 and 89, respectively) were interviewed . Questionnaires included detailed information on sexual behavior . Exfoliated cells were obtained from the glans penis and from the distal urethra of the penis . The specimens were analyzed for HPV DNA by use of a polymerase chain reaction-based assay that included a generic probe and 25 type-specific probes . Serum specimens were collected and analyzed for antibodies to Chlamydia trachomatis, Treponema pallidum, herpes simplex virus type II, and Neisseria gonorrhoeae . RESULTS: Limited education (adjusted odds ratio {OR} = 4.4; 95% confidence interval {CI} = 1.9-9.8; for no schooling versus secondary or higher education) and presence of antibodies to C . trachomatis (adjusted OR = 2.5; 95% CI = 1.5-4.4) in husbands were the only identified risk factors for cervical neoplasia in their wives . The prevalence of HPV DNA in the penis was 25.7% among husbands of case women and 18.9% among husbands of control women (adjusted OR = 1.2; 95% CI = 0.6-2.3) . Neither the lifetime number of female sexual partners (adjusted OR = 1.0; 95% CI = 0.4-2.6; for > 50 partners versus one to five) nor the lifetime number of female prostitutes as sexual partners (adjusted OR = 1.2; 95% CI = 0.7-2.0; for > or = 21 prostitutes versus one to five) was associated with the risk of cervical cancer . CONCLUSIONS: Our results are compatible with the hypothesis that in the population of Cali, whose women are at high risk of developing cervical cancer, exposure to HPV among young men is a common occurrence and is mediated by contacts with large numbers of female sexual partners and prostitutes . These widespread sexual practices limit the power of case-control studies to detect significant associations between men's sexual behavior and the cervical cancer risk in their sexual partners . HPV DNA detection in the penis of adult men is a poor reflection of lifetime exposure or of etiologically relevant exposure to HPV . The role of C . trachomatis in cervical carcinogenesis deserves further investigation . IMPLICATIONS: Further research is needed to elucidate the male's role in cervical carcinogenesis in populations at high risk for cervical cancer . HPV DNA prevalence surveys and studies of the natural history of HPV in young men will be of great value.

J Natl Cancer Inst, 1996 Aug 7, 88(15), 1060 - 7
Male sexual behavior and human papillomavirus DNA: key risk factors for cervical cancer in Spain; Bosch FX et al.; BACKGROUND: It is now established that certain types of human papillomaviruses (HPVs) are the sexually transmitted agents etiologically linked to cervical cancer . Studies assessing the contribution of the male's sexual behavior and genital HPV DNA status to the risk of development of cervical neoplasia in sexual partners have yielded inconsistent results . PURPOSE: This study evaluates the role of men's sexual behavior and the presence of HPV DNA in the penis on the development of cervical cancer in their sexual partners in Spain, a low-risk area for cervical neoplasia . METHODS: Husbands (n = 633) of women participating in two case-control studies of cervical neoplasia were interviewed to obtain information on lifestyle habits, including sexual practices . Cytologic samples were taken from the distal urethra and the surface of the glans penis of 183 husbands of case women and of 171 husbands of control women . These samples were analyzed by a polymerase chain reaction-based system using a generic probe and 25 type-specific probes for the detection and typing of HPV DNA . Serologic specimens were also obtained and analyzed for antibodies to Chlamydia trachomatis, Treponema pallidum, herpes simplex virus type II, and Neisseria gonorrhoeae . RESULTS: The presence of HPV DNA in the husbands' penis conveyed a fivefold risk of cervical cancer to their wives (adjusted odds ratio {OR} for HPV DNA positivity = 4.9; 95% confidence interval {CI} = 1.9-12.6) . The risk of cervical cancer was strongly related to HPV type (adjusted OR for HPV type 16 = 9.0; 95% CI = 1.1-77.5), to the husbands' number of extramarital partners (adjusted OR = 11.0; 95% CI = 3.0-40.0; for > or = 21 women versus one), and to the number of prostitutes as extramarital sexual partners (adjusted OR = 8.0; 95% CI = 2.9-22.2; for > or = 10 women versus none) . Presence of antibodies to C . trachomatis (adjusted OR = 2.6; 95% CI = 1.4-4.6) and an early age at first sexual intercourse of the husband (adjusted OR = 3.2; 95% CI = 1.7-5.9; for < or = 15 years versus > or = 21 years) were also associated with cervical neoplasia in the wife . After adjustment for these variables and for the wife's pack-years of smoking, the husband's smoking was moderately associated with cervical cancer in his wife (adjusted OR = 2.5; 95% CI = 1.4-4.4; for > or = 26.2 pack-years versus none) . CONCLUSIONS: The study supports the role of men as vectors of the HPV types that are related to cervical cancer . Life-time number of female sexual partners, number of female prostitutes as sexual partners, and detection of HPV DNA in the penis of husbands are all surrogate markers of exposure to HPV during marriage . IMPLICATIONS: Men who report multiple sexual partners or who are carriers of HPV DNA may be vectors of high-risk HPV types and may place their wives at high risk of developing cervical cancer . Prostitutes are an important reservoir of high-risk HPVs.

Enferm Infecc Microbiol Clin, 1996 Aug-Sep, 14(7), 441 - 3
{Variation of the incidence and antibiotic sensitivity of Neisseria gonorrhoeae in a 7-year period}; Nebreda T et al.; BACKGROUND: The aim of this study was to determine the incidence and antibiotic sensitivity of Neisseria gonorrhoeae versus penicillin, tetracycline, cefoxitin, ceftriaxone and spectinomycin from 1988 to 1994 in the province of Soria (Spain) . METHODS: From January 1988 to December 1994, clinical samples of 57 strains of N . gonorrhoeae were isolated . Auxotype, serotype, and minimum inhibitory concentration (MIC) studies were performed versus 5 antimicrobials and analysis of plasmids in the penicillinase producer strains (PPNG), was carried out in the Bacteriology Department of the Instituto Carlos III in Madrid (Spain) . RESULTS: The rate of incidence of the isolation of N . gonorrhoeae was similar from 1988 to 1990 (11.7 to 19.1 cases per 100,000 inhabitants) and decreased from 1991 to 1994 (6.4 to 0 cases per 100,000 inhabitants) . The strains belonged to 30 different auxotypes/serotypes indicating a great heterogeneity among them . The proportion of penicillinase producing N . gonorrhoeae (PPNG) increased over the study period . The first strains with high resistance to tetracycline (TRNG) were first isolated in the authors' area in 1991 and the proportion increased up to the end of the study . All the strains were sensitive to ceftriaxone, cefoxitin and spectinomycin . CONCLUSIONS: A decrease was observed in the rate of incidence of N . gonorrhoeae since 1991 with an increase in the proportion of NGPP and TRNG strains . Ceftriaxone and spectinomycin present good activity versus all the strains studied with their empiric use as treatment being possible in the province of Soria (Spain).

Genitourin Med, 1996 Aug, 72(4), 295 - 7
Mutation in DNA gyrase of norfloxacin-resistant clinical isolates of Neisseria gonorrhoeae; Tanaka M et al.; BACKGROUND AND OBJECTIVES: Recently a rapid decrease in the susceptibility of Neisseria gonorrhoeae isolates to fluoroquinolones has occurred and gonococcal fluoroquinolone resistance is now a significant problem in the treatment of gonorrhoea in Japan . Thus, in order to investigate the quinolone resistance mechanisms in clinical isolates of N gonorrhoeae we studied an alteration in the DNA gyrase subunit A (GyrA) which is well-known as a common mechanism of bacterial quinolone resistance . MATERIALS AND METHODS: Four clinical isolates of N gonorrhoeae resistant to norfloxacin and 5 strains susceptible to norfloxacin, including 2 clinical isolates and 3 WHO reference strains, were tested in this study . To identify mutations in the GyrA genes of gonococcal strains, polymerase chain reaction and direct DNA sequencing were performed . RESULTS: A single base change (serine codon TCC changed to phenylalanine codon TTC), which resulted in an amino acid change in GyrA at position 91, was identified in all 4 norfloxacin-resistant strains for which the MICs of norfloxacin ranged from 1.0 to 8.0 micrograms/ml, while no mutation within GyrA was detected in 5 norfloxacin-susceptible strains for which the MICs of norfloxacin ranged from 0.004 to 0.063 microgram/ml . CONCLUSIONS: The results from this study suggest that the serine-91 to phenylalanine substitution in GyrA is probably an essential mutation in fluoroquinolone resistance in clinical isolates of N gonorrhoeae.

Genitourin Med, 1996 Aug, 72(4), 253 - 7
Antimicrobial agents and gonorrhoea: therapeutic choice, resistance and susceptibility testing; Ison CA; INTRODUCTION: Neisseria gonorrhoeae, the causative agent of gonorrhoea is a particularly well adapted pathogen that has continued to evolve mechanisms to evade treatment with antimicrobial agents . THERAPEUTIC CHOICE: The choice of antibiotic for use in the first-line treatment of gonorrhoea should be made with knowledge of the susceptibility of the isolates of N gonorrhoeae to be encountered . RESISTANCE: High-level resistance to penicillin and tetracycline in N gonorrhoeae is plasmid-mediated and a major therapeutic problem . Penicillinase-producing N gonorrhoeae, first described in 1976, have now spread worldwide and tetracycline-resistant N gonorrhoeae, described in 1985, are becoming increasingly prevalent . Chromosomal resistance to penicillin is low-level and affects a range of antibiotics . High-level resistance to spectinomycin has been sporadic and has not limited its use whereas the emergence of resistance to ciprofloxacin will have a significant impact on its use for gonorrhoea . SUSCEPTIBILITY TESTING: A variety of methods are available including disc diffusion, breakpoint agar dilution technique, E-test and determination of the minimum inhibitory concentration (MIC) . The choice of methodology will depend on the number and type of isolates and the facilities available for testing . DISCUSSION: Surveillance programmes to monitor levels of antibiotic resistant isolates are essential to ensure therapeutic success.

J Obstet Gynaecol Res, 1996 Aug, 22(4), 331 - 40
The association of Chlamydia trachomatis/gonococcal infection and tubal factor infertility; Swasdio K et al.; OBJECTIVE: To determine the association of past Chlamydia trachomatis and past Neisseria gonorrhoeae infection with tubal factor infertility . METHODS: A cross-sectional study was conducted . Cases consisted of 55 primary infertile women with laparoscopy confirmed tubal damage (group A) and their husbands, consecutively attending the Infertility Unit at Maharaj Nakorn Chiang Mai Hospital between 1990 and 1992; and 58 primary infertile women with laparoscopy confirmed normal tubes (group B) and their husbands, consecutively attending the same hospital over the same period . Controls consisted of 59 postpartum women (group C) and their husbands omitted to the same hospital over the same period as cases . Past chlamydial and gonococcal infections were assessed by measuring serum IgG antibodies by enzyme immunoassay (EIA) . The EIA antigens consisted of purified elementary bodies of C . trachomatis serovar L1, or purified alpha pili of N . gonorrhoeae strain P9 . RESULTS: The prevalence of positive IgG antibody to gonococcal pili in sera from group A was 29.1%, significantly higher than the prevalence of 5.2% in group B or 3.4% in group C (p = 0.000) . The husbands of women in group A had a significantly higher prevalence of IgG antibody to gonococcal pili (36.4%) than the husbands of women in group B (8.6%) or group C (18.6%) (p = 0.002) . There was no significant difference in positive IgA antibody between case and control groups . After controlling for age, group A showed significantly higher prevalences of past gonorrhea (OR = 32.4, 95% CI 4.3, 242.2) and past chlamydial infection (OR = 3.2, 95% CI 1.2, 8.5) than group C . The husbands of women in group A also had higher prevalences of both types of infection than the husbands of women in group C; the odds ratios for past gonorrhea or chlamydial infections were 2.8 (95% CI 1.1, 6.9) and 2.9 (95% CI 1.2, 7.1), respectively . Neither infertile women with normal tubes (group B) nor their husbands showed any difference when compared with controls . CONCLUSION: These results suggest that in this region of northern Thailand there is an association between past gonorrhea and past chlamydial infection and tubal factor infertility.

Mol Microbiol, 1996 Aug, 21(3), 433 - 40
Questions about gonococcal pilus phase- and antigenic variation; Seifert HS; Pathogenic organisms inhabit one of several defined locations within a host where temperature, pH, and nutrients are relatively constant . While the microorganism must adapt to different environments within the host, the host immune system is the most formidable predator that can limit the growth of a pathogen . Neisseria gonorrhoeae (the gonococcus, Gc) is the causative agent of gonorrhoea, and has evolved several systems for varying the antigenicity of different surface antigens, presumably to help evade the effects of the human immune system . The On/Off/On phase variation of surface structure expression also alters the antigenic characteristics of the bacterial cell surface . Antigenic variation of the major subunit of the pilus, pilin, occurs by unidirectional, homologous recombination between a silent locus and the expression locus . The silent loci lie from 1 to 900 kb from the expression locus in the chromosome yet all can donate their sequences to the expression locus . The genetic composition of the pilin loci of two Gc strains has been elucidated, and the types of changes that lead to altered forms of the pilus have been extensively characterized . However, little is known about the precise molecular mechanisms used to allow high-frequency, non-reciprocal, chromosomal recombination between pilin loci or about what regulates the process of maintaining chromosome fidelity.

Pediatr Infect Dis J, 1996 Aug, 15(8), 662 - 7
Acceptability and usefulness of vaginal washes in premenarcheal girls as a diagnostic procedure for sexually transmitted diseases . The Child Protection Centre at the Winnipeg Children's Hospital; Embree JE et al.; OBJECTIVE: To assess the suitability of vaginal washes as specimens for sexually transmitted disease diagnosis and determine the usefulness of PCR technology for Chlamydia trachomatis diagnosis in prepubertal girls . STUDY DESIGN: Paired sets of vaginal secretions were collected with swabs and by vaginal wash from 138 prepubertal girls for evaluation because of alleged sexual abuse . Detection by culture of Neisseria gonorrhoeae and C . trachomatis was compared between the two sampling techniques . PCR techniques were also used to test 29 vaginal wash specimens for C . trachomatis . RESULTS: In the prepubertal girls N . gonorrhoeae was detected in two wash specimens but in only one swab specimen; C . trachomatis was detected by culture in both paired specimens from two children and by PCR in vaginal washes from both of the two children positive by culture; PCR identified two other infected children . CONCLUSIONS: A vaginal wash technique coupled with newer molecular amplification technology may be useful in the assessment of sexually abused children.

J Ethnopharmacol, 1996 Aug, 53(2), 57 - 63
Antineoplastic agents 338 . The cancer cell growth inhibitory . Constituents of Terminalia arjuna (Combretaceae); Pettit GR et al.; By means of bioassay-guided separation methods, the cancer cell growth inhibitory constituents residing in the bark, stem and leaves of the Mauritius medicinal plant Terminalia arjuna (Combretaceae) were examined . The cancer cell line active components were found to be gallic acid, ethyl gallate, and the flavone luteolin . Only gallic acid was previously known to occur in this plant . Luteolin has a well established record of inhibiting various cancer cell lines and may account for most of the rationale underlying the use of T . arjuna in traditional cancer treatments . Luteolin was also found to exhibit specific activity against the pathogenic bacterium Neisseria gonorrhoeae.

J Pediatr Adolesc Gynecol, 1996 Aug, 9(3), 129 - 32
The coincident diagnosis of pelvic inflammatory disease and pregnancy: are they compatible?
Acquavella AP, Rubin A, D'Angelo LJ.
OBJECTIVES: To determine the frequency of coincident diagnoses of pregnancy and pelvic inflammatory disease (PID) in adolescents seeking care at a large urban children's hospital . DESIGN: All inpatient medical records for the period from January 1, 1984 through December 31, 1993 were searched for dual diagnoses of pregnancy and PID (presumed secondary to endometritis, salpingitis, or both) . During this period, there were 1205 patients admitted for PID, 67 of whom were also pregnant . Ten of these 67 admissions were eliminated from this study because of incomplete or missing records, errors in diagnosis, or lack of proper examinations . The charts of the remaining 57 subjects were reviewed for demographics, physical findings, and laboratory studies . OUTCOME MEASURES: For the purposes of this study, a diagnosis of suspected PID was defined as lower abdominal tenderness, cervical motion tenderness, and adnexal tenderness ("major criteria"), as well as either a positive cervical specimen for Neisseria gonorrhoeae or Chlamydia trachomatis or adnexal fullness ("minor criteria") . RESULTS: The mean age of the 57 subjects was 16.8 years, and the mean gestational age was 6.7 weeks . Twenty-four (42.1%) of the subjects met the criteria for a concurrent diagnosis of PID and pregnancy; 13 had physical findings and a positive cervical specimen for either N . gonorrhoeae or C . trachomatis, and 11 subjects had the minor criteria of adnexal fullness . Twenty-six (45.6%) of the 57 subjects were primigravida, 35 (61.4%) had a history of a sexually transmitted disease, and 18 (31.6%) had been previously admitted to a hospital for PID . CONCLUSION: This study found that PID and pregnancy can coexist in adolescents . Therefore, physicians who treat adolescents must consider the possibility of PID in pregnant adolescents presenting with abdominal pain.

Am J Obstet Gynecol, 1996 Aug, 175(2), 435 - 41
Role of bacterial vaginosis-associated microorganisms in endometritis; Hillier SL et al.; OBJECTIVE: Our goal was to define the role of bacterial vaginosis and bacterial vaginosis-associated microorganisms in endometritis . STUDY DESIGN: Endometrial biopsies were obtained for histologic and microbiologic study from 178 consecutive women with suspected pelvic inflammatory disease, and 85 of them underwent laparoscopy to diagnose salpingitis . RESULTS: Histologic endometritis was confirmed in 117 (65%) of the women . Among women who underwent laparoscopy, salpingitis was present in 68% of those with and 23% of those without endometritis . Some but not all bacterial vaginosis-associated microorganisms were linked with endometritis . By logistic regression analysis, after adjustment for bacterial vaginosis and isolation of Neisseria gonorrhoeae and Chlamydia trachomatis, endometritis was associated with endometrial N . gonorrhoeae (odds ratio 5.7, 95% confidence interval 1.8 to 17.5), C . trachomatis (odds ratio 4.8, 95% confidence interval 1.3 to 18.2), anaerobic gram-negative rods (odds ratio 2.6, 95% confidence interval 1.1 to 5.7), and nonwhite race (odds ratio 2.3, 95% confidence interval 1.1 to 4.8) . CONCLUSIONS: The association of anaerobic gram-negative rods with endometritis, after adjustment for bacterial vaginosis, N . gonorrhoeae, and C . trachomatis, supports the role of these microorganisms in the etiology of histologic endometritis among women with clinically suspected pelvic inflammatory disease.

J Bacteriol, 1996 Aug, 178(16), 5020 - 3
Analysis of Fur binding to operator sequences within the Neisseria gonorrhoeae fbpA promoter; Desai PJ et al.; The gene encoding Neisseria gonorrhoeae periplasmic binding protein FbpA contains two regions whose sequences exhibit homology with the Escherichia coli ferric uptake regulator protein (Fur) consensus binding sequence . In this study, DNase I footprinting experiments were employed to characterize the operator sequences within the fbpA promoter region to which E . coli Fur binds . A 160-bp fragment encompassing the promotor region and the putative iron boxes of the fbpA promoter was incubated with Fur, DNaseI was added, and the products of these reactions were sequenced to identify nucleotide peaks that were protected . At 50 nM Fur, a protected region that spanned 33 bp and extended 19 bp upstream and 8 bp downstream of the -35 region of the fbpA promoter was observed . At higher concentrations of Fur (75 and 100 nM), an extension of this protected region upstream of the -35 region was observed . Introduction of a plasmid carrying an fbpA-cat transcriptional fusion in E . coli H1717 (Fur+) resulted in an 88% induction of chloramphenicol acetyltransferase expression under conditions of iron restriction; however, chloramphenicol acetyltransferase expression was not responsive to iron in E . coli H1745 (Fur-), indicating that transcriptional regulation of fbpA in response to iron occurs via the negative regulator Fur . The extent of the fbpA operator sequence (42 bp), as defined by our footprinting analysis, would suggest the binding of two Fur repressor dimers.

Infect Immun, 1996 Aug, 64(8), 3374 - 8
Functional characterization of a sialyltransferase-deficient mutant of Neisseria gonorrhoeae; Gill MJ et al.; Previous studies indicate that sialylation of lipopolysaccharide (LPS) by host CMP-N-acetylneuraminic acid (CMP-NANA) catalyzed by bacterial sialyltransferase rendered gonococci resistant to killing by phagocytes, to entry into epithelial cell lines, to killing by immune serum and complement, and to absorption of complement component C3 . These results have been confirmed by comparing a sialyltransferase-deficient mutant (strain JB1) with its parent (strain F62) in appropriate tests . In contrast to F62, JB1 was very susceptible to killing by human polymorphonuclear phagocytes in opsonophagocytosis tests and incubation with CMP-NANA did not decrease the level of killing . The inherent resistance of F62 in these tests was probably due to LPS sialylation by CMP-NANA and lactate present in the phagocytes . A JB1 variant expressing the invasion-associated Opa protein was as able to enter Chang human conjunctiva epithelial cells as an Opa-positive variant of F62, suggesting that the sialyltransferase is not required for Opa-mediated entry . After incubation with CMP-NANA, the number of F62 variant gonococci entering cells but not that of JB1 variant gonococci was drastically reduced . Both JB1 and F62 were killed by incubation with rabbit antibody to gonococcal major outer membrane protein, protein I, and human complement, but only F62 was rendered resistant to the killing by incubation with CMP-NANA . Finally, both JB1 and F62 absorbed similar amounts of complement component C3 and the binding was decreased by incubation with CMP-NANA only for the wild type, F62.

Infect Immun, 1996 Aug, 64(8), 2911 - 6
Identification of a transferrin-binding protein from Borrelia burgdorferi; Carroll JA et al.; Bacterial pathogens have evolved various strategies to acquire iron from the iron-restricted environment found in mammalian hosts . Borrelia burgdorferi should be no different with regard to its requirement for ferric iron, and previous studies have suggested that transferrin (Tf) may be a source of iron in vivo . By probing blots with Tf conjugated to horseradish peroxidase, we have identified an outer membrane protein (28 kDa) from B . burgdorferi B31 that bound holo-Tf but not apo-Tf . The 28-kDa protein bound human, rat, or mouse Tf and was produced only by low-passage (less than passage 5), virulent isolates of strain B31 . In addition, the Tf-binding protein (Tbp) from strain B31 retained the ability to bind Tf after treatment with 2% sodium dodecyl sulfate-1% beta-mercaptoethanol and heating to 100 degrees C for 5 min . These properties are remarkably similar to those of the Tbp of Staphylococcus aureus and Tbp2 from Neisseria meningitidis . B . burgdorferi Sh-2-82 produced an outer membrane protein different in size, i.e., 26 kDa, but with properties similar to those of to the protein from strain B31, suggesting variation in B . burgdorferi Tbps . The exact role of the 28-kDa protein in iron acquisition by B . burgdorferi remains to be determined.

J Bacteriol, 1996 Aug, 178(15), 4670 - 8
HmbR outer membrane receptors of pathogenic Neisseria spp.: iron-regulated, hemoglobin-binding proteins with a high level of primary structure conservation; Stojiljkovic I et al.; We have recently cloned and characterized the hemoglobin receptor gene from Neisseria meningitidis serogroup C . N . meningitidis cells expressing HmbR protein were able to bind biotinylated hemoglobin, and the binding was specifically inhibited by unlabeled hemoglobin and not heme . The HmbR-mediated hemoglobin binding activity of N . meningitidis cells was shown to be iron regulated . The presence of hemoglobin but not heme in the growth medium stimulated HmbR-mediated hemoglobin binding activity . The efficiency of utilization of different hemoglobins by the HmbR-expressing N . meningitidis cells was shown to be species specific; human hemoglobin was the best source of iron, followed by horse, rat, turkey, dog, mouse, and sheep hemoglobins, The phenotypic characterization of HmbR mutants of some clinical strains of N . meningitidis suggested the existence of two unrelated hemoglobin receptors . The HmbR-unrelated hemoglobin receptor was shown to be identical to Hpu, the hemoglobin-haptoglobin receptor of N . meningitidis . The Hpu-dependent hemoglobin utilization system was not able to distinguish between different sources of hemoglobin; all animal hemoglobins were utilized equally well . HmbR-like genes are also present in N . meningitidis serogroups A and B, Neisseria gonorrhoeae MS11 and FA19, Neisseria perflava, and Neisseria polysaccharea . The hemoglobin receptor genes from N . meningitidis serogroups A and B and N . gonorrhoeae MS11 were cloned, and their nucleotide sequences were determined . The nucleotide sequence identity ranged between 86.5% (for N . meningitidis serogroup B hmbR and MS11 hmbR) and 93.4% (for N . meningitidis serogroup B hmbR and N . meningitidis serogroup C hmbR) . The deduced amino acid sequences of these neisserial hemoglobin receptors were also highly related, with overall 84.7% conserved amino acid residues . A stop codon was found in the hmbR gene of N . gonorrhoeae MS11 . This strain was still able to use hemoglobin and hemoglobin-haptoglobin complexes as iron sources, indicating that some gonococci may express only the HmbR-independent hemoglobin utilization system.

J Bacteriol, 1996 Aug, 178(15), 4571 - 5
Cloning, complementation, and characterization of an rfaE homolog from Neisseria gonorrhoeae; Levin JC et al.; Neisseria gonorrhoeae WS1 is a spontaneous pyocin (a bacteriocin produced by Pseudomonas aeruginosa)-resistant mutant of N . gonorrhoeae FA19 that produces a truncated lipooligosaccharide (LOS) and is non-transformable . The LOS-specific mutation in WS1 was moved into a transformable background by transforming FA19 with chromosomal DNA from WS1 (generating strain JWS-1) . A clone (pJCL2) capable of restoring JWS-1 to wild-type LOS expression, as detected by its acquisition of reactivity with monoclonal antibodies and by its complemented sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile, was isolated . Sequential unidirectional deletion and DNA sequence analysis of pJCL2 identified an open reading frame, designated lsi-7, that could complement the defect in JWS-1 . Homology searches against various databases indicated that lsi-7 bad homology with several Escherichia coli genes involved in the phosphorylation of sugars . lsi-7 is adjacent to the lsi-6 gene, another gene involved in LOS biosynthesis . Complementation studies using Salmonella typhimurium lipopolysaccharide mutants showed lsi-6 and lsi-7 to be gonococcal homologs of S . typhimurium rfaD and rfaE, respectively . Reverse transcriptase PCR analysis demonstrated that lsi-6 and lsi-7 are part of the same transcriptional unit.

Proc Natl Acad Sci U S A, 1996 Jul 23, 93(15), 7985 - 90
Peptide methionine sulfoxide reductase contributes to the maintenance of adhesins in three major pathogens; Wizemann TM et al.; Pathogenic bacteria rely on adhesins to bind to host tissues . Therefore, the maintenance of the functional properties of these extracellular macromolecules is essential for the pathogenicity of these microorganisms . We report that peptide methionine sulfoxide reductase (MsrA), a repair enzyme, contributes to the maintenance of adhesins in Streptococcus pneumoniae, Neisseria gonorrhoeae, and Escherichia coli . A screen of a library of pneumococcal mutants for loss of adherence uncovered a MsrA mutant with 75% reduced binding to GalNAcbeta1-4Gal containing eukaryotic cell receptors that are present on type II lung cells and vascular endothelial cells . Subsequently, it was shown that an E . coli msrA mutant displayed decreased type I fimbriae-mediated, mannose-dependent, agglutination of erythrocytes . Previous work {Taha, M . K., So, M., Seifert, H . S., Billyard, E . & Marchal, C . (1988) EMBO J . 7, 4367-4378} has shown that mutants with defects in the pilA-pilB locus from N . gonorrhoeae were altered in their production of type IV pili . We show that pneumococcal MsrA and gonococcal PilB expressed in E . coli have MsrA activity . Together these data suggest that MsrA is required for the proper expression or maintenance of functional adhesins on the surfaces of these three major pathogenic bacteria.

Mol Gen Genet, 1996 Jul 19, 251(5), 509 - 17
Use of a non-selective transformation technique to construct a multiply restriction/modification-deficient mutant of Neisseria gonorrhoeae; Gunn JS et al.; A technique that allows for easy identification of transformants of Neisseria gonorrhoeae in the absence of selective pressure has been developed . A suicide vector that contains a gonococcal DNA uptake sequence was constructed to aid in DNA uptake . In this transformation procedure, a limiting number of cells is incubated with an excess amount of DNA, and the mixture is plated onto a non-selective medium . At least 20% of the resulting colonies contained cells that had been transformed . This strategy was utilized to construct specific deletions of the S.N goI, II, IV, V and VII restriction-modification (R/M) genes . All five deletions were successfully incorporated into the chromosome of FA19, producing strain JUG029 . Strain JUG029 could be transformed with non-methylated plasmid DNA while strain FA19 could not be transformed with such DNA . The development of a simple, non-selective transformation technique, coupled with the construction of a strain that is more permissive for DNA-mediated transformation, will aid in genetic manipulations of the gonococcus.

Biochem Biophys Res Commun, 1996 Jul 16, 224(2), 444 - 50
Determination of the size and degree of acetyl substitution of oligosaccharides from Neisseria meningitidis group A by ionspray mass spectrometry; Cescutti P et al.; The capsular polysaccharide produced by Neisseria meningitidis group A has the following structure: {formula: see text} {formula: see text} This polysaccharide was partially hydrolysed with acetic acid, and the oligomers obtained were separated by fast performance liquid chromatography . Six fractions were collected and characterised by ionspray mass spectrometry in the positive ion mode . This soft ionisation technique established the size of the obtained oligosaccharides and the degree of O-acetyl substitution for each fraction.

Srp Arh Celok Lek, 1996 Jul-Aug, 124(7-8), 193 - 6
{Antibiotic therapy in the treatment of inflammatory diseases in the minor pelvis}; Vasiljevic M et al.; Pelvic inflammatory diseases are usually caused by sexually transmitted microorganisms, as are Neisseria gonorrhoeae and Chlamydia trachomatis, either alone or associated with endogenous flora of the lower genital tract, as with other gram-positive and gram-negative anaerobic and aerobic bacteria {1, 2} . SUBJECT: The aim of the study was to estimate the effect of three broad-spectrum combinations of antimicrobial therapy in the treatment of pelvic inflammatory diseases in hospitalized patients . MATERIAL AND METHODS: We analysed the therapeutic success of some antimicrobial therapies in 154 patients with pelvic inflammatory disease, who were treated in the Narodni Front Hospital of Gynaecology and Obstetrics in Belgrade, during 1992 and 1993 . Three drug therapies were applied . The combination of Ceftriaxon plus Doxycycline was given to 51 women . Fifty five women were treated by a combination of Gentamycin plus Clidamycin, and 48 women were treated by a combination of Gentamycin and Metronidazole . RESULTS: The therapeutic success after the application of the three different antibiotic therapies was recorded in 139 of 154 women (90.26%) . Of 136 patients with uncomplicated pelvic inflammatory diseases, the therapeutic success was noted in 129 (94.85%) individuals, while of 18 women with tubo-ovarian abscess therapeutic success was recorded in 10 (55.56%) patients . Of 51 women treated by the combination of Ceftriaxon plus Doxycycline, the therapeutic success was observed in 46 (90.19%) patients . Fifty five women treated by the combination of Gentamycin plus Clindamycin, the therapeutic success was noted in 50 (90.19%) subjects . Of 48 women, treated by the combination of Gentamycin plus Metronidazole, the therapeutic success was found in 43 (89.58%) women . No statistically significant difference was found among the applied antibiotic therapies (p > 0.05) . Of 18 women with tubo-ovarian abscess 8 were operated on . Of these 8 women in 6 patients hysterectomy with bilateral salpingo-oophorectomy was performed and in two women unilateral salpingo-oophorectomy was carried out . DISCUSSION: Pelvic inflammatory diseases are often of polymicrobial aetiology . In 43 patients we found two types of bacteria in the cervical culture . The therapeutic success was achieved by these three antibiotic therapies . It was 90.26%, the therapeutic success in the treatment of pelvic inflammatory diseases by Ceftriaxon plus Doxycycline was noted in 90.19% of patients . The therapeutic success of antibiotic therapy with Gentamycin plus Clindamycin was obtained in 90.91% of patients . The success of antibiotic therapy with Gentamycin plus Metronidazole was recorded in 89.59% of patients . Our results are similar to those of other authors {3, 4, 6, 7} . No statistically significant difference was found among the applied antibiotic therapies . CONCLUSION: An early diagnosis and an aggressive treatment may prevent serious sequelae of this increasingly common sexually transmitted disease . The antibiotics should be of antimicrobial broad spectrum . Good effects can be best reached by a combined antibiotic therapy . Duration of parenteral administration of antibiotics should be several days and for at least 48 hours after the patient's defervescence.

FEMS Immunol Med Microbiol, 1996 Jul, 14(4), 211 - 20
Binding diversity of monoclonal antibodies to alpha(2-->8) polysialic acid conjugated to outer membrane vesicle via adipic acid dihydrazide; Devi SJ et al.; Murine monoclonal antibodies (mAbs) were generated using group B Neisseria meningitidis and Escherichia coli K1 polysaccharides (PSs) conjugated to outer membrane vesicle (OMV) via adipic acid dihydrazide, and were used to identify the immunodeterminants expressed on these capsular PSs . Ten mAbs representative of IgM and all subclasses of IgG were obtained which recognized diverse immunodeterminants on alpha(2-->8) polysialic acid (PSA) . The specificity of mAbs to different antigenic determinants was assessed by their differential binding to PSA attached to a solid phase by different methods and confirmed by absorption studies . Two mAbs from the E . coli K1 fusion were directed to the O-acetyl epitope and the rest reacted with both the PSs only when attached to a solid phase by certain means . The methods by which PSA was coated on the solid phase had an impact on the epitope expression and binding pattern . At the concentrations used, the O-acetyl-specific mAbs, IgG1 and IgG3 mAbs were not bactericidal against group B N . meningitidis, whereas other mAbs were . The conjugates B and K1 PSs present to the murine immune system different antigenic determinants, some of which elicit bactericidal antibodies.

Sex Transm Dis, 1996 Jul-Aug, 23(4), 270 - 2
A clinical isolate of Neisseria gonorrhoeae with in vitro resistance to erythromycin and decreased susceptibility to azithromycin; Ehret JM et al.; BACKGROUND AND OBJECTIVES: Erythromycin is a recommended treatment for penicillin-allergic pregnant women with gonorrhea, and azithromycin has been suggested as therapy for coexisting gonococcal and chlamydial infections . Although gonococcal resistance to erythromycin is not uncommon, decreased resistance to azithromycin is rare . A clinical isolate of Neisseria gonorrhoeae with in vitro resistance to erythromycin and decreased susceptibility to azithromycin is reported . STUDY DESIGN: This is a case report . RESULTS: Antimicrobial susceptibility testing of a clinical isolate of N . gonorrhoeae revealed a minimal inhibitory concentration (MIC) of 2 micrograms/ml to azithromycin and 32 micrograms/ml to erythromycin . Five hundred other urethral isolates were tested, resulting in an MIC for erythromycin ranging from 0.015 to 2 micrograms/ml . The range for azithromycin was 0.015 to 0.5 micrograms/ml . There was a strong correlation between erythromycin and azithromycin MICs (r = 0.73; P < 0.0001) . CONCLUSIONS: Continued national monitoring is needed to detect the appearance and early dissemination of new types of gonococcal resistance.

Antimicrob Agents Chemother, 1996 Jul, 40(7), 1720 - 1
Dose-ranging study of CP-99,219 (trovafloxacin) for treatment of uncomplicated gonorrhea; Hook EW 3rd et al.; Thirty-nine patients with uncomplicated gonorrhea were randomized to receive single, oral 50-, 100-, or 200-mg doses of trovafloxacin (CP-99,219), a new quinolone antibiotic . All 31 evaluable patients were cured of infection . Trovafloxacin was well tolerated . The trovafloxacin MICs at which 50 and 90% of 36 Neisseria gonorrhoeae isolates are inhibited were 0.002 and 0.004 mg/liter, respectively (MIC range, < 0.0005 to 0.008 mg/liter) . These preliminary studies suggest that trovafloxacin is effective for the treatment of uncomplicated gonorrhea at single oral doses as low as 50 mg.

J Clin Microbiol, 1996 Jul, 34(7), 1863 - 5
Auxotypes and serogroups of tetracycline-resistant Neisseria gonorrhoeae isolated in Malaysia; Koay AS et al.; Between 1992 and 1994, 253 tetracycline-resistant Neisseria gonorrhoeae (TRNG) strains were isolated and characterized by auxotype and serogroup (A/S) classes to study TRNG prevalence in different years . TRNG accounted for 28.1, 42.5, and 51.9% of the strains isolated in 1992, 1993, and 1994, respectively, showing a significant increase in each successive year (chi square = 26.7, P < 0.001) . There was no significant increase in penicillinase-producing TRNG, which accounted for 53.1, 53.8, and 63.2% of the TRNG isolates . The 253 TRNG isolates belonged to 53 A/S classes . Eighteen A/S classes not observed in 1992 were detected in 1993, and 11 A/S classes not observed in 1992 and 1993 were isolated in 1994, indicating dissemination of the tetracycline resistance gene among the N . gonorrhoeae strains in Malaysia . Its emergence and subsequent rapid spread are alarming . The plasmid is capable of self-transfer (S.A . Morse, S.R . Johnson, J.W . Biddle, and M.C . Roberts, J . Infect . Dis . 155:819-822, 1987), allowing further dissemination of tetracycline resistance.

J Bacteriol, 1996 Jul, 178(14), 4301 - 5
Interruption of the gpxA gene increases the sensitivity of Neisseria meningitidis to paraquat; Moore TD et al.; Antioxidant enzymes are thought to be important for the survival of pathogenic Neisseria species . We have further characterized the glutathione peroxidase homolog gene (gpxA), which we recently isolated from Neisseria meningitidis FAM20 (T.D.E . Moore and P.F . Sparling, Infect . Immun . 63:1603-1607, 1995) . GpxA was found to be produced constitutively in vivo . An isogenic, omega insertion mutant in the gpxA gene was constructed and characterized . The gpxA insertion mutant was much more sensitive to the oxidative stress caused by paraquat and slightly more sensitive to hydrogen peroxide . This is the first demonstration of a phenotype arising from a mutation of a glutathione peroxidase homolog gene in a prokaryotic organism . Protection of the cell by GpxA from the effects of oxidative stress caused by aerobic metabolism may contribute to the ability of Neisseria meningitidis to cause disease in the human host.

J Bacteriol, 1996 Jul, 178(14), 4224 - 32
Isolation and analysis of a fur mutant of Neisseria gonorrhoeae; Thomas CE et al.; The pathogenic Neisseria spp . produce a number of iron-regulated gene products that are thought to be important in virulence . Iron-responsive regulation of these gene products has been attributed to the presence in Neisseria spp . of the Fur (ferric uptake regulation) protein . Evidence for the role of Fur in neisserial iron regulation has been indirect because of the inability to make fur null mutations . To circumvent this problem, we used manganese selection to isolate missense mutations of Neisseria gonorrhoeae fur . We show that a mutation in gonococcal fur resulted in reduced modulation of expression of four well-studied iron-repressed genes and affected the iron regulation of a broad range of other genes as judged by two-dimensional polyacrylamide gel electrophoresis (PAGE) . All 15 of the iron-repressed spots observed by two-dimensional PAGE were at least partially derepressed in the fur mutant, and 17 of the 45 iron-induced spots were affected by the fur mutation . Thus, Fur plays a central role in regulation of iron-repressed gonococcal genes and appears to be involved in regulation of many iron-induced genes . The size and complexity of the iron regulons in N . gonorrhoeae are much greater than previously recognized.

Tidsskr Nor Laegeforen, 1996 Jun 30, 116(17), 2017 - 21
{Indications for laboratory testing of gonorrhea . New recommendations from the Working Group against Gonorrhea}; Aavitsland P; Culture testing for sexually transmitted Neisseria gonorrhoeae infection is widely used in Norwegian general practice . The gain from this testing is very low, since gonorrhoea is almost eradicated in Norway . This reduces the cost-effectiveness of the testing . In cooperation with a panel of medical microbiologists, gynaecologists, venerologists, general practitioners and public health specialists, we have reviewed the literature and drawn up a set of recommendations for the use of culture testing for gonococcal infection to suit the epidemiological situation in Norway . We emphasise clinical testing of men and women, partner notification and tests-of-cure . We do not recommend opportunistic screening for gonococci in any situation whatsoever.

Biochemistry, 1996 Jun 25, 35(25), 8429 - 38
Simultaneous detection of multiple nucleic acid targets in a homogeneous format; Nelson NC et al.; The acridinium ester 4-(2-succinimidyloxycarbonylethyl)phenyl-10-methylacridinium 9-carboxylate trifluoromethane sulfonate (AE), which reacts rapidly with alkaline hydrogen peroxide to produce light, has been used as a detection label in a number of assay procedures, including nucleic acid probe-based systems {Nelson et al . (1995) in Nonisotopic Probing, Blotting and Sequencing (Kricka, L . J., Ed.) pp 391-428, Academic Press, Inc., San Diego, CA} . We have synthesized a number of derivatives of this AE and characterized their chemiluminescent properties . These derivatives display significant differences in the kinetics of the chemiluminescence reaction as well as optimal pH for light production . These differences allow two or more derivatives to be simultaneously detected and quantitated in a single reaction vessel . Several of these derivatives have been covalently linked to nucleic acid probe molecules and have been further characterized in regard to chemiluminescence properties as well as hydrolysis of the ester bond in both single- and double-stranded conformations . On the basis of these properties, homogeneous assay formats utilizing DNA probes labeled with various AE derivatives were developed . Simultaneous detection and quantitation of Chlamydia trachomatis and Neisseria gonorrhoeae, the gag and pol regions of HIV, and wild-type and mutant HIV sequences was achieved with high sensitivity and discrimination.

Harefuah, 1996 Jun 16, 130(12), 811 - 4, 880, 879
{Diagnostic and therapeutic approach to sexually transmitted diseases}; Madjar S et al.; In an ongoing study we evaluated 71 males and 32 females attending our sexually transmitted diseases (STD) clinic . Intraurethral or endocervical swab specimens were cultured for Neisseria gonorrhea (NG), Ureaplasma urealyticum (UU), Mycoplasma homines (MH), Herpes simplex virus (HSV) and Chlamydia trachomatis (CT), using an ELISA technique and the polymerase chain reaction (PCR) . HIV antigen, hepatitis B (HBV) and Treponema pallidum (TP) seropositivity were tested by ELISA . Mean age was 33.4 and range 15-72 years . 83 patients (81%) used condoms only rarely, 35 (35%) had multiple sexual partners and 83 (81%) were treated empirically prior to evaluation . Dysuria and urethral discharge were found in 47 (45.6%), of whom 34 (33%) were males; the majority of females were asymptomatic . A specific etiology for STD was found in 53 patients (51.4%) and 1/3 had more than 1 pathogen . CT, UU, MH, HSV, NG and TV were found in 27, 24, 5, 3, 2 and 1, respectively . 8 patients were seropositive for HBV and 1 for TP; all were seronegative for HIV . CT was the most prevalent pathogen found . All patients with STD symptoms should be screened for all sexually transmitted pathogens, since many of them have more than 1 pathogen . STD clinics in Israel should be developed in conjunction with microbiology laboratories for better management of STD in the community.

Hybridoma, 1996 Jun, 15(3), 191 - 8
Generation and characterization of a human monoclonal IgM antibody that recognizes a conserved epitope shared by lipopolysaccharides of different gram-negative bacteria; Seifert M et al.; A hybridoma cell line secreting a human monoclonal antibody (humab) directed to an epitope in the lipid A region of lipopolysaccharides of Gram-negative bacteria was isolated . Peripheral blood lymphocytes (PBL) obtained from a healthy volunteer were immortalized by Epstein-Barr virus (EBV) transformation . Lymphoblastoid cell lines (LCL) secreting antibodies to the lipopolysaccharides of Gram-negative bacteria were determined by an enzyme-linked immunosorbent assay (ELISA) and subsequently fused with the human-mouse heteromyeloma cell line CB-F7 by polyethylenglycol (PEG)-mediated fusion . A hybridoma line producing a humab (LPD5H4), of the IgM/lambda isotype, which strongly reacted with the lipid A portion of Salmonella and E . coli spp . in ELISA, was established . The antibody was purified by hydrophobic interaction chromatography and gel filtration . Immunoblotting experiments showed a strong reactivity of the humab LPD5H4 with the lower molecular species of different rough and smooth lipopolysaccharide (LPS) types of the bacteria species Salmonella, E . coli, Klebsiella, and Neisseria meningitidis, whereas those of Pseudomonas spp . were negative . Binding of humab LPD5H4 to solid phase bound lipid A and different rough mutants of LPS could be inhibited by the corresponding antigens in solution . Competition assays with a murine monoclonal antibody to lipid A and with polymyxin B indicate that humab LPD5H4 recognizes its epitope in this extremely conserved part of the LPS molecule . In vitro tests demonstrated that the MAb is able to partially inhibit the LPS-induced release of TNF-alpha using isolated peripheral blood mononuclear cells (PBMC).

J Clin Microbiol, 1996 Jun, 34(6), 1551 - 2
Prevalence of fluorescent monoclonal antibody-nonreactive Neisseria gonorrhoeae in five North American sexually transmitted disease clinics; Smith KR et al.; We compared a direct fluorescent monoclonal antibody (DFA) test with alternative enzymatic and fermentation tests for identifying presumptive gonococcal isolates in a systematic sample from patients attending five sexually transmitted disease clinics in five cities . Fourteen (2.5%) of 556 isolates from three clinics were nonreactive with DFA confirmatory reagent and reactive by both the Quad-Ferm (BioMerieux Vitek Inc.) and the Rapid NH (Innovative Diagnostic Systems, Inc.) tests . The prevalence of DFA-nonreactive Neisseria gonorrhoeae isolates varies geographically and is independent of local methods for the identification of possible gonococci.

J Clin Microbiol, 1996 Jun, 34(6), 1548 - 50
The porA alleles are identical in subgroup III serogroup A Neisseria meningitidis strains isolated in China in the 1960s and 1980s; Malorny B et al.; The porA genes from serogroup A, subgroup III strains isolated in the People's Republic of China in 1966 and in 1984 and 1985 were amplified by PCR at an annealing temperature of 75 degrees C . The DNA sequences (5 strains) and the restriction patterns generated by MspI (14 strains) were identical, unlike the results reported by Peixuan et al . (Z . Peixuan, H . Xujing, and X . Li, J . Clin . Microbiol . 33:458-462, 1995) . Furthermore, PCR products which were amplified at an annealing temperature of 60 degrees C, as described by Peixuan et al., were heterogeneous in our study.

J Clin Microbiol, 1996 Jun, 34(6), 1468 - 73
Molecular subtyping of Neisseria meningitidis serogroup B: comparison of five methods; Swaminathan B et al.; In order to compare methods for subtyping Neisseria meningitidis serogroup B isolates, 96 isolates obtained from various locations in the United States and northwestern Europe were subtyped by five methods: monoclonal antibody (MAb)-based serotyping and serosubtyping, DNA macrorestriction analysis by pulsed-field gel electrophoresis (PFGE), multilocus enzyme electrophoresis (MEE), ribotyping, and PCR-restriction fragment length polymorphism of the internally transcribed spacer region of the rRNA operon (ITS PCR-RFLP) . All N . meningitidis serogroup B isolates were typeable by PFGE, MEE, ribotyping, and ITS PCR-RFLP . Only 44.8% of the isolates were completely typeable (both serotype and serosubtype determination) by MAb-based serotyping and serosubtyping . 60.4% of the isolates could be serotyped but not serosubtyped, and 90.6% of the isolates could be either serotyped or serosubtyped . Simpson's discrimination indices of diversity for the methods were as follows: PFGE, 99.7%; MEE, 99.4%; ribotyping, 98.8%; MAb serotyping, 75.8%; MAb serotyping and/or serosubtyping 97.5%; and ITS PCR-RFLP, 84.2% . The high degree of diversity observed by PFGE, MEE, and ribotyping can be explained by the fact that isolates were collected from different geographic locations at various times . PFGE, MEE, and ribotyping showed greater discriminatory abilities than MAb-based serotyping and serosubtyping or ITS PCR-RFLP.

J Clin Microbiol, 1996 Jun, 34(6), 1462 - 4
Detection of quinolone-resistant Neisseria gonorrhoeae; Kam KM et al.; The present National Committee for Clinical Laboratory Standards (NCCLS) guideline for testing Neisseria gonorrhoeae quinolone susceptibility defines only a susceptible category for ciprofloxacin, enoxacin, lomefloxacin, and ofloxacin, while susceptible, intermediate, and resistant categories are defined for fleroxacin . To further define the criteria for detection of quinolone resistance in gonococci, by standard disk diffusion and agar dilution methodologies recommended by the NCCLS, we tested 29 strains of quinolone-resistant N . gonorrhoeae (QRNG) recently isolated from ofloxacin-treated patients who were considered clinical failures . Regression analyses were performed on these results together with those of another 20 strains showing reduced susceptibility and 13 fully susceptible strains (ofloxacin MICs of < or = 0.25 microgram/ml) . With 5-micrograms ofloxacin disks, resistance in 27 (93.1%) of the QRNG strains (MICs of > 1 microgram/ml) was detected by the criterion of a zone diameter of < 22 mm, while in the remaining 2 (6.9%), the disks failed to detect resistance . A cluster of 15 highly resistant strains showed ofloxacin MICs of > 4 micrograms/ml and zone diameters of < 13 mm . When tested with 5-micrograms ciprofloxacin disks, the corresponding values for resistance and high-level resistance of these QRNG strains were < 25 mm (MICs of > 0.5 micrograms/ml) and < 15 mm (MICs of > 2 micrograms /ml), respectively . Six strains for which ofloxacin MICs were > or = 8 micrograms/ml showed no zones at all with both 5-micrograms ofloxacin and 5-micrograms ciprofloxacin disks . These QRNG strains are now firmly established in the Southeast Asia region, and it is important for clinical laboratories to recognize these clinically resistant strains and to monitor their spread.

Antimicrob Agents Chemother, 1996 Jun, 40(6), 1376 - 81
Antibacterial activities of epiroprim, a new dihydrofolate reductase inhibitor, alone and in combination with dapsone; Locher HH et al.; Epiroprim (EPM; Ro 11-8958) is a new selective inhibitor of microbial dihydrofolate reductase . EPM displayed excellent activity against staphylococci, enterococci, pneumococci, and streptococci which was considerably better than that of trimethoprim (TMP) . EPM was also active against TMP-resistant strains, although the MICs were still relatively high . Its combination with dapsone (DDS) was synergistic and showed as in vitro activity superior to that of the TMP combination with sulfamethoxazole (SMZ) . The EPM-DDS (ratio, 1:19) combination inhibited more than 90% of all important gram-positive pathogens at a concentration of 2 + 38 micrograms/ml . Only a few highly TMP-resistant staphylococci and enterococci were not inhibited . EPM was also more active than TMP against Moraxella catarrhalis, Neisseria meningitidis, and Bacteroides spp., but it was less active than TMP against all other gram-negative bacteria tested . Atypical mycobacteria were poorly susceptible to EPM, but the combination with DDS was synergistic and active at concentrations most probably achievable in biological fluids (MICs from 0.25 +/- 4.75 to 4 + 76 micrograms/ml) . EPM and the EPM-DDS combination were also highly active against experimental staphylococcal infections in a mouse septicemia model . The combination EPM-DDS has previously been shown to exhibit activity in Pneumocystis carinii and Toxoplasma models and, as shown in the present study, also shows good activity against a broad range of bacteria including many strains resistant to TMP and TMP-SMZ.

J Natl Med Assoc, 1996 Jun, 88(6), 353 - 6
Gonoccocal endocarditis; Thompson EC et al.; In the postantibiotic era, systemic complications from a gonococcal infection are rare . Females tend to have a higher frequency of gonococcal sepsis than males . In contrast, males have a higher rate of gonococcal endocarditis . This article describes a case of a previously healthy young male who presented with aortic insufficiency and blood cultures positive for Neisseria gonorrhoeae . Despite adequate antibiotic coverage, the patient's aortic insufficiency worsened, requiring aortic value replacement before discharge from the hospital . The patient's recovery was uneventful.

Bull Rheum Dis, 1996 Jun, 45(4), 6 - 8
Rheumatic diseases and inherited complement deficiencies; Ruddy S; Deficiencies of individual complement proteins may be accompanied by SLE or related syndromes . Deficiencies of the classic activation pathway are often involved . In cases of C4 and C2 deficiency, there is evidence that this association occurs more frequently than would be expected by chance . The clinical picture differs from classic SLE . There is an increased frequency of skin involvement, a decreased frequency of renal disease, low or absent levels of antibody to native DNA, and increased levels of anti-Ro (SS-A) . The mechanism for the association probably involves the effects of C3 and C4 on the precipitation of immune complex solubility, or on their processing through cell surface c4b/c3b receptors on phagocytes . Disseminated or recurrent Neisseria infections are common in patients lacking the constituents of the terminal MAC that are important in killing these organisms.

J Bacteriol, 1996 Jun, 178(12), 3544 - 9
The cryptic general secretory pathway (gsp) operon of Escherichia coli K-12 encodes functional proteins; Francetic O et al.; Systematic sequencing of the Escherichia coli K-12 chromosome (GenBank entry U18997) has revealed the presence of an apparently complete operon of genes (the gspC-0 operon) similar to genes coding for components of the main terminal branch of the general secretory pathway (e.g., the Klebsiella oxytoca pulC-0 pullulanase secretion operon) and to related genes required for type IV pilus biogenesis . For example, the last gene in the gsp operon, gspO (formerly hopD), encodes a protein which is similar to several type IV prepilin peptidases . Expression of gspO from lacZp promotes cleavage of two known prepilin peptidase substrates in E . coli K-12: Neisseria gonorrhoeae type IV prepilin and K . oxytoca prePulG protein . gspO also complements a mutation in the corresponding gene (pulO) of the pullulanase secretion operon when it is expressed from lacZp . Another gene in the gsp operon, gspG (formerly hopG), encodes a protein similar to prePulG, a component of the pullulanase secretion pathway . Expression of gspG from lacZp leads to production of a protein which (i) is recognized by PulG-specific antiserum (and by antiserum against the Pseudomonas aeruginosa PulG homolog XcpG {formerly XcpT}), (ii) is processed in cells expressing gspO, and (iii) restores secretion in cells carrying a pulG mutation . The chromosomal copies of gspG and gspO are apparently not expressed, probably because of very weak transcription from the upstream region, as measured by using a chromosomal gspC-lacZ operon fusion . Thus, the gsp operon of E . coli K-12 includes at least two functional genes which, together with the rest of the operon, are probably not expressed under laboratory conditions.

Arthritis Rheum, 1996 Jun, 39(6), 950 - 8
Molecular detection of bacterial DNA in venereal-associated arthritis; Li F et al.; OBJECTIVE: To evaluate the utility of polymerase chain reaction (PCR) amplification in detecting DNA from venereal-associated microorganisms in the synovial fluid of patients with inflammatory arthritis . METHODS: Oligonucleotide primers were developed for nested PCR based on Chlamydia, Ureaplasma, and Neisseria DNA sequences . PCR products were detected by gel electrophoresis and dot-blot hybridization . Primers specific for the target bacterial DNA were used to search for bacterial DNA in 61 synovial fluid specimens from patients with inflammatory arthritis, including several clinically associated with venereal infection . RESULTS: Five of the 61 synovial fluid specimens were positive for Neisseria gonorrhoeae DNA . Four of the 5 patients had clinical diagnoses of gonococcal arthritis; the other patient had an unexplained monarthritis . One specimen from a patient with a clinical diagnosis of gonococcal arthritis was negative for N gonorrhoeae . Three of the 61 specimens were positive for Chlamydia DNA . Two were derived from patients with clinical diagnoses of reactive arthritis or Reiter's syndrome, and 1 was from a patient with unexplained monarthritis . One of the 61 specimens was positive from Ureaplasma DNA; this sample was from a patient with a clinical diagnosis of Reiter's syndrome . In an additional patient with Reiter's syndrome, Ureaplasma DNA was also found in prostate biopsy tissue and a urine sample obtained after prostate massage (synovial fluid not available) . CONCLUSION: These data support the classification of these 3 venereal-associated arthritides as infectious processes, and suggest that PCR for bacterial DNA is a useful method for detecting infectious agents in synovial fluid.

J Infect Dis, 1996 Jun, 173(6), 1437 - 44
Risk factors for Chlamydia trachomatis pelvic inflammatory disease among sex workers in Nairobi, Kenya; Kimani J et al.; Among 302 female sex workers in Nairobi, Kenya, who were followed for 17.6 +/- 11.1 months, 146 had one or more infections with Chlamydia trachomatis; 102 had uncomplicated cervical infection only, 23 had C . trachomatis pelvic inflammatory disease (PID), and 21 had combined C . trachomatis and Neisseria gonorrhoeae PID . As determined by multivariate logistic regression analysis, risk factors for C . trachomatis PID included repeated C . trachomatis infection (odds ratio {OR}, 1.8; 95% confidence interval {CI}, 1.3-2.4; P = .0004), antibody to C . trachomatis heat-shock protein 60 (OR, 3.9; CI, 1.04-14.5; P = .04), oral contraceptive use (OR, 0.28; 95% CI, 0.08-0.99; P = .048), and number of episodes of nongonococcal nonchlamydial PID (OR, 1.7; 95% CI, 1.1-2.7; P = .02) . Among human immunodeficiency virus (HIV)-seropositive women, a CD4 lymphocyte count of <400/mm3 was an additional independent risk factor for C . trachomatis PID (OR, 21.7; 95% CI, 1.2-383; P = .036); among HLA-typed women, HLA-A31 was independently associated with C . trachomatis PID (OR, 5.6; 95% CI, 1.1-29.4; P = .043) . The results suggest an immune-mediated pathogenesis for C . trachomatis PID.

J Infect Dis, 1996 Jun, 173(6), 1422 - 7
Sialylation lessens the infectivity of Neisseria gonorrhoeae MS11mkC; Schneider H et al.; In a human challenge experiment, the infectivity of gonococci with sialylated lipooligosaccharide (LOS) was compared with the infectivity of gonococci with unsialylated LOS . Volunteers were intraurethrally inoculated with approximately 5000 sialylated or unsialylated piliated, non-opaque (P+Opa-, transparent) colony type gonococci, strain MS11mkC . Five (83%) of 6 volunteers inoculated with unsialylated gonococci became infected; however, only 1 of 5 volunteers became infected with sialylated gonococci . The unsialylated gonococcal infections, with a median incubation time of 62 h (range, 32-98), were similar to previously described experimental infections . Gonococci shed by infected volunteers showed a transition from the P+Opa- phenotype of the inoculation strain to the P+Opa+ (piliated, opaque) phenotype 12-60 h before onset of disease . The subject with sialylated gonococcus infection had an extended incubation period, showing a progressive increase in the number of organisms shed until he became symptomatic on day 6 after inoculation . These results show that gonococci with sialylated LOS are less infective than gonococci with unsialylated LOS.

Gene, 1996 May 24, 171(1), 133 - 4
Plasmids with erythromycin resistance and catechol 2,3-dioxygenase- or beta-galactosidase-encoding gene cassettes for use in Neisseria spp; Zhou D et al.; Four new plasmids containing the ermC' (encoding a methyltransferase which confers resistance to erythromycin), xylE-ermC' (xylE, encoding catechol 2,3-dioxygenase) and lacZ-ermC' cassettes have been constructed . The 10-bp gonococcal uptake sequence has been placed downstream from ermC' to facilitate the delivery of these cassettes into pathogenic Neisseria spp . Several restriction sites have been placed to flank the cassettes to allow their excision and directional cloning . These plasmids will provide valuable tools for constructing insertional mutants and transcriptional fusions in Neisseria spp . or other bacteria.

Cell, 1996 May 3, 85(3), 391 - 402
Modulation of Neisseria porin (PorB) by cytosolic ATP/GTP of target cells: parallels between pathogen accommodation and mitochondrial endosymbiosis; Rudel T et al.; PorB of the pathogenic Neisseria species belongs to the large family of pore-forming proteins (porins) produced by gram-negative bacteria . PorB is exceptional in that it is capable of translocating vectorially into membranes of infected target cells and functions in the infection process . Here we report on an unexpected similarity between Neisserial PorB and mitochondrial porins . Both porin classes interact with purine nucleoside triphosphates, which down-regulate pore size and cause a shift in voltage dependence and ion selectivity . Patch-clamp analyses indicate that PorB channel activity is tightly regulated in intact epithelial cells . In light of recent findings on the pivotal role of PorB in virulence and the prevention of phagosome lysosome fusion, these data provide important mechanistic clues on the intracellular pathogen accommodation reminiscent of mitochondrial endosymbiosis.

Pol Tyg Lek, 1996 May, 51(19-22), 293 - 4
{Deficiency of C2 component in a 9-year old girl}; Madalinski K et al.; The authors describe a 9 year-old girl with homozygous deficiency of C2 component, causing null activity of the classical pathway of complement . The girl suffered from several recurrent infections since birth, later developed signs of the Henoch-Schonlein disease . At the age of 1 1/2 years she was hospitalized due-to bacterial meningitis, most probably of Neisseria meningitis etiology.

Am J Obstet Gynecol, 1996 May, 174(5), 1618 - 21
Bacterial colonization of the vagina during pregnancy in four ethnic groups . Vaginal Infections and Prematurity Study Group; Goldenberg RL et al.; OBJECTIVE: Our purpose was determine whether there are ethnic differences in vaginal colonization with organisms reputed to be associated with preterm delivery . STUDY DESIGN: In seven urban medical centers in the United States from 1984 to 1989, 13,747 predominantly low-socioeconomic-status women at 23 to 26 weeks' gestation were grouped according to ethnic origin . Each had a vaginal culture for multiple organisms and Gram's stain and pH test to diagnose bacterial vaginosis . Multivariate analyses with adjustment for maternal age, insurance status, marital status, smoking history, education level, age at first sexual intercourse, and number of male partners in the past year were also performed to determine the odds ratio for a black woman versus a white woman having a positive test result . RESULTS: For nearly every organism studied, Asian-Pacific Islander and white women had the lowest percentages of positive test results whereas black women had the highest . When black and white women are compared, black women were more likely to be colonized with Chlamydia trachomatis, 16% versus 5%; Neisseria gonorrhoeae, 2.5% versus 0.4%; Bacteroides sp., 25% versus 14%; bacterial vaginosis, 23% versus 9%; and the four other organisms evaluated . Adjustment for potential confounders including certain health behaviors did not markedly change the risk of a black woman having a positive test result, with the odds ratios for colonization with each organism ranging from 1.8 to 6.4 . CONCLUSIONS: Highly significant differences exist in vaginal colonization patterns among women of different ethnic groups, with the highest rates of potentially pathogenic organisms observed in black women and the lowest in Asian-Pacific Islander women . Differences in health behaviors do not explain the variation in colonization rates.

Infection, 1996 May-Jun, 24(3), 258 - 60
Bioprosthetic valve endocarditis caused by Neisseria elongata subspecies nitroreducens; Meuleman P et al.; A new case of Neisseria elongata ssp . nitroreducens bacteremia and endocarditis in a 74-year-old woman who had undergone aortic valve replacement in 1992 is reported in detail . N . elongata ssp . nitroreducens differs from the other subspecies of N . elongata in the additional reduction of nitrate without gas formation . Like most Neisseria spp . except Neisseria meningitidis and Neisseria gonorrhoeae, this N . elongata ssp . nitroreducens is usually classified in the group of "non-pathogenic" Neisseria spp . This case report indicates that the presence of subspecies of this group is significant when isolated from normally sterile sites and can cause severe disease in susceptible individuals.

Med Microbiol Immunol (Berl), 1996 May, 185(1), 27 - 30
Avoiding artifacts in the infant rat model for bacterial meningitis: use of Sangur test strips for the rapid quantification of blood contamination in cerebrospinal fluid; Vogel U et al.; The infant rat model is widely used to study the pathogenesis of meningitis caused by a variety of gram-positive and gram-negative bacteria . However, the interpretation of published results concerning meningitis is difficult in many records because the fact that blood contamination of the cerebrospinal fluid (CSF) cannot be avoided during the traumatic puncture procedure has not been taken into consideration . Since bacterial invasion of the central nervous system develops following bacteremia in this model, blood contamination of the CSF leads to a falsification of the CSF bacterial counts . Here we present an evaluation of a rapid and quantitative test for CSF blood contamination using Sangur test strips . The procedure requires minimal amounts of CSF and allows direct calculation of the CSF bacterial load due to blood contamination and, thus, provides refined criteria for the presence of bacterial meningitis in the infant rat model . It is superior to the detection of erythrocytes using a hemocytometer since it is less time consuming . Furthermore, we demonstrate the value of this method for the experimental infection of rats with Neisseria meningitis.

Int J STD AIDS, 1996 May-Jun, 7(3), 221 - 3
Patients with genital warts: how are they managed by general practitioners?
Estcourt CS, Higgins SP, Hall J, Hillier VF, Chandiok S, Woolley PD.
This study was carried out to assess the management by general practitioners of patients with genital warts . An anonymized postal questionnaire was sent to 2060 general practitioners in the north west of England; 1260 (61.2%) completed questionnaires were returned . A total of 76.69% of GPs saw one or more cases per quarter . Of 468 (37.1%) GPs who managed patients in-house, 393 (83.9%) used podophyllin of concentration between 0.5 and 50%; 169 (43%) did not specify the podophyllin concentration and 112(23.9%) used podophyllotoxin . Of 395 GPs (31.3%) prescribing patient self-treatment, 259 (65.6%) prescribed podophyllin, but 60.23% did not specify the concentration; 134(33.9%) prescribed podophyllotoxin . Screening for selected sexually transmitted infections (STIs) was performed by 258/618 (41.7%) GPs . Only 74 (11.97%) screened for Chlamydia trachomatis and Neisseria gonorrhoeae and Trichomonas vaginalis . Partner notification was advised by 1138 (90.3%) and condom use by 1027 (81.5%) . Only 333 (26.4%) referred all patients to GUM departments . Many GPs manage patients in-house, most using podophyllin solutions which vary in concentration, are non-standardized and potentially hazardous . Most GPs who instigate patient self-treatment prescribe podophyllin which is not licensed for this usage . Podophyllotoxin 0.5% is a safe, effective alternative . Few non-referring GPs screened for the 3 common STIs, thus putting patients at risk of complications such as pelvic inflammatory disease . Non-referral of patients with genital warts to GUM departments may have costly medical and financial consequences.

Pathol Biol (Paris), 1996 May, 44(5), 341 - 6
{In vitro susceptibility of Pasteurella and related bacteria to five orally administered antibiotics}; Lion C et al.; Tetracyclines and beta-lactam antibiotics are usually recommended for the treatment of pasteurellosis following bite wounds . However other oral antimicrobial agents could be proposed . In vitro susceptibility of 94 Pasteurella strains {P . multocida (79), P . stomatis (11), P . dagmatis (2), P . canis (1), P . "SP" (1)}, 20 group EF-4 strains and 28 Neisseria weaveri strains (formerly group M-5), that are bacteria often isolated after animal-inflicted wounds, was studied towards five antibiotics: clarithromycin, azithromycin, pristinamycin, trimethoprim-sulfamethoxazole and ciprofloxacin . MICs were determined by the agar dilution method using HTM medium (Oxoid), and for pristinamycin using both HTM and Mueller-Hinton agar supplemented with 5% horse blood (BMH) . Most of Pasteurella isolates showed intermediate susceptibility to clarithromycin (63%) and to azithromycin (90.5%) with lower MICs for azithromycin . Fourty-two and thirty-two percent of Pasteurella strains were susceptible to pristinamycin respectively on HTM and on BMH . EF-4 and N . weaveri were more sensitive than Pasteurella to macrolides and to pristinamycin . Trimethoprim-sulfamethoxazole was active against all isolates, with higher MICs for EF-4 and N . weaveri . On all strains tested, the lowest MICs were observed for ciprofloxacin . Trimethoprim-sulfamethoxazole and ciprofloxacin could be proposed as a therapeutic alternative in case of pasteurellosis following animal bites.

J Antimicrob Chemother, 1996 May, 37 Suppl A, 115 - 22
Comparative efficacy and safety of single oral doses of sparfloxacin versus ciprofloxacin in the treatment of acute gonococcal urethritis in men; Moi H et al.; A double-blind, randomised, international multicentre study was conducted to compare the efficacy and safety of sparfloxacin 200 mg versus ciprofloxacin 250 mg as single oral-dose treatment for acute gonorrhoea in men . A total of 238 patients were included and 191 were evaluable for primary efficacy (eradication of Neisseria gonorrhoeae) . At follow-up, 99% (96/97) of the sparfloxacin-treated patients were culture-negative compared with 98% (92/94) in the ciprofloxacin group . Three of the four patients who harboured gonococci at follow-up admitted to having had unprotected sexual intercourse after treatment and were probably reinfections . The rate of post-gonococcal urethritis was 26% in both groups, although Chlamydia trachomatis was isolated in only 4% of patients at inclusion . Both drugs were well tolerated . Single oral doses of sparfloxacin 200 mg and ciprofloxacin 250 mg are equally effective in the treatment of acute gonorrhoea in men.

J Antimicrob Chemother, 1996 May, 37 Suppl A, 19 - 25
Comparative in-vitro activity of sparfloxacin against genital pathogens; Perea EJ et al.; The in-vitro activity of sparfloxacin against four pathogens commonly implicated in genital infections was compared with that of a number of other commonly administered antimicrobials . Sparfloxacin demonstrated excellent activity against Neisseria gonorrhoeae (MIC range of < or = 0.0002-5 mg/L for beta-lactamase producing strains, and < or = 0.0002-0.03 mg/L for non-beta-lactamase producing strains) . This activity was similar to that of lomefloxacin and ciprofloxacin and was greater than that of ofloxacin . Sparfloxacin was more active against Ureaplasma urealyticum (MIC90 1 mg/L) than the other three quinolones (MIC90 4 mg/L) . Sparfloxacin was much more active against Mycoplasma hominis (MIC90 0.06 mg/L) than the other quinolones (MIC90 1 mg/L) . Sparfloxacin showed the most potent inhibitory and bactericidal activity of the quinolones against Chlamydia trachomatis with MIC and MBC of 0.06 mg/L (ofloxacin MIC and MBC 1 mg/L; ciprofloxacin and lomefloxacin MIC and MBC 2 mg/L) . The results of this study and others performed by workers using different methods are consistently similar . Since sparfloxacin has broad activity against pathogens implicated in genital infections it may be a good therapeutic alternative for these syndromes.

Mol Microbiol, 1996 May, 20(3), 507 - 18
Limited variation and maintenance of tight genetic linkage characterize heteroallelic pilE recombination following DNA transformation of Neisseria gonorrhoeae; Hill SA; Genetic recombination impacts on neisserial biology in two ways: (i) specific loci undergo rearrangement at high frequency leading to the formation of many different alleles; and (ii) Neisseria, being naturally competent for DNA transformation, provide a means to disseminate the novel alleles throughout a population . In this study pilE was used as a model system to examine heteroallelic recombination following DNA transformation . When gonococci were transformed with chromosomal donor DNA containing different pilE alleles, the majority of pilE recombinants arose through allelic replacement . Co-conversion analysis across pilE showed that in approximately 85-90% of recombination events encompassing pilE and an adjacent opa locus, linkage was maintained (i.e . approximately 10-15% of recombination events terminated within the approximately 1000 base pair pilE/opaE interval) . In addition to those recombinants that arose through allelic replacement, a large pilus-minus subpopulation was also observed (approximately 10% of all recombinants), indicating that many recombination events did not yield recombinant pilEs that could be assembled into functional pili . PilE mosaics increased following transformation with plasmid donor DNAs carrying pilE with limited flanking-sequence homology, suggesting a potential role for flanking-sequence homologies in mosaic formation . Overall, the data support the view that horizontal transmission of chromosomal DNA between gonococci will favour the spread of intact alleles, as opposed to expanding the allelic repertoire through the formation of gene mosaics.

J Clin Microbiol, 1996 May, 34(5), 1143 - 6
Preservation of Neisseria gonorrhoeae at -20 degrees C; Harbec PS et al.; To explore the feasibility of preserving Neisseria gonorrhoeae at -20 degrees C, we studied its viability quantitatively and qualitatively for 12 and 18 months, respectively, in the following media: a gelatin-based medium used mainly to prepare dried gelatin discs (S . Yamai, Y . Obara, T . Nikkawa, Y Shimoda, and Y . Miyamoto, Br . J . Vener . Dis . 55:90-93, 1979), a simplified version (LSPQ preservation medium), and Trypticase soy broth with 10% (vol/vol) glycerol, a medium commonly used for preservation at -70 degrees C . The latter was studied for 4 months only . Four reference strains and two clinical isolates of N . gonorrhoeae were used . The storage temperature was rigorously preadjusted and monitored at -20 +/- 1 degree C during the entire project . After 12 months of storage, all strains remained viable in both gelatin-based media, whereas a significant loss of viability was observed in Trypticase soy broth-10% glycerol after only 4 months . After 18 months, five strains were still viable in both gelatin-based media and no significant difference was observed between antimicrobial susceptibility results and those of the original strains preserved at -70 degrees C . On the basis of these results, we believe that LSPQ preservation medium represents a good alternative for the storage of N . gonorrhoeae at -20 degrees C for at least a year . Furthermore, it is easy to prepare and use and can by stored at 4 to 8 degrees C for a year prior to use.

Trends Microbiol, 1996 May, 4(5), 185 - 91
Bacterial transferrin and lactoferrin receptors; Gray-Owen SD et al.; Pathogenic members of the Neisseriaceae and Pasteurellaceae express outer-membrane receptor proteins involved in the direct assimilation of iron from the host glycoproteins transferrin and lactoferrin . The critical requirement of iron for growth suggests that this function is an important component of colonization and infection . A model describing this novel process is presented.

Trends Microbiol, 1996 May, 4(5), 179 - 84
Iron acquisition in the pathogenic Neisseria; Genco CA et al.; Pathogenic Neisseria species need to obtain iron from the host to grow in vivo . Several iron-transport systems are known, and regulation of Neisseria iron-transport genes occurs via the transcriptional regulator Fur . There is evidence that the ability to transport iron is crucial to the survival of these organisms in vivo.

Sex Transm Dis, 1996 May-Jun, 23(3), 213 - 8
Gonococcal susceptibility to antimicrobials in Baltimore, 1988-1994 . What was the impact of ciprofloxacin as first-line therapy for gonorrhea?
Zenilman JM.
BACKGROUND: Single-dose, oral quinolones have been a recommended treatment option for gonorrhea since 1989 and have since been used as first-line therapy for gonorrhea in Baltimore . METHODS: The impact of this strategy on antimicobial susceptibility patterns in Neisseria gonorrhoeae was assessed utilizing data collected as part of the National Gonococcal Isolate Surveillance system . This system evaluates a systematically collected sample . Minimum inhibitory concentrations to penicillin, tetracycline, ceftriaxone, and ciprofloxacin were determined by agar dilution . RESULTS: Between January 1988 and September 1994, 1,846 gonococcal isolates were evaluated . The proportion of isolates with plasmid-mediated resistance (penicillinase-producing Neisseria gonorrhoeae or tetracycline-resistant Neisseria gonorrhoeae) increased from 22% in 1988 to 46% in 1992 and then dropped to 20% in 1994 . In contrast, the prevalence of chromosomally mediated resistant isolates ranged between 3% and 10% . Between 1988 and 1994, the geometric mean penicillin and tetracycline minimum inhibitory concentrations decreased slightly (penicillin: from 0.509 microgram/ml to 0.369 microgram/ml; tetracycline: from 1.01 micrograms/ml to 0.767 microgram/ml) . The mean ceftriaxone MIC increased from 0.005 microgram/ml in 1988 to 0.021 microgram/ml in 1992, and then abruptly decreased . Ciprofloxacin minimum inhibitory concentrations did not change substantially during the study period . Concurrent studies performed in this community suggest that quinolones were infrequently used for infections other than sexually transmitted ones during this time period . CONCLUSIONS: Single-dose quinolone therapy does not appear to foster development of resistant gonococcal isolates . However, resistance may develop as a result of complex ecological interactions with the community, underscoring the need for continued surveillance.

Obstet Gynecol, 1996 May, 87(5 Pt 1), 730 - 6
Laboratory evaluation of acute upper genital tract infection; Peipert JF et al.; OBJECTIVE: To evaluate the diagnostic test characteristics of common tests used to diagnose upper genital tract infection . METHODS: Subjects included women who either met the Centers for Disease Control and Prevention's minimal criteria for acute pelvic inflammatory disease or who had other signs of upper genital tract infection (i.e., atypical pelvic pain, abnormal uterine bleeding, or cervicitis) . The subjects were evaluated with a baseline interview, comprehensive laboratory testing, and either an endometrial biopsy or laparoscopy for definitive diagnosis of upper genital tract infection . Patients were considered positive for upper genital tract infection if they had any of the following findings: 1) histologic evidence of acute endometriosis or salpingitis, 2) laparoscopic visualization of purulent exudate in the pelvis without another source, or 3) positive testing for Neisseria gonorrhoeae or Chlamydia trachomatis from the endometrium, fallopian tubes, or pelvis . RESULTS: One hundred twenty women with adequate endometrial samples were evaluated between August 1993 and September 1995 . The median age of the study population was 24 years: 38% were white, 52% were smokers, 81% were insured by Medicaid or were uninsured, and 67% were single . Sensitivities for elevated white blood cell count (WBC), erythrocyte sedimentation rate, C-reactive protein, and increased vaginal white blood cells are 57, 70, 71, and 78%, respectively . If any one test is abnormal, the sensitivity is 100% and specificity is 18% . If all four tests are abnormal, sensitivity is 29% and specificity is 95% . CONCLUSION: Testing for increased vaginal white blood cells was found to be the most sensitive laboratory indicator for upper genital tract infection, whereas serum WBC was the most specific . No one diagnostic laboratory test is pathognomonic for upper genital tract infection . Combinations of positive tests can improve diagnostic specificity and positive predictive value, but with a diminution of sensitivity and negative predictive value . Combinations of negative tests can reliably exclude upper genital tract infection.

J Pediatr, 1996 May, 128(5 Pt 1), 649 - 53
Immunogenicity of heptavalent pneumococcal conjugate vaccine in infants; Anderson EL et al.; OBJECTIVE: To evaluate the safety, immunogenicity, and immunologic memory in young infants of a seven-valent (6B, 14, 19F, 23F, 18C, 4, 9V) pneumococcal vaccine conjugated to the outer membrane protein complex of Neisseria meningitidis . VACCINEES: Healthy 2-month-old infants 12- to 15-month-old control infants were recruited from participating private practices . METHODS: Infants (n = 25) were vaccinated at 2, 4, and 6 months of age with the conjugated pneumococcal vaccine, followed by a single dose of licensed pneumococcal polysaccharide vaccine (n = 20) at 12 to 15 months of age . Thirteen infants who had not received the investigational pneumococcal conjugate vaccine served as control subjects and were given a single dose of the licensed pneumococcal polysaccharide vaccine at 12 to 15 months of age . RESULTS: The investigational pneumococcal conjugate vaccine was well tolerated by infants . The vaccine was highly immunogenic in young infants, with significant increases in antibody to all seven serotypes after either two or three injections . At 12 to 15 months of age, infants who had been primed with the investigational pneumococcal conjugate vaccine had a brisk immunologic response to the booster injection of the licensed pneumococcal polysaccharide vaccine . Control infants, who received a single primary injection of the licensed pneumococcal polysaccharide vaccine, had negligible immunologic responses to four of the seven serotypes and low responses to the other three types . CONCLUSION: The investigational seven-valent pneumococcal conjugate vaccine administered to young infants was well tolerated and highly immunogenic and provided immunologic memory to an injection of the licensed pneumococcal polysaccharide vaccine.

J Infect Dis, 1996 May, 173(5), 1157 - 63
A noncovalent complex vaccine prepared with detoxified Escherichia coli J5 (Rc chemotype) lipopolysaccharide and Neisseria meningitidis Group B outer membrane protein produces protective antibodies against gram-negative bacteremia; Bhattacharjee AK et al.; Earlier studies showed that purified IgG from sera of rabbits immunized with a boiled Escherichia coli J5 (Rc chemotype) whole cell vaccine protected neutropenic rats against gram-negative bacterial sepsis . In the present study, de-O-acylated J5 lipopolysaccharide (J5 DLPS) as a noncovalent complex with Neisseria meningitidis group B outer membrane protein (GBOMP) elicited anti-J5 LPS antibodies in rabbits . IgG prepared from immune rabbit sera protected neutropenic rats against lethal challenge with Pseudomonas aeruginosa 12:4:4 (Fisher Devlin immunotype 6) . Sixteen of 26 rats treated with the postimmune serum IgG were protected compared with none of 20 rats treated with the control rabbit serum IgG (P < .001) . In vitro binding studies showed binding of anti-J5 IgG to several gram-negative bacteria . These results indicate that a subunit vaccine made of J5 DLPS as a noncovalent complex with GBOMP may protect against gram-negative bacteremia.

J Bacteriol, 1996 May, 178(9), 2527 - 32
Site-specific insertion of IS1301 and distribution in Neisseria meningitidis strains; Hilse R et al.; The insertion element IS1301 has been shown to mediate capsule phase variation in Neisseria meningitidis found in N . serogroup B by reversible insertional inactivation of the siaA gene . We have determined the target site specificity of this element by cloning and sequencing the insertion sites of 12 identical IS1301 copies found in N . meningitidis B1940 . A target consensus core of 5'-AYTAG-3' was identified, with the central TA being duplicated following insertion . Additional features around the target sites, including extended palindromic symmetry, stem-loop formation, and the high incidence of AT tracts, indicate that other factors, such as DNA secondary structure, are involved in target recognition . The left inverted repeat of an IS1016-like element acts as a hot spot for insertion, with one insertion element combination located upstream of their gene . According to further sequence analysis, we were able to place IS1301 in the IS5 subgroup within the IS4 family of elements . A survey of 135 Neisseria strains indicated the presence of IS1301 in 27.9 to 33.3% of N . meningitides serogroup B, C, and W135 strains and in 86.7% of serogroup Y strains . IS1301 did not occur in serogroup A strains, in Neisseria gonorrhoeae, or in apathogenic Neisseria spp.

Infect Immun, 1996 May, 64(5), 1621 - 30
Gonococcal opacity protein promotes bacterial entry-associated rearrangements of the epithelial cell actin cytoskeleton; Grassme HU et al.; Neisseria gonorrhoeae enters cultured human mucosal cells following binding of a distinct gonococcal opacity (Opa) outer membrane protein to cell surface proteoglycan receptors . We examined the route of internalization that is activated by Opa-expressing gonococci (strain VP1) . Microscopy of infected Chang epithelial cells showed that gonococcal uptake was insensitive to monodansylcadaverine (150 microM), which interferes with clathrin-mediated endocytosis . Similarly, indirect immunofluorescence staining for clathrin in infected cells showed distribution of cellular clathrin unaltered from the distribution in noninfected cells . The microtubule inhibitors colchicine (50 microM) and nocodazole (20 microM) but not the microtubule-stabilizing agent taxol (10 microM) caused a moderate (30 to 50%) reduction in gonococcal entry without affecting bacterial adherence . The most dramatic effects were obtained with the microfilament-disrupting agent cytochalasin D (3 microM), which totally blocked bacterial entry into the cells . Double immunofluorescence staining of gonococci and actin filaments in infected cells demonstrated bacterium-associated accumulations of F-actin as an early signal of bacterial entry . The recruitment of F-actin was transient and disappeared once the bacteria were inside the cells . Cytochalasin D disrupted the actin cytoskeleton architecture but did not prevent the recruitment of F-actin by the bacteria . Adherent, noninvasive gonococcal Opa variants lacked the ability to mobilize F-actin . Recombinant Escherichia coli expressing the gonococcal invasion-promoting Opa of gonococcal strain MS11 (Opa50) adhered to the epithelial cells in an Opa-dependent fashion but was not internalized and did not recruit detectable amounts of F-actin . Coinfection with the E . coli recombinant strain and gonococci resulted in specific entry of the diplococci, despite the presence of large numbers of adherent E . coli cells . Together, our results indicate that Opa-mediated gonococcal entry into Chang cells resembles phagocytosis rather than macropinocytosis reported for Salmonella spp . and sequentially involves gonococcal adherence to the cell surface, Opa-dependent and cytochalasin-insensitive recruitment of F-actin, and cytochalasin D-sensitive bacterial internalization.

FEMS Microbiol Lett, 1996 Apr 15, 138(1), 55 - 8
A peptidoglycan binding domain in the porin-associated protein (PAP) of Rhodospirillum rubrum FR1; Neumann U et al.; The porin-associated protein of Rhodospirillum rubrum FR1 was found to contain a peptidoglycan binding motif . A partial fragment of 179 amino acids, obtained by cleavage of PAP with trypsin, Asp-N protease, and CNBr, was sequenced . Substantial sequence homology was found of the C-terminal part (residues 126-179) of porin-associated protein with OmpA, the peptidoglycan-associated lipoprotein of several bacteria, protein F of Pseudomonas aeruginosa, and PIII of Neisseria gonorrhoeae, the latter being also a porin-associated protein . The 179 amino acid fragment comprised about 67% of the mass spectrometrically determined total mass of PAP of 27850 Da.

FEMS Microbiol Lett, 1996 Apr 1, 137(2-3), 159 - 64
Sequential action of factors involved in natural competence for transformation of Neisseria gonorrhoeae; Facius D et al.; We previously identified and genetically characterized several factors essential for the natural competence of transformation in Neisseria gonorrhoeae . Here we analyse the sequential action of these factors and dissect the overall transformation process into three distinct steps, (i) the sequence-specific uptake of transforming DNA into a DNase-resistant state, (ii) the transfer of DNA to the cytosol and (iii) the processing and recombination of the incoming with the resident DNA . While two pilus-associated factors, PilE and PilC, were previously implicated in the early DNA uptake event, we show here that three competence factors unrelated to pilus biogenesis, ComA, ComL and Tpc, are not essential for DNA uptake and rather act in a subsequent step . The respective mutants, however, lack the characteristic nucleolytic processing observed with the incoming DNA in both wild-type and non-transformable RecA-deficient N . gonorrhoeae, indicating that they are blocked in the processing and/or the delivery of DNA to the cytoplasm . A hypothetical model proposing a sequential action of the known gonococcal competence factors is presented.

J Adolesc Health, 1996 Apr, 18(4), 270 - 5
Risk for gonococcal and chlamydial cervicitis in adolescent females: incidence and recurrence in a prospective cohort study; Oh MK et al.; PURPOSE: This study attempted to determine the incidence and risk for gonococcal and chlamydial cervicitis among sexually active urban adolescent females . METHODS: The study design is a prospective cohort study . A cohort of 216 sexually active females were followed with repeat sexually transmitted diseases screening for 12-24 months . Subjects positive on any retest (FU) were compared with those who remained negative on all FU . Subjects were interviewed for history and screened for endocervical gonococcal and chlamydial infection . RESULTS: The number of visits per patient ranged from 2 to 9 (median, 3) . The initial Chlamydia trachomatis and Neisseria gonorrhoeae rates were 23.2 and 11.6%, respectively . The cumulative FU positive rates were 20.8% for C . trachomatis and 17.1% for N . gonorrhoeae . Although the initial gonococcal infection was a significant risk for a subsequent infection by C . trachomatis (p = .05) and N . gonorrhoeae (p = .001), the initial C . trachomatis status was not predictive of subsequent infections . The number of partners was not predictive of subsequent infections with either . In the entire study period, 86 patients had at least one episode of C . trachomatis and N . gonorrhoeae infection was confirmed in 52; 20 patients had recurrent cervicitis . During the study, 101 episodes of C . trachomatis and 68 episodes of N . gonorrhoeae infections were identified . Those with recurrent cervicitis (9.3%) were responsible for 33% of all cervicitis episodes identified during the study . CONCLUSIONS: Adolescents in our study were at high risk for cervicitis, particularly as a result of C . trachomatis . Risk for subsequent C . trachomatis cervicitis was the same among initially positive and negative groups . Our data underscore the importance of repeat screening for sexually transmitted infections and treatment of contacts of adolescent females.

Antimicrob Agents Chemother, 1996 Apr, 40(4), 1020 - 3
Quinolone-resistant Neisseria gonorrhoeae: correlation of alterations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV with antimicrobial susceptibility profiles; Deguchi T et al.; Fifty-five clinical strains of Neisseria gonorrhoeae were examined for mutations in the gyrA and parC genes and for antimicrobial susceptibility profiles . The MICs of quinolones for 31 strains with alterations in GyrA were significantly higher than the MICs for 24 strains without such alterations . Eleven strains with alterations in both GyrA and ParC were significantly more resistant to fluoroquinolones than those with alterations in GyrA alone . The MICs of cephalosporins for these strains were also significantly higher than those for other strains.

J Pediatr Adolesc Gynecol, 1996 Apr, 9(2), 79 - 80
Child sexual abuse examination: is there a need for routine screening for N . gonorrhoeae?
Muram D, Speck PM, Dockter M.
PURPOSE: To determine the yield of routine vaginal cultures for Neisseria gonorrhoeae from asymptomatic girls following sexual abuse . METHODS: Cultures were obtained from 865 prepubertal girls seen within 72 hours following an assault . Adult females (2743), age 18 or older, who were treated according to the same protocol constitute the control group . Data were analyzed using chi square analysis and comparison of proportions . RESULTS: It was found that 165 (4.6%) cultures were positive for N . gonorrhoeae . Of the 2743 adults, 153 were positive (5.6%) compared with 12 of the 865 prepubertal girls (1.4%) . The differences are statistically significant . All 12 prepubertal children who had positive vaginal cultures for N . gonorrhoeae revealed that they had signs of acute vulvovaginitis . CONCLUSION: Routine vaginal cultures in asymptomatic prepubertal girls may not be indicated as part of the initial evaluation.

Pathol Biol (Paris), 1996 Apr, 44(4), 249 - 53
{Kinetics of sialylation of Neisseria gonorrhoeae by radiolabelled cytidine 5'monophospho-N-acetyl neuraminic acid}; Goldner M et al.; Neisseria gonorrhoeae (Strain A) induces a sialyl-transferase when treated with a very low concentration of cytidine 5'monophospho-N-acetyl neuraminic acid, 2 x 10(-3) nmol.ml-1, a concentration which is insufficient to produce an adequate resistance to human serum complement . The sialyl-transferase activity was detected by measurement of fixed 14C radio-labeled sialyl groups . Without this stimulation, there was practically no transfer of sialyl groups . The gonococcal sialyl-transferase could be considered inducible.

Int J Pediatr Otorhinolaryngol, 1996 Apr, 35(2), 97 - 105
Microbial flora of the subglottis in intubated pediatric patients; Brown OE et al.; Infection of the intubated subglottis is felt to be one of the many factors involved in the pathogenesis of acquired cicatricial subglottic stenosis . The precise role of infection is unclear and the microbial flora has not been established . An analysis of subglottic culture material, from 22 intubated pediatric patients undergoing tracheotomy, has been performed to establish the nature of the subglottic microbial flora . Fifty-nine isolates were found, including 19 types of organisms . The number of isolates per patient ranged from one to eight, with an average of 2.7 isolates per patient . The most common isolates in the 22 patients were alpha-hemolytic Streptococcus viridans (17 isolates-77%), Neisseria sp . (6 isolates-27%), Pseudomonas sp . (5 isolates-22%), and coagulase negative Staphylococcus sp . (5 isolates-22%) . This data indicates that colonization of the subglottis in intubated pediatric patients is polymicrobial in nature . alpha-Hemolytic Streptococcus viridans and Neisseria sp . were most common, with a shift in cultured flora towards Pseudomonas sp . in patients intubated for more than 10 days . In view of this, antimicrobial therapy may be of benefit in preventing acquired cicatricial subglottic stenosis.

Mol Microbiol, 1996 Apr, 20(2), 241 - 6
The vacuolating cytotoxin of Helicobacter pylori; Cover TL; Helicobacter pylori, the causative agent of chronic superficial gastritis and duodenal ulcer disease in humans, produces a unique cytotoxin (VacA) that induces cytoplasmic vacuolation in eukaryotic cells . The structural organization and processing of the vacuolating cytotoxin are characteristic of a family of proteins exemplified by Neisseria gonorrhoeae IgA protease . Although only 50% of H . pylori isolates produce detectable cytotoxin activity in vitro, vacA homologues are present in virtually all isolates . Several families of vacA alleles have been identified, and there is a strong correlation between presence of specific vacA genotypes, cytotoxin activity, and peptic ulceration . Experiments in a mouse model of H . pylori-induced gastric damage indicate that the cytotoxin plays an important role in inducing gastric epithelial necrosis.

Genitourin Med, 1996 Apr, 72(2), 108 - 10
Comparison of DNA probe (Gen-Probe) with culture for the detection of Neisseria gonorrhoeae in an urban STD programme; Schwebke JR et al.; BACKGROUND AND OBJECTIVES: The DNA probe assay is an alternative to culture for the detection of Neisseria gonorrhoeae which does not depend on the viability of the organism . There have been few published studies comparing the two methodologies . The majority of these studies have shown the probe assay to be comparable to culture . The goal of this study was to compare the performance of the DNA probe assay with culture in a high prevalence setting with nearly optimal culture transport conditions . METHODS: Genital specimens for culture and for DNA probe assay were collected at the time of the routine visit for STD clinic patients . Results of the two methods were compared . Discrepant results were further analysed using the results of direct patient Gram stains . RESULTS: Satisfactory matched specimens were available for 999 patients . The overall sensitivity and specificity of the DNA probe as compared with culture was 90 and 96% . Resolved values achieved by factoring in the results of direct patient Gram stains did not significantly improve the performance of the probe . CONCLUSION: Culture remains the test of choice for the detection of gonorrhoea in a high prevalence setting where culture transport conditions are adequate.

Infect Immun, 1996 Apr, 64(4), 1240 - 5
Susceptibility of Neisseria gonorrhoeae to protegrins; Qu XD et al.; We developed a sensitive and quantitative radial diffusion method to ascertain the susceptibility of six strains of Neisseria gonorrhoeae to antimicrobial peptides derived from mammalian leukocytes . The test organisms included the well-characterized serum-resistant FA19 and serum-sensitive F62 strains plus four antibiotic-resistant clinical isolates . Although each N . gonorrhoeae strain was resistant to human neutrophil defensins, all six were exquisitely sensitive to protegrins, a family of small beta-sheet antimicrobial peptides recently identified in porcine leukocytes . Protegrin-treated N . gonorrhoeae became vacuolated and had striking membrane changes when viewed by transmission and scanning electron microscopy . Because low concentrations of protegrins can also inactivate Chlamydia trachomatis and human immunodeficiency virus, they show promise for development as topical agents to avert sexually transmitted diseases.

Nat Med, 1996 Apr, 2(4), 437 - 42
The maintenance of strain structure in populations of recombining infectious agents; Gupta S et al.; Using mathematical models that combine population genetic and epidemiological processes, we resolve the paradox that many important pathogens appear to persist as discrete strains despite the constant exchange of genetic material . We show that dominant polymorphic determinants (that is, those that elicit the most effective immune responses) will be organized into nonoverlapping combinations as a result of selection by the host immune system, thereby defining a set of discrete independently transmitted strains . By analysing 222 isolates of Neisseria meningitidis, we show that two highly polymorphic epitopes of the outer membrane protein PorA exist in nonoverlapping combinations as predicted by this general framework . The model indicates that dominant polymorphic determinants will be in linkage disequilibrium, despite frequent genetic exchange, even though they may be encoded by several unlinked genes . This suggests that the detection of nonrandom associations between epitope regions can be employed as a novel strategem for identifying dominant polymorphic antigens.

Mol Gen Genet, 1996 Mar 20, 250(5), 558 - 69
Construction of Hermes shuttle vectors: a versatile system useful for genetic complementation of transformable and non-transformable Neisseria mutants; Kupsch EM et al.; A versatile shuttle system has been developed for genetic complementation with cloned genes of transformable and non-transformable Neisseria mutants . By random insertion of a selectable marker into the conjugative Neisseria plasmid ptetM25.2, a site within this plasmid was identified that is compatible with plasmid replication and with conjugative transfer of plasmid . Regions flanking the permissive insertion site of ptetM25.2 were cloned in Escherichia coli and served as a basis for the construction of the Hermes vectors . Hermes vectors are composed of an E . coli replicon that does not support autonomous replication in Neisseria, e.g . ColE1, p15A, or ori(fd), fused with a shuttle consisting of a selectable marker and a multiple cloning site flanked by the integration region of ptetM25.2 . Complementation of a non-transformable Neisseria strain involves a three-step process: (i) insertion of the desired gene into a +Hermes vector; (ii) transformation of Hermes into a Neisseria strain containing ptetM25.2 to create a hybrid ptetM25.2 via gene replacement by the Hermes shuttle cassette; and (iii) conjugative transfer of the hybrid ptetM25.2 into the final Neisseria recipient . Several applications for the genetic manipulation of pathogenic Neisseriae are described.

J Immunol, 1996 Mar 15, 156(6), 2309 - 15
Molecular bases for inherited human complement component C6 deficiency in two unrelated individuals; Nishizaka H et al.; Deficiency of the sixth component of complement (C6D) is frequently associated with recurrent neisserial infections, especially meningitis caused by Neisseria meningitidis . We here report the molecular bases of C6D in two unrelated subjects, one African American (case 1) and the other Japanese (case 2) . Screening all 17 exons of the C6 gene and their boundaries by exon-specific PCR/single strand conformation polymorphism demonstrated aberrant single stranded DNA fragments in exon 12 of case 1 and exon 2 of case 2 . Nucleotide sequencing of the amplified DNA fragments revealed a homozygous single-base deletion (G1936) in exon 12 case 1 and a heterozygous single base deletion (C291/C292/C293/C294) in exon 2 of case 2 . Both mutations resulted in frame shifts and premature termination of the C6 polypeptide . Sequence-specific oligonucleotide probe hybridization and direct sequencing of exon 12 amplified from genomic DNA further supported the homozygosity of the mutation in case 1 . Case 2 is apparently compound heterozygote, but the putative mutation in the other allele of the C6 gene remains unknown . Both case 1 and case 2 were homozygous for the C6A allotype . These data indicate that at least three distinct mutational events can cause C6D, single nucleotide deletions in exons 2 and 12, and a mutation yet unidentified . Thus, similar to other complement protein deficiencies, the pathogenesis of C6D appears to be heterogeneous.

Chin Med Sci J, 1996 Mar, 11(1), 56 - 62
Sexually transmitted diseases: incidence and distribution; Wang Q; The incidence of sexually transmitted diseases (STDs) increased from 26.04 per 100000 in 1987 to 104.81 per 100000 in 1993 in selected areas of the country . Gonorrhea is by far the most common STD but its constituent ratio declined because of a rapid increase of nongonococcal uretheritis and genital warts during most recent years . The incidence of syphilis is relatively low and cases of congenital infection are noted . The wide spread of resistant Neisseria gonorrhoeae infection gives a challenge to the therapeutical and control strategies of STDs . Sexually transmitted Chlamydia trachomatis infections, an important cause of urethritis, cervicitis and pelvic inflammatory disease, is becoming common in our country . Attention has been drawn on viral hepatitis in their means of transmission by sexually behaviors, and also, on the homosexuals, assumed to be the high risk group to catch STDs . Coordinated national efforts to control STDs in China have been taken.

Sex Transm Dis, 1996 Mar-Apr, 23(2), 151 - 6
Etiologic study of genitourinary infections in women of childbearing age in Bobo-Dioulasso, Burkina Faso, 1992; Ledru S et al.; BACKGROUND AND OBJECTIVES: Genitourinary infections have a major impact on public health, especially in Africa . Relative distribution of the different pathogens is unknown in Bobo-Dioulasso . GOAL: To describe the etiology of genitourinary infections, to establish the sensitivity of Neisseria gonorrhoeae to antibiotics, and to provide epidemiologic and biologic evidence to optimize the treatment of genitourinary infections . STUDY DESIGN: Clinical and biologic diagnoses were performed on 223 women with genitourinary infections . RESULTS: Etiologies found were trichomoniasis (27.8%), chlamydia (26.9%), bacterial vaginosis (19.7%), candidiasis (16.6), and N . gonorrhoeae infection (10.9%) . Human immune deficiency virus antibodies were present in 42% of the patients . Spectinomycin or ceftriaxone should be recommended for the treatment of gonorrhoeae in Bobo-Dioulasso . CONCLUSIONS: The prevalence of Chlamydia trachomatis is higher than that of N . gonorrhoeae in Bobo-Dioulasso . This should be taken into account in clinical management of sexually transmitted diseases in this settingPIP: Genitourinary infections have a major impact upon public health, especially in Africa . This paper describes findings from a study conducted to describe the etiology of such infections in Bobo-Dioulasso, to establish the sensitivity of Neisseria gonorrhoeae to antibiotics, and to provide epidemiologic and biologic evidence to optimize the treatment of genitourinary infections . The findings are based upon clinical and biologic diagnoses among 223 women with genitourinary infections . Study found the following etiologies: trichomoniasis in 27.8%, chlamydia in 26.9%, bacterial vaginosis in 19.7%, candidiasis in 16.6%, and Neisseria gonorrhoeae infection in 10.9% . HIV antibodies were present in 42% of patients . The authors recommend spectinomycin or ceftriaxone for the treatment of gonorrhea in Bobo-Dioulasso . Moreover, that the prevalence of Chlamydia trachomatis is higher than that of Neisseria gonorrhoeae should be taken into account when managing STDs in this setting .

Sex Transm Dis, 1996 Mar-Apr, 23(2), 127 - 30
Evaluation of vaginal tampons for collection of specimens for detection of Chlamydial infection; Smith BL et al.; BACKGROUND AND OBJECTIVES: Previous studies have shown that self-inserted vaginal tampons can be used to obtain specimens for detection of Neisseria gonorrhoeae . There is a need to expand testing for infection with Chlamydia trachomatis to women who do not undergo regular gynecologic examinations . GOAL: To compare self-inserted tampons with clinician-obtained endocervical swabs for collection of samples to be examined for chlamydial infection . STUDY DESIGN: Women referred because of recently detected untreated chlamydial infection inserted a vaginal tampon . Two endocervical swabs were obtained after the tampon was removed . One swab was cultured for Chlamydia trachomatis . The other swab and the tampon were examined with the Testpack Chlamydia antigen detection test . RESULTS: Sixty-nine women were studied . Chlamydial infection was detected by culture in 75.4%, by Testpack (swab) in 63.8%, and by Testpack (tampon) in 52.2% . CONCLUSIONS: Tampon-obtained specimens evaluated in the Testpack Chlamydia test were relatively insensitive . More sensitive tests for the detection of Chlamydia trachomatis infection with self-obtained specimens should be evaluated.

Sex Transm Dis, 1996 Mar-Apr, 23(2), 103 - 8
Quinolone-resistant Neisseria gonorrhoeae in Hong Kong; Kam KM et al.; OBJECTIVE: To study the serologic characters and antibiotic susceptibilities of quinolone-resistant Neisseria gonorrhoeae in Hong Kong . STUDY DESIGN: Sixty-nine strains of Neisseria gonorrhoeae isolated from clinical failure cases after treatment with ofloxacin during the period January 1, 1992, to January 1, 1995, were studied . A panel of 14 monoclonal antibodies against protein I classified these strains into 21 serovars . The pattern of serovar distribution against varying minimum inhibitory concentrations of ofloxacin was compared with 143 strains isolated from a cohort of quinolone-susceptible, clinically responsive cases . Antibiotic susceptibilities tests were performed on quinolone-resistant strains to penicillin, tetracycline, ciprofloxacin, spectinomycin, and ceftriaxone . Epidemiologic information on location of contact was collected . RESULTS: Serologic characterization showed that Bop and Bpy were the dominant serovars among quinolone-resistant strains . Most IA and other IB serovars had declined in the selection process for quinolone resistance . Antibiotic susceptibility tests showed that 81.2%, 89.9%, and 78.3% of quinolone-resistant Neisseria gonorrhoeae strains were resistant to penicillin, tetracycline, and both, respectively, whereas 10 of 69 (14.5%) of such strains displayed high-level quinolone resistance (ofloxacin minimum inhibitory concentration > 8 micrograms/ml) . The quinolone-resistant strains remained fully susceptible to spectinomycin and ceftriaxone . CONCLUSIONS: Quinolone-resistant strains have become firmly established in Hong Kong . Serovar determination has documented shifts in the gonococcal population during the selection process for quinolone resistance . Places that use quinolones in the treatment of sexually transmitted diseases should be alert to the emergence of high-level quinolone-resistant Neisseria gonorrhoeae.

J Clin Microbiol, 1996 Mar, 34(3), 756 - 8
Neisseria elongata subsp . elongata, a case of human endocarditis complicated by pseudoaneurysm; Nawaz T et al.; We report an unusual case of endocarditis caused by Neisseria elongata subsp . elongata . The illness was complicated by a ruptured mycotic aneurysm of the right brachial artery, with compression of the brachial plexus . A cure was achieved after aneurysm resection and treatment with intravenous ceftriaxone and gentamicin.

Clin Infect Dis, 1996 Mar, 22(3), 477 - 84
Signs and symptoms of prevalent and incident cases of gonorrhea and genital chlamydial infection among female prostitutes in Kinshasa, Zaire; Alary M et al.; Most studies that have examined the clinical features of gonorrhea and chlamydial infection have been based on prevalent cases (cases of undetermined onset) . In our investigation, we compared signs and symptoms of incident (new) cases of these infections with those observed in prevalent cases (involving the same women) that were diagnosed at enrollment in a prospective study of female prostitutes in Kinshasa, Zaire . Neisseria gonorrhoeae or Chlamydia trachomatis was present at enrollment in 29.2% (225 of 771) of the women in the study . As they were followed during the study, 509 (66.0%) had at least one episode of gonorrhea or chlamydial infection . No symptom was significantly associated with these infections at enrollment or during follow-up . Clinical signs, such as endocervical mucopus (P < .001) and vaginal discharge (P = .001), were associated with both the prevalent and incident cases . However, none of these signs was simultaneously sensitive and specific for detection of these infections . The frequency of clinical signs was significantly reduced after successful treatment (all P values, < .05) . This study shows that a syndromic approach to screening for gonococcal and chlamydial infections in female prostitutes is as problematic for acute incident cases as for prevalent cases . Therefore, there is still an urgent need for simple, cheap, reliable tests that could be used in sexually transmitted disease intervention programs in developing countries.

Mol Microbiol, 1996 Mar, 19(5), 1095 - 105
A novel peptidoglycan-linked lipoprotein (ComL) that functions in natural transformation competence of Neisseria gonorrhoeae; Fussenegger M et al.; A novel peptidoglycan-linked lipoprotein (ComL) has been identified which is required for efficient transformation of Neisseria gonorrhoeae by species-related DNA . Although most mutations in comL appear to be lethal, transposon shuttle mutagenesis was successful in generating a single viable comL mutant of N . gonorrhoeae strain MS11 . This mutant, N457, exhibits a cratered and crinkled colony morphology and grows slower than wild-type MS11 . However, as indicated by electron microscopy, this retardation is due to a small bacterial size rather than to a decreased generation time of the mutant bacteria . Complementation of N457 with an intact comL gene via the Hermes shuttle system fully reconstitutes bacterial size, colony morphology, and transformation competence of the wild-type strain . comL is a single-copy gene and maps downstream of the previously described comA gene . It is transcribed in the opposite direction, probably using the same transcriptional terminator . ComL has a predicted size of 29 kDa and is synthesized in Escherichia coli under the control of its native promoter, which is highly conserved with the E . coli promoter consensus sequence . The 5' end of the coding sequence reveals a lipoprotein secretion signal shown to be functional by gene fusion with alkaline phosphatase (phoA') . In E . coli, cloned ComL can be labelled with {3H}-palmitic acid, thus demonstrating its lipoproteinaceous nature . Palmitoylated ComL appears to be covalently bound to the murein sacculus of E . coli and N . gonorrhoeae since it resists boiling in 4% sodium dodecyl sulphate and is released only by lysozyme treatment . Homologous counterparts of the comL gene are found in Neisseria meningitidis as well as in several nonpathogenic Neisseria species.

Ann Trop Paediatr, 1996 Mar, 16(1), 27 - 32
Ophthalmia neonatorum in the United Arab Emirates; Nsanze H et al.; Ophthalmia neonatorum in small babies is a common problem in neonatal units in Al Ain, but is infrequently associated with sexually transmitted disease agents . A study of 81 babies with ophthalmia neonatorum showed that 81.5% had bacterial or fungal infections . Staphylococcus aureus was the commonest causative organism followed by Escherichia coli and Pseudomonas aeruginosa while Chlamydia trachomatis and Neisseria gonorrhoeae were responsible for less than 5% of all cases . The disease was predominantly mild to moderately severe and easily treated with topical broad-spectrum antibiotic agents which are effective against Gram-positive and Gram-negative bacteria, including hospital-associated strains . The most effective antimicrobial agents were gentamicin topically or ceftriaxone systemicallyPIP: Neonatal conjunctivitis is common in many developing countries, often due to a sexually transmitted disease (STD) such as Chlamydia trachomatis and Neisseria gonorrhoea . The prevention of ophthalmia neonatorum (ON) depends considerably upon knowledge of the type and incidence of the causative organisms, just as the recommended treatment protocols for ON vary according to the local causative microorganisms and their sensitivity to available antimicrobial agents . ON is a common problem in the neonatal units of Al Ain, but infrequently associated with STD agents . A study found 81.5% of 81 babies with ON to have either bacterial or fungal infections . Staphylococcus aureus was the most common causative organism followed by Escherichia coli and Pseudomonas aeruginosa . Chlamydia trachomatis and Neisseria gonorrhoea were responsible for less than 5% of all cases . The disease was mainly mild to moderately severe and easily treated with topical broad-spectrum antibiotic agents effective against Gram-positive and Gram-negative bacteria, including hospital-associated strains . The most effective antimicrobial agents were gentamicin topically and ceftriaxone systemically .

Clin Ther, 1996 Mar-Apr, 18(2), 214 - 27; discussion 213
Recent advances in antibiotic regimens for the treatment of obstetric-gynecologic infections; Zambrano D; This paper reviews new information on antimicrobial agents for the treatment of obstetric-gynecologic infections . The bacteriology of these infections is complex, reflecting the bacteria that usually colonize the vagina and cervix . In general, these infections are polymicrobial in nature: the most frequently isolated microorganisms are gram-negative facultative aerobes, anaerobes, Chlamydia trachomatis, and Neisseria gonorrhoeae . Antibiotic regimens that do provide coverage of these pathogens showed unacceptably high failure rates . A review of the studies recently published confirmed the recommendations of the US Centers for Disease Control and Prevention for severe (inpatients) and mild-to-moderate (outpatients) pelvic inflammatory disease infections . In the case of severe infections, two regimens are recommended: a second-generation cephalosporin like cefoxitin or cefotetan, plus doxycycline or clindamycin/gentamicin . In the case of mild-to-moderate infection, a second- or third-generation cephalosporin plus doxycycline, or oral clindamycin plus an oral quinolone are recommended . Such studies produce high bacteriologic and clinical success rates . New studies indicate that gentamicin may be replaced with a monobactam such as aztreonam; this regimen leads to slightly better efficacy and less toxicity . The possibility of using clindamycin and a quinolone antibiotic is also discussed.

Mol Microbiol, 1996 Mar, 19(6), 1357 - 72
Tetrapac (tpc), a novel genotype of Neisseria gonorrhoeae affecting epithelial cell invasion, natural transformation competence and cell separation; Fussenegger M et al.; We characterized a novel mutant phenotype (tetrapac, tpc) of Neisseria gonorrhoeae (Ngo) associated with a distinctive rough-colony morphology and bacterial growth in clusters of four . This phenotype, suggesting a defect in cell division, was isolated from a mutant library of Ngo MS11 generated with the phoA minitransposon TnMax4 . The tpc mutant shows a 30% reduction in the overall murein hydrolase activity using Escherichia coli murein as substrate . Tetrapacs can be resolved by co-cultivation with wild-type Ngo, indicating that Tpc is a diffusible protein . Interestingly, Tpc is absolutely required for the natural transformation competence of piliated Ngo . Mutants in tpc grow normally, but show a approximately 10-fold reduction in their ability to invade human epithelial cells . The tpc sequence reveals an open reading frame of approximately 1 kb encoding a protein (Tpc) of 37 kDa . The primary gene product exhibits an N-terminal leader sequence typical of lipoproteins, but palmitoylation of Tpc could not be demonstrated . The ribosomal binding site of tpc is immediately downstream of the translational stop codon of the folC gene coding for an enzyme involved in folic acid biosynthesis and one-carbon metabolism . The tpc gene is probably co-transcribed from the folC promoter and a promoter located within the folC gene . The latter promoter sequence shares significant homology with E . coli gearbox consensus promoters . All three mutant phenotypes, i.e . the cell separation defect, the transformation deficiency and the defect in cell invasion can be restored by complementation of the mutant with an intact tpc gene . To some extent the tcp phenotype is reminiscent of iap in Listeria, lytA in Streptococcus pneumoniae and lyt in Bacillus subtilis, all of which are considered to represent murein hydrolase defects.

Sante, 1996 Mar-Apr, 6(2), 115 - 21
{Development and evaluation of screening algorithms for sexually transmitted diseases in pregnant women at Libreville, Gabon}; Bourgeois A et al.; The struggle against sexually transmitted diseases (STD) constitutes a priority of public health in developing countries: STD cause complications, particularly in pregnant women, and facilitate the transmission of HIV . One of the strategies in the struggle against STD is the diagnosis and the early treatment of these infections . The STD, and in particular infections of Neisseria gonorrhea and Chlamydia trachomatis, are difficult to diagnose in women without complementary analyses, which primary health care may not be able to supply . Health care provided to patients could be standardized and improved by considering the signs and symptoms . We studied the prevalence and risk factors of STD among 192 pregnant women consulting the health clinic in Libreville, Gabon, in September 1993 . The prevalence of STD was high (13.5% rate of cervical infection with gonorrhea or Chlamydia trachomatis) . We then evaluated the different diagnostic strategies or algorithms . Regardless of the type of examination (medical interview, simple clinical examination or examination with a speculum), the use of scores integrating risk factors, the clinical signs and symptoms outperformed hierarchical algorithms . This approach was more sensitive and specific and easy to perform . Use of this method may enable more effective screening of STD and also avoid most maternal and perinatal complications.

J Exp Med, 1996 Mar 1, 183(3), 1151 - 9
Neisserial porins induce B lymphocytes to express costimulatory B7-2 molecules and to proliferate; Wetzler LM et al.; The neisserial porins are the major protein components of the outer membrane of the pathogenic Neisseria (N . meningitidis and N . gonorrhoeae) . They have been shown to be able to enhance the immune response to poorly immunogenic substances (e.g., polysaccharides, peptides, glycolipids, etc.) . To explore the basis of their potent adjuvant activity, the effect of the neisserial porins on T-B cell interactions and T cell costimulation was examined . Neisserial porins increased the surface expression of the costimulatory ligand B7-2 (CD86) but did not affect the expression of B7-1 (CD80) . In addition, incubation with the neisserial porins increased the T lymphocyte costimulatory ability of B lymphocytes, which was inhibited by anti-B7-2 but not anti-B7-1 monoclonal antibodies . Upregulation of B7-2 on the surface of B lymphocytes may be the mechanism behind the immunopotentiating activity of neisserial porins.

Infect Immun, 1996 Mar, 64(3), 1039 - 42
Alteration of gonococcal protein expression in acidic culture; Pettit RK et al.; We grew Neisseria gonorrhoeae under acidic, neutral, and alkaline conditions and noted altered expression of at least 12 outer membrane proteins between 31 and 100 kDa in size . One protein whose expression was upregulated under acidic conditions was gonococcal heat shock protein 63 . These proteins may contribute to the pathogenesis of gonorrhea in the urogenital tract.

J Bacteriol, 1996 Mar, 178(5), 1437 - 44
Binding and surface exposure characteristics of the gonococcal transferrin receptor are dependent on both transferrin-binding proteins; Cornelissen CN et al.; Neisseria gonorrhoeae is capable of iron utilization from human transferrin in a receptor-mediated event . Transferrin-binding protein 1 (Tbp1) and Tbp2 have been implicated in transferrin receptor function, but their specific roles in transferrin binding and transferrin iron utilization have not yet been defined . We utilized specific gonococcal mutants lacking Tbp1 or Tbp2 to assess the relative transferrin-binding properties of each protein independently of the other . The apparent affinities of the wild-type transferrin receptor and of Tbp1 and Tbp2 individually were much higher than previously estimated for the gonococcal receptor and similar to the estimates for the mammalian transferrin receptor . The binding parameters of both of the mutants were distinct from those of the parent, which expressed two transferrin-binding sites . Tbp2 discriminated between ferrated transferrin and apotransferrin, while Tbp1 did not . Results of transferrin-binding affinity purification, and protease accessibility experiments were consistent with the hypothesis that Tbp1 and Tbp2 interact in the wild-type strain, although both proteins were capable of binding to transferrin independently when separated in the mutants . The presence of Tbp1 partially protected Tbp2 from trypsin proteolysis, and Tbp2 also protected Tbp1 from trypsin exposure . Addition of transferrin to wild-type but not mutant cells protected Tbp1 from trypsin but increased the trypsin susceptibility of Tbp2 . These observations indicate that Tbp1 and Tbp2 function together in the wild-type strain to evoke binding conformations that are distinct from those expressed by the mutants lacking either protein.

J Infect Dis, 1996 Mar, 173(3), 636 - 46
The pathogenesis of gonococcal urethritis in men: confocal and immunoelectron microscopic analysis of urethral exudates from men infected with Neisseria gonorrhoeae; Apicella MA et al.; Confocal and immunoelectron microscopic analysis of urethral exudates from 12 men with gonococcal urethritis showed that Neisseria gonorrhoeae can invade urethral epithelial cells . Studies with acridine orange stain demonstrated that the majority of organisms within urethral epithelial cells were viable at the time of fixation . Three-dimensional modeling of an infected epithelial cell using image analysis of 21 digitized confocal sections stained with YOYO-1 and DiIC 18(3) revealed that gonococcal invasion of these cells occurred in a polar fashion, most likely at the epithelial luminal surface . Serial immunoelectron micrographs showed evidence of membrane fusion with pedestal formation between the gonococcus and the epithelial cell, gonococci within vacuoles, and occasional gonococci free in the cytoplasm . Immunoelectron microscopy studies showed ruptured vacuoles at the cell surface releasing organisms . These studies demonstrate that urethral epithelial cells are invaded by gonococci during the course of infection in males.

Ann Emerg Med, 1996 Mar, 27(3), 299 - 304
Cost-effectiveness decision analysis of intramuscular ceftriaxone versus oral cefixime in adolescents with gonococcal cervicitis; Friedland LR et al.; STUDY OBJECTIVE: We compared the cost-effectiveness of two single-dose treatment strategies for adolescents with uncomplicated Neisseria gonorrhoeae cervicitis . METHODS: We used a cost-effectiveness decision- analysis model to compare the two methods: the standard, ceftriaxone 125 mg given by IM injection; and an alternative, cefixime 400 mg given orally . The effect of the costs associated with the risk of accidental needlestick during IM administration was also evaluated . Key baseline assumptions (with ranges, when tested) were from the literature or costs to our hospital . These included ceftriaxone, $8.60 per dose; cefixime, $4.67 per dose; ceftriaxone efficacy, 98% (range, 94.9% to 100%); cefixime efficacy, 97% (94.1% to 100%); and a 15% probability of pelvic inflammatory disease (PID) related to failed treatment . We included costs for PID necessitating hospitalization, disseminated gonococcal infection, infertility, and ectopic pregnancy . Assumptions related to accidental needlestick included the rate of needlesticks with the disposable syringe, 6.9 per 100,000 injections (range, 0 to 69); cost of accidental needlestick to hospital; risk of HIV seroconversion after needlestick exposure to HIV-infected blood, .36% (range, 0% to .86%); rate of HIV infection in 15- to 19-year-olds attending sexually transmitted diseases clinics, .4% (range, 0 to 5); and lifetime treatment costs for a person with HIV . RESULTS: At baseline values the model favored ceftriaxone ($45 per patient) or cefixime ($59 per patient) . However, over the range of efficacy of both drugs, two-way sensitivity analysis revealed no consistent cost advantage for either drug . The model was also insensitive to the economic effects associated with the risk of accidental needlestick during IM injection . CONCLUSION: over the range of efficacy by the 95% confidence intervals of both drugs, our analysis demonstrated no clear cost advantage for either . The economic effects of accidental needlestick do not change this conclusion . Compared with the IM alternative, oral cefixime is painless to the patient and simpler for the practitioner to administer . Oral cefixime also eliminates the psychologic effects associated with needlesticks in health care workers . For these reasons, we favor the use of oral cefixime for uncomplicated gonococcal cervicitis in adolescents.

Mol Gen Genet, 1996 Feb 25, 250(3), 277 - 85
Cloning and characterization of the Neisseria gonorrhoeae MS11 folC gene; Fussenegger M et al.; The gene coding for folylpoly-(gamma)-glutamate synthetase (FPGS)-dihydrofolate synthetase (DHFS) of Neisseria gonorrhoeae (Ngo) has been cloned by functional complementation of an Escherichia coli folC mutant (SF4) . The sequence encodes a 224-residue protein of 46.4 kDa . It shows 46% identity to the E . coli FPGS-DHFS and 29% identity to the FPGS of Lacto-bacillus casei . Sequence comparisons between the three genes reveal regions of high homology, including ATP binding sites required for bifunctionality, all of which may be important for FPGS activity . In contrast to L . casei FPGS, the E . coli and Ngo enzymes share some additional regions which may be essential for DHFS activity . The products of Ngo folC and flanking genes were monitored by T7 promoter expression . Interestingly, deletion of the upstream folI gene, which encodes a 16.5 kDa protein, abolishes the capacity of folC to complement E . coli SF4 to the wild-type phenotype . The ability to complement can be restored by folI provided in trans . Unlike folC mutants, gonococcal folI mutants are viable and display no apparent phenotype . Thus, in contrast to E . coli, Ngo folC is expressed at a sufficiently high level from its own promoter, in the absence of FolI This study provides the first insights into the genetic complexity of one-carbon metabolism in Nqo.

BMJ, 1996 Feb 3, 312(7026), 277 - 80
Asymptomatic gonorrhoea and chlamydial infection in rural Tanzanian men; Grosskurth H et al.; OBJECTIVE: To measure the prevalence of urethritis due to Neisseria gonorrhoeae and Chlamydial infection trachomatis in rural Tanzanian men DESIGN: About 500 men aged 15-54 years were selected from each of 12 rural communities by random cluster sampling; interviewed concerning past or present symptoms of sexually transmitted diseases; and asked to provide a first catch urine specimen, which was tested for pyuria with a leucocyte esterase dipstick test . Subjects with symptoms or with a positive result on testing were examined, and urethral swabs were taken for detection of N gonorrhoeae by gram stain and of C trachomatis by antigen detection immunoassay . SETTING: Mwanza region, north western Tanzania . SUBJECTS: 5876 men aged 15-54 years . MAIN OUTCOME MEASURES: Prevalence of urethral symptoms, observed urethral discharge, pyuria, urethritis ( > 4 pus cells per high power field on urethral smear), N gonorrhoeae infection (intracellular gram negative diplococci), and C trachomatis infection (IDEIA antigen detection assay) . RESULTS: 1618 (28%) subjects reported ever having a urethral discharge . Current discharge was reported by 149 (2.5%) and observed on examination in 207 (3.5%) . Gonorrhoea was found in 128 subjects (2.2%) and chlamydial infection in 39 (0.7%) . Only 24 of 158 infected subjects complained of urethral discharge at the time of interview (15%) . CONCLUSION: Infection with N gonorrhoeae and C trachomatis is commonly asymptomatic among men in this rural African population . This has important implications for the design of control programmes for sexually transmitted disease.

Diagn Microbiol Infect Dis, 1996 Feb, 24(2), 87 - 91
Quality control guidelines for amoxicillin, amoxicillin-clavulanate, azithromycin, piperacillin-tazobactam, roxithromycin, ticarcillin, ticarcillin-clavulanate, trovafloxacin (CP 99,219), U-100592, and U-100766 for various National Committee for Clinical Laboratory Standards susceptibility testing methods . Results from multicenter trials; Worth S et al.; Quality control guidelines for standardized antimicrobial susceptibility test methods are critical to the continuing accuracy of the tests . In this report, quality control limits were proposed for 22 organism-antimicrobial combinations with minimum inhibitory concentration (MIC) ranges of three or four log2 dilution steps . Disk diffusion zone diameter ranges were proposed for azithromycin compared with Neisseria gonorrhoeae ATCC 49226 and ticarcillin with and without clavulanic acid tested against Staphylococcus aureus ATCC 25923 . The data from five or six participating laboratories produced > or = 94.7% of results within proposed MIC limits, and 94.3%-99.0% of zones were found within suggested zone guidelines . These proposed quality control ranges should be validated by in-use results from clinical laboratories.

AIDS, 1996 Feb, 10(2), 207 - 12
Reported condom use is not associated with incidence of sexually transmitted diseases in Malawi; Taha TE et al.; OBJECTIVES: To establish frequency of reported condom use and validate reliability of self-reporting among urban women in Malawi . DESIGN: Cross-sectional survey in antenatal women in 1989 and 1993 . Prospective study in cohort first surveyed in 1989 . METHODS: A total of 6561 women in 1989 and 2460 women in 1993 answered questions about condom use and sexual activity, had a physical examination and were screened for HIV . A subset of women from the 1989 screening were administered a questionnaire and tested for syphilis, Neisseria gonorrhoeae and Trichomonas vaginalis infections every 6 months . RESULTS: Although between the two cross-sectional studies intermittent condom use increased from 6 to 15% (P < 0.001) with no difference according to HIV infection, consistent use was reported by less than 1% . In the prospective study, women reported a higher condom use at any visit than either group assessed cross-sectionally . Consistent condom use peaked at 62% in the first 6 months, but declined to as low as 8% in the second year of follow-up . Condom use at each visit, either intermittent or consistent, was higher in HIV-seropositive than HIV-seronegative women . Overall, the incidence of gonorrhea, trichomoniasis and syphilis did not decline in women reporting consistent condom use . CONCLUSIONS: In prospectively followed women reports of consistent condom use was substantially higher than in cross-sectional surveys, but rapidly decreased over time, irrespective of HIV status . The presence of new sexually transmitted diseases suggests that this population of urban women overreports condom use or underreports sexual activity, or both . Intensive and sustained education is needed to achieve consistent condom use . Biologic markers of sexual activity are useful in interpreting reported condom usePIP: To determine the frequency of reported condom use and validate the reliability of self-reporting among urban women in Malawi, 6561 women in 1989 and 2460 women in 1993 answered survey questions about condom use and sexual activity, had a physical examination, and were screened for HIV . A subset of women from the 1989 screening were administered a questionnaire and tested for syphilis, gonorrhea, and Trichomonas vaginalis infections every six months . The study populations consisted of consecutive women presenting for their first antenatal visit to Queen Elizabeth Hospital in Blantyre, Malawi . Intermittent condom use increased from 6% to 15% between the two cross-sectional studies, with no difference according to HIV infection; consistent condom use was reported by less than 1% . In the prospective study, women reported higher condom use at any visit than either group assessed cross-sectionally . Consistent condom use peaked at 62% in the first six months, but declined to as low as 8% during the second year of follow-up . Condom use at each visit, either intermittent or consistent, was higher among HIV-seropositive than HIV-seronegative women . Overall, the incidence of syphilis, gonorrhea, and trichomoniasis did not decline in women reporting consistent condom use . This incidence of new sexually transmitted diseases suggests that the studied population either overreports condom use or underreports sexual activity, or both .

Mol Microbiol, 1996 Feb, 19(4), 841 - 56
Allelic polymorphism and site-specific recombination in the opc locus of Neisseria meningitidis; Seiler A et al.; The opc gene is widespread in epidemic and endemic Neisseria meningitidis, but most strains of certain epidemic clones (ET-37 complex, Cluster A4) and a few random endemic isolates lack an opc gene . Four percent of the 1148 bp that contain opc plus the surrounding intergenic region was polymorphic (18 alleles), and many of the alleles contained a 230 bp insertion at a fixed location in the intergenic region . The presence or absence of the insertion reflects site-specific recombination . The alleles are stably inherited within clonal groupings for up to at least 50 years, with rare cases of horizontal genetic exchange . Most statistical methods indicated significant intragenic recombination events within this dataset.

J Clin Microbiol, 1996 Feb, 34(2), 479 - 81
Evaluation of in vitro spectra of activity of azithromycin, clarithromycin, and erythromycin tested against strains of Neisseria gonorrhoeae by reference agar dilution, disk diffusion, and Etest methods; Mehaffey PC et al.; The macrolide-azilide susceptibility testing (agar dilution, disk diffusion, Etest) criteria for 105 Neisseria gonorrhoeae strains were evaluated . In addition, the potencies of azithromycin, clarithromycin, and erythromycin were studied . The most active macrolide-azilide agent was azithromycin (MIC at which 90% of the isolates are inhibited {MIC90}, 0.5 microgram/ml) compared with clarithromycin (MIC90, 1.5 to 2 micrograms/ml) and erythromycin (MIC90, 2 to 4 micrograms/ml) . The Etest (AB Biodisk, Solna, Sweden) was observed to produce MIC results very similar to those of the reference agar dilution test (GC agar base), with 100% of the results within 1 log2 dilution step of the reference MICs . The disk diffusion test zone diameters for all three drugs correlated at an acceptable level (r = -0.81 to -0.92) with the reference agar dilution MICs . Interpretive criteria for susceptibility were proposed for azithromycin at a MIC of < or = 2 micrograms/ml and a disk diffusion test zone of > or = 25 mm . No category for resistance was proposed because of the paucity of strains for which MICs were > 2 micrograms/ml . These tentative criteria should be further validated by correlations with clinical trial data for gonococcal strains (as they emerge) that have azithromycin MICs above the proposed susceptible category range.

J Clin Microbiol, 1996 Feb, 34(2), 292 - 5
Genetic diversity of Neisseria gonorrhoeae IB-2 and IB-6 isolates revealed by whole-cell repetitive element sequence-based PCR; Poh CL et al.; Phenotypic characterization of 19 Neisseria gonorrhoeae serovar IB-2 and 8 serovar IB-6 isolates by the combined use of auxotypes, serological characterization, and penicillin susceptibility testing indicated intraserovar genetic diversity . In the present study, we applied whole-cell repetitive element sequence-based PCR (rep-PCR) analysis which allows a rapid assessment of the clonal relationships of IB-2 and IB-6 isolates . DNA templates were prepared by boiling cells harvested directly from plate cultures, eliminating the need for time-consuming phenol extraction . Six different rep-PCR profiles were established among the 19 IB-2 isolates . Rep-PCR typing results had a good correlation with pulsed-field gel electrophoresis patterns . It is slightly less discriminatory than BglII-generated macrorestriction pattern analysis by pulsed-field gel electrophoresis . It is capable of discriminating epidemiologically related from epidemiologically unrelated IB-2 isolates . It should serve as a rapid and useful subtyping tool for epidemiologic investigations in which there is a predominance of major serovar groups.

J Antimicrob Chemother, 1996 Feb, 37(2), 223 - 32
A Tn1545-like transposon carries the tet(M) gene in tetracycline resistant strains of Bacteroides ureolyticus as well as Ureaplasma urealyticum but not Neisseria gonorrhoeae; de Barbeyrac B et al.; The presence of the tet(M) determinant and conjugative transposons related to Tn1545 in urogenital pathogens, Bacteroides ureolyticus (10 strains), Neisseria gonorrhoeae (37 strains) and Ureaplasma urealyticum (81 strains), was studied by PCR analysis and hybridization assay . All tetracycline-resistant strains that hybridized with a probe specific for tet(M) gave the expected fragment when their DNA's were subject to PCR . The tet(M) gene and int-Tn, the gene encoding the protein required for the movement of Tn1545-like transposons, were always found together in tetracycline resistant strains of B . ureolyticus and U . urealyticum . In contrast, int-Tn was not detected in tet(M)-mediated tetracycline-resistant N . gonorrhoeae strains . These data suggest carriage of tet(M) by a Tn1545-like transposon in some tetracycline-resistant B . ureolyticus.

Epidemiol Infect, 1996 Feb, 116(1), 15 - 20
Characteristics of gonococci isolated from men with urethritis in Dubai; Al-Hattawi K et al.; Neisseria gonorrhoeae were collected from men attending out-patient clinics in Dubai . The susceptibility to a range of therapeutic antibiotics and their auxotype and serotype was determined . The plasmid content of all penicillinase-producing strains was also analysed . Thirty-six strains of N . gonorrhoeae were isolated from specimens collected from 79 patients over a 24-day period . Of the 36 isolates, 9(25%) were penicillinase-producing N . gonorrhoeae (PPNG) and 15 (42%) were chromosomally resistant N . gonorrhoeae (CMRNG) . CMRNG exhibited higher levels of resistance to cefuroxime, chloramphenicol, tetracycline and erythromycin than PPNG . All isolates were susceptible to ceftriaxone and spectinomycin . Three (8%) isolates showed reduced susceptibility (MIC, > or = 0.25 mg/l) to ciprofloxacin . Six isolates of PPNG carried the 4.4 MD and three the 3.2 MD penicillinase encoding plasmid . The total gonococcal population was phenotypically diverse, with 12 serovars, 6 auxotypes and 21 A/S classes . Gonorrhoea was found to be a major cause of urethritis in Dubai and the strains exhibited high levels of resistance to penicillin.

Am J Obstet Gynecol, 1996 Feb, 174(2), 659 - 66
Transferrin increases adherence of iron-deprived Neisseria gonorrhoeae to human endometrial cells; Heine RP et al.; OBJECTIVE: Our purpose was to study the effects of iron deprivation with and without human transferrin supplementation on the adherence and invasion of Neisseria gonorrhoeae to human endometrial cells . STUDY DESIGN: N . gonorrhoeae grown with our without iron was placed in media alone or media containing 2.5 mg/ml saturated human transferrin or unsaturated transferrin . N . gonorrhoeae was inoculated onto polarized human endometrial carcinoma cell (HEC 1-B) monolayers, and at various intervals monolayers were washed and incubated with media containing gentamicin or media alone . Colony-forming units per milliliter of N . gonorrhoeae associated with HEC 1-B cells were then determined . N . gonorrhoeae strains tested included both a transferrin receptor-positive (wild-type) and a transferrin receptor-negative mutant . Differences in percent of original inoculum remaining at varying time points were analyzed by the Mann-Whitney U test . Transmission electron microscopy using a primary endometrial cell line was used to verify findings . RESULTS: Iron-negative N . gonorrhoeae exhibited less adherence than did iron-positive N . gonorrhoeae . No difference in HEC 1-B adherence was seen when either saturated transferrin or unsaturated transferrin was added to the iron-positive N . gonorrhoeae . With iron-negative N . gonorrhoeae addition of either saturated transferrin or unsaturated transferrin significantly increased N . gonorrhoeae adherence although unsaturated transferrin did not permit growth of iron-negative N . gonorrhoeae in tissue culture media alone . Transmission electron microscopy confirmed increased adherence of iron-negative N . gonorrhoeae supplemented with unsaturated transferrin . An iron-negative N . gonorrhoeae mutant lacking the transferrin receptor exhibited no adherence regardless of addition of saturated transferrin or unsaturated transferrin . Invasion could not be quantitated reliably because of persistence of gentamicin effect . CONCLUSION: Iron and transferrin increased attachment of N . gonorrhoeae to human endometrial cells.

Infect Immun, 1996 Feb, 64(2), 644 - 52
Assembly and antigenicity of the Neisseria gonorrhoeae pilus mapped with antibodies; Forest KT et al.; The relationship between the sequence of Neisseria gonorrhoeae pilin and its quaternary assembly into pilus fibers was studied with a set of site-directed antibody probes and by mapping the specificities of antipilus antisera with peptides . Buried and exposed peptides in assembled pili were identified by competitive immunoassays and immunoelectron microscopy with polyclonal antibodies raised against 11 peptides spanning the pilin sequence . Pili did not compete significantly with pilin subunits for binding to antibodies against residues 13 to 31 (13-31) and 18-36 . Pilus fibers competed well with pilin protein subunits for binding to antibodies raised against peptides 37-56, 58-78, 110-120, 115-127, 122-139, and 140-159 and competed weakly for antibodies against residues 79-93 and 94-108 . Antibodies to sequence-conserved residues 37-56 and to semiconserved residues 94-108 preferentially bound pilus ends as shown by immunoelectron microscopy . The exposure of pilus regions to the immune system was tested by peptide mapping of antiserum specificities against sets of overlapping peptides representing all possible hexameric or octameric peptides from the N . gonorrhoeae MS11 pilin sequence . The immunogenicity of exposed peptides incorporating semiconserved residues 49-56 and 121-126 was revealed by strong, consistent antigenic reactivity to these regions measured in antipilus sera from rabbits, mice, and human and in sera from human volunteers with gonorrhea . The conservation and variation of antigenic responses among these three species clarify the relevance of immunological studies of other species to the human immune response against pathogens . Overall, our results explain the extreme conservation of the entire N-terminal one-third of the pilin protein by its dominant role in pilus assembly: hydrophobic residues 1-36 are implicated in buried lateral contacts, and polar residues 37-56 are implicated in longitudinal contacts within the pilus fiber.

FEMS Microbiol Lett, 1996 Jan 15, 135(2-3), 231 - 6
Characterization of conjugative transposon Tn5251 of Streptococcus pneumoniae; Provvedi R et al.; Tn5251 belongs to the Tn916-Tn1545 family of conjugative transposons (CT) and was found integrated into CT Tn5252, to form the composite element Tn5253 of Streptococcus pneumoniae . We show that Tn5251 is identical in structure and size to Tn916 . DNA sequence analysis of a 4,419-bp segment containing the tet(M) gene showed that only 73 nucleotides out of 4,419 were different in the two CT . Essentially all differences (66/73) were clustered in a 688-bp segment of tet(M), which was 90% identical to Tn916 and 100% identical to the tet(M) genes of Tn1545 from S . pneumoniae and pOZ101 from Neisseria gonorrhoeae . DNA sequence analysis of the Tn5251/Tn5252 junction fragments allowed us (i) to determine Tn5251 termini, (ii) to define the 6-bp coupling sequences flanking the CT, and (iii) to infer the structure of the integration site (attB) of Tn5251 into Tn5252 . Conjugal transfer of Tn5251 independent from Tn5253 could not be detected, even if we could show excision and formation of Tn5251 circular intermediates at a level of 5.4 copies per 10(6) chromosomes.

J Biol Chem, 1996 Jan 12, 271(2), 1166 - 73
Production and characterization of chimeric transferrins for the determination of the binding domains for bacterial transferrin receptors; Retzer MD et al.; Pathogenic bacteria in the Neisseriaceae and Pasteurellaceae possess outer membrane proteins that specifically bind transferrin from the host as the first step in the iron acquisition process . As a logical progression from prior studies of the ligand-receptor interaction using biochemical approaches, we have initiated an approach involving the production of recombinant chimeric transferrins to further identify the regions of transferrin involved in receptor binding . In order to prepare bovine/human hybrids, the bovine transferrin gene was cloned, sequenced, and compared with the existing human transferrin gene sequence . After identification of potential splice sites, hybrid transferrin genes were constructed using the polymerase chain reaction-based approach of splicing by overlap extension . Five hybrid genes containing sequences from both bovine and human transferrin were constructed . Recombinant transferrins were produced in a baculovirus expression vector system and affinity-purified using concanavalin A-Sepharose . The recombinant proteins were analyzed for reactivity against polyclonal and monoclonal antibodies and assessed for binding to Neisseria meningitidis transferrin receptor proteins in solid-phase binding assays and affinity isolation experiments . These experiments enabled us to localize the regions of human transferrin predominantly involved in binding to the N . meningitidis receptor to amino acid residues 346-588 . The construction of these chimeras provides unique tools for the investigation of transferrin binding to receptors from both human and bovine bacterial pathogens.

Arch Med Res, 1996 Winter, 27(4), 459 - 63
Flow cytometry evaluation of complement mediated bacterial membrane damage; Alvarado-Aleman FJ; The study of the complement-outer bacterial membrane interactions is gaining enthusiasm for applying a continuous development of more specialized techniques . In this paper a novel flow cytometry technique shows that highly fluorescent lucifer yellow-stained Neisseriae gonorrhoeae when exposed to the redox reaction of p-nitro blue chloride tetrazolium (NBT), as formazan precipitation takes place, a shift to lesser fluorescent channels of the histogram population occurs . That effect is labeled NBT-laser beam quenching (NBT-LBQ) . A significant difference by Kolmogorov-Smirnov summation curve analysis is found between complement heat-inactivated and its counterpart assessed with normal complement microorganisms . The operation of this NBT-LBQ effect by microbial flow cytometry casts an interesting potential for the evaluation of the outer membrane-complement-interaction of serum sensitive microorganisms.

Mol Microbiol, 1996 Jan, 19(2), 249 - 59
A physical and genetic map of Neisseria meningitidis B1940; Gaher M et al.; A physical map of the chromosome of Neisseria meningitidis B1940 has been constructed by one- and two-dimensional pulsed-field gel electrophoresis techniques . Complete macrorestriction maps for the enzymes Nhel (16 sites), Sgfl (13 sites), Sfil (11 sites) and l-Ceul (4 sites), as well as a partial restriction map for the restriction enzyme Spel (15 of c . 28 sites) could be established . Altogether, 59 restriction sites were mapped on a single circular chromosome of 2.3 Mbp . By restriction endonuclease digestion and Southern hybridization of cloned genetic markers, 39 genetic loci were assigned to this map . Comparison with the metabolic maps of Neisseria gonorrhoeae MS11-N198 and FA1090 revealed a high degree of conservation in the arrangement of gene loci among these two species, although four out of 24 genetic loci are located at different chromosomal positions, indicating several genomic rearrangements.

Mol Microbiol, 1996 Jan, 19(1), 159 - 69
Molecular characterization of hybrid Tbp2 proteins from Neisseria meningitidis; Legrain M et al.; Transferrin-binding protein 2 (Tbp2) from Neisseria is an outer membrane-associated extracellular lipoprotein that is involved in iron capture within the infected host . The analysis of tbp2 clones isolated from various bacterial strains revealed extensive divergences throughout the open reading frame (ORF), with predicted amino acid (aa) sequences displaying 47% to 83% identity . Such a variability is likely to have resulted from the selective pressure exerted by the host immune system, but raises questions regarding the existing constraints for conservation of protein function . Indeed, the neisserial Tbp2s include a large structured domain, extending throughout the N-terminal half of the protein (approximately 270-290 aa), which is extremely stable and whose conformational integrity is required for efficient binding to human transferrin (hTf) . In this work, a functional study of Tbp2s encoded by hybrid genes constructed by reassorting highly divergent tbp2 sequences in the region of the ORF encoding this structured domain was performed . The data demonstrate that the determinant intramolecular interactions allowing formation of a stable Tbp2 structure able to interact efficiently with hTf or/and that the Tbp2 residues involved in the interaction with hTf are not well conserved . However, a number of rearrangements appeared to generate genes encoding proteins which have retained structural stability and hTf-binding capacity . This suggested that despite the extreme aa sequence divergence and the conformational constraints, horizontal genetic exchanges, which are known to occur in neisserial populations, may have contributed significantly to the generation of sequence variation within tbp2 ORFs . The analysis of two tbp2 clones characterized in this work supports this hypothesis.

Zh Mikrobiol Epidemiol Immunobiol, 1996 Jan-Feb, (1), 49 - 53
{The immune status of patients with acute inflammatory diseases of adnexa uteri associated with different combinations of microorganisms}; Antonova LV et al.; The immune status of 130 patients with acute inflammatory diseases of uterine appendages was studied . As etiologically associated infective agents detected in these patients were opportunistic microorganisms (group 1), Neisseria gonorrhoeae in combination with opportunistic microorganisms (group 2) and Chlamydia trachomatis in combination with opportunistic microorganisms (group 3) . In all three groups of patients the response of T lymphocytes to PHA and hyperactivation of the B-cell element of immunity (an increase in the relative number of B-lymphocytes and the content of IgA and IgM in the serum) was found to be suppressed . In addition, in groups 1 and 3 an increase, and in group 2 a decrease in IgG were detected . In the mucus of the cervical canal in the patients of groups 1, 2 and 3 IgM was detected, while in patients of groups 2 and 3 a decrease in sigA was established.

J Clin Lab Anal, 1996, 10(4), 220 - 8
Polyclonal B-cell activation by Neisseria meningitidis capsular polysaccharides elicit antibodies protective against Trypanosoma cruzi infection in vitro; Oliveira TG et al.; A hyperimmune rabbit antiserum against group C Neisseria meningitidis agglutinated and lysed Trypanosoma cruzi metacyclic trypomastigotes in a complement-mediated reaction . Immunization of rabbits with the purified polysaccharide C from N . meningitidis and of human volunteers with the AC-polysaccharide vaccine against meningitis also resulted in antibody production cross-reactive with T . cruzi infective forms . The rabbit antibodies bound to parasites, lysed metacyclic forms, and recognized several components from lysates of cell-derived trypomastigotes . The sera from six human volunteers reacted with cell-cultured trypomastigotes in vitro, lysed these forms, and recognized glycoconjugates migrating diffusely on the top of immunoblots . One serum also reacted with the isolated mucin-like glycoconjugate carrying the Ssp-3 epitope from cell-derived trypomastigotes, but treatment with sialidase did not abolish this reactivity . The anti-AC human antiserum also protected against HeLa cell infection and markedly decreased the number of parasites liberated after cell burst . The polyclonal response that resulted from human immunization with N . meningitidis polysaccharides A and C comprised trypanolytic antibodies that recognized nonsialylated epitopes expressed on infective forms of the parasite . It is suggested that human AC vaccination could be potentially helpful as an adjuvant to a specific immunotherapy of Chagas disease, developed with native or recombinant antigens of the parasite.

Minerva Ginecol, 1996 Jan-Feb, 48(1-2), 19 - 23
{Acute pelvic inflammatory disease: comparison of therapeutic protocols}; Balbi G et al.; Acute pelvic inflammatory disease is a serious medical and economic consequence of sexually transmitted diseases among young women . The aim of the study is to compare the efficacy and safety of gentamycin plus clindamycin with that of ceftazidime plus doxycycline in the treatment of hospitalized patients with acute pelvic inflammatory disease . A total of 78 patients with acute PID, hospitalized in II Obstretic and Gynecologic Clinic of II University of Naples (Italy), entered and randomized into two treatment groups: gentamycin plus clindamycin (N = 40) and ceftazidime plus doxycycline (N = 36) . Patients were excluded if they were pregnant or were not over the age of 16 years of had a history of allergy to one of the drugs used in the Study of had hepatic disease or kidney trouble or had IUD . Acute PID was diagnosed by the following criteria: 1) lower abdominal pain; 2) cervical motion tenderness; 3) adnexal tenderness (all three should be present); plus at least one of the following additional criteria: a) temperature over 38 degrees C; b) leukocytosis (greater than 10.500 mm3); c) purulent material from the peritoneal cavity bt culdocentesis; d) inflammatory mass present on binomial pelvic examination and/or sonography; e) erythrocyte sedimentation rate > 15 mm/hr . Patients were enrolled into the study after obtaining informed consent, pretreatment and posttreatment cultures were obtained from the endocervix from Neisseria gonorrhoeae and Chlamydia trachomatis and aerobic-anaerobic bacteria . The study has shown that the acute PID has a polymicrobal origins . Both antibiotic regimens were very effective in the treatment of the PID: a complete recovery was obtained in over 90% of patients.

J Clin Microbiol, 1996 Jan, 34(1), 177 - 8
Use of gen-probe probe competition assay as a supplement to probes for direct detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urogenital specimens; Woods GL et al.; The potential for development of a cost-effective protocol for selective use of the Gen-Probe probe competition assay (PCA) in conjunction with PACE 2 for direct detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urogenital specimens that would not compromise patient care was investigated . To accomplish this, PCA data from testing performed over 12 months were retrospectively reviewed . Of 237 samples that were presumptively positive for C . trachomatis by initial probe assay and could be tested by PCA, positive PCA results were obtained for 100, 79, and 59%, respectively, of specimens that gave a signal of more than 1,500, 1,000 to 1,500, and less than 1,000 relative light units (RLU) . For the 141 specimens that were presumptively positive for N . gonorrhoeae and could be tested by PCA, positive PCA results were obtained for 99, 80, and 42%, respectively, of samples with a signal of more than 1,500, 1,000 to 1,500, and less than 1,000 RLU . These data indicate that PCA should be a routine supplement to Gen-Probe PACE 2 for specimens with an initial signal by probe assay of less than 1,500 RLU and may not be necessary for samples yielding a signal of more than 1,500 RLU.

Pathology, 1996 Jan, 28(1), 84 - 7
The frequency of co-infection with Neisseria gonorrhoeae and Chlamydia trachomatis in men and women in eastern Sydney; Tapsall JW et al.; The frequency of simultaneous infection with Neisseria gonorrhoeae and Chlamydia trachomatis in men and women in Eastern Sydney was examined prospectively over a 21 month period in patients from whom appropriate specimens were obtained . In men gonococcal disease (113 cases) and chlamydial infections (124 cases) occurred in approximately equal frequencies, but there were only 4 instances (3.5%) where C . trachomatis was detected at the same time as N . gonorrhoeae . Demographic data and phenotypic profiles of the gonococci isolated suggested that much of the gonorrhea occurred in homosexual men who are generally considered to have a low incidence of chlamydial disease . Gonococcal disease (34 cases) was far less common than chlamydial infection (112 cases) in women, but co-infection with chlamydia was noted in 6 (17.6%) of the cases of gonorrhea (p < 0.01) . The current recommendation for the routine addition of tetracyclines to gonococcal treatment regimens for men requires revision.

J Antimicrob Chemother, 1996 Jan, 37(1), 139 - 43
Tentative interpretive criteria and quality control parameters for in-vitro susceptibility testing of Neisseria gonorrhoeae to two fluoroquinolones (PD 131628 and grepafloxacin (OPC 17116)); Sewell DL et al.; The susceptibility of Neisseria gonorrhoeae to PD 131628 and grepafloxacin (OPC 17116) was evaluated by agar dilution and disc diffusion methods . A tentative susceptibility category for both fluoroquinolones included strains for which the MICs are < or = 0.06 mg/L and the zones of inhibition are > or = 38 mm for PD 131628 and > or = 37 mm for grepafloxacin . Quality control studies with N . gonorrhoeae ATCC 49226 suggested that agar dilution MIC limits were 0.002-0.008 mg/L and 0.004-0.03 mg/L for PD 131628 and grepafloxacin, respectively . The zone size limits were 50-58 mm for PD 131628 and 44-52 mm for grepafloxacin.

J Exp Med, 1996 Jan 1, 183(1), 323 - 7
Variation of gonococcal lipooligosaccharide structure is due to alterations in poly-G tracts in lgt genes encoding glycosyl transferases; Yang QL et al.; The lipooligosaccharide (LOS) expressed by gonococci spontaneously varies its structure at high frequency, but the underlying genetic mechanism has not been described . We have previously reported that the genes encoding the glycosyl transferases responsible for the biosynthesis of the variable alpha chain of the LOS of Neisseria gonorrhoeae are located in a locus containing five genes, lgtA, lgtB, lgtC, lgtD, and lgtE . Sequence analysis showed that lgtA, lgtC, and lgtD contained poly-G tracts within the coding frames, leading to the hypothesis that shifts in the number of guanosine residues in the poly-G tracts might be responsible for the high frequency variation in structure of gonococcal LOS . We now provide experimental evidence confirming this hypothesis.

Chest, 1996 Jan, 109(1), 280 - 2
Nonfebrile mitral valve endocarditis due to Neisseria subflava; Amsel BJ et al.; Native valve endocarditis normally presents with fever and only later in its course demonstrates dysfunction of the affected valve . We describe a case of endocarditis due to Neisseria subflava, a Gram-negative diplococcal saprophyte of the oral cavity, which was unsuspected clinically and found unexpectedly during a mitral valve operation performed for symptomatic prolapse with regurgitation.

J Infect Dis, 1996 Jan, 173(1), 77 - 82
Factors associated with cytomegalovirus infection among human immunodeficiency virus type 1-seronegative and -seropositive women from an urban minority community; Clarke LM et al.; Cytomegalovirus (CMV) seroprevalence and genital tract shedding in human immunodeficiency virus (HIV)-seronegative and HIV-seropositive women from an urban minority community were investigated . CMV seropositivity was high in both groups: 181 (95.2%) of 190 HIV-negative and 158 (90.3%) of 175 HIV-positive subjects . Cervicovaginal shedding was detected in 8 (4.4%) CMV-positive HIV-negative subjects and 31 (19.6%) HIV-positive subjects (odds ratio {OR}, 5.28; P < .001) . Multiple logistic regression analysis revealed that CMV shedding was independently associated with younger age (OR = 0.90; P < .001) and concurrent Chlamydia trachomatis or Neisseria gonorrhoeae infection (OR = 3.60; P = .08) . However, shedding was observed over a broad age range in HIV-positive subjects, with 54.8% of shedders being > or = 30 years old . Among HIV-positive subjects, CMV shedding was also associated with decreased CD4 cell counts (P = .04) and, compared with HIV-negative subjects, was significantly higher (P < .001) among subjects with CD4 cell counts < 500 x 10(6)/L (26.5% in subjects with counts < or = 200 and 22.1% in subjects with counts of 201-499 x 10(6)/L).

J Biol Chem, 1995 Dec 29, 270(52), 30874 - 80
Extracellular transport of VirG protein in Shigella; Suzuki T et al.; The ability of Shigella to spread within and between epithelial cells is a prerequisite for causing bacillary dysentery and requires the function encoded by the virG gene on the large plasmid . The outer membrane VirG (IcsA) protein is essential for bacterial spreading by eliciting polar deposition of filamentous actin (F-actin) in the cytoplasm of epithelial cells . Recent studies have indicated that an N-terminal 80-kDa VirG portion is exposed on the bacterial cell surface and released into the external medium, while the following 37-kDa C-terminal portion is embedded in the outer membrane, although little is known about the extracellular transport of the VirG protein . In this study, we attempted to elucidate the export pathway of VirG protein across the outer membrane and found that the C-terminal 37-kDa portion, termed VirG beta-core, serves as the self-transporter for the secretion of the preceding 80-kDa portion from the periplasmic side of the outer membrane to the external side . Indeed, foreign polypeptides such as MalE or PhoA covalently linked to the N terminus of VirG beta-core were transported to the external side of the outer membrane, and it was further shown that the folding structure of the passenger polypeptide at the periplasmic side of the outer membrane interferes with its translocation . Analysis of the secondary structure of VirG beta-core predicted that the critical structural property was a beta-barrel channel consisting of amphipathic anti-parallel transmembrane beta-strands, interspersed by hairpin turns and loops . These results thus strongly suggest that the secretion of VirG protein from Shigella is similar to the export system utilized by the IgA protease of Neisseria.

Ultrasound Obstet Gynecol, 1995 Dec, 6(6), 430 - 4
Transvaginal sonography in suspected pelvic inflammatory disease; Taipale P et al.; Transvaginal sonography was used in the evaluation of clinically suspected pelvic inflammatory disease (PID) . A total of 86 patients with PID were examined by transvaginal sonography within 1-3 days, 14 days and 90 days after hospitalization . The diagnosis of PID was confirmed in 37% of patients by laparoscopy, laparotomy or positive cervical culture of Chlamydia trachomatis or Neisseria gonorrhoea . In the remainder, the diagnosis was based on tenderness of the uterus, fever and lower abdominal pain that responded rapidly to antibiotics . At the time of admission, 31% of patients had a thick-walled (> or = 5 mm) echogenic tube raising the suspicion of pyosalpinx or pyo-ovary . During the following 3 months, 52% of patients developed tubes that were thin-walled (< 5 mm) and poorly echogenic, and hydrosalpinx was suspected . At the first transvaginal sonographic examination, 64% showed no adnexal mass, but 16% of these developed suspected hydrosalpinx during the following 3 months . Of the total of 86 patients, 6% exhibited a hydrosalpinx for the first time after 3 months . C-reactive protein concentrations and sedimentation rate values correlated positively with the ultrasonically determined volumes of pyosalpinx/pyoovaries, cul-de-sac fluid and ovaries . Transvaginal sonography appears to be very useful in the diagnosis of suspected PID and is recommended at least 3 months after symptoms have subsided for those patients with an adnexal mass at the time of the acute episode and for infertile patients.

Minerva Pediatr, 1995 Dec, 47(12), 545 - 7
{Pericarditis caused by Gemella morbillorum . Description of a case}; Condoluci C et al.; Pericardial diseases, in the pediatric age group are rare, and are usually due to viral infection or collagen diseases . Since the advent of the antibiotic era, bacterial pericardial has rarely been reported, and the bacteria most commonly involved are Staphylococcus aureus, Streptococcus pneumoniae, Haemophylus influenzae and Neisseria meningitidis; other pyogenic agents have rarely been isolated in pericarditis . Gemella morbillorum, also known as Streptococcus morbillorum, is a Gram-positive pyogen, usually a saprophite agent of the human gastrointestinal tract, rarely associated with human infections such as arthritis, endocarditis and meningitidis; to our knowledge, it has never been isolated in pericarditis . We report a case of pyogenic pericarditis in an 11-year-old boy, suffering from substernal chest pain for about one year, in which the clinical symptoms, the presence of acute phase proteins, the large amount of fluid within the pericardial space (echocardiographically estimated to be approximately 18 mm), the lack of improvement in clinical conditions and in laboratory values after therapy (diuretics, broad spectrum antibiotics and steroids) led to the indication for a pericardial drainage with the isolation of Gemella morbillorum . Antimicrobial therapy was then modified according to in vitro susceptibility of the Gemella morbillorum, with a decisive contribution to the recovery of the patient, although long-term steroid therapy (5 months) was necessary because of one recurrence of pericardial effusion . This case report shows how Gemella morbillorum, usually a saprophit microorganism, can become pathogenic and also underlines the importance of a correct etiologic diagnosis of pericarditis resistent to classical antibiotic therapy.

West Indian Med J, 1995 Dec, 44(4), 130 - 2
Antibiotic sensitivity of Neisseria gonorrhoeae isolates in Barbados: longitudinal surveillance 1990-1994; Levett PN; The antimicrobial susceptibility of 775 isolates of Neisseria gonorrhoeae, recovered in Barbados over a five-year period, was studied by disc diffusion . Sensitivity to penicillin declined from 56% in 1990 to 38% in 1994 . The prevalence of beta-lactamase production rose from 30% to 50% during this period and sensitivity to tetracycline declined from 96% to 46%, while co-trimoxazole and spectinomycin sensitivities varied . Ceftriaxone, norfloxacin and erythromycin were the only compounds tested to which sensitivity remained uniform . The results confirm the current recommendations by several agencies that ceftriaxone or an oral fluoroquinolone should be considered the drug of choice for the empirical treatment of gonorrhoea.

Mol Biotechnol, 1995 Dec, 4(3), 315 - 7
Direct transformation of Neisseria gonorrhoeae by gel-isolated DNA; Pritchard KH et al.; The naturally competent organism, Neisseria gonorrhoeae, can efficiently transform a marker carried on DNA purified in low-melting-temperature agarose without prior purification or dilution . Neither the agarose or buffer components inhibit transformation frequencies, but exposure to UV irradiation completely abrogates transformation.

J Clin Microbiol, 1995 Dec, 33(12), 3326 - 7
Comparison of commercial diagnostic tests for identification of serogroup antigens of Neisseria meningitidis; van der Ende A et al.; In the study that is described the sensitivities and specificities of three commercial tests and the standard Reference Laboratory test, used since 1961, to identify Neisseria meningitidis serogroups were compared . The tests marketed by Difco, Murex/Wellcome, and Sanofi/Pasteur showed overall sensitivities of 92, 95, and 100%, respectively, and specificities of 67, 88, and 82%, respectively . When limited to the common serogroups A, B, and C, the three tests yielded sensitivities of 93, 97, and 100%, respectively, and specificities of 98, 100, and 98%, respectively . However, determination of the uncommon serogroups X, W-135, Y, Z, and 29E with these tests is either unreliable or not possible.

J Clin Microbiol, 1995 Dec, 33(12), 3111 - 4
Ligase chain reaction for detection of Neisseria gonorrhoeae in urogenital swabs; Ching S et al.; The ligase chain reaction (LCR) is an in vitro nucleic acid amplification technique that exponentially amplifies targeted DNA sequences . In a multicenter study, we evaluated the use of a 4-h LCR-based assay for the diagnosis of Neisseria gonorrhoeae infection of the cervix and male urethra . The LCR results were compared with those of culture for N . gonorrhoeae by using selective media . This assay amplifies target sequences within the N . gonorrhoeae opacity gene . Discordant LCR-positive and culture-negative specimens were further evaluated by testing by another LCR assay which used N . gonorrhoeae-specific pilin probe sets . A total of 1,539 female endocervical specimens and 808 male urethral swab specimens were evaluated in the study . An expanded "gold standard" was defined to include all culture-positive as well as culture-negative, confirmed LCR-positive specimens . After resolution of discrepant samples, the sensitivities of the N . gonorrhoeae LCR assays for the female and male specimens were 97.3 and 98.5%, respectively, with specificities of 99.6 and 99.8%, respectively . Resolved culture sensitivities were 83.9 and 96.5% for the female and male specimens, respectively . The LCR assay for gonorrhea is a rapid, highly sensitive nonculture method for detecting gonococcal infection of the cervix and male urethra.

Genitourin Med, 1995 Dec, 71(6), 405 - 6
Prevalence of Trichomonas vaginalis in a male sexually transmitted disease clinic population by interview, wet mount microscopy, and the InPouch TV test; Borchardt KA et al.; OBJECTIVE--To determine the prevalence of trichomoniasis in male patients from their urine at a Sexually Transmitted Disease (STD) Clinic using the InPouch TV culture system . METHODS--Two hundred and four patients were examined for STD infections . Their ages ranged between 17 and 72 years . Depending on their clinical symptoms tests were ordered for Neisseria gonorrhoeae, chlamydia, and for syphilis . Each patient submitted a clean catch urine specimen for trichomonas testing . A 15 ml aliquot of urine was centrifuged and a drop of the sediment examined microscopically . The remainder was cultured in the InPouch TV test . Each pouch was examined at 24 h, 48 h, and 5 days . RESULTS--Twenty-four of the 204 patients (12%) were culture positive for Trichomonas vaginalis and only three of these were wet mount positive . CONCLUSION--The InPouch TV test demonstrated an epidemiological important infected male population that was not indicated by wet mount microscopy.

Genitourin Med, 1995 Dec, 71(6), 367 - 9
Serovar specific immunity to Neisseria gonorrhoeae: does it exist?
Ross JD, Moyes A, Young H.
OBJECTIVE--To determine whether the host immune response to gonorrhoea provides limited serovar specific protection from reinfection . SUBJECTS--508 episodes of gonorrhoea diagnosed at a city centre genitourinary medicine clinic including 22 patients with multiple infections over a 4 year period . METHODS--Patients with recurrent gonococcal infection were analysed with respect to the initial and subsequent serovars isolated . RESULTS--No significant difference was seen in the prevalence of serovars isolated following a repeat infection compared with those without repeat infections . The site of the initial infection did not appear to influence the subsequent serovar isolated . CONCLUSION--We found no evidence of serovar specific immunity in our population . It remains possible that populations with a higher prevalence of gonorrhoea and more frequent infections may have a quantitatively greater immune response.

Genitourin Med, 1995 Dec, 71(6), 363 - 6
Differentiation of Neisseria gonorrhoeae strains by polymerase chain reaction and restriction fragment length polymorphism of outer membrane protein IB genes; Lau QC et al.; OBJECTIVES--To employ polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis for the rapid differentiation of Neisseria gonorrhoeae protein IB (PIB) isolates and to compare its usefulness with the widely accepted auxotype/serovar classification scheme . METHODS--The outer membrane protein IB genes of 47 gonococcal isolates belonging to 10 different serovars were amplified by PCR . The approximately 1 kb DNA products were then digested separately with restriction enzymes CfoI and MspA1I, and electrophoresed on agarose gels . RESULTS--Cleavage of PIB genes by MspA1I and CfoI differentiated all the N gonorrhoeae strains into five and six PCR-RFLP profiles, respectively . PCR-RFLP was more discriminatory than auxotyping, which classifies the strains into only two auxotypes . Some strains belonging to common serovars could be further differentiated . A combination of PCR-RFLP analysis, auxotyping and serotyping further increased the discrimination of the strains into 34 subtypes . The PCR-RFLP method was easy to perform, reliable, reproducible, and consistent with published nucleotide sequence data . CONCLUSION--The PCR-RFLP method can augment auxotyping and serotyping or be used as a preliminary screening tool to differentiate N gonorrhoeae strains in areas where serotyping reagents are not easily available.

Arch Pediatr, 1995 Dec, 2(12), 1184 - 91
{Role of macrolides in the treatment of respiratory tract infections in children}; Floret D; Macrolides are antibiotics with high intracellular concentrations . They have a bacteriostatic activity but are also bactericides for concentrations five times greater than the minimal inhibitory concentration, concentrations in which they reach in the respiratory tract . They are usually active on Streptococcus, Neisseria, Moraxella catarrhalis, Listeria monocytogenes, Bordetella pertussis, Pasteurella multocida, Chlamydia, Mycoplasma pneumoniae, Legionella pneumophila and Helicobacter pylori . They have few secondary effects, some in relation with drug interactions . Their main indications are bronchopulmonary infections due to Mycoplasma pneumoniae, Chlamydia pneumoniae, Chlamydia trachomatis and Legionella pneumophila . They are also useful in whooping cough allowing the eradication of Bordetella pertussis in the rhinopharynx, thus limiting the dissemination of the infection in children . In amygdalitis and pharyngitis, macrolides are a good substitute in the case of allergy to penicillin . New generation of macrolides (roxithromycine, clarithromycine, dirithromycine, azithromycine) might open other interesting therapeutic perspectives.

Gene, 1995 Dec 1, 166(1), 101 - 4
Construction of a translational lacZ fusion system to study gene regulation in Neisseria gonorrhoeae; Silver LE et al.; A translational lacZ reporter system to study gene regulation in Neisseria gonorrhoeae (Ng) was developed . The pUC18-based vector pLES94 transforms Ng and recombines into the Ng chromosome at the site of the proAB genes . The vector contains a restriction site for cloning promoters that will result in a lacZ gene fusion . Initial cloning and characterization of promoters can be done in Escherichia coli . The vector contains both ApR and CmR genes, however the ApR gene is lost when the insert combines into the Ng chromosome . This system gives single copy expression of the fusion and does not result in the inactivation of the gene of interest.

J Bacteriol, 1995 Dec, 177(24), 7275 - 9
Genetic basis of Neisseria gonorrhoeae lipooligosaccharide antigenic variation; Danaher RJ et al.; Neisseria gonorrhoeae lipooligosaccharide (LOS) undergoes antigenic variation at a high rate, and this variation can be monitored by changes in a strain's ability to bind LOS-specific monoclonal antibodies . We report here the cloning and identification of a gene, lsi-2, that can mediate this variation . The DNA sequence of lsi-2 has been determined for N . gonorrhoeae 1291, a strain that expresses a high-molecular-mass LOS, and a derivative of this strain, RS132L, that produces a truncated LOS . In the parental strain, lsi-2 contains a string of 12 guanines in the middle of its coding sequence . In cells that had antigenically varied to produce a truncated LOS, the number of guanines in lsi-2 was altered . Site-specific deletions were constructed to verify that expression of a 3.6-kDa LOS is due to alterations in lsi-2.

J Bacteriol, 1995 Dec, 177(23), 6902 - 9
Identification of the gonococcal glmU gene encoding the enzyme N-acetylglucosamine 1-phosphate uridyltransferase involved in the synthesis of UDP-GlcNAc; Ullrich J et al.; In searching for the gonococcal sialyltransferase gene(s), we cloned a 3.8-kb DNA fragment from gonococcus strain MS11 that hybridized with the oligonucleotide JU07, which was derived from the conserved C terminus of the sialyl motif present in mammalian sialyltransferases . Sequencing of the fragment revealed four putative open reading frames (ORFs), one of which (ORF-1) contained a partial sialyl motif including the amino acid sequence VGSKT, which is highly conserved among sialyltransferases . The gene was flanked by two inverted repeats containing the neisserial DNA uptake sequence and was preceded by a putative sigma 54 promoter . Database searches, however, revealed a high degree of homology between ORF-1 and the N-acetylglucosamine 1-phosphate uridyltransferase (GlmU) of Escherichia coli and Bacillus subtilis and not with any known sialyltransferase . This homology was further established by the successful complementation of an orf-1 mutation by the E . coli glmU gene . Enzyme assays demonstrated that ORF-1 did not possess sialyltransferase activity but mimicked GlmU function catalyzing the conversion of N-acetylglucosamine 1-phosphate into UDP-N-acetylglucosamine, which is a key metabolite in the syntheses of lipopolysaccharide, peptidoglycan, and sialic acids.

Infect Immun, 1995 Dec, 63(12), 4790 - 4
Gonococcal infection in a nonhuman host is determined by human complement C1q; Nowicki S et al.; Human C1q displayed a dose-dependent protection of gonococcal cells (GC) from the bactericidal effect of newborn rat serum . All rat pups injected with C1q-preincubated GC developed bacteremia, while none of the animals injected with GC only were infected . After clearance of bacteremia at day 6, live GC could still be recovered from tested organs, including the liver . Preincubation of GC with higher concentrations of C1q was associated with increased morbidity . In contrast to human serum as a source of C1q, rat, rabbit, and mouse sera did not increase the in vivo virulence of Neisseria gonorrhoeae . C1q-deficient human serum, heat-inactivated C1q or human serum, type IV collagen, and complement C3 were inefficient in inducing infection . Experimental infection by C1q-preincubated GC was inhibited by anti-C1q antibodies in a dose-dependent fashion, demonstrating a causal effect of C1q function . This report demonstrates the novel finding that human C1q, a component of the human immune system with a general function for elimination of infection, may increase GC virulence and result in the development of disseminated infection in a nonhuman host.

Infect Immun, 1995 Dec, 63(12), 4634 - 41
Binding and accumulation of hemin in Neisseria gonorrhoeae; Desai PJ et al.; The ability to utilize hemin and hemin-containing compounds for nutritional iron (Fe) uptake has been documented for several pathogenic bacteria . Neisseria gonorrhoeae can utilize free hemin as a source of Fe for growth; however, little is known concerning the mechanisms involved in hemin transport . In this study we have characterized the binding and accumulation of hemin by N . gonorrhoeae and defined the specificity of the gonococcal hemin receptor . N . gonorrhoeae F62 was grown in a chemically defined medium containing the iron chelator Desferal, and hemin transport was initiated by the addition of {59Fe}hemin (4.0 or 8.0 microM; specific activity, 7.0 Ci/mol) . 59Fe uptake from radiolabeled hemin by N . gonorrhoeae was energy dependent, and 59Fe was shown to accumulate in the cell at a constant rate during logarithmic growth . However, we observed a decrease in the uptake of 59Fe from radiolabeled hemin when inorganic iron was present in the growth medium . Binding of 59Fe from radiolabeled hemin was inhibited by the addition of either cold hemin, hematoporphyrin, or hemoglobin, but not by ferric citrate . Although {14C}hemin was found to support the growth of N . gonorrhoeae, we did not detect the uptake of 14C from radiolabeled hemin . Extraction of the gonococcal periplasmic ferric binding protein (Fbp) from cultures grown with {59Fe}hemin indicated that a majority of the 59Fe was associated with the Fbp . Taken together, the results presented here indicate that hemin binds to a gonococcal outer membrane receptor through the protoporphyrin portion of the molecule and that following binding, iron is removed and transported into the cell, where it is associated with the gonococcal periplasmic ferric binding protein, Fbp.

Pediatrics, 1995 Dec, 96(6), 1090 - 4
The prevalence of sexually transmitted diseases in children and adolescents evaluated for sexual abuse in Cincinnati: rationale for limited STD testing in prepubertal girls; Siegel RM et al.; OBJECTIVE . To determine the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, syphilis, and human immunodeficiency virus (HIV) infection in sexually abused children and to develop selective criteria for sexually transmitted disease (STD) testing in these children in our community . DESIGN . Prospective . SETTING . University-affiliated children's hospital in Ohio . PARTICIPANTS . All children evaluated at our hospital for sexual abuse were eligible . Eight hundred fifty-five children were evaluated over a 1-year period . The study included 704 girls and 151 boys . Children ranged in age from 3 weeks to 18 years old . METHODS AND RESULTS . Standard STD testing (American Academy of Pediatrics recommendations) was defined as serum rapid plasma reagin test, examination for Trichomonas, N gonorrhoeae culture of the throat, rectum, and genitalia and C trachomatis culture of the rectum and genitalia . STD testing in this study was recommended in children with 1) a history of genital discharge or contact with the perpetrator's genitalia, 2) examination findings of genital discharge or trauma, and 3) all adolescents . HIV testing was obtained in children with risk factors for HIV infection, those with contact with a perpetrator with HIV risk factors, or if the family was concerned about HIV acquisition . A total of 423 children were tested for N gonorrhoeae, 415 for C trachomatis, 275 for syphilis, 208 for Trichomonas, and 140 for HIV . Twelve children were determined to have N gonorrhoeae infection, 11 had C trachomatis infection, and four had Trichomonas infection . Overall, the prevalence of STDs in prepubertal girls was 3.2% and 14.6% in pubertal girls . The prevalence of N gonorrhoeae in prepubertal girls with vaginal discharge was 11.1% and 0% in prepubertal girls without discharge (P < .001) . C trachomatis infection was diagnosed in 0.8% of prepubertal girls compared with 7.0% of pubertal girls (P < .001) . None of the children tested positive for syphilis or HIV and no males had a STD . CONCLUSIONS . In our community, N gonorrhoeae testing in prepubertal girls can be limited to those with a vaginal discharge on examination unless other risk factors are present . The prevalence C trachomatis and Trichomonas in prepubertal girls is low and may be omitted from routine evaluations . All pubertal girls evaluated for sexual abuse should be tested for STDs because of the high prevalence of asymptomatic infection in this patient population.

J Biol Chem, 1995 Nov 3, 270(44), 26045 - 8
The Neisseria transcriptional regulator PilA has a GTPase activity; Arvidson CG et al.; The pilA gene of Neisseria gonorrhoeae encodes the response regulator of a two-component regulatory system that controls pilin gene expression . Examination of the primary sequence of PilA indicates that the protein contains at least two functional domains . The N-terminal region has a proposed helix-turn-helix motif thought to be involved in DNA binding . This region also contains the residues that are presumed to form the acidic pocket involved in phosphorylation by PilB, the sensor kinase of the system . The C terminus of the protein has extensive homology to the G (GTP-binding) domains of the eukaryotic signal recognition particle (SRP) 54-kDa protein and the alpha subunit of the SRP receptor, or docking protein . This homology also extends to similar regions of the bacterial SRP homologs Ffh and FtsY . Here, we demonstrate that purified PilA has significant GTPase activity, and that this activity has an absolute requirement for MgCl2 and is sensitive to KCl and low pH . We also show that PilA has a strict specificity for GTP, and that GTP hydrolysis follows first order kinetics, with a maximum velocity (Vmax) of 1900 pmol of Pi produced per min per mg of protein and a Km for GTP of 9.6 microM at 37 degrees C.

Nature, 1995 Nov 2, 378(6552), 32 - 8
Structure of the fibre-forming protein pilin at 2.6 A resolution; Parge HE et al.; The crystallographic structure of Neisseria gonorrhoeae pilin, which assembles into the multifunctional pilus adhesion and virulence factor, reveals an alpha-beta roll fold with a striking 85 A alpha-helical spine and an O-linked disaccharide . Key residues stabilize interactions that allow sequence hypervariability, responsible for pilin's celebrated antigenic variation, within disulphide region beta-strands and connections . Pilin surface shape, hydrophobicity and sequence variation constrain pilus assembly to the packing of flat subunit faces against alpha 1 helices . Helical fibre assembly is postulated to form a core of coiled alpha 1 helices banded by beta-sheet, leaving carbohydrate and hypervariable sequence regions exposed to solvent.

Microb Pathog, 1995 Nov, 19(5), 285 - 97
Biochemical analysis of lactoferrin receptors in the Neisseriaceae: identification of a second bacterial lactoferrin receptor protein; Bonnah RA et al.; Bacterial transferrin receptors that have been described in the families Pasteurellaceae and Neisseriaceae are composed of two receptor proteins, transferrin binding proteins 1 and 2 (Tbp1 and Tbp2) . In contrast, bacterial lactoferrin receptors have only been described for human pathogens in the family Neisseriaceae, and were believed to consist of a single protein, Lbp1, which is highly homologous to Tbp1 . We describe a modified affinity isolation procedure that facilities isolation of a second lactoferrin receptor protein Lbp2 (a presumptive Tbp2 homologue) from Neisseria meningitidis, Moraxella catarrhalis and Moraxella bovis using immobilized lactoferrin . Antiserum specific for either the M . catarrhalis Tbp1+2 molecules, the M . catarrhalis Lbp1 molecule, or for a commercial preparation of human lactoferrin did not react on western blots with the same organisms' affinity purified Lbp2 . In addition, the M . catarrhalis Lbp2 could be isolated in a functional form without contaminating Lbp1 or Tbp1+2 . We also demonstrate that the bovine pathogen, M . bovis, produces functional transferrin and lactoferrin receptors specific for the bovine forms of these glycoproteins . A putative lbpB gene, recently speculated to reside immediately upstream of the N . meningitidis Lbp1 structural gene, lbpA, likely encodes the newly isolated Lbp2 protein from this bacterial species.

Antimicrob Agents Chemother, 1995 Nov, 39(11), 2442 - 5
Proposed criteria for interpretation of susceptibilities of strains of Neisseria gonorrhoeae to ciprofloxacin, ofloxacin, enoxacin, lomefloxacin, and norfloxacin; Knapp JS et al.; The susceptibilities of 45 strains of Neisseria gonorrhoeae, including 25 strains susceptible to ciprofloxacin (MICs, < or = 0.06 microgram/ml) and 20 strains exhibiting decreased susceptibilities to ciprofloxacin (MICs, > or = 0.125 microgram/ml), to ciprofloxacin, ofloxacin, enoxacin, lomefloxacin, norfloxacin, and nalidixic acid were determined by agar dilution and disk diffusion . On the basis of theoretical calculations of predicted susceptibilities at which infections may fail therapy (supported by observed failures of infections to respond to the therapeutic doses of enoxacin and ciprofloxacin), the Centers for Disease Control and Prevention has adopted the following agar dilution breakpoints for interpretation of resistance to these agents: MICs of > or = 1.0 microgram of ciprofloxacin, enoxacin, and norfloxacin per ml and MICs of > or = 2.0 micrograms of ofloxacin and lomefloxacin per ml . The corresponding disk diffusion breakpoints for these agents were as follows: ciprofloxacin, < or = 29 mm; ofloxacin, < or = 24 mm; enoxacin, < or = 31 mm; lomefloxacin, < or = 26 mm; and norfloxacin, < or = 32 mm . The Centers for Disease Control and Prevention recommends two strains as interim quality control strains for susceptibility testing of ciprofloxacin and ofloxacin . These are N . gonorrhoeae CDC-10,328 (MIC of ciprofloxacin, 0.125 to 0.25 microgram/ml {inhibition zone diameter range, 30 to 34 mm}; MIC of ofloxacin, 0.5 microgram/ml {inhibition zone diameter range, 27 to 32 mm}) and N . gonorrhoeae CDC-10,329 (MIC of ciprofloxacin, 1.0 to 2.0 micrograms/ml {zone inhibition diameter range, 21 to 26 mm}; MIC of ofloxacin 2.0 micrograms/ml {inhibition zone diameter range, 18 to 21 mm}).

Eur J Pediatr, 1995 Nov, 154(11), 896 - 900
Definition of a new score for severity of generalized Neisseria meningitidis infection; Nurnberger W et al.; Neisseria meningitidis infection may present as meningitis or as severe, fulminant sepsis . In order to classify individual patients early according to the expected course of the disease, we developed a score named Neisseria sepsis index {NESI} . The NESI was defined using the parameters heart rate, mean arterial blood pressure, base excess and presence of acute subcutaneous bleeding and/or skin necroses (minimal value {= no evidence for sepsis} NESI 0; maximum value {= most severe sepsis} NESI 8) . Seventeen patients with documented, systemic N . meningitidis infection were prospectively assessed for the terminal complement complex (TCC), serum tumour necrosis factor alpha (TNF alpha) levels (as laboratory parameters for severity of sepsis) and NESI score . The evaluation was immediately performed when the patients were admitted to the hospital . The 17 patients showed the following distribution of data: NESI 0 (n = 4), NESI 1 (n = 6), NESI 2 (n = 0), NESI 3 (n = 1), NESI 4 (n = 2), NESI 5 (n = 2), NESI 6 (n = 0), NESI 7 (n = 1), NESI 8 (n = 1) . Mortality was 4/17 patients, all had NESI > or = 5 . TCC values ranged from 647-6461 ng/ml (normal range: 130-360 ng/ml); and was not correlated to NESI . TNF alpha values ranged from 10-910 pg/ml and were correlated to NESI (r2 = 0.71, n = 17, P < 0.001) . In patients with fatal outcome, TNF alpha was 600 +/- 160 pg/ml (mean +/- SEM) and in surviving patients 130 +/- 50 pg/ml (mean +/- SEM) . TNF alpha was increased in 15/17 patients when compared to normal controls (< 27 pg/ml) . CONCLUSION . The NESI is based on few clinical, objective data, that are available in every hospital . NESI appears to offer an instrument: (1) for making decisions in regard to appropriate monitoring and treatment of vital organ function; and (2) for assessing the quality of care for this life-threatening infection.

Sex Transm Dis, 1995 Nov-Dec, 22(6), 380 - 2
Acute gonococcal sepsis in an HIV-infected woman; Jacoby HM et al.; This is a case report of a 35-year-old woman infected with the human immunodeficiency virus who presented with acute bacterial sepsis that proved to be secondary to Neisseria gonorrhoeae . Typical skin and joint findings developed only after the acute sepsis had resolved . Patients with disseminated gonococcal infection rarely have signs of acute bacterial sepsis . This case raises the question of whether HIV-infected patients are at an increased risk of contracting severe gonococcal disease.

Sex Transm Dis, 1995 Nov-Dec, 22(6), 335 - 41
Commonly used diagnostic criteria for pelvic inflammatory disease have poor sensitivity for plasma cell endometritis; Korn AP et al.; BACKGROUND: The majority of women with tubal damage do not have a history of acute pelvic inflammatory disease . The prevalence of upper genital tract inflammation was evaluated in women deemed not to have pelvic inflammatory disease by common diagnostic criteria . GOAL OF THIS STUDY: To compare clinical signs and laboratory tests used to diagnose pelvic inflammatory disease with endometrial biopsy histopathology . STUDY DESIGN: Endometrial biopsy and commonly used physical and laboratory tests were performed on 52 women with pelvic tenderness, 51 with vaginosis or cervicitis, and 22 control subjects who had no evidence of infection with Neisseria gonorrhoeae or Chlamydia trachomatis and who tested negative for bacterial vaginosis using vaginal swab Gram's stain . RESULTS: Thirty-six of 52 patients (69%) with pelvic tenderness, compared with 22 of 51 patients (43%) with vaginosis or cervicitis and two of 22 control subjects (9%), had plasma cell endometritis . The Centers for Disease Control and Prevention minimal diagnostic criteria for pelvic inflammatory disease had a sensitivity of 33% for plasma cell endometritis . CONCLUSIONS: The clinical diagnosis of pelvic inflammatory disease using published criteria correlates poorly with plasma cell endometritis.

J Clin Microbiol, 1995 Nov, 33(11), 3068 - 9
Comparison of Gram stain with DNA probe for detection of Neisseria gonorrhoeae in urethras of symptomatic males; Juchau SV et al.; The comparison of Gram-stained urethral smears with Gen-Probe for the detection of Neisseria Gonorrhoeae in the urethras of males with symptomatic urethritis revealed a 99.6% correlation between the two methods . A simple Gram stain would appear to be the method of choice for the detection of gonorrhea in symptomatic males, because it is much less expensive and much more rapid than the Gen-Probe method.

J Clin Microbiol, 1995 Nov, 33(11), 3049 - 53
Multiplex PCR for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Genitourinary specimens; Mahony JB et al.; We developed a multiplex PCR (M-PCR) assay for the simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae . M-PCR employed C . trachomatis-specific primers KL1-KL2 and N . gonorrhoeae-specific primers HO1-HO3 and produced products of 241 and 390 bp, respectively . PCR products were easily detected by agarose gel electrophoresis and confirmed by Southern hybridization using labelled oligonucleotide probes . M-PCR had a sensitivity of 10 fg of C . trachomatis and N . gonorrhoeae DNA (equivalent to 1 to 2 genome copies) . M-PCR detected the presence of C . trachomatis and N . gonorrhoeae DNA in 15 male urethral and 12 female endocervical specimens, 3 of which were positive for C . trachomatis, 18 of which were positive for N . gonorrhoeae and 6 of which were positive for both organisms . M-PCR was evaluated further by testing 200 male first void urine (FVU) specimens, of which 18 were positive by C . trachomatis PCR and Chlamydiazyme and 4 were positive by C . trachomatis PCR but negative by Chlamydiazyme . All 22 FVU specimens were positive by a confirmatory PCR using a second plasmid target and were positive by M-PCR . Ten of 11 men with cultures that were positive for N . gonorrhoeae had FVU specimens that were positive by both N . gonorrhoeae PCR and M-PCR . Two other men with negative N . gonorrhoeae urethral cultures had FVU specimens that were positive by N . gonorrhoeae PCR, by two confirmatory N . gonorrhoeae PCR assays using 165 rRNA and cytosine methyltransferase primers, and by M-PCR . The sensitivity of M-PCR for detecting C . trachomatis was 100% (22 of 22 specimens), compared with 81.8% (18 of 22 specimens) for enzyme immunoassay . Sensitivity of M-PCR for N . gonorrhoeae was 92.3% (12 of 13 specimens) compared with 84.6% (11 of 13 specimens) for urethral culture . The specificity of M-PCR was 100% for both C . trachomatis (178 of 13 specimens) and N . gonorrhoeae (187 of 187 specimens) . M-PCR testing of FVU specimens provided a sensitive and noninvasive method for detecting C . trachomatis and N . gonorrhoeae infection in men.

J Infect Dis, 1995 Nov, 172(5), 1371 - 4
Genital ulcers associated with human immunodeficiency virus-related immunosuppression in female sex workers in Abidjan, Ivory Coast; Ghys PD et al.; A cross-sectional study among female sex workers in Abidjan was conducted to study the association between sexually transmitted diseases and human immunodeficiency virus (HIV) infection and HIV-related immunosuppression . Among 1209 women tested for HIV, 962 (80%) were seropositive . HIV infection was independently associated with a longer duration of sex work, a lower price for intercourse, being an immigrant, and having a positive Treponema pallidum hemagglutination test (P < .05) . Genital ulcers (25% vs . 5%), genital warts (14% vs . 4%), Neisseria gonorrhoeae (32% vs . 16%), Trichomonas vaginalis (27% vs . 17%), and syphilis (27% vs . 17%) were more frequent (P < .05) in HIV-infected than -uninfected women . Among HIV-infected women, the proportions with a genital ulcer were 17%, 25%, and 36% for those with > 28%, 14%-28%, and < 14% CD4 cells, respectively (P < .001) . This study suggests that genital ulcers are an opportunistic disease in female sex workers in Abidjan.

J Bacteriol, 1995 Nov, 177(22), 6390 - 400
The physical map of the chromosome of a serogroup A strain of Neisseria meningitidis shows complex rearrangements relative to the chromosomes of the two mapped strains of the closely related species N . gonorrhoeae; Dempsey JA et al.; A physical map of the chromosome of N . meningitidis Z2491 (serogroup A, subgroup IV-1) has been constructed . Z2491 DNA was digested with NheI, SpeI, SgfI, PacI, BglII, or PmeI, resulting in a limited number of fragments that were resolved by contour-clamped homogeneous electric field (CHEF) electrophoresis . The estimated genome size for this strain was 2,226 kb . To construct the map, probes corresponding to single-copy genes or sequences were used on Southern blots of chromosomal DNA digested with the different mapping enzymes and subjected to CHEF electrophoresis . By determining which fragments from different digests hybridized to each specific probe, it was possible to walk back and forth between digests to form a circular macrorestriction map . The intervals between mapped restriction sites range from 10 to 143 kb in size . A total of 117 markers have been placed on the map; 75 represent identified genes, with the remaining markers defined by anonymous cloned fragments of neisserial DNA . Comparison of the arrangement of genetic loci in Z2491 with that in gonococcal strain FA1090, for which a physical map was previously constructed, revealed complex genomic rearrangements between the two strains . Although gene order is generally conserved over much of the chromosome, a region of approximately 500 kb shows translocation and/or inversion of multiple blocks of markers between the two strains . Even within the relatively conserved portions of the maps, several genetic markers are in different positions in Z2491 and FA1090.

MMWR Morb Mortal Wkly Rep, 1995 Oct 20, 44(41), 761 - 4
Fluoroquinolone resistance in Neisseria gonorrhoeae--Colorado and Washington, 1995; A role for bacteriophages in the evolution and transfer of bacterial virulence determinants; Department of Molecular and Cell Biology, University of New England, Armidale, New South Wales, AustraliaA virulence-associated region in the genome of Dichelobacter nodosus has been shown to contain an integrase gene which is highly related to the integrases of Shigella flexneri phage Sf6 and coliphages P4 and phi R73, together with open reading frames (vapB, C and D) related to genes borne on plasmids in Neisseria gonorrhoeae, Escherichia coli, Actinobacillus actinomycetemcomitans and Treponema denticola . Similar to P4 and phi R73, the vap region is bracketed by putative bacteriophage att sites and is adjacent to a tRNA gene, which suggests that the vap region has been derived by the integration of a bacteriophage, or a plasmid carrying a bacteriophage-related integrase gene . Many similarities in genes and genes clusters encoding virulence determinants have been found in distantly related bacteria . These genes are often located on plasmids in one organism but on the chromosome in others, implying that transmission of the genes has been followed by integration . Thus, the events which have generated the vap regions of D . nodosus may represent a common mechanism for transfer of virulence determinants . A number of genes involved in the virulence of bacterial pathogens are found on integrated bacteriophages, and we suggest that others will prove to be associated with tRNA genes and/or integrase genes derived from bacteriophages . The use of tRNA genes as integration sites for many bacteriophages and plasmids may favour intergeneric transmission, as tRNA genes are highly conserved.

Antimicrob Agents Chemother, 1995 Oct, 39(10), 2367 - 70
Emergence of in vitro resistance to fluoroquinolones in Neisseria gonorrhoeae isolated in Japan; Tanaka M et al.; To investigate emerging fluoroquinolone resistance in Neisseria gonorrhoeae isolated in Japan, we compared the in vitro antimicrobial susceptibilities of 79 gonococcal isolates from 1992 through 1993 to 14 fluoroquinolones and 14 other antibiotics with those of 27 isolates from between 1981 and 1984 . The MICs at which 90% of the isolates were inhibited by nine fluroquinolones, including norfloxacin, enoxacin, ofloxacin, ciprofloxacin, tosufloxacin, lomefloxacin, fleroxacin, levofloxacin, and sparfloxacin, for isolates from 1992 to 1993 were 8- or 16-fold higher than those for isolates from 1981 to 1984 . Furthermore, the MICs at which 90% of the isolates were inhibited by five fluroquinolones, including OPC-17116, T-3761, DU-6859a, AM-1155, and Q-35, that have recently been synthesized but have not yet been introduced for clinical use in Japan for isolates from 1992 to 1993 were also 2- to 16-fold higher than those for isolates from 1981 to 1984 . The gonococcal isolates from 1992 to 1993 showed no significant decreases in susceptibility to beta-lactams, tetracyclines, macrolides, and spectinomycin, compared with those for isolates from 1981 to 1984 . Our data indicate that the incidence of gonococcal strains with decreased susceptibilities to fluoroquinolones is increasing in Japan.

Eur J Clin Microbiol Infect Dis, 1995 Oct, 14(10), 914 - 8
Epidemiology of penicillin-resistant Neisseria gonorrhoeae in Frankfurt, Germany; Schafer V et al.; In the present study 150 Neisseria gonorrhoeae strains from prostitutes in Frankfurt, Germany, were investigated for their in vitro susceptibility to eight antimicrobial agents by the agar dilution test . While 21.3% of gonococci were highly resistant to penicillin (MICs > or = 2 mg/l), 49.4% of the organisms were moderately sensitive (MICs = 0.125-0.5 mg/l) and only 29.3% of strains were fully susceptible to penicillin (MICs < or = 0.06 mg/l) . No resistance to other antibiotics tested was observed, with the exception of doxycycline, to which overall resistance was 9.7% . Genotypic analysis of Neisseria gonorrhoeae by the restriction enzyme technique showed that most penicillin-resistant strains could be grouped into two main bacterial clones, whereas penicillin-susceptible isolates displayed a remarkable heterogeneity . This observation suggests that a restricted number of resistant Neisseria gonorrhoeae strains are likely to spread within the prostitute population of Frankfurt, Germany.

J Ethnopharmacol, 1995 Oct, 48(2), 85 - 8
Antigonorrhoeal activity of plants used in Guatemala for the treatment of sexually transmitted diseases; Caceres A et al.; Plants popularly used in Guatemala for the treatment of gonorrhoea were macerated in 50% alcohol and the tincture tested for in vitro activity against Neisseria gonorrhoeae using strains isolated from symptomatic patients and confirmed by standard bacteriological procedures . From 46 plants investigated, 13 (28.3%) showed evident inhibition zones (> 9 mm), seven (15.2%) showed small activity (6.1-8.9 mm) and 26 (56.5%) showed no activity; nine of these plants inhibited five strains of N . gonorrhoea freshly isolated . The most active plants of American origin were: bark of Bixa orellana fruits of Parmentiera edulis, leaf of Diphysa robinioides, Eupatorium odoratum, Gliricidia sepium, Physalis angulata, Piper aduncum and Prosopis juliflora, root of Casimiroa edulis, and whole Clematis dioica.

J Clin Microbiol, 1995 Oct, 33(10), 2587 - 91
Evaluation of nucleic acid-based test (PACE 2C) for simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical specimens; Iwen PC et al.; A nucleic acid-based test (Gen-Probe PACE 2C System) was evaluated for the ability to detect Chlamydia trachomatis and Neisseria gonorrhoeae from endocervical specimens in a single assay . Three swab samples, randomized for collection order, were obtained from each patient and tested by N . gonorrhoeae and C . trachomatis culture and by the PACE 2C probe assay . Fifty of 395 specimens were culture positive for N . gonorrhoeae (17 specimens), C . trachomatis (26 specimens), or both (7 specimens), of which PACE 2C testing detected 48 specimens . The PACE 2C assay was positive for 56 specimens, including 8 specimens not positive by culture . Of the total of 10 discrepancies between culture and PACE 2C results, resolution testing yielded four false-negative culture, four false-positive PACE 2C, and two false-negative PACE 2C results . The sensitivity, specificity, and positive and negative predictive values for PACE 2C after reevaluation were 96.3, 98.8, 92.9 and 99.4%, respectively . The overall sensitivities for C . trachomatis and N . gonorrhoeae culture were 89.2 and 88.9%, respectively . The prevalence rate for C . trachomatis was 9.4%, and that for N . gonorrhoeae was 6.8% . The Gen-Probe PACE 2C System is a reliable alternative for screening endocervical specimens for both C . trachomatis and N . gonorrhoeae in a single assay.

Child Abuse Negl, 1995 Oct, 19(10), 1303 - 10
Sexually transmitted diseases in children and evidence of sexual abuse; Argent AC et al.; During the period June 1989 to March 1991, laboratory evidence of sexually transmitted diseases (STDs) was found in 107 patients at the Red Cross War Memorial Children's Hospital in Cape Town . Data was available on 96 patients aged 23 months to 14 years (mean 75.9 months) . Vaginal discharge was the most frequent presenting symptom (76%), particularly in those less than 5 years of age (90%) . Although a history of abuse was not given on presentation in 62%, evidence of abuse was subsequently elicited in 67% of patients . Neisseria gonorrhoeae was the most common sexual pathogen (61 isolates, 8 penicillin resistant), followed by G vaginalis (17 isolates), Trichomonas vaginalis (7 infections), and T pallidum (9 TPHA positive, 5 with VDRL 1:4 or higher) . Chlamydia trachomatis was demonstrated by immunofluorescence in 14 children . Multiple STDs were demonstrated in 10 patients . Although evidence of CSA was not found in all patients with STDs it was likely that the vast majority of patients had acquired these infections by CSA . Symptomatic prepubertal children with G vaginalis isolates should be investigated for CSA . Chlamydial immunofluorescence tests did not assist the diagnosis of CSA in children and should not be used, as they have no medicolegal significancePIP: This study examines the nature and extent of sexually transmitted diseases related to sexual abuse among patients seeking treatment during June 1989-March 1991 at the Red Cross War Memorial Children's Hospital in Rondebosch, Cape Town, South Africa . 107 patients aged 23-174 months (about 2-15 years of age) were identified by laboratory testing as having a sexually transmitted disease . Analysis pertained to 96 patients . 10 children had multiple infections . 60 patients were assessed after 2-3 months, of whom 20% (12 cases) indicated evidence of child sexual abuse . 47% of the 36 patients who were not followed-up had evidence of child sexual abuse . Age was unrelated to follow-up or sexual abuse . Genital symptoms were evident at the initial visit among 87 patients (91%), of whom 65% had no prior history of abuse . 26% had a history of sexual abuse . 8 patients had a history of sexual abuse but no genital symptoms . 79% had genital symptoms of vaginal discharge . 39 patients aged under 5 years had a vaginal discharge, and 10 had a history of sexual abuse . 29 cases were determined not to be related to sexual abuse . There were 61 children infected with Neisseria gonorrhoea, 17 children with Gardnerella vaginalis, 14 children with Chlamydia trachomatis, 9 children with Trichomonas vaginalis, and 9 children with Treponema pallidum (positive cultures for syphilis, but without clinical features) . Sexual child abuse was found to be higher than previously reported at this center . The approach in this study was to deny sexual abuse until confirmation of a sexually transmitted disease . Almost 33% of children in this study had no physical evidence of sexual abuse . The hospital policy is to teach children and caregivers safety skills and awareness and to develop a rapport with parents . Ceftriaxone was administered as therapy, since most children had gonorrhea, and there was resistance to penicillin and poor follow-up . The authors find that all children with Gardnerella vaginalis should be investigated for child sexual abuse . There was an express need for follow-up of patients due to the almost 10% with multiple infections .

J Obstet Gynecol Neonatal Nurs, 1995 Oct, 24(8), 759 - 67
Pelvic inflammatory disease: clinical overview; Kottmann LM; Acute pelvic inflammatory disease (PID) adversely affects the health of 1 million American women annually . The best understood etiologic organisms are Chlamydia trachomatis and Neisseria gonorrhea; however, information regarding the contributions of facultative and causative anaerobes continues to emerge . Chronic PID presents a diagnostic and management challenge to health care providers . Because of morbidity and sequelae the importance of early detection of, and intervention for, upper tract pelvic infections cannot be overemphasized . Nursing interventions in PID prevention, early assessment, accurate diagnosis, and thorough treatment can have a positive impact on women's health.

J Clin Pathol, 1995 Oct, 48(10), 919 - 23
Comparison of three methods for culture confirmation of Neisseria gonorrhoeae strains currently circulating in the UK; Turner A et al.; AIMS--To establish the current sensitivity of two commercial kits for culture confirmation of Neisseria gonorrhoeae strains circulating in the UK . METHODS--A total of 544 strains were studied (199 gonococci from male patients attending hospitals in the county of Avon, 204 unselected N gonorrhoeae isolates from male patients in Leeds, 20 strains referred to the Gonococcus Reference Unit because of difficulties with the Syva fluorescent antibody test (FAT), and 121 strains collected over a four year period which had not reacted with serotyping antibodies) . Strains were tested by sugar utilisation in cysteine trypticase base agar (CTA test), the Phadebact Monoclonal GC Test and Syva MicroTrak Neisseria gonorrhoeae Culture Confirmation Test . The auxotype and serovar of each strain were also determined . RESULTS--The sugar utilisation test confirmed the identity of 99% (197/199) of gonococci from Avon and 97% (198/204) of those from Leeds . The Syva FAT confirmed 76% (151/199) of isolates from Avon and 84% (171/204) of those from Leeds . The Phadebact test confirmed all but one isolate from the 403 strains from both cities . Half of the 20 referred FAT negative isolates also give a negative result in the Syva FAT; however, only 10% of the remainder gave a strong reaction in our laboratory . All serotyping antibody negative strains were negative in the FAT, although all these and all of the 20 strains that give a negative result in the FAT gave positive reactions in the other culture confirmation tests . Typing tests revealed a greater diversity amongst the FAT negative strains from Leeds than those from Avon . CONCLUSIONS--Considerable differences in the sensitivity of the MicroTrak but not with the Phadebact or CTA tests were found for the identification of isolates from two geographically distinct areas of the UK . Our results suggest that the Syva FAT would not be suitable, if used alone, for culture confirmation in Avon or Leeds.

Microbiology, 1995 Oct, 141 ( Pt 10), 2367 - 77
Pilus biogenesis gene, pilC, of Neisseria gonorrhoeae: pilC1 and pilC2 are each part of a larger duplication of the gonococcal genome and share upstream and downstream homologous sequences with opa and pil loci; Jonsson AB et al.; Pili of Neisseria gonorrhoeae mediate attachment of the bacteria to target cells and undergo both phase and antigenic variation . PilC is a 110 kDa minor pilus-associated protein involved in pilus biogenesis and attachment . The expression of PilC is turned on and off at high frequency and is controlled by frameshift mutations in a run of G residues positioned in the region encoding the signal peptide . Most strains of N . gonorrhoeae carry two copies of pilC . The DNA sequence of pilC1 of strain MS11 is presented and compared to the sequence of the 3' end of pilC2 . These two genes are highly homologous, but not identical . The putative transcriptional terminator of pilC1 contains a pair of inverted uptake sequences for gonococcal DNA (5'-GCCGTCTGAA-3') . An 88 bp sequence located upstream of the pilC1 gene has also been reported to precede several opa genes of N . gonorrhoeae . Shorter regions positioned both downstream and upstream of pilC1 can also be found in silent pil loci as well as close to opa genes . The pilC genes are part of a duplication of a larger DNA region extending more than 2 kb downstream of the coding region.

Infect Immun, 1995 Oct, 63(10), 4181 - 4
Molecular characterization of FrpB, the 70-kilodalton iron-regulated outer membrane protein of Neisseria meningitidis; Pettersson A et al.; The structural gene encoding the 70-kDa outer membrane protein FrpB of Neisseria meningitidis was cloned and sequenced . A mutant lacking FrpB was constructed . No difference in iron utilization between the mutant and the parental strain was observed . A minor effect of the mutation on serum resistance was observed . A topology model for FrpB in the outer membrane is proposed.

Hosp Pract (Off Ed), 1995 Sep 15, 30(9), 61 - 6
Pelvic inflammatory disease: an ongoing epidemic; Augenbraun MH et al.; In the absence of a practical screening test, diagnosis is based on physical and laboratory findings, a strategy compromised by low sensitivity and specificity--and by asymptomatic cases . Antibiotic combinations must be directed against a wide range of pathogens, including Neisseria gonorrhoeae and Chlamydia trachomatis.

J Antimicrob Chemother, 1995 Sep, 36(3), 557 - 60
The potential effect on Neisseria gonorrhoeae of the use of clindamycin vaginal cream in the empirical treatment of vaginal discharge; Adu-Sarkodie YA et al.; The minimum inhibitory concentrations (MIC) of clindamycin for 62 consecutive isolates of Neisseria gonorrhoeae were found to be 0.03-4 mg/L; the MIC50 and MIC90 were 0.125 and 2.0 mg/L respectively . Seven women treated with clindamycin vaginal cream had cervical mucus samples taken after seven days treatment . The concentrations of clindamycin achieved in the cervical mucus were 30-150 times higher (141-337 mg/L) than the highest MIC of the 62 N . gonorrhoeae isolates . Clindamycin vaginal cream is being used increasingly in Genitourinary Medicine clinics and General Practice for the treatment of bacterial vaginosis . This study shows that clindamycin vaginal cream achieves intra cervical concentrations that are high enough to inhibit N . gonorrhoeae . Empirical use of this therapy should be preceded by urethral and cervical swabs for N . gonorrhoeae in any woman at risk of gonorrhoeae.

Diagn Microbiol Infect Dis, 1995 Sep-Oct, 23(1-2), 11 - 13
Antigonococcal activity of 11 drugs used for therapy or prophylaxis of malaria; Mehaffey PC et al.; Antibacterial agents are often used in malarial endemic areas for antimalarial prophylaxis (such as doxycycline and clindamycin) . Gonococcal infections may coexist in the same geographic area, thus becoming suppressed by compounds directed toward malarial parasites . We tested 11 drugs with activity for Plasmodium species against 105 Neisseria gonorrhoeae strains . Traditional or investigational antimalarials such as arteflene (Ro 42-1611), chloroquine, primaquine, pyrimethamine, quinacrine, and quinine were observed to be inactive . Fansidar (sulfadoxine-pyrimethamine) and mefloquine possess marginal action in a minority of gonococcus strains (< 10%) . Doxycycline {minimum inhibitory concentration inhibiting 90% of tested strains (MIC 90) 2 micrograms/ml} and azithromycin (MIC 90, 0.5 microgram/ml) among the antibacterials were very active, indicating a dual role as antimalarial and antigonococcal agents . Thus, the gonorrhea and sexually transmitted disease epidemiologic data from geographic regions where doxycycline or newer macrolides may be used for malarial prophylaxis or therapy could be significantly altered.

Mol Microbiol, 1995 Sep, 17(5), 855 - 63
High adhesiveness of encapsulated Neisseria meningitidis to epithelial cells is associated with the formation of bundles of pili; Marceau M et al.; Pili are indispensable in adhesion of encapsulated Neisseria meningitidis (MC) to eukaryotic cells . Intrastrain variability with respect to the degree of adhesion is the result of pilin antigenic variation . We have localized the region responsible for this variability to the 20-amino-acid hypervariable domain of pilin . The replacement of an aspartic acid, located in the hypervariable region of a low-adhesive variant by a lysine restored high adhesiveness . To assess whether hyperadhesiveness conferred by some pilin variants was related to the generation of a new pilus-associated ligand, high- and low-adhesive variants were purified . In a first step, low- and high-adhesive pilins were fused to maltose binding protein (MBP) . These hybrid proteins bound epithelial cells with the same affinity . Truncated MBP pilin fusions identified a cell-binding domain within the 77 residues of the N-terminal end of mature pilin . This region of the protein is common to low- and high-adhesive derivatives used in this work, thus eliminating the possibility that high adhesiveness conferred by some pilin variants was because of the generation of a new pilus-associated ligand . Electron-microscopic examination showed that low-adhesive derivatives expressed long and distinct pili and adhered as single cells . In contrast, pili of derivatives expressing high-adhesive pilins, either wild type or mutagenized from the low-adhesive variant, formed large bundles which bound bacteria and caused them to grow as colonies on infected monolayers . These data demonstrate that aggregative pili promote high adhesiveness of encapsulated MC.

Mol Microbiol, 1995 Sep, 17(6), 1133 - 42
Single amino acid substitutions in the N-terminus of Vibrio cholerae TcpA affect colonization, autoagglutination, and serum resistance; Chiang SL et al.; The toxin-coregulated pilus (TCP) of Vibrio cholerae O1 is required for successful infection of the host . TcpA, the structural subunit of TCP, belongs to the type IV family of pilins, which includes the PilE pilin of Neisseria gonorrhoeae . Recently, single amino acid changes in the N-terminus of PilE were found to abolish autoagglutination in gonococci . As type IV pilins demonstrate some similarities in function and amino acid sequence, site-directed mutagenesis and allelic exchanges were used to create corresponding mutations in TcpA . All four mutant strains demonstrated autoagglutination defects, and all were highly defective for colonization in the infant mouse model . These results support the previously proposed correlation between autoagglutination and colonization . Finally, all four mutants are serum sensitive, indicating that TcpA plays a role in serum resistance, a phenotype previously attributed to TcpC . As the mutations have similar effects in N . gonorrhoeae and V . cholerae, our results support the idea that type IV pilins have similar functions in a variety of pathogenic bacteria.

Mol Microbiol, 1995 Sep, 17(6), 1073 - 83
Uptake and nuclear transport of Neisseria IgA1 protease-associated alpha-proteins in human cells; Pohlner J et al.; Pathogenic Neisseria species, the causative agents of gonorrhoea and bacterial meningitis, encode a family of polymorphic exo-proteins which are autoproteolytically processed into several distinct extracellular components, including an IgA1 protease and an alpha-protein . IgA1 protease, a putative virulence determinant, is a sequence-specific endopeptidase known to cleave human IgA1, but additional target proteins have been postulated . The physical linkage of IgA1 protease and alpha-protein suggests a functional relationship of both precursor components . Previous work has shown that alpha-protein is essential neither for extracellular transport nor for the proteolytic activity of IgA1 protease . Intriguingly, alpha-proteins carry amino acid sequences reminiscent of nuclear location signals of viral and eukaryotic proteins . Here we demonstrate the functionality of these nuclear location signal sequences in transfected eukaryotic cells . Chimeric alpha-proteins show nuclear transport and selectively associate with nucleolar structures . More importantly, native purified alpha-proteins are capable of entering certain human primary cells from the exterior via an endocytotic route and accumulate in the nuclei . The neisserial alpha-proteins share several features with eukaryotic transcription factors, such as the formation of dimers via a heptad repeat sequence . We propose a role for alpha-proteins in the regulation of host-cell functions . As the alpha-proteins are covalently connected with IgA1 protease they may also serve as carries for the IgA1 protease into human cells where additional proteolytic targets may exist . Neisseria meningitidis, which locally colonizes the nasopharyngeal mucosa of many human individuals without apparently causing symptoms, secretes this nucleus-targeted factor in large quantities.

Mol Microbiol, 1995 Sep, 17(6), 1057 - 71
Pilus biogenesis and epithelial cell adherence of Neisseria gonorrhoeae pilC double knock-out mutants; Rudel T et al.; The phase-variable PilC proteins of pathogenic Neisseria species have recently been implicated in both assembly and cellular adherence functions of the type 4 pili of these pathogens . We describe here the cloning of full-length pilC1 and pilC2 genes and the complete sequencing of the pilC2 gene of Neisseria gonorrhoeae MS11 . Sequential inactivation of both genes by gene replacement in piliated (P+) variants of N . gonorrhoeae MS11 led initially to a non-piliated (P-) phenotype; however, spontaneous P+ variants could be derived from some pilC1,2 double mutants which produced morphologically intact pili . Purified pili from pilC1,2 mutants revealed no detectable PilC protein . Instead, a novel protein about 70 kDa in size appeared in the pili preparations of P+ mutants; this protein exhibited no immunological cross-reactivity with PilC1 or PilC2 . We propose that this novel factor replaces the function of PilC in pilus biogenesis . Using isogenic N . gonorrhoeae strains which produce identical PilE (pilin) proteins we demonstrate that pili associated with the 70 kDa protein do not confer gonococcal adherence to human epithelial cells, in contrast to pili assembled in the presence of PilC1 or PilC2.

Minerva Chir, 1995 Sep, 50(9), 793 - 8
{Venereal perihepatitis: Fitz-Hugh-Curtis syndrome}; Durastante V et al.; A case of Fitz-Hugh-Curtis syndrome (venereal perihepatitis) is reported . This syndrome is observed almost exclusively in women as a complication of genital gonococcal or chlamydial infections . A sudden, acute biliary-type pain generally characterizes the disease; only a few cases present symptoms related to associated genital infection . The real clinical incidence of venereal perihepatitis is quite high; in fact, a lot of the emergency admitted patients diagnosed with biliary colic or acute cholecystitis as a matter of fact suffer from this syndrome . If haematological investigations, ultrasonography and cholangiography do not confirm a suspected biliary lithiasis, it will be needed to investigate the genital tract . After clinical and ultrasound examinations, neisseria gonorrhoeae and chlamydia trachomatis must be sought in vaginal and cervical secretions and serum antichlamydial antibodies level is to be sought too . Through these examinations, the venereal perihepatitis can usually be diagnosed . In uncertain cases laparoscopy can be useful: in fact, it can reveal the typical violin-string-like adhesions between the anterior liver capsule and the anterior abdominal wall, and, in the same session, it allow to resect them . Tetracycline, doxycycline and, more recently, ofloxacine gave good results in the syndrome's treatment.

Neurol Neurochir Pol, 1995 Sep-Oct, 29(5), 687 - 93
{C-reactive protein (CRP) and its significance in purulent meningitis}; Przyjalkowski W et al.; CRP level was determined in the cerebrospinal fluid in 40 cases of bacterial meningitis . Similar determination in serum was done in 32 of these patients . Aetiological verification was possible in 90% of cases . Meningitis caused by Str . pneumonia and Neisseria meningitides prevailed (52.5% and 27.5% respectively) . The control group comprised 20 subjects . For CRP demonstration immunochemical and turbidimetric methods were used . CRP in CSF was raised in 62.5% of the study cases while in the serum it was raised in all of them . CRP detection in serum in acute phase of central nervous system infection is diagnostically important since CRP increase suggests a purulent process.

Vaccine, 1995 Sep, 13(13), 1213 - 9
Neisseria gonorrhoeae IgA1 proteases share epitopes recognized by neutralizing antibodies; Lomholt H et al.; The antigenic diversity among IgA1 proteases of 61 Neisseria gonorrhoeae strains isolated during a period of 23 years and on four continents was examined in enzyme neutralization assays employing rabbit antisera raised against selected IgA1 proteases . The antigenic analyses were compared with results of iga gene-region RFLP patterns and enzyme cleavage specificity for substrate IgA1 . Type 1 IgA1 proteases were antigenically uniform while six different antigenic types were detected among type 2 enzymes . Extensive cross-reactions of antibodies against the different antigenic types suggested only minor differences in relevant epitopes . Epitopes previously found to be common to all Neisseria meningitidis IgA1 proteases were also shared by all N . gonorrhoeae IgA1 proteases in the collection . Human sera from patients with gonorrhoea showed broadly cross-reactive neutralizing activity at titers comparable to those of sera from immunized rabbits . In conclusion, N . gonorrhoeae IgA1 proteases show a remarkable lack of diversity of epitopes recognized by enzyme-neutralizing antibodies . If future studies confirm that cleavage of IgA1 is an important step in gonococcal infections, Neisseria IgA1 proteases may be attractive vaccine candidates.

Int J STD AIDS, 1995 Sep-Oct, 6(5), 345 - 7
Epidemic spread of plasmid-mediated tetracycline resistant Neisseria gonorrhoeae in Zaire; Van Dyck E et al.; A cohort of 650 prostitutes from Kinshasa, Zaire, was followed at monthly intervals for sexually transmitted diseases as part of an HIV intervention project . Neisseria gonorrhoeae isolates, obtained during a period of 30 months, were auxotyped, serotyped and tested for antimicrobial susceptibility . Among 1085 gonococcal isolates tested, 725 (67%) produced beta-lactamase (PPNG) and 323 (30%) showed plasmid-mediated resistance to tetracyline (TRNG) . Over time, the prevalence of PPNG varied between 60 and 73%, while the level of TRNG increased from 11 to 45%PIP: During May 1988-October 1990 in Zaire, Neisseria gonorrhoeae isolates were obtained from 650 initially HIV-negative prostitutes in Kinshasa who were followed monthly for 30 months . After conservation of the gonococci, the N . gonorrhoeae isolates were then transported to the Institute of Tropical Medicine in Antwerp, Belgium, to test for antimicrobial resistance, especially tetracycline resistant isolates of N . gonorrhoeae . Among the 1085 isolates, 67% were resistant to penicillin (i.e., penicillinase producing N . gonorrhoeae {PPNG}) . 30% exhibited plasmid-mediated resistance to tetracycline (TRNG) . 37% were resistant to thiamphenicol . Thiamphenicol resistance was more common in non-TRNG isolates than TRNG isolates (49% vs . 8%; p 0.0001) . The frequency of TRNG among PPNG isolates was higher than it was among non-PPNG isolates (37% vs . 16%; p 0.001) . PPNG prevalence ranged from 60% to 73% . TRNG prevalence increased steadily from 11% to 45% during the 30-month period . Both TRNG and PPNG isolates were significantly associated with the auxotype/serovar class Pro-/IA-6 (p 0.0001 and p = 0.0002, respectively) . They were also associated with growth inhibition by 0.25 mM phenylalanine (p 0.0001 and p = 0.001, respectively) . The number of different TRNG auxotype/serovar classes ranged from 6 to 13 . It has been suggested that tetracycline use to control gonorrhea in the US and in the Netherlands increased the frequency and spread of TRNG . Only spectinomycin and ciprofloxacin were used to treat gonorrhea in this study . Yet, tetracycline was prescribed for genital Chlamydia trachomatis infection, which many of the prostitutes had . Also, males self-medicate for urethritis with tetracycline . Populations with a high incidence of gonococcal infections may experience an epidemic spread of TRNG .

Int J STD AIDS, 1995 Sep-Oct, 6(5), 305 - 12
Preventing pelvic infection after abortion; Stevenson MM et al.; PIP: Induced abortion is one of the most frequent surgical procedures in the UK . Even though it is considered safe, it sometimes has complications and long-term sequelae . Pelvic inflammatory disease (PID) is the most prevalent complication and can lead to chronic pelvic pain, pain during intercourse, infertility, and a higher risk of ectopic pregnancy . Chlamydia trachomatis is perhaps the leading etiologic agent for PID among women who have undergone induced abortion and who develop PID . Gonorrhea is another major etiologic agent for PID . Strategies used to try to reduce pelvic infection revolve around administration of antibiotic prophylaxis based on demographic features and on the presence of certain organisms in the genital tract that may increase their risk (e.g., C . trachomatis and Neisseria gonorrhoeae) and universal antibiotic prophylaxis for all women undergoing abortion . Most of the literature suggests that antibiotic prophylaxis does provide some protection against PID but does not clearly indicate who should be screened and for which pathogens and who should be treated and with which antibiotics . Demographic features useful for identifying who should receive antibiotic prophylaxis are: a history of PID, single status, nulliparity, and youth (especially reliable for chlamydial infection) . Screening for bacterial vaginosis involves diagnosis based on 3 of 4 criteria: characteristic vaginal discharge, positive amine test, raised vaginal pH, and the presence of clue cells on microscopy of wet or stained preparations of vaginal discharge . Since C . trachomatis is the most important pathogen, drugs sensitive to it should be administered: tetracyclines and erythromycin . Screening women seeking abortion for sexually transmitted diseases (STDs) provides an opportunity to educate them about STDs and treatment compliance and to contact their partners for investigation, treatment, and contact-tracing to reduce the STD-infected pool in the community .

Clin Lab Med, 1995 Sep, 15(3), 665 - 84
Update on laboratory diagnosis of sexually transmitted diseases; Woods GL; Several new techniques for diagnosis of the sexually transmitted diseases commonly seen in the United States have been developed during the past decade . The rapid, nonculture techniques for direct detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and herpes simplex virus are reviewed, and their advantages and disadvantages are compared with conventional diagnostic methods . In addition, new tests for diagnosis of syphilis, vaginitis, and vaginosis are briefly discussed.

Zh Mikrobiol Epidemiol Immunobiol, 1995 Sep-Oct, (5), 37 - 40
{The effect of the conditions of the controlled cultivation of Neisseria meningitidis on the lipopolysaccharide yield}; Stukalova NV et al.; The preparations of lipopolysaccharide (LPS) were obtained from N.meningitidis cells grown in synthetic nutrient medium in a fermenter . Different N.meningitidis strains, groups A, B, C, were found to give different yield of the preparation of LPS: the maximum yield was obtained from N.meningitidis cells C 133 and the minimum yield, from N.meningitidis cells B 125 . The comparison of group B strains 125 and 150 showed that the former gave a higher yield of LPS . The influence of the phase of growth on the yield of the preparation of LPS obtained from N.meningitidis cells 125 was shown to be manifested under the conditions of oxygen excess; in the exponential phase the highest yield of the preparation was observed . Oxygen dissolved in the medium was found to influence the yield of the preparation of LPS: with the same content of glucose in the medium the yield of LPS was higher under the conditions of the limited concentration of oxygen; with the same concentration of oxygen in the medium the yield of LPS increased with the increase of the concentration of glucose.

Obstet Gynecol, 1995 Sep, 86(3), 360 - 6
The inflammatory Papanicolaou smear: what does it mean?
Eckert LO, Koutsky LA, Kiviat NB, Krone MR, Stevens CE, Eschenbach DA.
OBJECTIVE: To determine the correlation between inflammation detected on Papanicolaou smear and specific lower genital tract agents, and, based on these findings, to develop recommendations for follow-up tests and treatment of young women with inflammation on smears . METHODS: A high-risk population of 779 randomly selected women attending a sexually transmitted disease (STD) clinic and a low-risk population of 1050 consecutive women presenting for annual examination at a university student health center underwent a standardized history and gynecologic examination . Univariate and multivariate analyses, focusing on the association between dense inflammation on Papanicolaou smear and specific lower genital tract pathogens or findings on cervical examination, were done for each population . RESULTS: Dense inflammation was present on the Papanicolaou smear of 256 (33%) of the 779 women in the STD clinic and 200 (19%) of 1050 students . Dense inflammation on Papanicolaou smear was independently associated with mucopus, cervical ectopy, cervical infection with Neisseria gonorrhoeae, Chlamydia trachomatis, herpes simplex virus (HSV), and vaginal infection by Trichomonas vaginalis in the STD population; in the student population, it was associated with cervical ectopy, C trachomatis, and mucopus . CONCLUSION: Although dense inflammation on Papanicolaou smear was a common finding in both the high- and low-risk populations, about half of the inflammation detected in the high-risk setting was associated with a specific microbial organism (C trachomatis, N gonorrhoeae, HSV, or T vaginalis), whereas less than 10% of the dense inflammation detected in the low-risk setting was linked with a specific pathogen (C trachomatis) . In both settings, a substantial population of sexually active women had dense inflammation associated with cervical ectopy but none of the specific organisms evaluated in this study.

J Med Microbiol, 1995 Sep, 43(3), 208 - 15
Utility of ribotyping, restriction endonuclease analysis and pulsed-field gel electrophoresis to discriminate between isolates of Neisseria gonorrhoeae of serovar IA-2 which require arginine, hypoxanthine or uracil for growth; Li H et al.; Neisseria gonorrhoeae isolates that require arginine--i.e., either citrulline (C), or ornithine (O)--uracil (U) and hypoxanthine (H) have generally been considered to be similar when characterised by auxotype, serovar and plasmid content . The MICs of penicillin, tetracycline, erythromycin, spectinomycin, cefoxitin and ceftriaxone for 552 isolates belonging to serovar IA-2 with these phenotypes were found to be similar . Therefore, restriction fragment length polymorphism analysis of rRNA genes (ribotyping), restriction enzyme (RE) analysis of chromosomal DNA, and pulsed-field gel electrophoresis (PFGE) were evaluated to determine whether these isolates could be distinguished by molecular methods . A subset of 27 isolates of N . gonorrhoeae that were OUH-requiring, CUH-requiring or OH-requiring, belonged to serovar IA-2 and carried a 2.6-MDa plasmid, were selected for further study . Based on the RE analysis of SmaI-digested genomic DNA, the 27 isolates fell into a single RE pattern, five ribotypes and 17 PFGE profiles which did not correlate with the specific arginine-requiring subtypes of these isolates . Each ribotype varied by the presence of only a single fragment, which was of a different size in each pattern, and 17 (63%) of the 27 isolates belonged to ribotype I . PFGE yielded the highest level of discrimination with 17 different profiles.

J Med Microbiol, 1995 Sep, 43(3), 201 - 7
Differentiation of Neisseria gonorrhoeae IB-3 and IB-7 serovars by direct sequencing of protein IB gene and pulsed-field gel electrophoresis; Poh CL et al.; Serotyping of Neisseria gonorrhoeae based on the differential recognition by a panel of six monoclonal antibodies (MAbs) against Protein IB (PIB) is a valuable tool for studying the epidemiology of gonorrhoea . However, the predominance of certain serovars in specific geographic regions necessitates the development of new MAbs or new genotyping approaches . Nucleotide and amino-acid sequence analysis of PIB from strains within the IB-3 and IB-7 serovars revealed strain differences within the same serovar . Based on the generation of PIB nucleotide and deduced amino-acid sequences that centred on amino-acid residues 196 and 237, eight serovar IB-3 strains and nine serovar IB-7 strains were separately subdivided into five groups . Intra-serovar differences were also established by pulsed-field gel electrophoresis (PFGE) of macro-restriction fragments generated by SpeI- and NheI-cleavage of genomic DNA . There was good correlation between the results based on PIB gene PCR-sequencing and those based on PFGE analysis of macro-restriction fragment patterns . These data demonstrate the high precision of PFGE analysis and indicate that this approach can be used as a rapid epidemiological subtyping system for major serovars of N . gonorrhoeae.

Curr Microbiol, 1995 Sep, 31(3), 146 - 51
Surface antigen analysis of group B Neisseria meningitis outer membrane by monoclonal antibodies: identification of bactericidal antibodies to class 5 protein; Danelli MG et al.; Twenty-four monoclonal antibodies (mAbs) against group B Neisseria meningitidis surface antigens were analyzed by immunoenzymatic assays and by a bactericidal test . Two mAbs were specific to polysaccharide B and one to lipopolysaccharide . The others were specific to polysaccharide B and one to lipopolysaccharide . The others were directed against outer membrane proteins ranging in molecular mass from 25 to 200 kDa . The outer membrane protein epitopes recognized by the mAbs were not conformational and were located on the outer surface of the microorganism . Linear epitopes on the class 5 protein, exposed on the surface of the membrane, were able to induce bactericidal antibodies to the homologous strain . The susceptibility of Neisseria meningitidis to these antibodies was unchanged when this organism was cultivated under conditions of iron depletion . These results demonstrate that peptides derived from class 5 proteins are potentially important in synthetic peptide or in recombinant protein vaccines containing linear bactericidal epitopes.

Sex Transm Dis, 1995 Sep-Oct, 22(5), 310 - 1
High-level quinolone resistance in Neisseria gonorrhoeae: a report of two cases; Tapsall JW et al.; BACKGROUND AND OBJECTIVES: Fluoroquinolones are widely used oral agents for treating Neisseria gonorrhoeae . Resistance to these agents is sporadic and usually at a low level . Two instances of high-dose ciprofloxacin regimens failing in the treatment of gonococcal infection, caused by strains with high-level quinolone resistance, are reported . STUDY DESIGN: This is a case report . CONCLUSION: High-level resistance to quinolone antibiotics resulting in treatment failure was observed in two distinct gonococcal isolates from patients infected in the Philippines (ciprofloxacin; minimal inhibitory contribution = 16 mg/l) . Continued monitoring of the quinolone sensitivity of Neisseria gonorrhoeae is appropriate and prudent.

Sex Transm Dis, 1995 Sep-Oct, 22(5), 289 - 95
Risk factors for infection in women undergoing testing for Chlamydia trachomatis and Neisseria gonorrhoeae in Manitoba, Canada; Jolly AM et al.; BACKGROUND AND OBJECTIVES: Despite sharing common modes of transmission, characteristics of individuals infected with Chlamydia trachomatis differ in several respects from those with Neisseria gonorrhoeae infection . Further characterization of women at high risk for chlamydial infection is needed to deliver appropriate and effective preventive, diagnostic, and therapeutic care to this population . GOAL OF THIS STUDY: The demographic and socioeconomic characteristics of women with laboratory confirmed chlamydia, gonorrhea, or coinfection were compared with those of control women who tested negative for both pathogens . STUDY DESIGN: A random sample of 400 women in Manitoba, Canada, who had undergone testing for sexually transmitted diseases at a public health laboratory in 1988 were studied . After linkage with medical insurance and census databases, logistic regression analysis was used to compare age, ethnicity, urban status, and mean income (using postal codes) of women with gonorrhea alone, chlamydia alone, and coinfection, with the same data for women who tested negative for both organisms . RESULTS: Young age, North American Indian status, urban residence, and low mean income according to postal code were significantly associated with gonococcal and chlamydial infection in the study population, compared with women who tested negative for both infections . Young age, Indian status, and urban residence also were associated with gonorrhea infection alone . Only young age and Indian status were associated with chlamydial infection . Mean incomes of women with chlamydial infection alone and control subjects were higher than those of women with gonorrhea and gonorrhea and chlamydia coinfection . CONCLUSIONS: Differences in the demographic and socioeconomic characteristics of women with gonorrhea, chlamydia, and coinfection suggest the existence of multiple reservoirs of infection due to these agents in the study population . The preventive, diagnostic, and therapeutic strategies of sexually transmitted disease control programs must be adapted to the individual needs of identified high-risk groups.

Infect Dis Clin North Am, 1995 Sep, 9(3), 783 - 804
Antimicrobial prophylaxis of infection; Dickinson GM et al.; Antimicrobial agents are used to prevent infections in a variety of clinical circumstances . In certain instances, the precise indications for prophylaxis remain controversial, and the preferred regimens undergo alterations based upon evolving clinical experience, changing patterns of microbial susceptibility, and innovations in medical and surgical practice . This article outlines the general principles underlying the use of antimicrobial prophylaxis and presents recommendations for the use of such prophylaxis in three areas: (1) surgery involving contaminated, clean-contaminated, and clean procedures; (2) prevention of infections due to specific pathogens, including Neisseria meningitidis, Hemophilus influenzae, Streptococcus pneumoniae, and Streptococcus pyogenes; and (3) prevention of infective endocarditis.

APMIS, 1995 Sep, 103(9), 609 - 27
Fastidious gram-negative bacteria: meeting the diagnostic challenge with nucleic acid analysis; Tonjum T et al.; The extended panorama of fastidious Gram-negative bacteria (FGNB) as opportunistic etiological agents of infectious diseases in immunocompromised patients is largely due to improved medical expertise and technology . The heightened awareness of infectious diseases due to FGNB species mandates comprehensive classification and identification systems as a basis for rapid and reliable diagnostics . The most useful approaches are combinations of nucleic acid techniques such as hybridization, genetic transformation, amplification and base sequence analysis with selected conventional criteria . Among these approaches, the widely distributed feature of natural competence in these organisms facilitates the use of the biological method of genetic transformation as a valuable addition to the more common nucleic acid techniques . We describe the development of the taxonomy of FGNB through the last four decades, with particular emphasis on the families Neisseriaceae, Moraxellaceae, and Pasteurellaceae.

Proc Natl Acad Sci U S A, 1995 Aug 15, 92(17), 7986 - 90
Role of pili and the phase-variable PilC protein in natural competence for transformation of Neisseria gonorrhoeae; Rudel T et al.; The Gram-negative bacterial pathogen Neisseria gonorrhoeae is naturally competent for transformation with species-related DNA . We show here that two phase-variable pilus-associated proteins, the major pilus subunit (pilin, or PilE) and PilC, a factor known to function in the assembly and adherence of gonococcal pili, are essential for transformation competence . The PilE and PilC proteins are necessary for the conversion of linearized plasmid DNA carrying the Neisseria-specific DNA uptake signal into a DNase-resistant form . The biogenesis of typical pilus fibers is neither essential nor sufficient for this process . DNA uptake deficiency of defined piliated pilC1,2 double mutants can be complemented by expression of a cloned pilC2 gene in trans . The PilC defect can also be restored by the addition of purified PilC protein, or better, pili containing PilC protein, to the mutant gonococci . Our data suggest that the two phase-variable Pil proteins act on the bacterial cell surface and cooperate in DNA recognition and/or outer membrane translocation.

Oral Microbiol Immunol, 1995 Aug, 10(4), 247 - 55
Microbial alterations in supragingival dental plaque in response to a triclosan-containing dentifrice; Zambon JJ et al.; A total of 325 subjects were entered into a double-blind, stratified 2-treatment clinical study that examined the effects of a dentifrice containing 0.3% triclosan, 2% Gantrez copolymer and 0.243% sodium fluoride on supragingival dental plaque and gingivitis . A subset of 159 subjects including 72 men and 87 women participated in the microbiological component of this study, which was designed to detect shifts in supragingival bacterial species in response to triclosan . Subjects were divided into two groups: one performed normal oral hygiene with the triclosan/copolymer dentifrice and a control group used a placebo dentifrice without triclosan . At baseline, 3 and 6 months during treatment and at 6, 12, 18 and 24 weeks post-treatment, supragingival dental plaque was collected from the buccal and lingual surfaces of the 4 first molar teeth and assayed for: 1) bacterial morphotypes by phase-contrast microscopy; 2) select bacterial groups and bacterial species by culture; and 3) target periodontal pathogens including Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis and Prevotella intermedia by immunofluorescence microscopy . There were few statistically significant differences between treatment groups in any of the 90 microbiological parameters measured at the 7 different time points . The control group demonstrated significantly higher levels of cultivable Neisseria and higher proportions at the 3-month treatment period of P . gingivalis-infected subjects and mean cells . After 6 months of treatment, the triclosan group demonstrated higher levels of fusiforms . Analysis of triclosan resistance data failed to detect a shift towards increased proportions of bacteria resistant to triclosan, and both treatment groups demonstrated triclosan-resistant strains, predominantly Veillonella dispar . This study confirms the microbiological safety of triclosan-containing dentifrices and suggests that continued use can be associated with beneficial alterations in the bacterial composition of supragingival dental plaque.

S Afr Med J, 1995 Aug, 85(8), 775 - 80
Characterisation of penA and tetM resistance genes of Neisseria gonorrhoeae isolated in southern Africa--epidemiological monitoring and resistance development; Chalkley LJ et al.; OBJECTIVE: To investigate penA and tetM resistance gene variation of Neisseria gonorrhoeae in order to define gene types for epidemiological monitoring and resistance development . DESIGN: Isolates of N . gonorrhoeae which were susceptible and resistant to penicillin and/or tetracycline were selected . Strains comprised South African isolates (22 from Bloemfontein, 13 from Transvaal, 20 from the Cape) and 15 Botswana and 4 Namibia isolates . The penA genes (2 kb) of all strains and tetM genes (765 bp) of 11 high-level tetracycline-resistant strains were amplified and restricted with HpaII . RESULTS AND CONCLUSIONS: Twelve different HpaII fingerprint patterns were obtained from the 74 isolates analysed for penicillin-binding protein (PBP) 2 gene (penA) alterations . Focusing on the transpeptidase domain, 25 isolates (3 whole gene patterns, minimal inhibitory concentrations (MICs) < or = 0.03-0.125 micrograms/ml) had restriction sites equivalent to those previously described for a susceptible strain . Of the remaining 9 PBP 2 gene groups, 25 strains fell into a designated group E . Penicillin/ penicillin + clavulanic acid MICs determined on these group E isolates gave a range of 0.125-2.0 micrograms/ml, although MICs against 4 strains were < or = 0.03 micrograms/ml . MICs of penicillin/penicillin + clavulanic acid for the 24 isolates that contained altered PBP 2 transpeptidase gene regions not designated group E were only < or = 0.03-0.125 micrograms/ml . The lack of a HpaII restriction site at nucleotide 1934 in the PBP 2 gene of group E strains was indicative of a small terminal region of N . cinerea DNA . This gene block, which was found in all the southern African areas studied, appears to predispose isolates to increased penicillin resistance . The 25.2 MDa conjugative plasmid carrying the tetM resistance determinant was readily demonstrated in 11 Botswana/Namibia isolates exhibiting high-level resistance to tetracycline (MICs > or = 16 micrograms/ml) . The tetM gene was shown to be of the American type.

Am J Obstet Gynecol, 1995 Aug, 173(2), 534 - 43
Determinants of cervical ectopia and of cervicitis: age, oral contraception, specific cervical infection, smoking, and douching; Critchlow CW et al.; OBJECTIVE: Our purpose was to assess determinants of cervical ectopia and cervicitis, specifically after adjustment for cervical infection . STUDY DESIGN: A cross-sectional study was conducted with colposcopic, cytologic, and microbiologic examination of 764 randomly selected women attending a sexually transmitted disease clinic and 819 consecutive college students undergoing routine annual examination . RESULTS: After we controlled for potential confounders, cervical ectopia was positively associated with oral contraception and Chlamydia trachomatis infection and negatively associated with aging in both populations, with recent vaginal douching in patients with sexually transmitted diseases, and with current smoking in college students . Oral contraception wa also associated with the radius of ectopia, and among users of oral contraception ectopia was associated with duration of oral contraception . Cervicitis (evaluated by Gram stain, Papanicoloau smear, and colposcopy) was associated with cervical infection by C . trachomatis and cytomegalovirus (both populations) and with gonorrhea and cervical herpes simplex virus infection (patients with sexually transmitted diseases) . Cervicitis was independently associated with ectopia but not with oral contraception after we adjusted for these four cervical infections . However, oral contraception was associated with edema and erythema of the zone of ectopia among women without cervical infection . CONCLUSIONS: Oral contraception, aging, cervical infection, smoking, and douching have effects on cervical ectopia that may influence the acquisition, transmission, or effects of sexually transmitted agents . Ectopia is associated with young age, oral contraception, and cervical infection; cervicitis is associated with ectopia and cervical infection by C . trachomatis, Neisseria gonorrhoeae, herpes simplex virus, and cytomegalovirus . In women without cervical infection, edema and erythema of the zone of ectopia are associated with oral contraception.

Am J Ophthalmol, 1995 Aug, 120(2), 143 - 50
Anterior chamber contamination after uncomplicated phacoemulsification and intraocular lens implantation; Samad A et al.; PURPOSE: To determine the frequency of anterior chamber contamination occurring during uncomplicated cataract surgery with intraocular lens implantation using phacoemulsification through a scleral tunnel incision . METHODS: In a prospective study, anterior chamber aspirates of one eye each of 103 consecutive ambulatory patients who underwent uncomplicated cataract surgery with lens implantation were cultured . The anterior chamber aspirate was withdrawn immediately upon the completion of surgery . Conjunctival cultures of the same eye were taken immediately before surgery, after the eye and periocular area had been prepared and draped . Multiple use topical medications used preoperatively on all patients were cultured at the end of the surgical day . RESULTS: Intraocular aspirates yielded positive cultures in five specimens (5%), four of which were identified as Staphylococcus epidermidis . Quantification disclosed colony counts ranging between 100 and 200 colony forming units per milliliter . Results of conjunctival cultures were positive in ten specimens (10%) . Staphylococcus epidermidis was the most common isolate, identified in seven of the ten positive cultures . Positive intraocular and conjunctival culture results were not present concurrently in any patient . Microorganisms were recovered from the multiple use topical medications on nine of the 26 successive surgical days . Neisseria species was the most frequent isolate (7,44%), followed by S . epidermidis (5,31%) . The frequency of contamination of the anterior chamber was independent of wound width (chi 2 = 0.869) and operative time (chi 2 = 4.77) . CONCLUSION: Bacterial contamination of the anterior chamber was detected in five (5%) of the patients . This reduced incidence of contamination compared with that of previous studies may be related to the preoperative preparation, the surgical technique, or both . Contamination of the multiple use topical medications and bulbar conjunctiva define possible sources of pathogens that may enter the anterior chamber . The absence of any clinical ocular infection in all patients attests to the small inoculum sizes, as well as the ability of the anterior chamber to clear small bacterial loads.

J Exp Med, 1995 Aug 1, 182(2), 511 - 7
Adherence of pilus- Opa+ gonococci to epithelial cells in vitro involves heparan sulfate; Chen T et al.; Neisseria gonorrhoeae attaches to host epithelial cells via pili and opacity-associated (Opa) outer membrane proteins . Pilus- gonococci (Gc) of strain MS11 adhere to both human and nonhuman cells, but only when particular Opa proteins are expressed; OpaA+ variants adhere best, OpaC+ variants are next best, and the seven other Opa+ variants adhere poorly or not at all . The adherence of OpaA+ Gc to Chinese hamster ovary (CHO) cells is inhibited by heparin or heparan sulfate (HS), but not by chondroitin sulfate . OpaA+ Gc do not adhere to CHO cells devoid of HS proteoglycans; low concentrations of heparin restore OpaA+ Gc adherence to these HS-deficient CHO cells and high concentrations inhibit it . 3H-heparin binding to whole Gc parallels their adherence abilities (OpaA+ > OpaC+ > OpaH+ >> Opas B, D, E, F, G, I = Opa- = 0) . Opa proteins separated by SDS-PAGE also bind 3H-heparin . These data suggest that adherence of pilus-, Opa+ Gc involves HS-proteoglycan of eukaryotic cells.

Genitourin Med, 1995 Aug, 71(4), 234 - 8
The virgin population of Neisseria gonorrhoeae in Stockholm has decreased and antimicrobial resistance is increasing; Backman M et al.; AIMS--To investigate the evolution of chromosomal and plasmid mediated resistance for ampicillin and tetracycline of N gonorrhoeae strains in Stockholm during 1982-1993 . METHODS--A total of 404 gonococcal strains isolated in 1982, 1987, 1990, 1992, 1993 were analysed for minimal inhibitory concentrations (MIC) of ampicillin and tetracycline and for plasmid content . MIC values were determined by the agar dilution method and plasmid preparations were performed using alkaline lysis . To detect additional gonococcal strains with tet(M) plasmids all strains isolated in 1988-1989 and 1991, in all 234 isolates, were analysed retrospectively for MIC values of tetracycline . If an MIC value of > or = 4.0 mg/l was recorded plasmid analysis was performed . RESULTS--Increased proportions of chromosomally mediated resistance to tetracycline (p < 0.001) as well as plasmid mediated resistance to both ampicillin (p < 0.02) and tetracycline were found in the later part of the study . In 1991 the first gonococcus with tet(M) plasmid was isolated in Sweden . The proportion of strains with chromosomally mediated resistance for ampicillin did not change during the study period . The proportion of gonococcal strains with the 39 kb conjugative plasmid was increased in the later part of the study . CONCLUSIONS--The increased proportion of N gonorrhoeae strains with resistance to ampicillin and tetracycline is most likely due to importation of strains from areas with high prevalence of antibiotic resistant gonococci . The proportion of N gonorrhoeae strains with tet(M) plasmids is low in Sweden, but might increase in the same way as the proportion of PPNG strains has increased during 1982-1993.

Genitourin Med, 1995 Aug, 71(4), 207 - 11
Should screening of genital infections be part of antenatal care in areas of high HIV prevalence? A prospective cohort study from Kigali, Rwanda, 1992-1993 . The Pregnancy and HIV (EGE) Group; Leroy V et al.; OBJECTIVE--To study the prevalence and incidence of genital infections and their association with HIV-1 infection among pregnant women in Kigali, Rwanda . SUBJECTS AND METHODS--HIV+ and HIV- pregnant women were followed prospectively during the last three months of pregnancy . At enrolment, syphilis test (RPR) on blood sample, Chlamydiae trachomatis ELISA test on cervical smear, laboratory gonococcal culture, trichomonas and candida direct examination, CD4 lymphocyte count were performed . At each monthly follow-up clinic visit until delivery, genital infections were screened in the presence of clinical signs and symptoms . RESULTS--The HIV seroprevalence rate was 34.4% (N = 1233), 384 HIV+ women and 381 HIV- women of same parity and age were enrolled . Prevalence of genital infections at enrolment was generally higher in HIV+ women than in HIV- women: syphilis, 6.3% versus 3.7% (p = 0.13); Neisseria gonorrhoea, 7.0% versus 2.4% (p = 0.005); Trichomonas vaginalis, 20.2% versus 10.9% (p = 0.0007); Chlamydia trachomatis, 3.4% versus 5.5% (p = 0.21); Candida vaginalis, 22.3% versus 20.1% (p = 0.49) . Until delivery, the relative risk of acquiring genital infections was also higher in HIV+ women than in HIV- women: 1.0 for syphilis (95% CI: 0.5-2.2), 3.7 for Neisseria gonorrhoea (1.0-13.3), 2.6 for Trichomonas vaginalis (1.5-4.6) and 1.6 for Candida vaginalis (1.1-2.4) . CONCLUSION--In the context of high HIV-1 seroprevalence among pregnant women, prenatal care should include at least once screening for genital infections by clinical examination with speculum and a syphilis testing in Africa.

Clin Pediatr (Phila), 1995 Aug, 34(8), 419 - 23
Gonococcal and chlamydial genitourinary infections in symptomatic and asymptomatic adolescent women; Biro FM et al.; To determine the prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae endocervical infections in a group of adolescent women, gynecologic histories and evaluations were done on symptomatic and asymptomatic adolescent women presenting for pelvic examinations in an urban, hospital-based, adolescent clinic . C . trachomatis and N . gonorrhoeae cultures and three nonculture diagnostic tests for chlamydia (two ELISA assays and one DNA-probe) were performed on each patient . Patients were 12 to 21 years of age (mean 17.0); 53% were African-American, and 47% were Caucasian . Two hundred twenty-eight women were asymptomatic and 249 women had symptoms . There were 64 cases of chlamydia and 19 cases of gonorrhea; an additional 11 patients had both chlamydia and gonorrhea . Approximately one third of patients with chlamydia, gonorrhea, or both had asymptomatic disease an important reservoir of infection.

Clin Pediatr (Phila), 1995 Aug, 34(8), 415 - 8
The management of prepubertal children with gonorrhea; Christian CW et al.; To determine whether test-of-cure cultures are necessary for prepubertal children diagnosed with Neisseria gonorrhoeae, we examined the records of all 66 patients < 10 years old seen at Children's Hospital of Philadelphia over a 7.5-year period (1987-1994) diagnosed with gonorrhea . Ninety-eight percent had genital discharge on examination . All children with genital gonorrhea were symptomatic, but only 10% of children with rectal gonorrhea and 20% with pharyngeal infection were symptomatic . Seventy-seven percent of children were treated with ceftriaxone . Of these, 72% returned for test-of-cure cultures . Ninety-three percent of children had complete resolution of physical symptoms at test-of-cure, and all follow-up cultures were negative for N . gonorrhoeae . Our data suggest that most prepubertal children with gonorrhea are symptomatic at initial presentation and are cured after recommended treatment with ceftriaxone . The Centers for Disease Control and Prevention recommendations for obtaining test-of-cure cultures in young children with gonorrhea are unnecessary, potentially harmful, and should be revised.

Zhonghua Liu Xing Bing Xue Za Zhi, 1995 Aug, 16(4), 217 - 9
{The study of Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum infection status by polymerase chain reaction}; Kong FR et al.; The study involved 355 specimens of STD clinic patients collected from Beijing, Shantou and Wuhan, for the detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum by polymerase chain reaction . The results showed that the positive rate of Neisseria gonorrhoeae being the highest in the patients attending STD clinics from the three cities . The detection rate of Neisseria gonorrhoeae in male patients was higher than in females . The positive rates of Chlamydia trachomatis and Ureaplasma urealyticum in the three cities differed from each other greatly . Polymerase chain reaction was suitable for clinical Jetection and epidemiological study of the three kinds of STD pathogens.

Infect Immun, 1995 Aug, 63(8), 3125 - 30
The inflammatory cytokine response to Chlamydia trachomatis infection is endotoxin mediated; Ingalls RR et al.; Chlamydia trachomatis is a major etiologic agent of sexually transmitted diseases . Although C . trachomatis is a gram-negative pathogen, chlamydial infections are not generally thought of as endotoxin-mediated diseases . A molecular characterization of the acute immune response to chlamydia, especially with regard to the role of its lipopolysaccharide (LPS), remains to be undertaken . We extracted 15 mg of LPS from 5 x 10(12) C . trachomatis elementary bodies (EB) for analysis of structure and biological activity . When methylated lipid A was subjected to high-pressure liquid chromatography followed by mass spectrometry, the majority of the lipid A was found to be pentaacyl . The endotoxin activities of whole C . trachomatis EB and purified LPS were characterized in comparison with whole Salmonella minnesota R595 and with S . minnesota R595 LPS and lipooligosaccharide from Neisseria gonorrhoeae . Both C . trachomatis LPS and whole EB induced the release of tumor necrosis factor alpha from whole blood ex vivo, and C . trachomatis LPS was capable of inducing the translocation of nuclear factor kappa B in a Chinese hamster ovary fibroblast cell line transfected with the LPS receptor CD14 . In both assays, however, C . trachomatis was approximately 100-fold less potent than S . minnesota and N . gonorrhoeae . The observation that C . trachomatis is a weak inducer of the inflammatory cytokine response correlates with the clinical observation that, unlike N . gonorrhoeae infection, genital tract infection with C . trachomatis is often asymptomatic . The ability of specific LPS antagonists to completely inhibit the tumor necrosis factor alpha-inducing activity of whole C . trachomatis EB suggests that the inflammatory cytokine response to chlamydia infection may be mediated primarily through LPS . This implies that the role of other surface protein antigens, at least in terms of eliciting the proinflammatory cytokine response, is likely to be minor.

Antimicrob Agents Chemother, 1995 Aug, 39(8), 1864 - 5
Susceptibilities of Neisseria gonorrhoeae to the glycylcyclines; Whittington WL et al.; To assess the activities of two glycylcyclines, N,N-dimethylglycylamido (DMG) derivatives of minocycline (MINO) and 6-demethyl-6-deoxytetracycline (DMDOT), 203 gonococcal isolates recovered at six sexually transmitted disease clinics in the western United States were evaluated . Antimicrobial susceptibilities to tetracycline HCl, doxycycline, MINO, DMG-DMDOT, and DMG-MINO were determined by agar dilution tests . DMG-DMDOT and DMG-MINO were more active than tetracycline HCl, doxycycline, or MINO regardless of the presence of Tet M or of chromosomal mutations mediating tetracycline resistance (P < 0.001).

FEMS Microbiol Lett, 1995 Jul 15, 130(1), 37 - 44
Evaluation of the chick embryo for the determination of relative virulence of Neisseria meningitidis; Pine L et al.; The chick embryo model was evaluated as a method to compare virulence between selected strains of Neisseria meningitidis . Inoculation of 13-day-chick embryos via the egg yolk distinguished strains having an LD50 of 10(3) colony forming units (CFU) or greater (low virulence) from those having an LD50 of approximately 10(1) or less (high virulence) . A strain of serogroup B and a spontaneous nonpiliated strain of group C were found to be of relatively high virulence while a strain of N . lactamica, a serogroup A carrier strain, and certain nongroupable strains were found to be of low virulence . Strains having an LD50 of 10(2) were not differentiated from either of these . Alternatively, inoculation of the chorioallantoic membrane (CAM) of 9-day-old chick embryos statistically differentiated most strains of N . meningitidis although inoculation via this route was less sensitive.

Ann Acad Med Singapore, 1995 Jul, 24(4), 579 - 83
Antimicrobial therapy of non-viral sexually transmitted diseases--an update; Chan RK; Azithromycin is an azalide antibiotic with important properties which allow it to be used as a single-dose treatment for genital Chlamydia trachomatic infections . A single 1 g dose is as effective as a standard seven-day course of doxycycline . Ofloxacin 400 mg bid for seven days is also effective against Chlamydia trachomatis . Both azithromycin 2 g and ofloxacin are also effective against uncomplicated gonorrhoea . Neisseria gonorrhoeae continues to be sensitive to third generation cephalosporins, e.g . ceftriaxone 125 mg . Oral single dose cephalosporins offer ease of administration and safety, e.g . cefixime (400 mg), cefuroxime axetil (1 g) and cefpodoxime proxetil (200 mg) . The fluoroquinolones, e.g . ciprofloxacin (500 mg) and ofloxacin (400 mg), are being increasingly used as first-line medications, however, caution is recommended as the development of resistance is anticipated and already being detected in many areas . Syphilis continues to be sensitive to penicillin . This should be administered parenterally . Coexistent human immunodeficiency virus infection may make standard therapy inadequate, and closer follow-up is recommended . Therapy with non-penicillin antibiotics is still inadequately studied . Chancroid is treated with ceftriaxone, ciprofloxacin, azithromycin, or erythromycin . In some areas, resistance to tetracyclines and TMP-SMX has made these drugs ineffective as first-line treatments . Bacterial vaginosis is effectively treated with a single dose of metronidazole 1 g or 500 mg bid over seven days . Similar regimens are also effective against trichomoniasis . Vulvovaginal candidiasis can be treated with topical imidazole preparations or oral antifungal medications.

Ann Acad Med Singapore, 1995 Jul, 24(4), 515 - 8
Comparing the efficacy of pefloxacin and ciprofloxacin in the treatment of acute uncomplicated gonococcal urethritis in males; Leow YH et al.; The aim of this study was to compare the efficacy of single-dose pefloxacin 400 mg and ciprofloxacin 250 mg in the treatment of acute uncomplicated gonococcal urethritis in males . One hundred and twenty male patients with uncomplicated gonococcal urethritis were assigned alternately to receive single oral doses of either pefloxacin 400 mg or ciprofloxacin 250 mg . Forty-one out of 43 patients (95.3%) of the pefloxacin group and 46 of 47 (97.9%) of the ciprofloxacin group were cured of gonorrhoeae . The rates of post-gonococcal urethritis were 57.7% and 53.3% in the pefloxacin and ciprofloxacin groups respectively . There was a high incidence of penicillinase-producing gonococci (34.2%) . High level resistance to pefloxacin (minimum inhibitory concentration {MIC} >1.0 mg/l) resulting in clinical failure on 400 mg stat dose was noted in 1 isolate . It also showed decreased susceptibility to ciprofloxacin (MIC 0.25 mg/l) . Another isolate showed high-level resistance (MIC 0.06 mg/l) to ciprofloxacin 250 mg stat dose with concomitant decreased susceptibility to pefloxacin (MIC >1.0 mg/l) . Ciprofloxacin 250 mg stat dose is still useful for the treatment of uncomplicated gonococcal urethritis in males . The cure rate of 95.3% with pefloxacin at 400 mg stat dose is acceptable, but needs to be monitored with caution . The emergence of a more resistant strain of Neisseria gonorrhoeae to fluoroquinolones calls for vigilance in the monitoring of antimicrobial susceptibility.

Indian J Public Health, 1995 Jul-Sep, 39(3), 93 - 9
Sexually transmitted disease services in Madras: could their role in AIDS prevention be strengthened?
Kantharaj K, Mertens TE, Smith GD, Mugrditchian D, Van Dam CJ, Radhakrishnan KM.
A baseline evaluation of the quality of STD case management was conducted in five areas of Madras city in 1992, using HIV prevention indicators recommended by the World Health Organization . Eighty-four interviews and 108 observations of private and public clinic practitioners were conducted . Sixty-one percent of interviewed doctors reported making only specific "clinical" diagnoses and 17% reported making only the WHO recommended syndrome-based diagnoses while 22% reported making both types of diagnosis . The adequacy of treatment was compared against various standards, including the Indian National Guidelines for STD management . Almost half of the health care providers (HCP) reported using a treatment effective against the two main pathogens that may cause male urethritis, while 20% reported using a treatment that was not effective against either . For male ulcers only 12% of HCPs reported using treatment effective against both syphilis and chancroid . Seventy-nine percent of the HCP reported that they advised their patients to use condoms, but in 30% only of observed consultations, condoms were promoted for STD or HIV/AIDS prevention . As information concerning the relative prevalence of pathogens in different areas is unlikely to be available, there is an urgent need for the syndromic approach to STD treatment be adopted by health care providersPIP: During November-December 1992, in Madras, India, interviews were conducted with 84 public and private physicians who treated at least five sexually transmitted disease (STD) patients each week in Royapuram, Washerman, Evr High Road, T . Nagar, and Adyar regions of the city . Field workers also observed 108 medical consultations of 48 of the physicians interviewed . Researchers aimed to use the findings of this baseline survey to optimize STD services in order to reduce the population at risk of HIV/AIDS . 61% of the physicians interviewed made specific clinical diagnoses . 17% made only syndrome-based diagnoses . 22% used both types of diagnoses . 48% used a treatment effective against the two main pathogens for male urethritis, Neisseria gonorrhoeae and Chlamydia trachomatis . 20% used a treatment that was not effective against either of these pathogens . Only 12% used a treatment effective against chancroid and syphilis for men with genital ulcers . 29% did not use a treatment effective against chancroid or syphilis . For female genital ulcers, only 10.7% of physicians used an effective treatment against chancroid or syphilis . 21.4% provided treatment effective against neither chancroid nor syphilis in female STD patients with genital ulcers . 79% claimed to counsel their STD patients to use condoms, but only 30% were observed actually promoting condoms for STD or HIV/AIDS prevention . Physicians instructed only 6% of STD patients how to use condoms . Condoms were given to only one STD patient . These findings highlight the need for physicians and other health care providers to adopt the syndromic approach to STD treatment and for developing and evaluating innovative and effective programs of patient education in order to reduce the risk of HIV infection .

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