Neisseria

J Immunol, 2000 Dec 15, 165(12), 7125 - 32
Evidence for an accessory protein function for Toll-like receptor 1 in anti-bacterial responses; Wyllie DH et al.; Members of the Toll-like receptor (TLR) family are components of the mammalian anti-microbial response, signaling with a domain closely related to that of IL-1 receptors . In this report the expression and function of TLR1, a TLR of unknown function, are examined . TLR1 is expressed by monocytes, as demonstrated using a novel mAb . Monocytes also express TLR2 . TLR1 transfection of HeLa cells, which express neither TLR1 nor TLR2, was not sufficient to confer responsiveness to several microbial extracts . However, cotransfection of TLR1 and TLR2 resulted in enhanced signaling by HeLa cells to soluble factors released from Neisseria meningitidis relative to the response with either TLR alone . This phenomenon was also seen with high concentrations of some preparations of LPS . The N . meningitidis factors recognized by TLR1/TLR2 were not released by N . meningitidis mutant in the LpxA gene . Although LpxA is required for LPS biosynthesis, because cooperation between TLR1 and TLR2 was not seen with all LPS preparations, the microbial component(s) TLR1/2 recognizes is likely to be a complex of LPS and other molecules or a compound metabolically and chemically related to LPS . The functional IL-1R consists of a heterodimer; this report suggests a similar mechanism for TLR1 and TLR2, for certain agonists . These data further suggest that mammalian responsiveness to some bacterial products may be mediated by combinations of TLRs, suggesting a mechanism for diversifying the repertoire of Toll-mediated responses.

Bioinformatics, 2000 Oct, 16(10), 932 - 40
Finding pathogenicity islands and gene transfer events in genome data; Lio P et al.; MOTIVATION: There is a growing literature on wavelet theory and wavelet methods showing improvements on more classical techniques, especially in the contexts of smoothing and extraction of fundamental components of signals . G+C patterns occur at different lengths (scales) and, for this reason, G+C plots are usually difficult to interpret . Current methods for genome analysis choose a window size and compute a chi(2) statistics of the average value for each window with respect to the whole genome . RESULTS: Firstly, wavelets are used to smooth G+C profiles to locate characteristic patterns in genome sequences . The method we use is based on performing a chi(2) statistics on the wavelet coefficients of a profile; thus we do not need to choose a fixed window size, in that the smoothing occurs at a set of different scales . Secondly, a wavelet scalogram is used as a measure for sequence profile comparison; this tool is very general and can be applied to other sequence profiles commonly used in genome analysis . We show applications to the analysis of Deinococcus radiodurans chromosome I, of two strains of Helicobacter pylori (26695, J99) and two of Neisseria meningitidis (serogroup B strain MC58 and serogroup A strain Z2491) . We report a list of loci that have different G+C content with respect to the nearby regions; the analysis of N . meningitidis serogroup B shows two new large regions with low G+C content that are putative pathogenicity islands . AVAILABILITY: Software and numerical results (profiles, scalograms, high and low frequency components) for all the genome sequences analyzed are available upon request from the authors.

Epidemiol Infect, 2000 Oct, 125(2), 285 - 98
Genetic characterization of a new variant within the ET-37 complex of Neisseria meningitidis associated with outbreaks in various parts of the world; Jelfs J et al.; A new variant within the electrophoretic type (ET)-37 complex of Neisseria meningitidis, ET-15, first detected in Canada in 1986, has been associated with severe clinical infections and high mortality rates in several European countries, Israel and Australia . To ascertain the genetic and epidemiological relationships of ET-15 strains from different geographical areas, 72 ET-15 isolates from 10 countries were compared to 13 isolates representing other clones of the ET-37 complex . The 85 strains were analysed by pulsed-field gel electrophoresis (PFGE) using 2 restriction endonucleases and Southern hybridization with 10 genetic markers . Four ET-15 strains and 4 other strains of the ET-37 complex were further examined using an additional restriction enzyme and a total of 18 genetic markers . PFGE fingerprints of the ET-15 strains were closely related . Strains within each country were even more closely related, suggesting single introductions of the clone . Physical mapping of genes in ET-15 and other strains of the ET-37 complex demonstrated that large genetic rearrangements of the genome have occurred in association with the appearance of the ET-15 variant.

J Infect Dis, 2001 Jan 15, 183(2), 351 - 354 Epub 2000 Dec 08.
Potency of a genetically detoxified mucosal adjuvant derived from the heat-labile enterotoxin of Escherichia coli (LTK63) is not adversely affected by the presence of preexisting immunity to the adjuvant; Ugozzoli M et al.; The objective of the current studies was to evaluate whether the potency of a genetically detoxified mucosal adjuvant, derived from heat-labile enterotoxin of Escherichia coli (LTK63), was adversely affected by preexisting immunity . Studies of mice and pigs have involved consecutive intranasal immunization with LTK63 and 2 different vaccines (influenza virus hemagglutinin and a protein-polysaccharide conjugate of Neisseria meningitidis group C) . The antibody responses to the vaccines plus LTK63 in naive animals were compared with the responses achieved in animals that previously had been immunized with the alternate vaccine plus LTK63 . The data showed that the responses of both animal models to intranasal immunization were not adversely affected by the presence of preexisting immunity to the LTK63 adjuvant.

Int Microbiol, 1999 Sep, 2(3), 133 - 6
Interactions between encapsulated Neisseria meningitidis and host cells; Nassif X; A major feature of Neisseria meningitidis is its ability to invade human brain meninges . To access the meninges, the bacteria must cross the blood-brain barrier (BBB), which is one of the tightest barriers in the body . Therefore, N . meningitidis must have evolved some type of sophisticated means to bypass the physical properties of this cellular barrier . As N . meningitidis is encapsulated when present in the bloodstream, this review will focus on the mechanisms that encapsulated N . meningitidis has developed to interact with host cells and will suggest ways in which these mechanisms may be helpful for crossing the BBB.

Int Microbiol, 1998 Mar, 1(1), 59 - 63
Multicolonization of human nasopharynx due to Neisseria spp; Saez Nieto JA et al.; The colonization due to Neisseria spp . in the nasopharynx of forty healthy adults was studied by using a selective medium that allows the differentiation of Neisseria species and inhibits the rest of pharyngeal microbiota . The medium detected a variety of colonial morphology types and some metabolic characteristics of the isolates . We demonstrated the multicolonization by several Neisseria spp . in the same individual, and we isolated several strains of the same species, after analysis by pulsed-field gel electrophoresis (PFGE) patterns obtained from the different colonial types previously identified as the same species . The forty adults studied were colonized by 112 forms of Neisseria spp., and twelve colonization patterns were obtained: one species (45%), two (45%), three (7.5%) and four (2.5%) . N . perflava-N . sicca, either alone or in combination with other species was the most frequent isolate (92.5%) . The analysis of PFGE patterns obtained from different colonial types revealed the multicolonization by several strains of the same species in some individuals . This fact was found in N . perflava-N . sicca (50%) and N . mucosa (2.5%).

Indian J Pediatr, 2000 Oct, 67(10), 709 - 11
Factors predicting occult bacteremia in young children; Jamuna R et al.; A febrile child without a definite localizing sign of infection may be in initial phase of bacteremia which unless treated would result in systemic complication . These instances are referred to as "Occult bacteremia" . The common pathogens isolated in these children are Streptococcus pneumoniae, Hemophilus influenzae and Neisseria meningitidis . A hundred consecutive children in the age group of 3-36 months attending pediatric outpatient department and casualty were clinically evaluated using AIOS (acute illness observation scale) score and were subjected to complete blood counts, smear for malarial parasites, ESR and blood culture . In the 19-month study period, 4 instances of occult bacteremia were identified . Streptococcus pneumoniae was cultured in 3 cases and H . influenzae in one . A febrile and toxic child in the age group of 3-36 months has a high risk of occult bacteremia . High fever of temperature > or = 102 degrees F, ESR > or = 15 mm/hour, and total leukocyte count > or = 15,000/mm3, in a child with AIOS score of > or = 10 may be considered for more detailed investigations and early intervention with antimicrobial therapy.

Microbiol Mol Biol Rev, 2000 Dec, 64(4), 672 - 93
Molecular properties of bacterial multidrug transporters; Putman M et al.; One of the mechanisms that bacteria utilize to evade the toxic effects of antibiotics is the active extrusion of structurally unrelated drugs from the cell . Both intrinsic and acquired multidrug transporters play an important role in antibiotic resistance of several pathogens, including Neisseria gonorrhoeae, Mycobacterium tuberculosis, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Vibrio cholerae . Detailed knowledge of the molecular basis of drug recognition and transport by multidrug transport systems is required for the development of new antibiotics that are not extruded or of inhibitors which block the multidrug transporter and allow traditional antibiotics to be effective . This review gives an extensive overview of the currently known multidrug transporters in bacteria . Based on energetics and structural characteristics, the bacterial multidrug transporters can be classified into five distinct families . Functional reconstitution in liposomes of purified multidrug transport proteins from four families revealed that these proteins are capable of mediating the export of structurally unrelated drugs independent of accessory proteins or cytoplasmic components . On the basis of (i) mutations that affect the activity or the substrate specificity of multidrug transporters and (ii) the three-dimensional structure of the drug-binding domain of the regulatory protein BmrR, the substrate-binding site for cationic drugs is predicted to consist of a hydrophobic pocket with a buried negatively charged residue that interacts electrostatically with the positively charged substrate . The aromatic and hydrophobic amino acid residues which form the drug-binding pocket impose restrictions on the shape and size of the substrates . Kinetic analysis of drug transport by multidrug transporters provided evidence that these proteins may contain multiple substrate-binding sites.

J Clin Microbiol, 2000 Dec, 38(12), 4430 - 8
Comparison of sequencing of the por gene and typing of the opa gene for discrimination of Neisseria gonorrhoeae strains from sexual contacts; Viscidi RP et al.; Typing of gonococcal strains is a valuable tool for the biological confirmation of sexual contacts . We have developed a typing method based on DNA sequencing of two overlapping por gene fragments generated by a heminested PCR . We compared sequencing of the por gene (POR sequencing) and typing of the opa gene (OPA typing) for the characterization of strains from 17 sexual partnerships . Both methods were highly discriminatory . A different genotype was detected in 15 of the 17 epidemiologically unconnected couples by POR sequencing and in 16 of the 17 couples by OPA typing with restriction enzyme HpaII . Within partnerships, identical genotypes were obtained from 16 of the 17 known sex contacts by POR sequencing and from 15 of the 17 by OPA typing . Compared to OPA typing, which relies on interpretation of bands in a gel, DNA sequence data offer the advantage of being objective and portable . As costs for sequencing decline, the method should become affordable for most laboratory personnel who wish to type gonococcal strains.

EMBO J, 2000 Dec 1, 19(23), 6408 - 18
Components and dynamics of fiber formation define a ubiquitous biogenesis pathway for bacterial pili; Wolfgang M et al.; Type IV pili (Tfp) are a unique class of multifunctional surface organelles in Gram-negative bacteria, which play important roles in prokaryotic cell biology . Although components of the Tfp biogenesis machinery have been characterized, it is not clear how they function or interact . Using Neisseria gonorrhoeae as a model system, we report here that organelle biogenesis can be resolved into two discrete steps: fiber formation and translocation of the fiber to the cell surface . This conclusion is based on the capturing of an intermediate state in which the organelle is retained within the cell owing to the simultaneous absence of the secretin family member and biogenesis component PilQ and the twitching motility/pilus retraction protein PilT . This finding is the first demonstration of a specific translocation defect associated with loss of secretin function, and additionally confirms the role of PilT as a conditional antagonist of stable pilus fiber formation . These findings have important implications for Tfp structure and function and are pertinent to other membrane translocation systems that utilize a highly related set of components.

J Chromatogr A, 2000 Oct 13, 894(1-2), 311 - 8
Determination of free carrier protein in protein-polysaccharide conjugate vaccines by micellar electrokinetic chromatography; Lamb DH et al.; Protein-polysaccharide conjugate vaccines offer the prospect of reducing morbidity and mortality due to bacterial pneumonia and meningitis but, because of their size and heterogeneity, are often a challenge to characterize by traditional analytical methods . Vaccines consisting of Streptococcus pneumoniae, or Neisseria meningiditis polysaccharide covalently linked to formaldehyde-inactivated diphtheria toxoid carrier protein were resolved from non-conjugated toxoid by micellar electrokinetic chromatography . Separation was achieved using alkaline sodium borate solutions containing sodium dodecyl sulfate in excess of the critical micellar concentration . No sample pretreatment was required prior to analysis . Diphtheria toxoid peak migration times were highly reproducible . Measurement of absolute toxoid peak area showed poor precision, but good precision was observed when peak area was normalized against an internal standard (myoglobin) . Good linearity was observed over useful ranges of both protein content and injection time.

Acad Emerg Med, 2000 Dec, 7(12), 1437 - 9
Follow-up program for emergency department patients with gonorrhea or chlamydia; Kelly JJ et al.; OBJECTIVES: To study the performance of a centralized regional follow-up program organized by a municipal department of health (DH) for female patients presenting to the emergency department (ED) with Neisseria gonorrhoeae and/or Chlamydia trachomatis, who are not diagnosed or treated at the time of presentation . METHODS: This was a retrospective observational study of female patients seen in the ED with positive cervical specimens, and their subsequent treatment and follow-up by the DH . Medical records were reviewed to determine the female patients seen in the ED who had positive specimens for N . gonorrhoeae or C . trachomatis . The DH followed up those not treated in the ED . Analysis of how long it took for these patients to be treated and the proportion lost to follow-up was performed . RESULTS: Of 2,121 specimens, 342 were positive for N . gonorrhoeae or C . trachomatis . Of the 342, 154 (45%) were recognized and appropriately treated in the ED . One hundred fifty-nine of the 342 (46.5%) patients were discharged from the ED without treatment but were contacted by the DH and appropriate treatment was provided . The DH could not locate 23 (6.7%) patients, and four (1.2%) refused treatment . One died before treatment . Only 21 of the 159 were treated within nine days . Median time to treatment was 36 days . CONCLUSIONS: Centralized laboratory analysis and follow-up by the DH for N . gonorrhoeae and C . trachomatis identified many female patients undiagnosed and untreated in the ED . The DH follow-up program provided appropriate treatment to most female patients.

Curr Infect Dis Rep, 2000 Feb, 2(1), 44 - 50
The 1998 CDC Sexually Transmitted Diseases Treatment Guidelines; Workowski KA; Health care providers who evaluate persons who have or are at risk for sexually transmitted diseases (STDs) should be aware of current national guidelines for STD treatment and should provide assessment and treatment according to these recommendations . The Centers for Disease Control and Prevention STD Treatment Guidelines were developed by using an evidence-based approach incorporating available scientific evidence, specialist knowledge, and consultation with professional organizations and other agencies with diverse perspectives on STD treatment . The guidelines provide recommendations about new antimicrobial agents for the treatment of primary and recurrent genital herpes, a novel patient-applied therapy for genital warts, additional parenteral alternative regimens for the treatment of pelvic inflammatory disease, oral therapy for vaginal candidiasis, and alternative regimens for treatment of chlamydia in pregnancy . Expanded sections in the guidelines also address the management of urethritis, recognition of the acute retroviral syndrome, and the emergence of quinolone-resistant Neisseria gonorrhoeae . Prevention of sexually transmitted hepatitis A and hepatitis B through the use of preexposure vaccinations is discussed.

Int J STD AIDS, 2000 Nov, 11(11), 731 - 3
Evaluation of a peer education programme for female sex workers in Bali, Indonesia; Ford K et al.; The objective of this paper is to evaluate a peer education programme for female sex workers in Bali, Indonesia . Sex workers participated in face-to-face interviews and STD exams in August-September 1998 . In October 1998 one woman from each of 30 clusters was selected to be a peer educator and received a 2-day training on AIDS, STDs, condom use, and condom negotiation . After training, the peer educators were visited twice a week by field workers to answer questions and offer support . All sex workers received group education every 2 months . In January-February 1999, the sex workers again participated in face-to-face interviews and examinations . One month after peer education training, only 50% of the peer educators were still working in the clusters where they were trained . To evaluate the impact of the peer educators, sex workers in clusters where a peer educator continued to work were compared with sex workers in clusters where women did not continue to work (n=189) . In clusters where women continued to work, there were higher levels of AIDS knowledge (P < 0.05), STD knowledge (P < 0.05) and condom use (82 vs 73%, P=0.15) . The prevalence of Neisseria gonorrhoeae infection was also lower in clusters with a peer educator (39% vs 55%, P=0.05) than in clusters without a peer educator.

Commun Dis Intell, 2000 Sep, 24(9), 269 - 71
Surveillance of antibiotic resistance in Neisseria gonorrhoeae in the WHO Western Pacific Region, 1999 . The WHO Western Pacific Region Gonococcal Antimicrobial Surveillance Programme; Neisseria gonorrhoeae recJ mutants show defects in recombinational repair of alkylated bases and UV-induced pyrimidine dimers; Department of Biological Sciences, Northern Illinois University, DeKalb 60115, USA . sahill@niu.edu

Neisseria gonorrhoeae lacks several common DNA repair pathways found in other organisms . As recent evidence had indicated that gonococci use recombinational repair to repair UV-induced DNA lesions, this study examined whether the gonococcal RecJ homologue contributes in this repair capacity . The recJ gene from strain MS11 was cloned and sequenced and was found to show a considerable degree of identity to its Escherichia coli homologue . A N . gonorrhoeae delta recJ mutant was constructed and tested for recombinational proficiency as well as for defects in DNA repair . In the absence of the RecJ exonuclease, DNA transformation and pilin switching occurred at wild type levels, indicating that the efficiency of recombination remained unimpaired . In contrast, N . gonorrhoeae delta recJ mutants showed extreme sensitivity to low levels of UV irradiation and to exposure to DNA-alkylating reagents {e.g . ethyl methanesulfonate (EMS) and methyl methanesulfonate (MMS)} . Complementation of the gonococcal recJ mutant in cis restored resistance to low-level UV, indicating that the gonococcal RecJ protein is involved in recombinational repair, and can act independently of other single-strand-specific exonucleases . Furthermore, transformation competence was not required for RecJ-dependent DNA repair . Overall, the data show that N . gonorrhoeae recJ mutants present a unique phenotype when compared to their E . coli recJ counterparts, and further support the contention that RecORJ-dependent recombinational repair is a major DNA repair pathway in the genus Neisseria.

Commun Dis Intell, 2000 May, 24(5), 113 - 7
Annual report of the Australian Gonococcal Surveillance Programme, 1999; Tapsall J; The primary aim of the Australian Gonococcal Surveillance Programme (AGSP) is to monitor the antibiotic susceptibility of Neisseria gonorrhoeae . In 1999 the AGSP examined 3,740 isolates of gonococci from all States and Territories . The rates and sites of infection and antibiotic susceptibility patterns varied considerably between regions, reflecting the considerable differences between non-urban and urban gonorrhoea in Australia . Resistance to the penicillin and quinolone groups of antibiotics was highest in urban centres . Although penicillins remained suitable for use in many parts of non-urban Australia, enhanced surveillance is required as levels of resistance increase . Endemic transmission of quinolone-resistant gonococci (QRNG) in homosexually active men increased substantially in New South Wales and Victoria where more then 90% of all QRNG were found . QRNG in other centres continued to be isolated mostly from overseas travellers and at a low frequency . All isolates remained sensitive to spectinomycin and ceftriaxone . A further increase in the number of gonococcal isolates from homosexually active men was recorded in New South Wales and Victoria . Strains examined in South Australia, New South Wales and Victoria were predominantly from male patients and rectal and pharyngeal isolates were common . In other centres the male to female ratio of cases was lower, and most isolates were from the genital tract in rates similar to those occurring in previous years . The impact of non-culture based detection methods will adversely affect the ability of the AGSP to monitor trends in gonococcal disease in future years.

Am J Obstet Gynecol, 2000 Nov, 183(5), 1243 - 8
Levels of vaginal secretory leukocyte protease inhibitor are decreased in women with lower reproductive tract infections; Draper DL et al.; OBJECTIVE: Secretory leukocyte protease inhibitor contributes resistance to primary human immunodeficiency virus infection in the oral cavity . However, the levels of this inhibitor in the genital tract of women with sexually transmitted diseases or vaginitis are not well described . The objective was to determine vaginal inhibitor levels in women with symptomatic and asymptomatic genital infections . STUDY DESIGN: We tested 207 nonpregnant women for Neisseria gonorrhoeae, Trichomonas vaginalis, Chlamydia trachomatis, Candida species, and bacterial vaginosis by standard methods . A second group of symptom-free pregnant women (N = 231) was also studied . Secretory leukocyte protease inhibitor was measured by enzyme-linked immunosorbent assay, and results were compared by nonparametric methods . RESULTS: Vaginal levels of secretory leukocyte protease inhibitor in both groups were significantly lower in women with any sexually transmitted disease than in those without infection (P<.0001) . Patients with bacterial vaginosis and those with bacterial vaginosis with yeast vaginitis also had decreased levels (P<.025) . CONCLUSIONS: Levels of secretory leukocyte protease inhibitor in vaginal fluid are decreased in women with lower genital tract infection . This may represent a common mechanism of increasing susceptibility to infection with human immunodeficiency virus.

Infect Immun, 2000 Dec, 68(12), 7166 - 71
fbpABC gene cluster in Neisseria meningitidis is transcribed as an operon; Khun HH et al.; The neisserial fbpABC locus has been proposed to constitute a single transcriptional unit . To confirm this operonic arrangement, transcription assays using reverse transcriptase PCR amplification were conducted with Neisseria meningitidis . The presence of fbpAB and fbpBC transcripts obtained by priming cDNA synthesis with an fbpC-sequence-specific oligonucleotide indicates that fbpABC is organized as a single expression unit . The ratio of fbpA to fbpABC mRNA was approximately between 10- to 20-fold, as determined by real-time quantitative PCR.

Infect Immun, 2000 Dec, 68(12), 7028 - 38
Molecular variation among type IV pilin (bfpA) genes from diverse enteropathogenic Escherichia coli strains; Blank TE et al.; Typical enteropathogenic Escherichia coli (EPEC) strains produce bundle-forming pili (BFP), type IVB fimbriae that have been implicated in EPEC virulence, antigenicity, autoaggregation, and localized adherence to epithelial cells (LA) . BFP are polymers of bundlin, a pilin protein that is encoded by the bfpA gene found on a large EPEC plasmid . Striking sequence variation has previously been observed among type IV pilin genes of other gram-negative bacterial pathogens (e.g., Pseudomonas and Neisseria spp.) . In contrast, the established sequences of bfpA genes from two distantly related prototype EPEC strains vary by only a single base pair . To determine whether bundlin sequences vary more extensively, we used PCR to amplify the bfpA genes from 19 EPEC strains chosen for their various serotypes and sites and years of isolation . Eight different bfpA alleles were identified by sequencing of the PCR products . These alleles can be classified into two major groups . The alpha group contains three alleles derived from strains carrying O55, O86, O111, O119, O127, or O128 somatic antigens . The beta group contains five alleles derived from strains carrying O55, O110, O128ab, O142, or nontypeable antigens . Sequence comparisons show that bundlin has highly conserved and variable regions, with most of the variation occurring in the C-terminal two-thirds of the protein . The results of multilocus enzyme electrophoresis support the hypothesis that bfpA sequences have spread horizontally across distantly related clonal lineages . Strains with divergent bundlin sequences express bundlin protein, produce BFP, and carry out autoaggregation and LA . However, four strains lack most or all of these phenotypes despite having an intact bfpA gene . These results have important implications for our understanding of bundlin structure, transmission of the bfp gene cluster among EPEC strains, and the role of bundlin variation in the evasion of host immune system responses.

Infect Immun, 2000 Dec, 68(12), 6988 - 96
Identification of discrete domains within gonococcal transferrin-binding protein A that are necessary for ligand binding and iron uptake functions; Boulton IC et al.; The availability of free iron in vivo is strictly limited, in part by the iron-binding protein transferrin . The pathogenic Neisseria spp . can sequester iron from this protein, dependent upon two iron-repressible, transferrin-binding proteins (TbpA and TbpB) . TbpA is a TonB-dependent, integral, outer membrane protein that may form a beta-barrel exposing multiple surface loops, some of which are likely to contain ligand-binding motifs . In this study we propose a topological model of gonococcal TbpA and then test some of the hypotheses set forth by the model by individually deleting three putative loops (designated loops 4, 5, and 8) . Each mutant TbpA could be expressed without toxicity and was surface exposed as assessed by immunoblotting, transferrin binding, and protease accessibility . Deletion of loop 4 or loop 5 abolished transferrin binding to whole cells in solid- and liquid-phase assays, while deletion of loop 8 decreased the affinity of the receptor for transferrin without affecting the copy number . Strains expressing any of the three mutated TbpAs were incapable of growth on transferrin as a sole iron source . These data implicate putative loops 4 and 5 as critical determinants for receptor function and transferrin-iron uptake by gonococcal TbpA . The phenotype of the DeltaL8TbpA mutant suggests that high-affinity ligand interaction is required for transferrin-iron internalization.

Infect Immun, 2000 Dec, 68(12), 6526 - 34
Effect of alpha-oligosaccharide phenotype of Neisseria gonorrhoeae strain MS11 on invasion of Chang conjunctival, HEC-1-B endometrial, and ME-180 cervical cells; Minor SY et al.; The genes encoding the glycosyltransferases responsible for the addition of the five sugars in the alpha-oligosaccharide (alpha-OS) moiety of lipooligosaccharide (LOS) have been identified . Disruption of these glycosyltransferase genes singly or in combination results in corresponding truncations in LOS . In the present work we show that sequential deletion of the terminal four sugar residues of gonococcal alpha-OS had no discernible effect on the invasion of human conjunctival, endometrial, and cervical cell lines . However, deletion of the proximal glucose, which resulted in the complete deletion of alpha-OS, significantly impaired invasion of the gonococci into all three cell lines . The effect of deleting alpha-OS on invasion was independent of and additive to the known invasion-promoting factor OpaA . These data suggest that the proximal glucose residue of the alpha-OS chain of LOS is required for efficient invasion of gonococci into host mucosa.

Mem Inst Oswaldo Cruz, 2000 Nov-Dec, 95(6), 853 - 4
Characterization of Neisseria gonorrhoeae strains isolated from patients with conjunctivitis; Sosa J et al.; The conjunctivitis produced by Neisseria gonorrhoeae is the less frequently reported clinical form of gonococcal infection . We aim to phenotypically characterize N . gonorrhoeae isolated from conjunctivae sites . A total of six cases of this disease were notified in the Camaguey province, Cuba . All the strains isolated were penicillin-producing, showed the serogroup WI and exhibited the same antimicrobial susceptibility pattern and plasmid profile (2.6-3 . 2-24.5) . The results contribute to the characterization of N . gonorrhoeae strains circulating in our environment.

Arch Pathol Lab Med, 2000 Nov, 124(11), 1649 - 52
Performance characteristics of the Becton Dickinson ProbeTec System for direct detection of Chlamydia trachomatis and Neisseria gonorrhoeae in male and female urine specimens in comparison with the Roche Cobas System; Chan EL et al.; OBJECTIVE: The Becton Dickinson BDProbeTec ET System is a new semiautomated system using strand displacement amplification technology that simultaneously amplifies and detects Chlamydia trachomatis and Neisseria gonorrhoeae DNA . The strand displacement amplification products are hybridized with a fluorescent detector probe and are captured by a chemiluminescent assay in a microwell format . An amplification control is also included to monitor assay inhibition . This study evaluated the performance of the BDProbTec ET system in detecting C trachomatis and N gonorrhoeae in male and female urine specimens, calculated its ability to process large volumes of specimens, and determined the inhibition rate . MATERIALS AND METHODS: Eight hundred twenty-five male and 399 female urine specimens were tested for both C trachomatis and N gonorrhoeae with the BDProbeTec ET system, and results were compared with those of the Roche Amplicor Cobas system . All urine specimens were processed on both assays on the same day they were received, according to the manufacturers' instructions . Discrepant results were resolved by in-house polymerase chain reaction assays . Internal or amplification controls were also used in each specimen assay to monitor inhibition . The throughput of the BDProbTec ET system was further tested with 150 urine specimens on an 8-hour shift for 2 days . RESULTS: The overall sensitivity, specificity, positive predicative value, and negative predicative value for for detection of chlamydia were 95.3%, 99.3%, 95.9%, and 99.2% for strand displacement amplification and 95.9%, 98.3%, 90.6%, and 99.3% for the Roche Amplicor system . For detection of gonorrhea, these values were 100%, 99.7%, 88.2%, and 100% and 96.7%, 98.9%, 69%, and 99.9%, respectively . The overall inhibition rates for both strand displacement amplification and Roche Amplicor were less than 3.5% . The BDProbTec ET system was able to produce 150 results each for chlamydia and gonorrhea and the internal control within the 8-hour shift . CONCLUSIONS: The performance characteristics of the BDProbeTec ET assay are similar to those of the Roche Amplicor polymerase chain reaction for detection of chlamydia and gonorrhea in male and female urine specimens . The system was able to produce 300 results in an 8-hour shift.

Lett Appl Microbiol, 2000 Oct, 31(4), 294 - 8
The influence of different concentrations of melatonin on the cell surface hydrophobic characteristics of Neisseria meningitidis; Uberos J et al.; The cell surface hydrophobicity of micro-organisms is a characteristic that has been associated with the colonization of mammalian epithelia and with their capacity to induce diseases . Melatonin is a hormone produced by the pineal gland that affects the immune response mechanism . This study investigated, as an expression of the virulence of Neisseria meningitidis, how its hydrophobic characteristics were affected by exposure to increasing concentrations of melatonin . An increase in the cell surface hydrophobicity of N . meningitidis was found at concentrations of 1 mmol l(-1), while lower concentrations of melatonin did not significantly affect this particular cell surface characteristic of the micro-organism . It may be concluded that melatonin clearly influences the cell surface hydrophobicity of N . meningitidis, a circumstance that should be taken into account in future studies to determine whether this hormone plays a role in the variable pathogenicity of the bacteria in different hosts.

Rev Soc Bras Med Trop, 2000 Sep-Oct, 33(5), 451 - 64
{Gonorrhea}; Penna GO et al.; Gonorrhea is a common bacterial infection caused by Neisseria gonorrhoeae, a Gram-negative diplococcus that is transmitted almost exclusively by sexual contact or perinatally . It primarily affects the mucous membranes of the lower genital tract and less frequently those of the rectum, oropharynx, and conjunctivae . Ascending genital infection in women leads to the predominant complication, acute salpingitis, one of the most common causes of female infertility in the world . Since the 1990s, a remarkable surge of information ensued regarding the pathogenesis of gonorrhea and its agent . Gonorrhea has proven difficult to control in most populations and remains a prime example of the influence that social, behavioral, and demographic factors can have on the epidemiology of an infectious disease . The management of gonorrhea and other sexually transmitted infections requires both treatment of the patient as an individual and of his or her sexual partner(s) as a public health measure to interrupt the onward spread of infection and prevent long-term complications.

Clin Cornerstone, 2000, 3(1), 1 - 11
Diagnosis and treatment of common sexually transmitted diseases in women; Johnson RA; Chlamydia trachomatis and Neisseria gonorrhoeae are 2 very common sexually transmitted organisms, whose clinical manifestations in women can range from an asymptomatic carrier state to active pelvic inflammatory disease with known serious sequelae, including chronic pelvic pain, infertility, and ectopic pregnancy . The economic and clinical burden of these 2 infectious organisms are significant in the sexually active population . New developments in diagnosis and treatment of these infections raise great hope that substantial reduction in morbidity and disease prevalence rates can be achieved . Herpes simplex virus is probably better publicized and more feared in the sexually active population, and is far more prevalent than previously recognized; fortunately, however, it is not generally associated with significant morbidity . This article will review the current diagnoses and treatments of these conditions and consider some of the issues surrounding the impact of screening asymptomatic sexually active individuals . The treatment guidelines will emphasize the 1998 Guidelines for Treatment of Sexually Transmitted Diseases from the Centers for Disease Control and Prevention.

J Clin Microbiol, 2000 Nov, 38(11), 4076 - 9
Diagnostic performance of the Roche AMPLICOR PCR in detecting Neisseria gonorrhoeae in genitourinary specimens from female sex workers in Cotonou, Benin; Mukenge-Tshibaka L et al.; The objective of this study was to evaluate the diagnostic performance of the Roche multiplex AMPLICOR Chlamydia trachomatis/Neisseria gonorrhoeae PCR test for the detection of Neisseria gonorrhoeae infection in female urine specimens and wet and dry endocervical swabs . Endocervical swabs and urine specimens were collected from 342 female sex workers from Cotonou, Benin, and were tested using the AMPLICOR C . trachomatis/N . gonorrhoeae test (Roche Diagnostic Systems, Inc., Branchburg, N.J.) with internal control detection . Endocervical swabs were also cultured on Thayer-Martin medium . A series of alternate standards that included a combination of all the tests but not the test being evaluated was used to assess the performance of the test with each type of specimen . The sensitivity, specificity, and positive and negative predictive values for the urine were 53.8, 98.9, 93.5, and 87.5%, respectively . Corresponding figures for the wet swab were 91.5, 100, 100, and 97.4%, respectively . Those for the dry swab were 96.3, 96.2, 88.5, and 98.8%, respectively . Based on this study, the AMPLICOR PCR assay showed a low sensitivity for detection of N . gonorrhoeae infection in urine specimens, whereas the test was found to be highly sensitive and specific with endocervical specimens.

Hautarzt, 2000 Sep, 51(9), 666 - 9
{Antibiotic susceptibility of Neisseria gonorrhoeae isolates in Berlin}; Wagner J et al.; BACKGROUND AND OBJECTIVE: Because of the increasing resistance of Neisseria gonorrhoeae, we studied the actual resistance of isolates in Berlin . PATIENTS/METHODS: 85 Neisseria gonorrhoeae isolates were collected between 1995 and 1997 . Susceptibility testing was performed for penicillin G, tetracycline, spectinomycin, ceftriaxone, ciprofloxacin and azithromycin by agar dilution . RESULTS: 18.8% isolates were resistant or intermediately resistant to penicillin G (including 6 PPNG) . 12.9% isolates were resistant, 43.5% intermediately resistant to tetracycline . One strain was resistant against ciprofloxacin, 4 isolates showed increased MIC values (0.06-0.5 mg/l), whereas 78 isolates were fully susceptible (< 0.007 mg/l) . All isolates were susceptible to spectinomycin, ceftriaxone, and azithromycin . CONCLUSIONS: Penicillin G and tetracycline should be given only in cases of proven sensibility . Resistance against ciprofloxacin may occur, especially in isolates acquired in south-east Asia . Ceftriaxone, spectinomycin and azithromycin were active against all isolates . The actual resistance situation should be monitored.

Int J Cancer, 2000 Nov 1, 88(3), 342 - 50
Acetaldehyde production by non-pathogenic Neisseria in human oral microflora: implications for carcinogenesis in upper aerodigestive tract; Muto M et al.; Many epidemiological studies have identified chronic alcohol consumption as a significant risk factor for cancer of the upper aerodigestive tract (UAT) in human . Although acetaldehyde, the first metabolite from ethanol by alcohol dehydrogenase (ADH), is regarded as a carcinogen, how systemic production of acetaldehyde particularly affects the UAT remains unclear . In our study, we searched for the regional source of acetaldehyde in UAT, especially the involvement of bacteria in the human normal oral microflora . Here we demonstrate that, among the bacterial species identified from the human oral cavity, genus Neisseria had extremely high ADH activity and produced significant amounts of acetaldehyde when cultured with medium containing ethanol in vitro . The ability to produce acetaldehyde was more than 100-fold higher than that produced by any other genera we studied . Furthermore, alcohol ingestion influences the bacterial composition of the oral microflora, resulting in an increased proportion of Neisseria . Although Neisseria present in normal oral microflora is generally non-pathogenic, these findings suggest that this microbe can be a regional source of carcinogenic acetaldehyde and thus potentially play an important role in alcohol-related carcinogenesis in human UAT.

J Immunol, 2000 Nov 1, 165(9), 4803 - 13
The ability of B cells and dendritic cells to present antigen increases during ontogeny; Muthukkumar S et al.; The immune response to polysaccharide (PS) Ags in mice is delayed during ontogeny even when administered in a thymus-dependent (TD) form . In this study, Neisseria meningitidis group C PS-tetanus toxoid conjugate (MCPS-TT) vaccine was used to examine whether the delay in the development of Ab responses to TD PS conjugate vaccines in neonatal mice is due to defective Ag presentation . The results show that B cells and dendritic cells (DC) from 3- and 7-day-old mice were severely defective in presenting TT and MCPS-TT to Ag-specific T cell clones . The ability of these cells to present Ag reaches adult levels by 4 wk . The development of anti-MCPS and anti-TT Abs in neonatal mice parallels the functional ability of their APC to present Ag . DC from neonatal mice expressed very low levels of MHC class II, costimulatory molecules B7.1, B7.2, and CD11c but high levels of monocyte-specific markers F4/80 and CD11b and granulocyte marker, Ly6G . Significant changes in the expression of these markers were observed as the age of the mice increased . MHC class II, B7.1 and B7.2, and CD11c all increased with age, reaching adult levels between 3 and 4 wk, concurrent with the function of APC . These results demonstrate that one reason neonates fail to produce high titers of anti-PS Abs even when immunized in a TD form is that their B cells and DC are not fully functional.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 3184 - 5
Antimicrobial resistance of Neisseria gonorrhoeae and emerging ciprofloxacin resistance in the Netherlands, 1991 to 1998; de Neeling AJ et al.; Surveillance of antibiotic resistance in Neisseria gonorrhoeae showed a decrease in the percentage of beta-lactamase-producing isolates but an increase in intermediately penicillin-resistant strains and strains resistant to a high level of tetracycline . MICs for the ciprofloxacin-resistant isolates that emerged increased, and these isolates had mutations in gyrA and parC similar to those observed in the Far East.

Infect Immun, 2000 Nov, 68(11), 6321 - 8
Staphylococcal fibronectin binding protein interacts with heat shock protein 60 and integrins: role in internalization by epithelial cells; Dziewanowska K et al.; We reported previously that internalization of Staphylococcus aureus by nonprofessional phagocytes involves an interaction between fibronectin (Fn) binding protein (FnBP) and the host cell, resulting in signal transduction, tyrosine kinase activity, and cytoskeletal rearrangement (K . Dziewanowska, J . M . Patti, C . F . Deobald, K . W . Bayles, W . R . Trumble, and G . A . Bohach, Infect . Immun . 67:4673-4678, 1999) . The goal of the present study was to identify the host molecules responsible for uptake of the organism through an interaction with FnBP . First, Fn was required for internalization . Addition of small amounts of exogenous Fn stimulated the uptake of S . aureus by HEp-2 cells, which are deficient in Fn synthesis . Fn antibodies blocked internalization of the organism by MAC-T cell monolayers, a bovine epithelial cell line which expresses Fn . Second, a monoclonal antibody (MAb) specific for beta(1) integrins dramatically reduced S . aureus invasion, suggesting that the formation of a Fn bridge linking the host cell beta(1) integrin and FnBP precedes internalization . However, ligand blotting of cell membrane proteins with a functional fragment of FnBP consistently identified an additional approximately 55-kDa receptor on both human and bovine epithelial cells . This protein was purified and identified by N-terminal microsequencing as heat shock protein 60 (Hsp60) . The interaction between FnBP and Hsp60 also occurred when the whole cells were used . Cell membrane localization of Hsp60 was confirmed by biotinylation with an agent nonpermeable to the cell membrane . Pretreatment of epithelial cells with a MAb specific for eukaryotic Hsp60 significantly reduced internalization of S . aureus . Combined, these results suggest that the FnBP binds directly to both Hsp60 and Fn and is linked to beta(1) integrins through a Fn bridge . The simultaneous involvement of Fn and two host cell ligands, beta(1) integrins and Hsp60, suggests that FnBP is a multifunctional adhesin that mediates internalization in a manner similar to that proposed for OpaA, the Neisseria gonorrhoeae FnBP homolog (J . P . M . van Putten, T . D . Duensing, and R . L . Cole, Mol . Microbiol . 29:369-379, 1998).

Infect Immun, 2000 Nov, 68(11), 6215 - 22
Neisseria gonorrhoeae porin modifies the oxidative burst of human professional phagocytes; Lorenzen DR et al.; A hallmark of infection with the gram-negative bacterium Neisseria gonorrhoeae is the local infiltration and subsequent activation of polymorphonuclear neutrophils . Several gonococcal outer membrane proteins are involved in the interaction with and the activation of these phagocytes, including gonococcal porin, the most abundant protein in the outer membrane . Previous work suggests that this porin plays a role in various cellular processes, including inhibiting neutrophils activation and phagosome maturation in professional phagocytes . Here we investigated the ability of porin to modify the oxidative metabolism of human peripheral blood neutrophils and monocytes in response to particulate stimuli (including live gonococci) and soluble agents . The activation of the oxidative metabolism was determined by chemiluminescence amplified with either luminol or lucigenin . We found that treatment of the phagocytes with porin inhibits the release of reactive oxygen species measured as luminol-enhanced chemiluminescence in response to zymosan, latex particles, and gonococci . The engulfment of these particles was not, however, affected by porin treatment . Similar effects of porin on the chemiluminescence response were observed in cytochalasin B-treated neutrophils exposed to the soluble chemotactic peptide N-formylmethionyl-leucyl-phenylalanine . This indicates that porin selectively inhibits granule fusion with those cellular membranes that are in direct contact with porin, namely, the phagosomal and plasma membranes . This porin-induced downregulation of oxidative metabolism may be a potent mechanism by which gonococci modulate oxygen-dependent reactions by activated phagocytes at inflammation sites.

Diagn Microbiol Infect Dis, 2000 Oct, 38(2), 109 - 13
Antimicrobial activity of gemifloxacin (SB-265805), a newer fluoroquinolone, against clinical isolates of Neisseria gonorrhoeae, including fluoroquinolone-resistant isolates; Tanaka M et al.; Antimicrobial activity of gemifloxacin (SB-265805), a newly developed fluoroquinolone, to Japanese isolates of Neisseria gonorrhoeae was compared with those of various fluoroquinolones, including norfloxacin, ciprofloxacin, tosufloxacin, levofloxacin, sparfloxacin, and trovafloxacin . Among the fluoroquinolones tested, gemifloxacin was most active against N . gonorrhoeae isolates . The MIC90 values of gemifloxacin for 94 N . gonorrhoeae isolated from 1992 through 1993 and 100 isolated from 1996 through 1997 were 0.03 and 0.125 microg/ml, respectively . On the other hand, MIC90 values of the other fluoroquinolone for the 1992-1993 isolates and the 1996-1997 isolates ranged from 0.125 to 2 microg/ml and from 0.5 to 8 microg/ml, respectively . Gemifloxacin was also the most potent fluoroquinolone against 31 ciprofloxacin-resistant isolates with the ciprofloxacin MIC of 1 to 16 microg/ml, for which the gemifloxacin MIC50 and MIC90 values were 0.25 and 2 microg/ml, respectively . Moreover, the activity of gemifloxacin against fluoroquinolone-resistant gonococcal isolates containing multiple amino acid substitutions in both GyrA and ParC proteins was superior to those of the other compounds.

FEBS Lett, 2000 Sep 1, 480(2-3), 147 - 50
LEAP-1, a novel highly disulfide-bonded human peptide, exhibits antimicrobial activity; Krause A et al.; We report the isolation and characterization of a novel human peptide with antimicrobial activity, termed LEAP-1 (liver-expressed antimicrobial peptide) . Using a mass spectrometric assay detecting cysteine-rich peptides, a 25-residue peptide containing four disulfide bonds was identified in human blood ultrafiltrate . LEAP-1 expression was predominantly detected in the liver, and, to a much lower extent, in the heart . In radial diffusion assays, Gram-positive Bacillus megaterium, Bacillus subtilis, Micrococcus luteus, Staphylococcus carnosus, and Gram-negative Neisseria cinerea as well as the yeast Saccharomyces cerevisiae dose-dependently exhibited sensitivity upon treatment with synthetic LEAP-1 . The discovery of LEAP-1 extends the known families of mammalian peptides with antimicrobial activity by its novel disulfide motif and distinct expression pattern.

J Clin Periodontol, 2000 Oct, 27(10), 722 - 32
Microbial composition of supra- and subgingival plaque in subjects with adult periodontitis; Ximenez-Fyvie LA et al.; BACKGROUND, AIMS: The purpose of the present study was to compare and relate the microbial composition of supra and subgingival plaque in 23 adult periodontitis subjects (mean age 51 +/- 14 years) . METHODS: A total of 1,170 samples of supra and subgingival plaque were collected from the mesial aspect of every tooth (up to 28 supra and 28 subgingival samples) from each subject and evaluated for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization . Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth . The counts (levels), % DNA probe count (proportion) and % of sites colonized (prevalence) of each species in supra and separately in subgingival plaque were computed for each subject . Significance of differences between supra and subgingival plaque for each species was sought using the Wilcoxon signed ranks test and adjusted for multiple comparisons . RESULTS: All 40 taxa were detected in both supra and subgingival plaque . Actinomyces species were the most prevalent taxa in both habitats . 75 to 100% of supra and 62 to 100% of subgingival sites were colonized by at least one of the 5 Actinomyces species . Supragingival samples exhibited significantly higher counts of Actinomyces naeslundii genospecies 1, Actinomyces israelii, Actinomyces odontolyticus, Neisseria mucosa, Streptococcus gordonii, Capnocytophaga ochracea and Capnocytophaga sputigena when compared with mean counts in subgingival samples taken from the same tooth surfaces . Subgingival plaque samples presented significantly higher counts of Prevotella nigrescens, Prevotella intermedia, Bacteroides forsythus and Porphykromonas gingivalis . Subgingival samples exhibited a significantly higher proportion of "red" and "orange complex" species, while supragingival plaque exhibited higher proportions of "green" and "purple" complex species as well as Actinomyces species . Suspected periodontal pathogens could be detected in supragingival plaque from sites where subgingival samples were negative for the same species . CONCLUSIONS: The data indicate that supragingival plaque can harbor putative periodontal pathogens, suggesting a possible role of this environment as a reservoir of such species for the spread or reinfection of subgingival sites.

EMBO J, 2000 Oct 16, 19(20), 5332 - 43
Targeting of the pro-apoptotic VDAC-like porin (PorB) of Neisseria gonorrhoeae to mitochondria of infected cells; Muller A et al.; Infection of cell cultures with Neisseria gonorrhoeae results in apoptosis that is mediated by the PorB porin . During the infection process porin translocates from the outer bacterial membrane into host cell membranes where its channel activity is regulated by nucleotide binding and voltage-dependent gating, features that are shared by the mitochondrial voltage-dependent anion channel (VDAC) . Here we show that porin is selectively and efficiently transported to mitochondria of infected cells . Prevention of porin translocation also blocked the induction of apoptosis . Mitochondria of cells treated with porin both in vitro and in vivo were depleted of cytochrome c and underwent permeability transition . Overexpression of Bcl-2 blocked porin-induced apoptosis . The release of cytochrome c occurred independently of active caspases but was completely prevented by Bcl-2 . Our data suggest that the Neisseria porin can, like its eukaryotic homologue, function at the mitochondrial checkpoint to mediate apoptosis.

Rev Esp Salud Publica, 2000 Jul-Aug, 74(4), 419 - 24
{Survey of carriers of Neisseria meningitidis in the health area of Gran Canary}; Garcia Rojas A et al.; BACKGROUND: A) To ascertain the rate of carriers and the Types of Neisseria Meningitidis circulating in the population resident in the health jurisdiction of Gran Canaria . B) to ascertain the pattern of distribution of such carriers . METHODS: A descriptive transversal design was made, with a random sampling in multiple stages and by conglomerates . A minimum sample size was determined at 707 individuals for an expected prevalence of 8.6%, with a rate of reliability of 95.6% and a precision of 0.02 . Assuming that 15% if the individuals would not be willing to co-operate, the sample size was increased to 831 individuals, distributed in each conglomerate in proportion to the existing population . This size was distributed in turn into four groups by age and sex, in proportion to their significance in each basic health care zone selected at random . The individuals in the sample were identified from among those who attended the blood extraction units, and after they had passed the criteria of exclusion, their co-operation was requested as volunteers in the study . If they accepted, a questionnaire was filled out with a number of variables of epidemiological interest and a pharyngeal smear was taken . Since the Primary Care units were selected on a simple random basis, and the same method was used to select the individuals within the units, the estimate of the prevalence was made by means of an unbiased estimator . RESULTS: A total of 828 samples were obtained, that is, 99.6% of the number foreseen . With the exception of three, all of the individuals selected participated voluntarily in the study, a circumstance rendering it highly representative . All of the strains obtained corresponded to N . Meningitidis Serogroup B, except for one identified as N . Meningitidis Serogroup C Sero/Subtype 4:P1.2,5 . The strains of N . Meningitidis serogroup B identified corresponded to 25 different sero-subtypes . The prevalence determined after having studied the sample was 6.45%, the variance = 0.0275 and the standard error = 1.66 . We can conclude with a 95% degree of reliability that the prevalence of carriers of N . Meningitidis in the Gran Canaria health jurisdiction is estimated to be between 3.2% and 9.7% . CONCLUSIONS: A clear predominance of N . Meningitidis serogroup B strains among carriers has been verified . There are no statistically significant differences in the prevalence observed among the different age groups nor between sexes.

Am J Public Health, 2000 Oct, 90(10), 1615 - 8
Risk and prevalence of treatable sexually transmitted diseases at a Birmingham substance abuse treatment facility; Bachmann LH et al.; OBJECTIVES: We evaluated the prevalence of gonorrhea, chlamydia, trichomoniasis, and syphilis in patients entering residential drug treatment . METHODS: Data on sexual and substance abuse histories were collected . Participants provided specimens for chlamydia and gonorrhea ligase chain reaction testing . Trichomonas vaginalis culture, and syphilis serologic testing . RESULTS: Of 311 patients, crack cocaine use was reported by 67% and multisubstance use was reported by 71% . Sexually transmitted disease (STD) risk behaviors were common . The prevalence of infection was as follows: Chlamydia trachomatis, 2.3%; Neisseria gonorrhoeae, 1.6%; trichomoniasis, 43%; and syphilis, 6% . CONCLUSIONS: STD counseling and screening may be a useful adjunct to inpatient drug treatment.

J Biol Chem, 2001 Jan 26, 276(4), 2719 - 24 Epub 2000 Oct 11.
Fe3+ coordination and redox properties of a bacterial transferrin; Taboy CH et al.; The Fe(3+) binding site of recombinant nFbp, a ferric-binding protein found in the periplasmic space of pathogenic Neisseria, has been characterized by physicochemical techniques . An effective Fe(3+) binding constant in the presence of 350 microm phosphate at pH 6.5 and 25 degrees C was determined as 2.4 x 10(18) m(-1) . EPR spectra for the recombinant Fe(3+)nFbp gave g' = 4.3 and 9 signals characteristic of high spin Fe(3+) in a strong ligand field of low (orthorhombic) symmetry . (31)P NMR experiments demonstrated the presence of bound phosphate in the holo form of nFbp and showed that phosphate can be dialyzed away in the absence of Fe(3+) in apo-nFbp . Finally, an uncorrected Fe(3+/2+) redox potential for Fe-nFbp was determined to be -290 mV (NHE) at pH 6.5, 20 degrees C . Whereas our findings show that nFbp and mammalian transferrin have similar Fe(3+) binding constants and EPR spectra, they differ greatly in their redox potentials . This has implications for the mechanism of Fe transport across the periplasmic space of Gram-negative bacteria.

Arch Pediatr, 2000 Sep, 7(9), 927 - 32
{Kingella kingae osteoarticular infections in children . A report of a series of eight new cases}; Abuamara S et al.; Kingella kingae is a Gram-negative bacillus which belongs to the Neisseriaceae family . Its involvement in osteoarticular infections is relatively recent . METHODS AND RESULTS: We report eight cases of Kingella kingae osteoarticular infections that have been diagnosed at the paediatric surgical centre of Rouen University Hospital since October 1995 . Six boys and two girls (mean age: 30.6 months) presented with osteomyelitis in six cases and arthritis in two . Only 75% of patients had a fever at time of diagnosis . The biological findings were slightly modified . All samples were obtained from blood, bone or joint fluid . These samples were systematically inoculated into a blood culture tube . Positive Kingella kingae culture was achieved in seven local samples and in one blood culture . All children received two antibiotics via intravenous injection while waiting for the bacteriologic results . Later, the antibiotic treatment (amoxycillin) was given per os . The mean duration of treatment was 33 days . Patients were given intravenous treatment for a period of only ten days . Six patients were followed up for a period of more than 18 months and outcome was always uneventful . DISCUSSION: Kingella kingae is usually present in the nasopharyngeal mucosa and spreads in the blood due to various infections . Different types of Kingella kingae infection have been reported with a large frequency of osteoarticular infection . CONCLUSION: This type of infection does not present any unusual characteristics as compared to other osteoarticular infections . Because of its antibiotic sensitivity treatment duration could be reduced . Kingella kingae is a fragile microbe and its culture is often difficult; therefore, it is important to use blood culture tubes to inoculate joint fluid and bone samples.

Sex Transm Infect, 2000 Aug, 76(4), 303 - 6
Syndromic management of vaginal discharge among women in a reproductive health clinic in India; Vishwanath S et al.; OBJECTIVES: To examine the performance of the syndromic approach in the management of vaginal discharge among women attending a reproductive health clinic in New Delhi, India . METHODS: Women who sought services from the clinic and who had a complaint of vaginal discharge were interviewed, underwent a pelvic examination, and provided samples for laboratory investigations of bacterial vaginosis, candidiasis, syphilis, trichomoniasis, and Chlamydia trachomatis and Neisseria gonorrhoeae infections . Data analysis focused on the prevalence of infection and on the performance of the algorithm recommended by the national authorities for the management of vaginal discharge . RESULTS: The most common infection among 319 women was bacterial vaginosis (26%) . At least one sexually transmitted infection was detected in 21.9% of women . The prevalence of C trachomatis infection was 12.2%; trichomoniasis 10%; syphilis 2.2%; N gonorrhoeae was not isolated . An algorithm based on risk assessment and speculum assisted clinical evaluation was not helpful in predicting cervical infections associated with C trachomatis (sensitivity 5% and PPV 9%) . This algorithm was sensitive (95%) though not specific (22%) in selecting women for metronidazole therapy effective against bacterial vaginosis or trichomoniasis, and overtreatment was a problem (PPV 38%) . The sensitivity, specificity, and PPV of this algorithm for the treatment of candidiasis were 46%, 98%, and 88% respectively . The cost per case assessed using the algorithm was $2 and the cost per infection correctly treated was $4.25 . CONCLUSIONS: The prevalence of cervical infection associated with C trachomatis was high among these "low risk" women . The syndromic approach is not an efficient tool for detecting this condition, and alternative approaches to evaluation and intervention are required . The syndromic management of vaginal discharge among women seeking family planning and other reproductive health services should focus on vaginal infections, thus enhancing quality of care and addressing women's concerns about their health.

Prim, Care Update Ob Gyns . 2000 Sep 1, 7(5), 200 - 206
Infections and infertility; Rhoton-Vlasak A; Infertility affects 10-15% of all couples . Pelvic infections are an important cause of infertility, primarily as a result of tubal damage . Damage to the fallopian tubes from infections may be due to adhesions, tubal mucosal damage, or tubal occlusion that interferes with normal ovum transport . The infections most commonly related to infertility include gonorrhea, chlamydia, and pelvic inflammatory disease . Tuberculosis also is a common cause of infertility in Third World nations . Sequelae resulting from these infections include ectopic pregnancy, infertility, chronic pelvic pain, hydrosalpinx, and tuboovarian abscess . Neisseria gonorrhoeae and Chlamydia trachomatis are the primary causes of pelvic inflammatory disease . Chlamydial infections may be asymptomatic, and the resulting salpingitis is often referred to as silent pelvic inflammatory disease . Polymicrobial infection with other organisms such as anaerobes or facultative aerobes may be initiated by gonorrhea, chlamydia, or both . Early recognition of infection, prompt institution of appropriate antibiotic therapy, and proper follow-up are important to prevent the sequelae of pelvic inflammatory disease . Surgical intervention may be needed to treat immediate or long-term sequelae of infection . Prevention of pelvic infections should be a high priority . Fortunately, treatment options such as tubal microsurgery and assisted reproductive technologies offer couples reproductive options even when infertility occurs as the result of a previous pelvic infection.

J Clin Microbiol, 2000 Oct, 38(10), 3825 - 6
InTray GC medium versus modified Thayer-Martin agar plates for diagnosis of gonorrhea from endocervical specimens; Beverly A et al.; A new self-contained medium system for the cultivation of Neisseria gonorrhoeae was compared to modified Thayer-Martin medium for the diagnosis of gonorrhea from endocervical specimens . There was no difference in the ability of the two methods to support the growth of N . gonorrhoeae.

J Clin Microbiol, 2000 Oct, 38(10), 3544 - 9
Multicenter evaluation of AMPLICOR and automated COBAS AMPLICOR CT/NG tests for Neisseria gonorrhoeae; Martin DH et al.; The fully automated COBAS AMPLICOR CT/NG and semiautomated AMPLICOR CT/NG tests were evaluated in a multicenter trial for their ability to detect Neisseria gonorrhoeae infections . Test performance compared to that of culturing was evaluated for 2,192 matched endocervical swab and urine specimens obtained from women and for 1, 981 matched urethral swab and urine specimens obtained from men . Culture-negative, PCR-positive specimens that tested positive in a confirmatory PCR test for an alternative target sequence within the N . gonorrhoeae 16S rRNA gene were considered to be true positives . The overall prevalences of gonorrhea were 6.6% in women and 20.1% in men . The COBAS AMPLICOR and AMPLICOR formats yielded concordant results for 98.8% of the specimens and exhibited virtually identical sensitivities and specificities . The results that follow are for the COBAS AMPLICOR format . With the infected patient as the reference standard, the resolved sensitivities of PCR were 92.4% for endocervical swab specimens and 64.8% for female urine specimens . There were no significant differences in these rates between women with and without symptoms . Among symptomatic men, COBAS AMPLICOR sensitivities were 94.1% for urine and 98.1% for urethral swabs; for asymptomatic men, the results were 42.3 and 73.1%, respectively . In comparison, the sensitivities of culturing were 84.8% for endocervical specimens, 92.7% for symptomatic male urethral specimens, and only 46.2% for urethral specimens obtained from asymptomatic men . When PCR results were analyzed as if only a single test had been performed on a single specimen type, the resolved sensitivity was always higher . The resolved specificities of PCR were 99.5% for endocervical swab specimens, 99.8% for female urine specimens, 98.9% for male urethral swab specimens, and 99.9% for male urine specimens . The internal control revealed that 2.1% of specimens were inhibitory when initially tested . Nevertheless, valid results were obtained for 99.2% of specimens because 60.0% of the inhibitory specimens were not inhibitory when a second aliquot was tested . The COBAS AMPLICOR CT/NG test for N . gonorrhoeae exhibited high sensitivity and specificity with urethral swab and urine specimens from men and endocervical swab specimens from women and thus is well suited for diagnosing and screening for N . gonorrhoeae infection.

MMWR Morb Mortal Wkly Rep, 2000 Sep 22, 49(37), 833 - 7
Fluoroquinolone-resistance in Neisseria gonorrhoeae, Hawaii, 1999, and decreased susceptibility to azithromycin in N . gonorrhoeae, Missouri, 1999; Role of the male partner in the lower genitourinary tract infection of female; Department of Dermatology, Postgraduate Institute of Medical Education & Research, ChandigarhWe studied the relationships of selected microbes and the role of consorts in the causation of vaginal discharge which may be due to cervicitis or vaginitis . A total of 93 consecutive patients in the reproductive age group with symptoms of vaginal discharge along with their sexual partners were studied . Samples were collected from the cervix and posterior fornix of the female patients and from the urethra and sub-prepucial area of the male partner for culture of Neisseria gonorrhoeae, Gardnerella vaginalis, Mycoplasma hominis, ureaplasma, candida, aerobic and anaerobic organisms . Apart from cultures, KOH and Gram stain of the discharge were made . Predominant pathogen isolated was Ureaplasma urealyticum from 40 (43.01%) females and 23 (24.7%) males . The next common pathogenic organisms isolated were candida species from 11 (11.8%) females and 5 (5.4%) males and Chlamydia trachomatis in 3 (3.2%) females and 1 (1.1%) male . Various organisms were more frequently isolated from the 29 of 43 (67.4%) couples who had had sexual intercourse 7 days prior to the recruitment as compared to 14 of 43 (32.6%) who had had coitus more than 7 days prior to their recruitment . This may be due to the spontaneous disappearance or decrease in the number of organisms to the level that they could be detected by culture . In our study, 6 (6.5%) of male partners carrying pathogenic organisms were asymptomatic, indicating that their screening and treatment is vital.

Carbohydr Res, 2000 Aug 18, 328(1), 3 - 16
Polymer-supported and chemoenzymatic synthesis of the Neisseria meningitidis pentasaccharide: a methodological comparison; Yan F et al.; Neisseria meningitidis trisaccharide {GlcNAc{(1-->3)Galbeta(1-->4)Glc-R}, tetrasaccharide {Galbeta(1-->4)GlcNAcbeta(1--> 3)Galbeta(1-->4)Glc-R}, and a pentasaccharide {Neu5Acalpha(2-->3)Galbeta(1-->4)GlcNAcbeta(1-->3)G albeta(1-->4)Glc-SPh} were prepared via conventional chemical synthesis, polymer-supported synthesis, and chemoenzymatic methods, starting from D-lactose . The polymer polyethyleneglycol monomethylether (MPEG) and the linker dioxyxylene (DOX) were used with a lactose-bound acceptor to improve the purification process . Several enzymes (LgtA, GalE-LgtB fusion, and CMP-Neu5Ac synthetase/sialyltransferase fusion) were used for syntheses of these oligosaccharides . Excellent stereo- and regioselectivities as well as high yield (> 90% from Gal(1-->4)Glc-SPh) of the pentasaccharide were obtained . Both of the convenient processes are suitable for efficient preparation of target oligosaccharides.

Mol Microbiol, 2000 Sep, 37(6), 1389 - 404
Helicobacter pylori inhibits phagocytosis by professional phagocytes involving type IV secretion components; Ramarao N et al.; Gastric infections by Helicobacter pylori are characteristically associated with an intense inflammation and infiltration of mainly polymorphonuclear lymphocytes (PMNs) and monocytes . The inflammatory response by infiltrated immune cells appears to be a primary cause of the damage to surface epithelial layers and may eventually result in gastritis, peptic ulcer, gastric cancer and/or MALT-associated gastric lymphoma . Our analysis of the interaction between H . pylori and PMNs and monocytes revealed that H . pylori inhibits its own uptake by these professional phagocytes . To some degree, this effect resembles antiphagocytosis by Yersinia enterocolitica . Increasing numbers of bacteria associated per cell are more efficient at blocking their own engulfment . In H . pylori, bacterial protein synthesis is necessary to block phagocytic uptake, as shown by the time and concentration dependence of the bacteriostatic protein synthesis inhibitor chloramphenicol . Furthermore, H . pylori appears broadly to inhibit the phagocytic function of monocytes and PMNs, as infection with H . pylori abrogates the phagocytes' ability to engulf latex beads or adherent Neisseria gonorrhoeae cells . This antiphagocytic phenotype depends on distinct virulence (vir) genes, such as virB7 and virB11, encoding core components of a putative type IV secretion apparatus . Our data indicate that H . pylori exhibits an antiphagocytic activity that may play an essential role in the immune escape of this persistent pathogen.

Curr Biol, 2000 Sep 21, 10(18), 1143 - 6
Type IV pilus of Myxococcus xanthus is a motility apparatus controlled by the frz chemosensory system; Sun H et al.; Although flagella are the best-understood means of locomotion in bacteria {1}, other bacterial motility mechanisms must exist as many diverse groups of bacteria move without the aid of flagella {2-4} . One unusual structure that may contribute to motility is the type IV pilus {5,6} . Genetic evidence indicates that type IV pili are required for social gliding motility (S-motility) in Myxococcus, and twitching motility in Pseudomonas and Neisseria {6,7} . It is thought that type IV pili may retract or rotate to bring about cellular motility {6,8}, but there is no direct evidence for the role of pili in cell movements . Here, using a tethering assay, we obtained evidence that the type IV pilus of Myxococcus xanthus functions as a motility apparatus . Pili were required for M . xanthus cells to adhere to solid surfaces and to generate cellular movement using S-motility . Tethered cells were released from the surface at intervals corresponding to the reversal frequency of wild-type cells when gliding on a solid surface . Mutants defective in the control of directional movements and cellular reversals (frz mutants) showed altered patterns of adherence that correlate reversal frequencies with tethering . The behavior of the tethered cells was consistent with a model in which the pili are extruded from one cell pole, adhere to a surface, and then retract, pulling the cell in the direction of the adhering pili . Cellular reversals would result from the sites of pili extrusion switching from one cell pole to another and are controlled by the frz chemosensory system.

Nature, 2000 Sep 7, 407(6800), 98 - 102
Pilus retraction powers bacterial twitching motility; Merz AJ et al.; Twitching and social gliding motility allow many gram negative bacteria to crawl along surfaces, and are implicated in a wide range of biological functions . Type IV pili (Tfp) are required for twitching and social gliding, but the mechanism by which these filaments promote motility has remained enigmatic . Here we use laser tweezers to show that Tfp forcefully retract . Neisseria gonorrhoeae cells that produce Tfp actively crawl on a glass surface and form adherent microcolonies . When laser tweezers are used to place and hold cells near a microcolony, retractile forces pull the cells toward the microcolony . In quantitative experiments, the Tfp of immobilized bacteria bind to latex beads and retract, pulling beads from the tweezers at forces that can exceed 80 pN . Episodes of retraction terminate with release or breakage of the Tfp tether . Both motility and retraction mediated by Tfp occur at about 1 microm s(-1) and require protein synthesis and function of the PilT protein . Our experiments establish that Tfp filaments retract, generate substantial force and directly mediate cell movement.

Sex Transm Dis, 2000 Sep, 27(8), 480 - 2
Analysis of the antibiotic sensitivity of Neisseria gonorrhoeae in Guangzhou, Peoples Republic of China; Wenling C et al.; BACKGROUND: Sexually transmitted diseases began to re-emerge in China in the mid 1980s . During the last one and a half decades, Neisseria gonorrhoeae infection has become one of the three most common sexually transmitted diseases in China . At present, resistant strains of N . gonorrhoeae are increasing each year . This study was undertaken to better understand the sensitivity of five antibiotics to N . gonorrhoeae isolates in Guangzhou, China . GOAL: To determine the frequency and diversity of antibiotic resistance, particularly to penicillin and tetracycline, on gonococcal strains in Guangzhou . STUDY DESIGN: Strains of N . gonorrhoeae isolates from 203 patients with uncomplicated urethral gonococcal infections from Guangzhou, China were reviewed from September 1997 to August 1998 . All strains were characterized with five different antimicrobials for sensitivity, including penicillin, tetracycline, spectinomycin, ciprofloxacin, and ceftriaxone . RESULTS: Penicillin resistance was present in 121 of 203 isolated strains (59.6%) . The plasmid-mediated strains and chromosome-mediated strains among the penicillin-resistant strains that were resistance to penicillin were 5.8% and 94.2%, respectively . Plasmid-mediated strains resistant to penicillin and tetracycline were each 3.4% . Most isolated strains were resistant to ciprofloxacin, accounting for 60.6% . All strains were sensitive to spectinomycin and ceftriaxone . CONCLUSION: N . gonorrhoeae isolates exhibited a high rate of resistance to penicillin and ciprofloxacin . Spectinomycin highly effective for penicillin-producing N . gonorrhoeae, tetracycline-resistant N . gonorrhoeae and the highly resistant strains of ciprofloxacin.

Structure Fold Des, 2000 Sep 15, 8(9), 981 - 92
Crystal structure of Omp32, the anion-selective porin from Comamonas acidovorans, in complex with a periplasmic peptide at 2.1 A resolution; Zeth K et al.; BACKGROUND: Porins provide diffusion channels for salts and small organic molecules in the outer membrane of bacteria . In OmpF from Escherichia coli and related porins, an electrostatic field across the channel and a potential, originating from a surplus of negative charges, create moderate cation selectivity . Here, we investigate the strongly anion-selective porin Omp32 from Comamonas acidovorans, which is closely homologous to the porins of pathogenic Bordetella and Neisseria species . RESULTS: The crystal structure of Omp32 was determined to a resolution of 2.1 A using single isomorphous replacement with anomalous scattering (SIRAS) . The porin consists of a 16-stranded beta barrel with eight external loops and seven periplasmic turns . Loops 3 and 8, together with a protrusion located within beta-strand 2, narrow the cross-section of the pore considerably . Arginine residues create a charge filter in the constriction zone and a positive surface potential at the external and periplasmic faces . One sulfate ion was bound to Arg38 in the channel constriction zone . A peptide of 5.8 kDa appeared bound to Omp32 in a 1:1 stoichiometry on the periplasmic side close to the symmetry axis of the trimer . Eight amino acids of this peptide could be identified, revealing specific interactions with beta-strand 1 of the porin . CONCLUSIONS: The Omp32 structure explains the strong anion selectivity of this porin . Selectivity is conferred by a positive potential, which is not attenuated by negative charges inside the channel, and by an extremely narrow constriction zone . Moreover, Omp32 represents the anchor molecule for a peptide which is homologous to proteins that link the outer membrane to the cell wall peptidoglycan.

J Bacteriol, 2000 Oct, 182(19), 5586 - 91
Pathogenic neisseriae can use hemoglobin, transferrin, and lactoferrin independently of the tonB locus; Desai PJ et al.; Redundant TonB systems which function in iron transport from TonB-dependent ligands have recently been identified in several gram-negative bacteria . We demonstrate here that in addition to the previously described tonB locus, an alternative system exists for the utilization of iron from hemoglobin, transferrin, or lactoferrin in Neisseria meningitidis and Neisseria gonorrhoeae . Following incubation on media containing hemoglobin, N . meningitidis IR3436 (tonB exbB exbD deletion mutant) and N . gonorrhoeae PD3401 (tonB insertional mutant) give rise to colonies which can grow with hemoglobin . Transfer of Hb(+) variants (PD3437 or PD3402) to media containing hemoglobin, transferrin, and/or lactoferrin as sole iron sources resulted in growth comparable to that observed for the wild-type strains . Transformation of N . meningitidis IR3436 or N . gonorrhoeae PD3401 with chromosomal DNA from the Hb(+) variants yielded transformants capable of growth with hemoglobin . When we inactivated the TonB-dependent outer membrane hemoglobin receptors (HmbR or HpuB) in the Neisseria Hb(+) variants, these strains could not grow with hemoglobin; however, growth was observed with transferrin and/or lactoferrin . These results demonstrate that accumulation of iron from hemoglobin, transferrin, and lactoferrin in the pathogenic neisseriae can occur via a system that is independent of the previously described tonB locus.

J Infect Dis, 2000 Oct, 182(4), 1169 - 76 Epub 2000 Sep 08.
Diversity and prevalence of PorA types in Neisseria meningitidis serogroup B in the United States, 1992-1998; Sacchi CT et al.; Two hundred eighty-one sporadic Neisseria meningitidis serogroup B isolates, collected through active laboratory-based surveillance, were selected to be analyzed by PorA variable region (VR) typing to determine the prevalence of PorA types in the United States . A substantial number of distinct VR types were identified, 31 in VR1 and 41 in VR2 . A total of 73 different PorA types were found, and 76 . 7% of these types comprise nonprototype sequences in VR1, VR2, or both . The most prevalent PorA types were P1.7,16-20 (previously P1.7, 16i), P1.22,14, P1.22-1,14 (previously P1.22a,14), P1.7,16, P1.7-1,1 (previously P1.7d,1), P1.19,15, and P1.17,16-3 (previously P1.B,16d) . No correlation was observed between the PorA types and geographic origin of the isolates . These data may aid in the design of an efficacious outer membrane protein-based vaccine by identifying the most appropriate PorA types for vaccine formulation . Studies are needed to fully evaluate the extent of cross-protection in humans among the variants and prototypes in each PorA VR family.

Mol Microbiol, 2000 Sep, 37(5), 1146 - 58
Molecular models accounting for the gene conversion reactions mediating gonococcal pilin antigenic variation; Howell-Adams B et al.; The pilus antigenic variation (Av) system of Neisseria gonorrhoeae is one of several high-frequency variation systems that utilize gene conversion to switch between numerous forms of an antigen on the cell surface . We have tested three predictions of the first models that explain the movement of DNA during pilin Av: (i) Av requires two recombinations at short regions of identity, (ii) circular intermediates exist that carry pilE/pilS hybrid loci and (iii) these pilE/pilS hybrid loci target the pilS sequences to a recipient pilE gene . We confirm that normal pilin Av utilizes recombination at very short regions of DNA sequence identity and that these recombination events can occur independent of homologous recombination functions . We have isolated covalently closed circular DNA molecules carrying hybrid pilin loci, but propose that an alternative hybrid molecule is the intermediate of pilin Av . Our most striking finding is that transformation of isolated pilE/pilS hybrid loci targets the pilS sequences of the hybrid to a recipient pilE at frequencies much higher than normal recombination frequencies . These results show that the different steps of a model that explains pilin Av can be separately tested to support the validity of these novel models that account for the high-frequency gene conversions that mediate pilin Av.

Mol Microbiol, 2000 Sep, 37(5), 1075 - 86
Iron availability regulates DNA recombination in Neisseria gonorrhoeae; Serkin CD et al.; The pilus of Neisseria gonorrhoeae (the gonococcus Gc), the causative agent of gonorrhoea, promotes attachment of the gonococcus to the host epithelium and is essential for the establishment of disease . The ability of N . gonorrhoeae to infect previously exposed individuals is partially due to pilus antigenic variation . In addition, variation of the pilus has been proposed to function in the adaptation of the gonococcus to host environments . Previously, we described the development of a competitive reverse transcriptase (RT)-PCR assay that quantifies the frequency of pilin antigenic variation within a gonococcal population . Using this assay, the effect of different biologically relevant environmental conditions on the frequency of pilin antigenic variation was tested . Of the environmental conditions examined in vitro, only limited iron affected a significant change in the frequency of antigenic variation . Further investigation revealed that an observed increase in pilin antigenic variation reflected an increase in other DNA recombination and DNA repair processes within iron-starved cultures . In addition, this low iron-induced increase was determined to be independent of changes in RecA expression and was observed in a Fur mutant strain . As gonococci encounter conditions of low iron during infection, these data suggest that iron-limitation signals for increased recombinational events that are important for gonococcal pathogenesis.

Mol Microbiol, 2000 Aug, 37(4), 839 - 55
Identification of transcription activators that regulate gonococcal adaptation from aerobic to anaerobic or oxygen-limited growth; Lissenden S et al.; Analysis of the Neisseria gonorrhoeae DNA sequence database revealed the presence of two genes, one encoding a protein predicted to be 37 . 5% identical (50% similar) in amino acid sequence to the Escherichia coli FNR protein and the other encoding a protein 41% and 42% identical (54 and 51% sequence similarity) to the E . coli NarL and NarP proteins respectively . Both genes have been cloned into E . coli and insertionally inactivated in vitro . The mutated genes have been transformed into gonococci and recombined into the chromosome . The fnr mutation totally abolished and the narP mutation severely diminished the ability of gonococci to: (i) grow anaerobically; (ii) adapt to oxygen-limited growth; (iii) initiate transcription from the aniA promoter (which directs the expression of a copper-containing nitrite reductase, AniA, in response to the presence of nitrite); and (iv) reduce nitrite during growth in oxygen-limited media . The product of nitrite reduction was identified to be nitrous oxide . Immediately upstream of the narL/narP gene is an open reading frame that, if translated, would encode a homologue of the E . coli nitrate- and nitrite-sensing proteins NarX and NarQ . As transcription from the aniA promoter was not activated during oxygen-limited growth in the presence of nitrate, the gonococcal two-component regulatory system is designated NarQ-NarP rather than NarX-NarL . As far as we are aware, this is the first well-documented example of a two-component regulatory system working in partnership with a transcription activator in pathogenic neisseria . A 45 kDa c-type cytochrome that was synthesized during oxygen-limited, but not during oxygen sufficient, growth was identified as a homologue of cytochrome c peroxidases (CCP) of other bacteria . The gene for this cytochrome, designated ccp, was located, and its regulatory region was cloned into the promoter probe vector pLES94 . Transcription from the ccp promoter was repressed during aerobic growth and induced during oxygen-limited growth and was totally FNR dependent, suggesting that the gonococcal FNR protein is a transcription activator of at least two genes . However, unlike AniA, synthesis of the CCP homologue was insensitive to the presence of nitrite during oxygen-limited growth.

J Clin Microbiol, 2000 Sep, 38(9), 3489 - 91
Characterization of Neisseria gonorrhoeae strains with decreased susceptibility to fluoroquinolones isolated in Greece from 1996 to 1999; Mavroidi A et al.; Of the 331 Neisseria gonorrhoeae strains isolated in Greece from 1996 to 1999, 39 (11.8%) exhibited decreased susceptibility to quinolones due to gyrA and parC mutations . Conventional typing and pulsed-field gel electrophoresis showed that 34 of these isolates were clonally related . Epidemiological data indicated that the epidemic clone was sustained in a group of high-frequency transmitters.

Trop Med Int Health, 2000 Jul, 5(7), A32 - 6
Interactions between HIV infection and other sexually transmitted diseases; Mabey D; Mechanisms and implications of the interaction between HIV and other STDs such as syphilis, Herpes simplex, Chlamydia and Neisseria gonorrhoeae infection are presented and relevant case reports and trials described.

Yonsei Med J, 2000 Jun, 41(3), 381 - 6
Emerging antimicrobial resistance, plasmid profile and pulsed-field gel electrophoresis pattern of the endonuclease-digested genomic DNA of Neisseria gonorrhoeae; Lee K et al.; Resistant gonococci are very prevalent in many countries, particularly in Asia . This study was conducted to determine the trend of resistance, the effect of decreasing the ciprofloxacin susceptibilities of gonococci on the prevalence of penicillinase-producing N . gonorrhoeae (PPNG), and to compare the epidemiology of strains with the previous studies . A total of 602 strains of gonococci were isolated from prostitutes in 1997-1999 . Antimicrobial susceptibility was tested by NCCLS disk diffusion and agar dilution methods . For epidemiologic analysis, plasmid analysis and pulsed-field gel electrophoresis (PFGE) were performed . The proportion of PPNG remained high (79%), and the strains with decreased susceptibility to ciprofloxacin increased significantly from 67% in 1997 to 84% in 1999 . Compared to our previous study, the PFGE patterns were similar, while the proportion of strain with the 3.2-MDa plasmid markedly decreased . In conclusion, a rapid increase in ciprofloxacin-nonsusceptible strains may suggest difficulties in the treatment of gonococcal infections in the near future with the drug . The recent decrease of PPNG with the 3.2-MDa plasmid may suggest that there is an epidemiological change in gonococcal infections, and the prevalence of related PFGE patterns suggests the dissemination of a few clones among the high risk populations.

Microbes Infect, 2000 Jun, 2(7), 821 - 7
Do pathogenic neisseriae need several ways to modify the host cell cytoskeleton?
Pujol C, Eugene E, Morand P, Nassif X.
Neisseria meningitidis and Neisseria gonorrhoeae are human pathogens which have to interact with mucosa and/or cellular barriers for their life cycle . Even though they both give rise to dramatically different diseases, most of the mechanisms mediating cellular interactions are common to N . meningitidis and N . gonorrhoeae . This suggests that bacterial cell interactions may be essential not only for pathogenesis but also for other aspects of the bacterial life cycle that are common to both N . meningitidis and N . gonorrhoeae . Opacity proteins and pili are two major components identified as transducing signals to host cells, thus leading to cytoskeleton modifications . This manuscript will review the recent developments concerning the mechanisms mediating cellular interactions of pathogenic Neisseria and will tentatively put them into the perspective of pathogenesis and bacterial life cycle.

J Urol, 2000 Sep, 164(3 Pt 1), 847 - 51
In vitro selection of fluoroquinolone-resistant Neisseria gonorrhoeae harboring alterations in DNA gyrase and topoisomerase IV; Yasuda M et al.; PURPOSE: We attempted to select increasingly fluoroquinolone-resistant strains of Neisseria gonorrhoeae in vitro and to assess whether selected mutants harbored alterations in the GyrA subunit of DNA gyrase and the ParC subunit of DNA topoisomerase IV, which were analogous to those in fluoroquinolone-resistant clinical isolates . MATERIALS AND METHODS: A fluoroquinolone-susceptible strain was exposed to norfloxacin in vitro . Selected mutants were sequentially exposed to norfloxacin, and this procedure was repeated . For 11 mutants, minimum inhibitory concentrations (MICs) of antimicrobial agents were determined, and mutations in the region corresponding to the quinolone resistance-determining region (QRDR) of the Escherichia coli gyrA gene and the analogous region of the parC gene were analyzed . RESULTS: Mutants obtained in one step exhibited significantly increased MICs of norfloxacin, ofloxacin and ciprofloxacin and had a single amino acid change in GyrA . Two-step mutants exhibited significantly higher norfloxacin MICs . Three of four two-step selected strains had single amino acid changes in both GyrA and ParC . Three-step mutants exhibited further increases in fluoroquinolone MICs and were assigned to the ciprofloxacin-resistant category . Two had a double amino acid change in GyrA, and one had a double GyrA change and a single amino acid change in ParC . CONCLUSION: We selected fluoroquinolone-resistant strains that carried GyrA and ParC alterations analogous to those in clinical isolates . The serial accumulation of changes in the QRDR of GyrA and the analogous region of ParC was associated with a stepwise increase in fluoroquinolone resistance, although the development of additional alterations in other regions of GyrA and ParC or other mechanisms of fluoroquinolone resistance also might contribute to the enhancement in fluoroquinolone resistance . The clinical emergence of fluoroquinolone-resistant strains may be due to in-vivo stepwise selection of strains with genetic alterations in GyrA and ParC, as observed here in the in-vitro selection of fluoroquinolone-resistant mutants.

Antimicrob Agents Chemother, 2000 Sep, 44(9), 2543 - 4
In vitro susceptibilities of 400 Spanish isolates of Neisseria gonorrhoeae to gemifloxacin and 11 other antimicrobial agents; Berron S et al.; The in vitro activity of gemifloxacin versus those of 11 other antimicrobial agents against 400 strains of Neisseria gonorrhoeae was determined by microdilution with supplemented GC agar . A total of 37.5% of the strains were beta-lactamase positive . A total of 70 and 6.4% of the beta-lactamase-negative strains exhibited intermediate and high-level penicillin resistance, respectively . Ceftriaxone and gemifloxacin were the most active drugs (MICs at which 90% of isolates are inhibited, 0.01 versus 0.007 microg/ml, respectively), with 100% of strains inhibited by 0.12 microg/ml.

J Infect Dis, 2000 Sep, 182(3), 848 - 55 Epub 2000 Aug 15.
Intranasal immunization with gonococcal outer membrane preparations reduces the duration of vaginal colonization of mice by Neisseria gonorrhoeae; Plante M et al.; Nasal immunization was studied to determine if it could elicit an immune response capable of preventing vaginal colonization by Neisseria gonorrhoeae or of reducing its duration in the estradiol-treated mouse model . Nasal administration of gonococcal outer membrane (OM) preparations induced the development of systemic and vaginal immune responses that were directed mainly against a limited number of gonococcal OM proteins . The impact of nasal immunization on vaginal colonization by N . gonorrhoeae was evaluated by use of an experimental model, in which mice were treated with estradiol to prolong the infection . Bacterial clearance was significantly faster for mice immunized intranasally with N . gonorrhoeae OM preparations (4.0+/-2.5 days) than for control mice (8.5+/-4.3 days) . The estradiol-treated mouse model may serve as a useful tool for the evaluation of potential gonococcal vaccine candidates.

Sex Transm Dis, 2000 Aug, 27(7), 401 - 10
Improvement of clinical algorithms for the diagnosis of Neisseria gonorrhoeae and Chlamydia trachomatis by the use of Gram-stained smears among female sex workers in Accra, Ghana; Deceuninck G et al.; BACKGROUND: Screening for cervical infection is difficult in developing countries . Screening strategies must be improved for high-risk women, such as female sex workers . GOAL: To evaluate the sensitivity and specificity of screening algorithms for cervical infection pathogens among female sex workers in Accra, Ghana . STUDY DESIGN: A cross-sectional study among female sex workers was conducted . Each woman underwent an interview and a clinical examination . Biologic samples were obtained for the diagnosis of HIV, syphilis, bacterial vaginosis, yeast infection, Trichomonas vaginalis, Neisseria gonorrhoeae, and Chlamydia trachomatis infection . Signs and symptoms associated with cervicitis agents were identified . Algorithms for the diagnosis of cervical infection were tested by computer simulations . RESULTS: The following prevalences were observed: HIV, 76.6%; N . gonorrhoeae, 33.7%; C . trachomatis, 10.1%; candidiasis, 24.4%; T . vaginalis, 31.4%; bacterial vaginosis, 2.3%; serologic syphilis, 4.6%; and genital ulcers on clinical examination, 10.6% . The best performance of algorithms were reached when using a combination of clinical signs and a search for gram-negative diplococci on cervical smears (sensitivity, 64.4%; specificity, 80.0%) . CONCLUSIONS: In the algorithms, examination of Gram-stained genital smears in female sex workers without clinical signs of cervicitis improved sensitivity without altering specificity for the diagnosis of cervical infection.

Infect Immun, 2000 Sep, 68(9), 5354 - 63
Neisseria gonorrhoeae elicits membrane ruffling and cytoskeletal rearrangements upon infection of primary human endocervical and ectocervical cells; Edwards JL et al.; Neisseria gonorrhoeae is a strict human pathogen that is, primarily, transmitted by close sexual contact with an infected individual . Gonococcal infection of the male urogenital tract has been well studied in experimental human models and in urethral cell culture systems . Recent studies, using tissue culture cell systems, have suggested a role for the cervical epithelium in gonococcal infection of females; however, the nature of gonococcal infection of the normal uterine cervix remains controversial . To address this enigma, we have developed two primary human cervical epithelial cell systems from surgical biopsies . Gonococcal infection studies and electron microscopy show that N . gonorrhoeae is capable of infecting and invading both the endo- and the ectocervix . Invasion was found to occur primarily in an actin-dependent manner, but it does not appear to require de novo protein synthesis by either the bacterium or the host cervical cell . Membrane ruffles appear to be induced in response to gonococci . Consistent with membrane ruffling, gonococci were found residing within macropinosomes, and a concentrated accumulation of actin-associated proteins was observed to occur in response to gonococcal infection . Electron microscopy of clinically derived cervical biopsies show that lamellipodia formation and cytoskeletal changes, suggestive of membrane ruffles, also occur in the cervical epithelium of women with naturally acquired gonococcal cervicitis . These studies demonstrate the ability of N . gonorrhoeae to infect and invade both the endo- and the ectocervix of the normal uterine cervix . Gonococcal induced ruffling is a novel finding and may be unique to the cervical epithelium.

Infect Immun, 2000 Sep, 68(9), 5241 - 6
Gonococcal nitric oxide reductase is encoded by a single gene, norB, which is required for anaerobic growth and is induced by nitric oxide; Householder TC et al.; The gene encoding a nitric oxide reductase has been identified in Neisseria gonorrhoeae . The norB gene product shares significant identity with the nitric oxide reductases in Ralstonia eutropha and Synechocystis sp . and, like those organisms, the gonococcus lacks a norC homolog . The gonococcal norB gene was found to be required for anaerobic growth, but the absence of norB did not dramatically decrease anaerobic survival . In a wild-type background, induction of norB expression was seen anaerobically in the presence of nitrite but not anaerobically without nitrite or aerobically . norB expression is not regulated by FNR or NarP, but a functional aniA gene (which encodes an anaerobically induced outer membrane nitrite reductase) is necessary for expression . When aniA is constitutively expressed, norB expression can be induced both anaerobically and aerobically, but only in the presence of nitrite, suggesting that nitric oxide, which is likely to be produced by AniA as a product of nitrite reduction, is the inducing agent . This was confirmed with the use of the nitric oxide donor, spermine-nitric oxide complex, in an aniA null background both anaerobically and aerobically . NorB is important for gonococcal adaptation to an anaerobic environment, a physiologically relevant state during gonococcal infection . The presence of this enzyme, which is induced by nitric oxide, may also have implications in immune evasion and immunomodulation in the human host.

Eur J Clin Microbiol Infect Dis, 2000 Jun, 19(6), 443 - 51
Novel genus-specific PCR-based assays for rapid identification of Neisseria species and Neisseria meningitidis; Lansac N et al.; This study presents the development of polymerase chain reaction (PCR)-based tests for the identification and detection of Neisseria species and Neisseria meningitidis . Currently, isolating and identifying these pathogens using conventional biochemical methods require 48-72 h . To improve speed and accuracy in diagnosing Neisseria infections, simple PCR-based tests that are specific for the genus Neisseria and the species Neisseria meningitidis have been developed . The genus-specific and species-specific DNA sequences were chosen by selecting and analyzing available database sequences . Neisseria-specific and Neisseria meningitidis-specific primer pairs were derived from the genes asd (coding for the aspartate beta-semialdehyde dehydrogenase) and ctrA (coding for a conserved outer membrane protein), respectively . Both the Neisseria-specific and Neisseria meningitidis-specific PCR assays were specific (they amplified only DNA from the target genus or species, out of 84 bacterial species tested) . In addition, the Neisseria-specific assay amplified DNA from 321 of 322 strains tested representing 13 species of Neisseria, while the Neisseria meningitidis-specific assay amplified DNA from all 256 strains tested representing nine serogroups of Neisseria meningitidis . These PCR assays, which can be combined in multiplex, have been adapted to ensure that they are simple and can be performed within approximately 90 min . The tests provide new diagnostic tools for identifying Neisseria infections.

Am J Obstet Gynecol, 2000 Aug, 183(2), 340 - 4; discussion 345-6
Detection of Neisseria gonorrhoeae and Chlamydia trachomatis colonization of the gravid cervix; Asbill KK et al.; OBJECTIVES: The aims of the study were to determine whether a Gram stain of cervical mucus can accurately rule out infection with Neisseria gonorrhoeae or Chlamydia trachomatis and to compare a diagnostic test that is based on the polymerase chain reaction with a deoxyribonucleic acid probe in the detection of these organisms . STUDY DESIGN: Gravid patients were screened for N gonorrhoeae and C trachomatis with a deoxyribonucleic acid probe, Gram stain, and analysis with the polymerase chain reaction . A normal, noninfected sample was defined by <10 polymorphonuclear leukocytes per high-power field on the Gram stain . Standard statistical methods were used to compare results of the Gram stain and the deoxyribonucleic acid probe, as well as to compare results of deoxyribonucleic acid probe hybridization and polymerase chain reaction analysis . A P value of <.05 was considered statistically significant . RESULTS: Patient enrollment totaled 519 . The prevalence of infection as determined by deoxyribonucleic acid probe hybridization was 1.4% for N gonorrhoeae (7/518) and 6.8% for C trachomatis (35/518) . The cervical Gram stain predicted the absence of infection in 17% (90/518) of patients, with a negative predictive value of 99% for N gonorrhoeae and 97% for C trachomatis . African American race, age <20 years, and unmarried status were all predictors of the presence of C trachomatis or N gonorrhoeae cervicitis . For the patients who lacked these risk factors (n = 74), the Gram stain had 100% negative predictive value . Analysis with the polymerase chain reaction detected 8 additional patients with C trachomatis and 105 additional patients with N gonorrhoeae, in comparison with deoxyribonucleic acid probe hybridization . CONCLUSION: The cervical Gram stain can accurately predict the absence of N gonorrhoeae and C trachomatis in gravid women . Analysis with the polymerase chain reaction indicates that N gonorrhoeae and C trachomatis are significantly more prevalent in this population than previously reported.

J Bacteriol, 2000 Sep, 182(17), 5005 - 8
Substrate ambiguity of 3-deoxy-D-manno-octulosonate 8-phosphate synthase from Neisseria gonorrhoeae revisited; Sheflyan GY et al.; Homogeneous, recombinant 3-deoxy-D-manno-octulosonate 8-phosphate synthase from Neisseria gonorrhoeae is shown to catalyze the formation of 3-deoxy-D-manno-octulosonate 8-phosphate from phosphoenolpyruvate and D-arabinose 5-phosphate as determined from (1)H-nuclear magnetic resonance analysis of the product . This enzyme does not catalyze the condensation of D-erythrose 4-phosphate and phosphoenolpyruvate to form 3-deoxy-D-ribo-heptulosonate 7-phosphate, as was previously reported (P . S . Subramaniam, G . Xie, T . Xia, and R . A . Jensen, J . Bacteriol . 180:119-127, 1998).

Int J Syst Evol Microbiol, 2000 Jul, 50 Pt 4, 1421 - 4
Pelczaria aurantia ATCC 49321T (=DSM 12801T) is a strain of Kocuria rosea (Flügge 1886) Stackebrandt et al . 1995; Schumann P et al.; Phylogenetic and chemotaxonomic analyses of Pelczaria aurantia ATCC 49321T (= DSM 12801T) indicate that this species is very closely related to Kocuria rosea . The DNA-DNA reassociation value of 87.1% determined for the type strains of the two species supports this finding . The results of phylogenetic analysis of the 16S rDNA of a subculture of the original strain of Pelczaria aurantia, deposited at the National Institutes of Health, Bethesda, MD, USA, as 'Neisseria aurantia', are identical to those for strain ATCC 49321T and indicate that Pelczaria aurantia ATCC 49321T is an authentic subculture of the original culture described by Poston (1993) . On the basis of these findings it is concluded that P . aurantia ATCC 49321T and K . rosea DSM 20447T are members of the same taxon . The taxonomic consequences of this union are discussed.

Braz J Infect Dis, 2000 Jun, 4(3), 144 - 50
Detecting polysaccharide antigen of Neisseria meningitidis group C in cerebrospinal fluid by dot-ELISA assay; Correia Barbosa SF et al.; Cerebrospinal fluid (CSF) samples from 210 patients (200 with clinical evidence of bacterial meningitis, 10 with other clinical neurologic disease) were tested by a Dot-ELISA assay for detection of polysaccharide antigen of N . meningitidis group C . CSF samples were treated with EDTA 0.1 M, at pH 7.5 and heated to 90>C for 10 min . Polyclonal antiserum was purified by use of ethanol fractionation . The results were compared to those using bacterial culture (BC), latex agglutination (LA), counterimmunoelectrophoresis (CIE), and direct microscopy (DM) methods . Test results showed a correlation of 93.3%, 94.3%, 91.0% and 69.5% respectively, and sensitivity of 0.947 and specificity of 0.930 . This study suggests that the dot-ELISA assay of CSF is a useful alternative technique for the diagnosis of group C meningitis.

Mol Microbiol, 2000 Jul, 37(1), 207 - 15
Repeat-associated phase variable genes in the complete genome sequence of Neisseria meningitidis strain MC58; Saunders NJ et al.; Phase variation, mediated through variation in the length of simple sequence repeats, is recognized as an important mechanism for controlling the expression of factors involved in bacterial virulence . Phase variation is associated with most of the currently recognized virulence determinants of Neisseria meningitidis . Based upon the complete genome sequence of the N . meningitidis serogroup B strain MC58, we have identified tracts of potentially unstable simple sequence repeats and their potential functional significance determined on the basis of sequence context . Of the 65 potentially phase variable genes identified, only 13 were previously recognized . Comparison with the sequences from the other two pathogenic Neisseria sequencing projects shows differences in the length of the repeats in 36 of the 65 genes identified, including 25 of those not previously known to be phase variable . Six genes that did not have differences in the length of the repeat instead had polymorphisms such that the gene would not be expected to be phase variable in at least one of the other strains . A further 12 candidates did not have homologues in either of the other two genome sequences . The large proportion of these genes that are associated with frameshifts and with differences in repeat length between the neisserial genome sequences is further corroborative evidence that they are phase variable . The number of potentially phase variable genes is substantially greater than for any other species studied to date, and would allow N . meningitidis to generate a very large repertoire of phenotypes through expression of these genes in different combinations . Novel phase variable candidates identified in the strain MC58 genome sequence include a spectrum of genes encoding glycosyltransferases, toxin related products, and metabolic activities as well as several restriction/modification and bacteriocin-related genes and a number of open reading frames (ORFs) for which the function is currently unknown . This suggests that the potential role of phase variation in mediating bacterium-host interactions is much greater than has been appreciated to date . Analysis of the distribution of homopolymeric tract lengths indicates that this species has sequence-specific mutational biases that favour the instability of sequences associated with phase variation.

FEBS Lett, 2000 Aug 4, 478(3), 260 - 6
Acid sphingomyelinase is involved in CEACAM receptor-mediated phagocytosis of Neisseria gonorrhoeae; Hauck CR et al.; The interaction with human phagocytes is a hallmark of symptomatic Neisseria gonorrhoeae infections . Gonococcal outer membrane proteins of the Opa family induce the opsonin-independent uptake of the bacteria that relies on CEACAM receptors and an active signaling machinery of the phagocyte . Here, we show that CEACAM receptor-mediated phagocytosis of Opa(52)-expressing N . gonorrhoeae into human cells results in a rapid activation of the acid sphingomyelinase . Inhibition of this enzyme by imipramine or SR33557 abolishes opsonin-independent internalization without affecting bacterial adherence . Reconstitution of ceramide, the product of acid sphingomyelinase activity, in imipramine- or SR33557-treated cells restores internalization of the bacteria . Furthermore, we demonstrate that CEACAM receptor-initiated stimulation of other signalling molecules, in particular Src-like tyrosine kinases and Jun N-terminal kinases, requires acid sphingomyelinase . These studies provide evidence for a crucial role of the acid sphingomyelinase for CEACAM receptor-initiated signalling events and internalization of Opa(52)-expressing N . gonorrhoeae into human neutrophils.

MMWR Morb Mortal Wkly Rep, 2000 Jun 23, 49(24), 538 - 42
Gonorrhea--United States, 1998; {Prevalence of sexually transmitted diseases (STD) and HIV-infection among attendees of STD Outpatient Clinic et al.; van der Snoek EM, Chin-A-Lien RA, de Ridder MA, Willems PW, Verkooyen RP, van der Meijden WI.

Academisch Ziekenhuis Rotterdam-Dijkzigt, afd . Dermato-venerologie, RotterdamOBJECTIVE: To gain an impression as to the possible trends in the prevalence of VD and HIV between 1993 and 1998 . DESIGN: Retrospective . METHOD: Data of age, gender, ethnic background, sexual preference, intravenous drug use and a diagnosis of VD or HIV infection for persons attending the VD Outpatient Clinic of the Department of Dermato-Venereology of the Dijkzigt University Hospital in Rotterdam, the Netherlands, for the first time were compared for the years 1993 and 1998 . RESULTS: The number of patients examined was 3099 (1894 men and 1205 women) in 1993 and 3696 (1878 men and 1818 women) in 1998 . No diagnosis of venereal disease was made in 58.9% and 56.2% of them, respectively . The prevalence of HIV infection decreased from 1.3% in 1993 to 0.6% in 1998 . This was caused mainly by a decline in the prevalence of HIV-antibodies among men with homosexual contacts . In the total population, the prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis decreased significantly (from 5.4% to 2.5% and from 9.3% to 7.5%, respectively), but there was no similar decrease among the men with homosexual contacts.

Proc Natl Acad Sci U S A, 2000 Aug 1, 97(16), 9070 - 5
Neisseria meningitidis porin PorB interacts with mitochondria and protects cells from apoptosis; Massari P et al.; Neisserial porins are strong immune adjuvants and B cell activators . The effect of neisserial porin PorB on activation-induced cell death was investigated, as a potential additional mechanism of the porin's immunopotentiating ability . Neisserial porins interact with target cells to localize intracellularly in the mitochondrial compartment without negatively affecting cellular survival . Pretreatment with Neisseria meningitidis PorB porin decreased or abrogated the mitochondrial damage induced by apoptotic stimuli . In addition, end stage determinants of apoptosis, including DNA breakdown, were diminished by PorB . Immunoprecipitation experiments revealed that PorB interacts with the mitochondrial porin VDAC (voltage-dependent anion channel) . The mechanism of the antiapoptotic effect of neisserial porins could be explained by the protein-protein interaction of PorB with VDAC, similar to the interaction of VDAC with antiapoptotic Bcl-2 proteins, resulting in an enhancement of cell survival and continued activation of B cells.

J Clin Microbiol, 2000 Aug, 38(8), 2914 - 6
Use of the neisserial lipoprotein (Lip) for subtyping Neisseria gonorrhoeae; Trees DL et al.; The pathogenic Neisseria species N . meningitidis and N . gonorrhoeae possess an outer membrane lipoprotein, designated Lip, which is present in all strains tested . The predicted protein sequence of Lip consists of a consensus AAEAP amino acid repeat . The objective of this study was to determine the feasibility of using the Lip repeat number and sequence for subtyping of Neisseria gonorrhoeae . The lip genes of each isolate were amplified by PCR and sequenced to determine the repeat number and sequence . Among the 46 strains we examined, eight different Lip repeat numbers were identified, with lengths of 11 (1 strain), 12 (14 strains), 13 (2 strains), 14 (10 strains), 15 (5 strains), 16 (10 strains), 17 (3 strains), and 20 (1 strain) repeats . Analysis indicated differences in the sequences within the repeats that resulted in amino acid alterations in repeat classes that contained multiple strains . Among the 46 isolates examined, we were able to identify 17 unique Lip subtyping patterns.

Mil Med, 2000 Jul, 165(7), 549 - 51
Neisseria gonorrhoeae screening in pregnancy; Ries AM; All patients undergoing prenatal care at the U.S Naval Hospital in Yokosuka, Japan, have a gonorrhea culture performed at their intake visit . The objective of this study was to determine the prevalence of Neisseria gonorrhoeae in this military population and to propose an effective antenatal screening strategy based on the data . A retrospective 10-month review of patient data was performed . In addition, the guidelines for antenatal N . gonorrhoeae screening by the American College of Obstetricians and Gynecologists, the Centers for Disease Control and Prevention, the U.S . Department of Health and Human Services, and the World Health Organization were reviewed . During the study period, none of the screened patients (N = 518) had a gonococcal infection . The screening program at this facility will be modified based on the low-risk nature of the pregnant population . Specifically, high-risk patients will be screened selectively in the third trimester.

Adolesc Med, 2000 Jun, 11(2), 401 - 25
Serious infections of the central nervous system: encephalitis, meningitis, and brain abscess; Rajnik M et al.; Central nervous system infections in adolescents range from the diffuse cerebritis of encephalitis to the regional inflammation of meningitis, and very focal disease of brain abscess . Clinical presentations reflect this wide spectrum, with encephalitis primarily characterized by altered mental status, meningitis by fever, headache, and neck stiffness, and brain abscess manifesting localizing findings . Encephalitis and viral meningitis are frequently caused by the seasonal enteroviruses and arboviruses, while most adolescent bacterial meningitis is due to Neisseria meningitidis and Streptococcus pneumoniae . The microbiology of brain abscess reflects underlying host risk factors . Gram-positive cocci are seen in patients with congenital heart disease, while respiratory flora including anaerobes are associated with sinus or otic disease . Lumbar puncture to characterize and culture the CSF remains the optimal test for the diagnosis and management of encephalitis and meningitis, while CT-guided needle biopsy may be both diagnostic and therapeutic for brain abscesses . New diagnostic tests include the use of PCR . A variety of safe and effective treatment regimens exists for most bacterial infections as well as for some herpesvirus infections . New vaccines are under study to further control bacterial meningitis.

Adolesc Med, 2000 Jun, 11(2), 225 - 49
Upper respiratory tract infections in adolescents; White CB et al.; While upper respiratory tract infections (URTIs) cause much infectious morbidity in infants and young children, adolescents are not immune to infections . Adolescents experience two to four episodes of viral nasopharyngitis annually . In addition to group A streptococcus (GAS), pharyngitis may occur with other streptococci, Arcanobacterium haemolyticum, Epstein-Barr virus, Neisseria gonorrhoeae, and other pathogens . Uvulits, typically in association with GAS, occurs occasionally . Peritonsillar abscess is the most common deep neck space infection seen in adolescents, but retropharyngeal and parapharyngeal abscesses also occur, causing major morbidity . Adolescents experience fewer cases of otitis media than younger children . Rhinosinusitis occurs commonly in adolescents, occasionally leading to chronic sinusitis and serious sequelae such as osteomyelitis . This article reviews the major URTIs likely to be encountered by physicians caring for adolescents . For each entity there is a brief description of the epidemiology, morbidity, microbiology, clinical and laboratory features, treatment, and prevention.

Acta Paediatr, 2000 Jun, 89(6), 661 - 5
Neisseria meningitidis: evolution of penicillin resistance and phenotype in a children's hospital in Barcelona, Spain; Latorre C et al.; Neisseria meningitidis is the most prevalent micro-organism involved in paediatric bacterial meningitis in the Barcelona area in children over 3 mo of age and it is an important cause of morbidity and mortality in Spain . A total of 498 strains of N . meningitidis, obtained between the years 1986 and 1997 from children with sepsis and/or meningitis, were characterized according to their serogroup and penicillin resistance; their distribution in serotypes and subtypes was studied from 1990 . A decreasing tendency in the number of annual isolates was observed in this period . Most isolates belonged to serogroups B (403 strains) and C (77 strains) . Serogroup C accounted for 1.8% of the strains in 1986 and 57.1% in 1997 . The most prevalent phenotype between 1990 and 1996 was B:4:P1.15 . but C:2b:P1.2,5 was the most prevalent in 1997 . Overall penicillin-resistance rates ranged from 9.1% in 1986 (when a non-susceptible strain was isolated for the first time in the Hospital Sant Joan de Deu, Barcelona, Spain) to 71.4% in 1997, and it was more common among strains belonging to serogroup C (52% of resistant strains) than to serogroup B (22.1 % of resistant strains) . The penicillin-resistance level was low, MIC always < or = 0.5 microg/ml . The present increase in N . meningitidis group C isolates, mainly C:2b:P1.2,5, and the availability of preventive measures for this highly pathological and resistant phenotype, argues strongly for the establishment of an epidemiological monitoring system . Detection of penicillin resistance should be standardized worldwide in order to unify data from all laboratories . CONCLUSION: A shift between serogroups B and C is observed in Barcelona from 1986 to 1997, as well as a rapid distribution of decreased penicillin susceptibility.

Clin Infect Dis, 2000 Jul, 31(1), 34 - 41 Epub 2000 Jul 26.
Immunogenicity of a heptavalent pneumococcal conjugate vaccine in Apache and Navajo Indian, Alaska native, and non-native American children aged <2 years; Miernyk KM et al.; High rates of invasive pneumococcal disease have been described among infants living in various Native American communities . In this study, we evaluated the immunogenicity of a 7-valent pneumococcal vaccine consisting of serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F covalently linked to the outer membrane protein complex of Neisseria meningitidis in Apache and Navajo Indian, Alaska Native, and non-Native American children . The vaccine was administered at ages 2, 4, and 6 months; a booster dose was given at age 15 months . Levels of serotype-specific immunoglobulin G (IgG) were measured by a standardized enzyme-linked immunosorbent assay . The responses after 3 primary doses of vaccine were similar in all 3 groups of children, except for those to serotypes 14 and 23F . One month after the booster dose, geometric mean concentrations (GMCs) of serotype-specific IgG antibodies increased significantly in all 3 groups of children, compared with GMCs of IgG antibodies to pneumococcal serotypes before the booster dose.

Sex Transm Dis, 2000 Jul, 27(6), 358 - 62
Prevalence of sexually transmitted infections among clients of female commercial sex workers in Thailand; Tabrizi SN et al.; BACKGROUND: Clients of commercial sex workers are considered at high risk for the acquisition and transmission of sexually transmitted diseases (STDs) . Identification and treatment of infections in this group could help to reduce the transmission of STDs . GOAL: To ascertain the prevalence of sexually transmitted organisms in male clients of female sex workers in Thailand by analysis of seminal fluid collected after intercourse . STUDY DESIGN: Used condoms were collected from 291 male clients attending a brothel in Hat Yai, Thailand during a 7-day period . Nucleic acid was extracted from seminal fluid and tested by polymerase chain reaction for the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, herpes simplex virus (HSV) and HIV sequences . RESULTS: Overall, 17 (6%), 47 (16%), and 2 (1%) of specimens were positive for C trachomatis, N gonorrhoeae and T vaginalis respectively . HSV sequences were found in 24 (8%) of the specimens: 14 specimens (5%) with HSV type 1, and 11 specimens (4%) with HSV type 2 . HIV RNA was detected in two samples (1%) . Overall, 75 specimens (26%) were positive for one or more infections, and more than one pathogen was detected in 16 specimens (5%) . CONCLUSION: This study reports a high rate of STDs among clients of female sex workers in Thailand . Consequently, this population is a significant risk for transmitting STDs to commercial sex workers and to other noncommercial partners . Strategies that target this population of men are needed to reduce STD and HIV transmission.

Diagn Microbiol Infect Dis, 2000 Jul, 37(3), 195 - 201
Development of gemifloxacin in vitro susceptibility test methods for gonococci including quality control guidelines . The Quality Control Study Group; Jones RN et al.; Gemifloxacin (formerly SB-265805 or LB20304a) is a new fluoronapthyridone with documented activity against Gram-positive and -negative organisms . The activity of gemifloxacin was tested against 150 Neisseria gonorrhoeae strains, using reference agar dilution, standardized disk diffusion, and Etest (AB BIODISK, Solna, Sweden) methods . Gemifloxacin was very potent against ciprofloxacin (CIPRO)-susceptible strains (MIC(90,) 0.008 microg/ml) but was significantly less active against the CIPRO-resistant gonococci (MIC(90,) 0.12 microg/ml) . Etest and reference agar dilution MIC results showed excellent correlation (r = 0.96), and 98.7% MICs were within +/- one log(2) dilution . Agar dilution MICs were also compared to zone diameters obtained using gemifloxacin 5-microg disks; and complete intermethod categorical agreement (100%) was achieved applying breakpoints proposed as follows: < or =0.25 microg/ml (zone, > or =25 mm) for susceptible and > or =1 microg/ml (zone, < or =21 mm) for resistant . Gemifloxacin MIC and disk diffusion te quality control (QC) ranges were established for N . gonorrhoeae ATCC 49226 . Data were collected from > or = seven laboratories, three GC agar medium lots for both agar MICs and disk methods, and two lots each of the 5- and 10-microg disks . The proposed MIC QC range was 0.002 to 0.016 microg/ml and the calculated mm zone ranges (median +/- 0.5x average mm range) for both disks were similar, but contained only 88.1 to 91.9% of participant results . To achieve the acceptable > or = 95% of all study results within range, a 43 to 54 mm limits (5-microg disks) were necessary . The excellent broad-spectrum activity and a low reported adverse effects profile of gemifloxacin shows a potential for treatment of fluoroquinolone-resistant gonorrhea.

Immunopharmacology, 2000 Aug, 49(1-2), 149 - 57
Host recognition and target differentiation by factor H, a regulator of the alternative pathway of complement; Pangburn MK; Factor H is responsible for recognition of host cells and tissues and mediates discrimination among microbial pathogens during activation of the alternative pathway of complement (AP) . Its unique structure of 20 SCR domains arranged in a flexible chain permits a variety of functional sites to interact with complement proteins and surface markers in a biological example of single-molecule combinatorial chemistry . In addition to the complement regulatory site located in the N-terminal four SCR domains, two other sites bind complement protein C3b and three sites appear to recognize a variety of polyanions that serve as host markers . Recent studies indicate that cooperativity among several C3b- and polyanion-binding sites influences the biological functions of factor H and that the degree of influence of each site varies on different cells . The engagement of one or more of the host marker recognition sites enables factor H to control activation of the AP . The absence of host-like markers allows AP activation, but many common pathogens have developed receptors for factor H or mimics of host markers of varying degrees of authenticity allowing them to escape detection by this innate defense system . Organisms using one or more of these evasive techniques include Neisseria gonorrhoeae, Streptococcus pyogenes, Yersinia enterocolitica, Trypanosoma cruzi, and the HIV virus.

Bioorg Khim, 2000 May, 26(5), 323 - 9
{Induction of anti-meningitis immunity using the synthetic peptides . I . The immunoreactive synthetic fragments of porin A from Neisseria meningitidis}; Koroev DO et al.; Fourteen peptides corresponding to sequences of all the exposed and some of the transmembrane protein regions of porin A from the outer membrane of Neisseria meningitidis strain B:15:P1.7,16 were synthesize . Mice of various lines were immunized with the free peptides not conjugated with any protein carrier . It was shown that the majority of the peptides possess immunogenic properties . Two peptides were identified binding to antibodies present in the serum of mice after meningitis . Protective properties of a number of the synthesized peptides were studied, and three peptide sequences inducing mice protection from an experimental infection with N . meningitidis were identified.

Infect Immun, 2000 Aug, 68(8), 4725 - 35
Antigenic and sequence diversity in gonococcal transferrin-binding protein A; Cornelissen CN et al.; Neisseria gonorrhoeae is a gram-negative pathogen that is capable of satisfying its iron requirement with human iron-binding proteins such as transferrin and lactoferrin . Transferrin-iron utilization involves specific binding of human transferrin at the cell surface to what is believed to be a complex of two iron-regulated, transferrin-binding proteins, TbpA and TbpB . The genes encoding these proteins have been cloned and sequenced from a number of pathogenic, gram-negative bacteria . In the current study, we sequenced four additional tbpA genes from other N . gonorrhoeae strains to begin to assess the sequence diversity among gonococci . We compared these sequences to those from other pathogenic bacteria to identify conserved regions that might be important for the structure and function of these receptors . We generated polyclonal mouse sera against synthetic peptides deduced from the TbpA sequence from gonococcal strain FA19 . Most of these synthetic peptides were predicted to correspond to surface-exposed regions of TbpA . We found that, while most reacted with denatured TbpA in Western blots, only one antipeptide serum reacted with native TbpA in the context of intact gonococci, consistent with surface exposure of the peptide to which this serum was raised . In addition, we evaluated a panel of gonococcal strains for antigenic diversity using these antipeptide sera.

Clin Exp Rheumatol, 2000 May-Jun, 18(3), 383 - 6
Musculoskeletal infections in intravenous drug addicts: report of 34 cases with analysis of microbiological aspects and pathogenic mechanisms; Belzunegui J et al.; OBJECTIVE: To analyse characteristics of musculoskeletal infections in Spanish intravenous (i.v.) drug addicts . METHODS: A retrospective analysis of 34 medical records was carried out and the aetiologic agent was confirmed in all cases . RESULTS: The mean age was 26 years and the sex ratio was 25M/9F . Twenty-four patients were HIV-positive and 10 HIV-negative . There were no differences between the 2 subgroups . Septic arthritis, seen in 27 cases (79%), was the most common type of infection . Axial involvement was present in 23 cases (67%) . The organisms isolated included Staphylococcus aureus (25 cases), Candida albicans (3 cases), Mycobacterium tuberculosis (2 cases), and Neisseria gonorrhoeae, Salmonella typhi, Staphylococcus epidermidis and Streptococcus pyogenes in one case each . Candida albicans affected the sternocostal joints . Laboratory data showed non-specific findings . The outcome was mostly good and only one patient died . CONCLUSION: Human immunodeficiency virus does not affect characteristics of musculoskeletal infections in i.v . drug addicts . Staphylococcus aureus is the most frequently isolated pathogen in this population.

Child Abuse Negl, 2000 Jun, 24(6), 861 - 6
{Isolation of Gardnerella vaginalis in the diagnosis of sexual abuse in children}; Fernandez JP et al.; INTRODUCTION: The study of sexual transmitted diseases is more and more frequent in patient with suspicion of sexual abuse, and this help to the final medical diagnosis . Organisms like Neisseria gonorrhoeae and Treponema indicate a sure sexual contact . However the role of the rest of organisms in the sexual abuse is more difficult of specifying . OBJECTIVE: To know the role of the Gardnerella vaginalis in the diagnosis of sexual abuse in patient with this possible diagnosis . METHODS: Retrospective study of 135 girls with suspicion of abuse, visited from May of 1997 to February of 1998 . Vaginal swabs were taken from 45 of 135 girls evaluated . RESULTS: In five patients the vaginal culture were positive to Gardnerella . The characteristics of these five patients are described (age, aggressor, type and duration of the abuse, physical exploration and final diagnosis) . All our patients were the pubertal or prepubertal girls . In these patients the final diagnosis settled with the clinical history and the physical exploration, without keeping in mind the result of the culture . CONCLUSION: It is necessary to discard sexual abuse in all prepubertal girl with a positive culture with Gardnerella vaginalis, being difficult to specify the value of the Gardnerella in the adolescent patients.

Rev Cubana Med Trop, 1999 May-Aug, 51(2), 133 - 4
{First report of Neisseria polysaccharea in Cuba}; Guzman Hernandez D et al.; The National Laboratory of Reference of Pathogenic Neisserias of "Pedro Kouri" Tropical Medicine Institute studied 58 strains which had been previously identified as Neisseria meningiditis and isolated from nasopharyngeal carriers from 1985-1998 . They were applied the knapp detection scheme plus the glutamiltransferase production test . The latter and the polyscharide production based on 5% sacarose allowed to make a differential diagnosis between N . meningitidis and Neisseria polyccharea . It was possible to identify a N . polysaccharia strain, a species that is reported for the first time in Cuba.

Rev Cubana Med Trop, 1999 May-Aug, 51(2), 116 - 9
{Antimicrobial susceptibility of Neisseria gonorrhoeae strains determined by disk diffusion}; Llanes Caballero R et al.; The Gonoccocus Laboratory of "Pedro Kouri" Tropical Medicine Institute carried out a study of in vitro susceptibility of Neisseria gonorrhoeae to penicillin, tetracycline, cefuroxime ceftriaxone, cefotaxine and ciprofoxacin by means of a disk diffusion method with the culture medium agar base GC plus supplement . In the first phase, the method was standardized and the reference N . gonorrhoeae ATCC 49226 strain was used whereas in the second phase, 50 gonococcal strains isolated in 8 provinces during 1995 and 1996 were examined . The results of such standardization confirmed that the antimicrobial susceptibility values were within the allowable limits . 52 and 34% of strains were resistant to penicillin and tetracycline respectively and all of them showed susceptibility to the rest of evaluated antimicrobial drugs . We recommend the use of the disk diffusion method for surveillance of gonococci resistance to these drugs in our country.

Rev Cubana Med Trop, 1999 May-Aug, 51(2), 99 - 105
{Validation of an ELISA for the measurement of human IgG against serogroup B Neisseria meningitidis protein}; Ferriol Marchena X et al.; An indirect ELISA test was optimised in which B meningoccocus outer membrane proteins included in the Cuban Vaccine known as VA-MENGOC-BC were used as capture antigents . Specific antibodies in serum samples from vaccinated individuals were detected using a human anti-IgG conjugate, i.e . alklayne phosphatase, thus the reaction developed with a specific substrate called p-nitrophenilphophate . Standard serum was obtained from a reference standard, the gage curve range was 625 to 20,000/mL . The control serum was selected from the most interesting areas for the samples, hence, assay characteristics were determined . Intra-assay, inter-assay and total inaccuracies were lower than 10% in the most linear area of the curve . Detectability was 700/mL . Recovery, paralelism and linearity studies showed an under 10% inaccuracy.

Syst Appl Microbiol, 2000 Apr, 23(1), 9 - 14
Organization of the ribosomal operon 165-235 gene spacer region in representatives of Neisseria gonorrhoeae; Van Looveren M et al.; Ribosomal rRNA gene fragments (rDNA) encompassing part of the 16S rDNA, the 16S-23S rDNA spacer region and part of the 23S rDNA of 229 Neisseria gonorrhoeae strains were enzymatically amplified using conserved primers . The fragments of approximately 1200 bp were subjected to restriction analysis with HinfI . This revealed 13 patterns (patterns I-XIII) of which patterns I (78 strains), II (32 strains), III (38 strains) and IV (56 strains) were the most abundant, comprising 89.1% of the strains . The obtained restriction patterns consisted of 3 to 8 bands, ranging in size from 32 to 854 bp . The sum of the obtained bands was about 1200 bp for patterns I, II, III, IV, V, IX, and XIII . However, for patterns VI, VII, VIII, X, XI and XII, the sum of the bands well exceeded the estimated size of approximately 1200 bp . We demonstrated that this results from sequence divergence in the 4 rRNA operons, present in the genome of N . gonorrhoeae, giving rise to patterns that are a combination of several other patterns.

Ann Emerg Med, 2000 Jul, 36(1), 28 - 32
Opportunistic urine ligase chain reaction screening for sexually transmitted diseases in adolescents seeking care in an urban emergency department; Embling ML et al.; STUDY OBJECTIVE: Neisseria gonorrhoeae and Chlamydia trachomatis are the most common bacterial sexually transmitted diseases (STDs) in sexually active youth and many infections are asymptomatic or unrecognized . This study used ligase chain reaction assays for determination of prevalence of gonococcal and chlamydial infections in adolescents seeking care at an urban emergency department . METHODS: An unlinked prevalence study was performed with ligase chain reaction tests on voided urine specimens from a convenience sample of adolescents 14 years or older who sought care at the Children's Hospital of Alabama ED . Demographic data and data on care provided in the ED were determined from retrospective chart review of those patients whose urine specimens were tested . RESULTS: Of 282 urine specimens screened, 13.5% (38) yielded positive findings on ligase chain reaction testing for either or both pathogens (20 {7%} positive for N gonorrhoeae, 23 {8%} positive for C trachomatis) . In the context of acute care, gonorrhea or chlamydial infection was diagnosed in 5 (1.8%) . STD prevalence did not vary significantly by age . Only 39% (15/38) of patients with infections detected by ligase chain reaction testing received potentially effective antibiotics as a result of their urgent care evaluation . CONCLUSION: Many adolescents use the ED for nonurgent care and unsuspected STDs are often missed . Urine ligase chain reaction testing is a sensitive, noninvasive means of detecting STDs by which unsuspected adolescent STD cases can be detected in an ED setting.

Presse Med, 2000 May 27-Jun 3, 29(19), 1048 - 52
{Current therapeutics in infectious dermatology}; Lascaux AS et al.; NEW AGENTS: Among new treatments used for infectious dermatology diseases, new agents for genital herpes, valaciclovir and famciclovir, have greatly simplified therapeutic schemes . Cidofovir has also been shown to be effective against aciclovir-resistant cutaneous and mucosal herpetic lesions and for the treatment of molluscum contagiosum . NEW ADMINISTRATION ROUTES: For genital papillomavirus infections, trials using systemic or intralesional administered interferon have not provided conclusive evidence but imiquimode appears to be quite promising . Itaconazole and fluconazole are effective for onchomycoses . NEW POSSIBILITIES: Ivermectine is effective against scabies, but must be reserved for particularly severe forms . Finally, the emergence of Neisseria gonorrhoeae strains resistant to fluoroquinolones is disquieting.

Int J STD AIDS, 2000 Jun, 11(6), 402 - 5
The prevalence of urethral infections amongst asymptomatic young men in Hat Yai, southern Thailand; Chandeying V et al.; The aim of this study was to survey sexual behaviour and estimate the prevalence of urethral infections amongst male vocational college students . A cross-sectional survey was performed among 479 young men attending 2 vocational colleges in Hat Yai, southern Thailand . Polymerase chain reaction (PCR) tests of first-void urine (FVU) samples were used to detect infection with Chlamydia trachomatis, Neisseria gonorrhoeae, Ureaplasma urealyticum, Mycoplasma genitalium and Mycoplasma hominis . Girlfriends were the usual sexual partners for 89% of men with only 11% regularly patronizing sex workers . Condom usage was low . The prevalence of any urethral infection was 15.9% with: C . trachomatis 4%, N . gonorrhoeae 0.2%, U . urealyticum 10.9%, M . genitalium 2.3% and M . hominis 1.3% . Infection with more than one organism was found in 2% of men . While the prevalence of infection with chlamydia or gonorrhoea was relatively low, the prevalence of 'any urethral infection' was moderately high and suggests that unprotected sexual intercourse is commonly occurring . As girlfriends were the most usual sexual partners, they must be at significant risk of pelvic infection . There is a need for programmes targeting this group of people.

Annu Rev Biochem, 1999, 68, 89 - 125
Inorganic polyphosphate: a molecule of many functions; Kornberg A et al.; Inorganic polyphosphate (poly P) is a chain of tens or many hundreds of phosphate (Pi) residues linked by high-energy phosphoanhydride bonds . Despite inorganic polyphosphate's ubiquity--found in every cell in nature and likely conserved from prebiotic times--this polymer has been given scant attention . Among the reasons for this neglect of poly P have been the lack of sensitive, definitive, and facile analytical methods to assess its concentration in biological sources and the consequent lack of demonstrably important physiological functions . This review focuses on recent advances made possible by the introduction of novel, enzymatically based assays . The isolation and ready availability of Escherichia coli polyphosphate kinase (PPK) that can convert poly P and ADP to ATP and of a yeast exopolyphosphatase that can hydrolyze poly P to Pi, provide highly specific, sensitive, and facile assays adaptable to a high-throughput format . Beyond the reagents afforded by the use of these enzymes, their genes, when identified, mutated, and overexpressed, have offered insights into the physiological functions of poly P . Most notably, studies in E . coli reveal large accumulations of poly P in cellular responses to deficiencies in an amino acid, Pi, or nitrogen or to the stresses of a nutrient downshift or high salt . The ppk mutant, lacking PPK and thus severely deficient in poly P, also fails to express RpoS (a sigma factor for RNA polymerase), the regulatory protein that governs > or = 50 genes responsible for stationary-phase adaptations to resist starvation, heat and oxidant stresses, UV irradiation, etc . Most dramatically, ppk mutants die after only a few days in stationary phase . The high degree of homology of the PPK sequence in many bacteria, including some of the major pathogenic species (e.g . Mycobacterium tuberculosis, Neisseria meningitidis, Helicobacter pylori, Vibrio cholerae, Salmonella typhimurium, Shigella flexneri, Pseudomonas aeruginosa, Bordetella pertussis, and Yersinia pestis), has prompted the knockout of their ppk gene to determine the dependence of virulence on poly P and the potential of PPK as a target for antimicrobial drugs . In yeast and mammalian cells, exo- and endopolyphosphatases have been identified and isolated, but little is known about the synthesis of poly P or its physiologic functions . Whether microbe or human, all species depend on adaptations in the stationary phase, which is truly a dynamic phase of life . Most research is focused on the early and reproductive phases of organisms, which are rather brief intervals of rapid growth . More attention needs to be given to the extensive period of maturity . Survival of microbial species depends on being able to manage in the stationary phase . In view of the universality and complexity of basic biochemical mechanisms, it would be surprising if some of the variety of poly P functions observed in microorganisms did not apply to aspects of human growth and development, to aging, and to the aberrations of disease . Of theoretical interest regarding poly P is its antiquity in prebiotic evolution, which along with its high energy and phosphate content, make it a plausible precursor to RNA, DNA, and proteins . Practical interest in poly P includes many industrial applications, among which is the microbial removal of Pi in aquatic environments.

J Formos Med Assoc, 2000 Apr, 99(4), 300 - 4
Microbiologic features of adult community-acquired bacterial meningitis in Taiwan; Fang CT et al.; BACKGROUND AND PURPOSE: Community-acquired bacterial meningitis (CABM) is a life-threatening disease that requires prompt initiation of appropriate antibiotic therapy . The purpose of this study was to determine the causative microorganisms of CABM and their antimicrobial susceptibility patterns at a major teaching hospital in Taipei from 1993 to 1998 . METHODS: A review of medical records and microbiologic data was used to identify cases of CABM and causative pathogens . Antimicrobial susceptibility testing for bacterial isolates was performed by the disk diffusion method . RESULTS: Among the 48 adult patients with a diagnosis of CABM during the study period, the causative pathogens were identified in 36 cases . Unlike reports from other countries, Klebsiella pneumoniae was the leading causative pathogen (33%), followed by Streptococcus pneumoniae (28%), Listeria monocytogenes (11%), Neisseria meningitidis (6%), Staphylococcus aureus (6%), streptococci (6%), and Pseudomonas aeruginosa (6%) . The incidence of CABM due to K . pneumoniae increased during the study period (p = 0.012, Poisson regression), while the incidence of CABM due to other pathogens remained stable . All of the CABM-associated K . pneumoniae isolates were susceptible to cefotaxime but 25% of the CABM-associated S . pneumoniae strains were not susceptible to penicillin G . CONCLUSIONS: Penicillin G alone was not an appropriate empiric therapy for adult CABM because a high percentage of cases were due to K . pneumoniae or penicillin nonsusceptible S . pneumoniae . While the recommendations for the initial empiric regimen for CABM due to S . pneumoniae in Taiwan remain to be developed, third-generation cephalosporins appear to be an appropriate initial empiric regimen for the treatment of CABM due to K . pneumoniae.

J Pediatr Adolesc Gynecol, 2000 May, 13(2), 90 - 1
Monitoring std prevalence and reproductive health care among high-risk adolescent women
Rietmeijer CA, Oh MK, Bull SS, Brown PR, Wang SA, Mertz KJ.
Background: Urban adolescent women, particularly those in socially disadvantaged situations are at high risk for infections with Chlamydia trachomatis (CT), and Neisseria gonorrhoeae (GC) and their sequelae . Non-invasive screening technologies, including ligase chain reaction (LCR) and polymerase chain reaction (PCR) urine tests for CT and GC allow for provision of screening services in settings frequented by high-risk adolescents where such services have not been traditionally provided . In 1999, the CDC lead a multi-site project monitoring STD prevalence and reproductive health service, collecting a standard set of variables in women <20 yrs in juvenile justice system, drug treatment centers, and school-based clinics . This report provides preliminary data from Alabama and Colorado . Methods: Adolescent women who present for intake/services at selected facilities are enrolled in the project . In collaboration with CDC, sites developed a set of standard variables to be collected, including demographics, risk behaviors, symptoms, test performed, and test results . Dependent on services already performed, sites added CT and/or GC urine PCR/LCR and pregnancy testing to existing screening protocols.Results: In first 3-months, 533 adolescent women were enrolled in this on-going study; 379 in juvenile institutions, 38 in drug treatment centers, and 116 in school-based clinics . Mean age was 16 yrs, over 90% were sexually experienced, nearly 50% did not use condom with the last sex, less than 40% reported "current" birth control use, over 2/3 had sexual intercourse in past 3 months, many with multiple partners . Prevalence rates for CT and GC are summarized in the table . Conclusions: Non-invasive (urine) LCR/PCR tests allow for easy monitoring of selected STDs among adolescent women in high-risk settings . These findings may contribute to a sentinel surveillance system among high-risk populations and, in turn, may provide the rationale for the development of health care/intervention priorities at policy level.

Mol Immunol, 2000 Feb-Mar, 37(3-4), 133 - 40
Expression of the recombinant human immunoglobulin J chain in Escherichia coli; Symersky J et al.; Selective transport of polymeric (p) immunoglobulins (Ig) of IgA and IgM isotypes into external secretions by pIg receptor-mediated mechanism depends on the incorporation of joining (J) chain into the polymers . Until now, availability of a free J chain for immunological and biophysical studies has been limited to preparations of denatured J chain forms with moderate yield . Here we report that a recombinant J chain (rJ) can be over-expressed as a soluble fusion protein with thioredoxin using a modified vector pET32 in Escherichia coli . An intact J chain was released by digestion with IgA1 protease from Neisseria gonorrhoeae and isolated in a good yield with immunological and biochemical properties similar to those of J chain obtained by chemical cleavage from pIgA.

Ann Periodontol, 1999 Dec, 4(1), 20 - 31
Non-plaque-induced gingival lesions; Holmstrup P; The origin of gingival inflammation is occasionally different from that of routine plaque-associated gingivitis, and such non-plaque-associated types of gingivitis often present characteristic clinical features . Examples of such forms of gingivitis are specific bacterial, viral, and fungal infections . Specific bacterial infections of gingiva may be due to Neisseria gonorrhea, Treponema pallidum, streptococci, and other organisms . The most important viral infections of gingiva are herpes simplex virus type 1 and 2 and varicella-zoster virus . Fungal infections may be caused by several fungi, the most important of these being Candida species including C . albicans, C . glabrata, C . krusei, C . tropicalis, C . parapsilosis, and C . guillermondii . Gingival histoplasmosis is a granulomatous disease caused by the fungus Histoplasma capsulatum and, as for the other specific infections of gingiva, a confirmed diagnosis may require histopathologic examination and/or culture . Atypical gingivitis may also occur as gingival manifestations of dermatological diseases, the most relevant of these being lichen planus, pemphigoid, pemphigus vulgaris, erythema multiforme, and lupus erythematosus . Non-plaque induced gingival inflammation can be caused by allergic reactions to dental restorative materials, toothpastes, mouthwashes, and foods . In addition, gingival inflammation may result from toxic reactions, foreign body reactions, or mechanical and thermal trauma.

Infect Immun, 2000 Jul, 68(7), 4368 - 9
Expression of AniA, the major anaerobically induced outer membrane protein of Neisseria gonorrhoeae, provides protection against killing by normal human sera; Cardinale JA et al.; Anaerobically grown Neisseria gonorrhoeae has previously been shown to have elevated serum resistance in the absence of exogenous CMP-N-acetylneuraminic acid or detectable sialylation . We hypothesized that the anaerobically induced gonococcal outer membrane protein AniA might have a role in this phenomenon, as it is the only known gonococcal protein that is absent under aerobic conditions . An N . gonorrhoeae F62 derivative, RUG7035, in which aniA is under control of the tac promoter, was used to examine the effect of AniA expression on serum resistance . In this study, we found that expression of AniA enhanced the serum resistance of N . gonorrhoeae and may account for these earlier observations.

Infect Immun, 2000 Jul, 68(7), 3894 - 9
The lipopolysaccharide structures of Salmonella enterica serovar Typhimurium and Neisseria gonorrhoeae determine the attachment of human mannose-binding lectin to intact organisms; Devyatyarova-Johnson M et al.; Mannose-binding lectin (MBL) is an important component of the innate immune system . It binds to the arrays of sugars commonly presented by microorganisms and activates the complement system independently of antibody . Despite detailed knowledge of the stereochemical basis of MBL binding, relatively little is known about how bacterial surface structures influence binding of the lectin . Using flow cytometry, we have measured the binding of MBL to a range of mutants of Salmonella enterica serovar Typhimurium and Neisseria gonorrhoeae which differ in the structure of expressed lipopolysaccharide (LPS) . For both organisms, the possession of core LPS structures led to avid binding of MBL, which was abrogated by the addition of O antigen (Salmonella serovar Typhimurium) or sialic acid (N . gonorrhoeae) . Truncation of the LPS within the core led to lower levels of MBL binding . It was not possible to predict the magnitude of MBL binding from the identity of the LPS terminal sugar alone, indicating that the three-dimensional disposition of LPS molecules is probably also of importance in determining MBL attachment . These results further support the hypothesis that LPS structure is a major determinant of MBL binding.

Microbiology, 2000 Jun, 146 ( Pt 6), 1321 - 32
Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa; Semmler AB et al.; Transposon mutagenesis was used to identify a new locus required for twitching motility in Pseudomonas aeruginosa . Four Tn5-B21 mutants which lacked twitching motility and a fifth which exhibited impaired motility were found to map to the same KPN:I restriction fragment at approximately 40 min on the P . aeruginosa genome . Cloning and sequencing studies showed that all five transposon insertions occurred within the same 2.8 kb ORF, which was termed fimV . The product of this gene has a putative peptidoglycan-binding domain, predicted transmembrane domains, a highly acidic C terminus and anomalous electrophoretic migration, indicating unusual primary or secondary structure . The P . aeruginosa genome also possesses a paralogue of fimV . Homologues of fimV were also found in the sequenced genomes of the other type-IV-fimbriated bacteria Neisseria gonorrhoeae, Neisseria meningitidis, Legionella pneumophila and Vibrio cholerae, but not in those of other bacteria which lack type IV fimbriae . A fimV homologue was also found in the genome sequence of Shewanella putrefaciens, along with many other homologues of type IV fimbrial genes, indicating that this bacterium is also likely to produce type IV fimbriae . Wild-type twitching motility was restored to fimV mutants by complementation in a dosage-dependent manner . Overexpression of fimV resulted in an unusual phenotype where the cells were massively elongated and migrated in large convoys at the periphery of the colony . It is suggested that FimV may be involved in remodelling of the peptidoglycan layer to enable assembly of the type IV fimbrial structure and machinery.

Mol Microbiol, 2000 Jun, 36(5), 1059 - 70
Gonococcal lipooligosaccharide is a ligand for the asialoglycoprotein receptor on human sperm; Harvey HA et al.; In the present study, we show that Neisseria gonorrhoeae lipooligosaccharide (LOS) can bind to the asialoglycoprotein receptor (ASGP-R) on human sperm . This work demonstrates the presence of ASGP-R on human sperm . Binding of purified ASGP-R ligand decreased in the presence of gonococci . Binding of purified iodinated gonococcal LOS identified a protein of molecular weight corresponding to that of human ASGP-R . The presence of excess unlabelled LOS blocked binding of iodinated gonococcal LOS . Binding of wild-type gonococcal LOS to sperm was higher than that of mutant LOS lacking the galactose ligand for ASGP-R . These data suggest that the ASGP-R on human sperm cells recognizes and binds wild-type gonococcal LOS . This interaction may contribute to the transmission of gonorrhea from infected males to their sexual partners.

J Antimicrob Chemother, 2000 Jun, 45(6), 777 - 82
A multiplex polymerase chain reaction to differentiate beta-lactamase plasmids of neisseria gonorrhoeae; Palmer HM et al.; In penicillinase-producing Neisseria gonorrhoeae (PPNG), resistance to penicillin may be mediated by one of several related plasmids of different sizes . These include the Asian, African and Rio/Toronto plasmids . Identification of these plasmids provides useful epidemiological information, but has necessitated plasmid purification and gel analysis . We have developed a rapid, simple multiplex polymerase chain reaction (PCR) which discriminates between the beta-lactamase resistance plasmids that are frequently found in strains of N . gonorrhoeae . Amplicons of 1191, 958 and 650 bp were produced from strains containing the African, Asian and Rio/Toronto plasmids, respectively, whilst no products resulted from non-PPNG strains harbouring the cryptic, conjugative or tetracycline resistance plasmids . PCR analysis of 123 strains of PPNG identified 60 strains with African, 16 strains with Asian and 47 strains with Rio/Toronto plasmids and showed complete agreement with the standard plasmid analysis.

J Infect Dis, 2000 Jun, 181(6), 2080 - 2 Epub 2000 May 26.
Gonorrhea among men who have sex with men: outbreak caused by a single genotype of erythromycin-resistant Neisseria gonorrhoeae with a single-base pair deletion in the mtrR promoter region; Xia M et al.; During 1995-1997, an outbreak of 66 cases of gonorrhea caused by an erythromycin-resistant (Ery(r); MIC >/=1.0 microgram/mL) prototrophic (proto) auxotype IB-1 serovar of Neisseria gonorrhoeae occurred in King County, Washington; 65 cases involved men who have sex with men (MSM), which accounted for approximately 37% of infections among MSM during this period . Isolates from 19 of these 65 cases of infection were analyzed by DNA sequencing of the polymerase chain reaction-amplified promoter region of the mtrR gene and by pulsed-field gel electrophoresis (PFGE) analysis of genomic DNA after NheI and SpeI digestion . Eighteen of the 19 isolates had a 1-bp A/T deletion in a 13-bp inverted repeat of the mtrR promoter region and shared a single PFGE type . Among MSM who provided data about sexual behavior, 37 (64%) of 58 MSM infected by the proto/IB-1 Ery(r) strain reported having >2 sex partners during the past 60 days, compared with 32 (30%) of 106 MSM infected by other strains (P<.001) . This clonal outbreak of gonorrhea illustrates the ongoing need for behavioral preventive interventions among MSM.

J Clin Microbiol, 2000 Jun, 38(6), 2416 - 8
Reproducibility problems with the Abbott laboratories LCx assay for Chlamydia trachomatis and Neisseria gonorrhoeae; Gronowski AM et al.; This study demonstrates that significant reproducibility problems can occur during routine use of the Abbott Laboratories LCx assay for Chlamydia trachomatis and Neisseria gonorrhoeae . These problems can go undetected by the quality control procedures outlined in the manufacturer's package insert . We outline here procedures for detecting and preventing contamination and reproducibility problems.

Microbiology, 2000 May, 146 ( Pt 5), 1151 - 6
Additive effects of a two-amino-acid insertion and a single-amino-acid substitution in dihydropteroate synthase for the development of sulphonamide-resistant Neisseria meningitidis; Qvarnstrom Y et al.; Sulphonamide resistance in some clinical isolates of Neisseria meningitidis is associated with an insertion in the chromosomal folP gene leading to the addition of two amino acids, serine and glycine, in the drug target enzyme dihydropteroate synthase (DHPS) . Removal of the insertion resulted in a markedly higher Km for the substrate p-aminobenzoic acid and a markedly lower Km for 2-amino-4-hydroxy-6-(hydroxymethyl)-7,8-dihydropteridine pyrophosphate . In the same isolates an additional important difference, compared to wild-type enzymes, was found at amino acid position 68, which is a proline in most DHPS enzymes, but is serine in one and leucine in another clinical isolate of sulphonamide-resistant N . meningitidis . The alteration at position 68 was found to affect mainly the level of sulphonamide resistance and had only a minor effect on the Km for the substrates . Introduction of the serine-glycine dipeptide at position 194 and a proline to serine substitution at position 68 in DHPS from normal, susceptible N . meningitidis failed to produce a functional sulphonamide-resistant enzyme . The conclusion of this study is that it is not possible to change a normal chromosomally encoded DHPS of N . meningitidis to a sulphonamide-resistant one simply by an insertion of serine and glycine as seen in clinical isolates . It is likely that the resistance gene found in clinical isolates has evolved in another bacterial species where a combination of other amino acid changes may have contributed to produce a functionally resistant enzyme . This new resistance gene may have then been introduced into N . meningitidis by natural transformation.

J Inorg Biochem, 2000 Apr, 79(1-4), 219 - 23
Engineering outer-membrane proteins in Pseudomonas putida for enhanced heavy-metal bioadsorption; Valls M et al.; Metallothioneins (MTs) are small, cysteine-rich proteins with a strong metal-binding capacity that are ubiquitous in the animal kingdom . Recombinant expression of MT fused to outer-membrane components of gram-negative bacteria may provide new methods to treat heavy-metal pollution in industrial sewage . In this work, we have engineered Pseudomonas putida, a per se highly robust microorganism able to grow in highly contaminated habitats in order to further increase its metal-chelating ability . We report the expression of a hybrid protein between mouse MT and the beta domain of the IgA protease of Neisseria in the outer membrane of Pseudomonas cells . The metal-binding capacity of such cells was increased three-fold . The autotranslocating capacity of the beta domain of the IgA protease of Neisseria, as well as the correct anchoring of the transported protein into the outer membrane, have been demonstrated for the first time in a member of the Pseudomonas genus.

FEBS Lett, 2000 May 26, 474(1), 33 - 7
Identification of key amino acid residues in Neisseria polysaccharea amylosucrase; Sarcabal P et al.; Amylosucrase from Neisseria polysaccharea catalyzes the synthesis of an amylose-like polymer from sucrose . Sequence alignment revealed that it belongs to the glycoside hydrolase family 13 . Site-directed mutagenesis enabled the identification of functionally important amino acid residues located at the active center . Asp-294 is proposed to act as the catalytic nucleophile and Glu-336 as general acid base catalyst in amylosucrase . The conserved Asp-401, His-195 and His-400 residues are critical for the enzymatic activity . These results provide strong support for the predicted close structural and functional relationship between the sucrose-glucosyltransferases and enzymes of the alpha-amylase family.

Int J STD AIDS, 2000 May, 11(5), 284 - 7
Rectal gonorrhoea in homosexual men: source of infection; McMillan A et al.; The objective of this retrospective study was to determine the possible source of infection in homosexual men with rectal gonorrhoea: the probable source of rectal gonorrhoea was identified in 46/155 cases . Although the urethra was the site of infection in 33 (72%) of these contacts, only pharyngeal gonorrhoea was identified in 9 (20%) men . In 25/26 cases, there was concordance in the auxo/serotypes of Neisseria gonorrhoeae between contacts with urethral gonorrhoea and the index men with rectal gonorrhoea . Eleven out of 12 pharyngeal isolates were of the same auxo/serotype as the index cases . This study supports the hypothesis that rectal gonorrhoea in homosexual men can be acquired from the oropharynx . Because infection at this site is an independent risk factor for acquisition of HIV, screening for rectal and pharyngeal gonorrhoea should be offered to men who have sex with men, even when there is no history of unprotected receptive anal intercourse.

J Antimicrob Chemother, 2000 Apr, 45 Suppl 1, 67 - 70
Anti-gonococcal activity of gemifloxacin against fluoroquinolone-resistant strains and a comparison of agar dilution and Etest methods; Jones RN et al.; Gemifloxacin is a novel quinolone with excellent activity against Gram-positive and some Gram-negative pathogens . Its activity was tested against 150 Neisseria gonorrhoeae strains, including 50 ciprofloxacin-resistant isolates, using reference agar dilution and Etest methods . Gemifloxacin was found to be highly potent against ciprofloxacin-susceptible strains (MIC(90) 0.008 mg/L), but was 16-fold less potent against ciprofloxacin-resistant gonococci . The order of quinolone potency against these fluoroquinolone-resistant mutants was: gemifloxacin (MIC(90) 0.12 mg/L) > trovafloxacin (0.25 mg/L) > moxifloxacin = grepafloxacin (0.5 mg/L) > ciprofloxacin (1 mg/L) . Etest and reference agar dilution MIC results showed excellent correlation (r = 0.96) and >98% of MICs were within +/-1 log(2) dilution step (essential agreement) . The excellent potency of gemifloxacin indicates its potential for the treatment of infections with quinolone-resistant N . gonorrhoeae.

J Infect Dis, 2000 May, 181(5), 1652 - 60 Epub 2000 May 15.
A typing system for neisseria gonorrhoeae based on biotinylated oligonucleotide probes to PIB gene variable regions; Thompson DK et al.; The porin proteins PIA and PIB of Neisseria gonorrhoeae are serotyping antigens for the serovar classification system and leading candidates for gonococcal vaccine development . Although serotyping has been a useful tool, this method can be insensitive to critical sequence changes in the por gene, including those in surface-exposed variable regions (VRs) . A sensitive and specific typing system for N . gonorrhoeae has been developed that uses biotin-labeled oligonucleotide probes with chemiluminescence detection to type PIB gene VRs . The PIB VR types of geographically and temporally diverse gonococcal strains and sexual contact isolates were determined . por VR typing discriminated between most unrelated isolates and provided information about individual VR type that was not apparent from serovar designations . PIB VR typing avoids limited monoclonal antibody availability, interlaboratory variation, and the requirement for culture-based surveillance associated with gonococcal serotyping, and provides useful information about the molecular epidemiology of individual por gene VRs.

Sex Transm Dis, 2000 May, 27(5), 278 - 83
Neisseria gonorrhoeae MS11mkC opacity protein expression in vitro and during human volunteer infectivity studies; Schmidt KA et al.; BACKGROUND AND OBJECTIVES: Neisseria gonorrhoeae MS11mkC harbors 11 independently expressed opacity (Opa) protein genes with distinct in vitro expression frequencies . In experimental infections in which human male volunteers were inoculated with transparent (Opa), piliated (P+) strains, the authors associate onset of symptoms with recovery of opaque (Opa+) gonococci . GOALS: In vitro and recovered (Opa) protein expression rates were compared to determine if the human host influences Opa expression . STUDY DESIGN: Opa expression was determined using Western immunoblot analysis; Opa sizes were determined using a scanning densitometer . RESULTS: Seven of 10 Opa proteins were identified in gonococci recovered from all of the volunteers at frequencies consistent with in vitro results (Opa C, 29.5 kDa; Opa K, 30 kDa; Opa G, 31 kDa; Opa I, 32 kDa; Opa J, 33 kDa; Opa D, 34 kDa; and Opa H, 37 kDa) (P > or = 0.01, Fisher exact test) . Opa B (30.5 kDa) was identified at lower than expected frequencies, whereas Opa E (31.2) and F (31.5) were identified at higher' than expected frequencies . When recovered gonococci were reanalyzed for in vitro expression frequencies, they were consistent with preinfection frequencies . CONCLUSIONS: The host may influence the prevalence of some Opa proteins.

Sex Transm Dis, 2000 May, 27(5), 243 - 8
Effect of coinfection with STDs and of STD treatment on HIV shedding in genital-tract secretions: systematic review and data synthesis; Rotchford K et al.; OBJECTIVE: To determine whether coinfection with sexually transmitted diseases (STD) increases HIV shedding in genital-tract secretions, and whether STD treatment reduces this shedding . DESIGN: Systematic review and data synthesis of cross-sectional and cohort studies meeting predefined quality criteria . MAIN OUTCOME MEASURES: Proportion of patients with and without a STD who had detectable HIV in genital secretions, HIV load in genital secretions, or change following STD treatment . RESULTS: Of 48 identified studies, three cross-sectional and three cohort studies were included . HIV was detected significantly more frequently in participants infected with Neisseria gonorrhoeae (125 of 309 participants, 41%) than in those without N gonorrhoeae infection (311 of 988 participants, 32%; P = 0.004) . HIV was not significantly more frequently detected in persons infected with Chlamydia trachomatis (28 of 67 participants, 42%) than in those without C trachomatis infection (375 of 1149 participants, 33%; P = 0.13) . Median HIV load reported in only one study was greater in men with urethritis (12.4 x 104 versus 1.51 x 104 copies/ml; P = 0.04) . In the only cohort study in which this could be fully assessed, treatment of women with any STD reduced the proportion of those with detectable HIV from 39% to 29% (P = 0.05), whereas this proportion remained stable among controls (15-17%) . A second cohort study reported fully on HIV load; among men with urethritis, viral load fell from 12.4 to 4.12 x 104 copies/ml 2 weeks posttreatment, whereas viral load remained stable in those without urethritis . CONCLUSION: Few high-quality studies were found . HIV is detected moderately more frequently in genital secretions of men and women with a STD, and HIV load is substantially increased among men with urethritis . Successful STD treatment reduces both of these parameters, but not to control levels . More high-quality studies are needed to explore this important relationship further.

Am J Obstet Gynecol, 2000 May, 182(5), 1048 - 9
The papanicolaou smear: inadequate screening test for bacterial vaginosis during pregnancy; Greene JF 3rd et al.; OBJECTIVE: Our purpose was to evaluate the ability of the Papanicolaou smear to identify bacterial vaginosis in comparison with the Amsel clinical criteria . STUDY DESIGN: We retrospectively identified 159 pregnant women screened for bacterial vaginosis with the Amsel criteria who had a contemporaneous Papanicolaou smear and negative results on screening for Chlamydia trachomatis and Neisseria gonorrhoeae . Bacterial vaginosis was identified in 45 women . We used the McNemar chi(2) test to determine discrepancies between the two screening methods for the detection of bacterial vaginosis . RESULTS: Compared with the Amsel criteria, the sensitivity and specificity of the Papanicolaou smear for yielding a diagnosis of bacterial vaginosis were 49% (95% confidence interval, 36%-64%) and 93% (95% confidence interval, 86%-97%), respectively, with a positive predictive value of 73% and a negative predictive value of 82% . The detection of bacterial vaginosis by Papanicolaou smear was significantly different from that by Amsel criteria (P = . 01) . CONCLUSION: The Papanicolaou smear is not a reliable screening test for bacterial vaginosis during pregnancy.

DNA Res, 2000 Apr 28, 7(2), 75 - 81
Genomic, transcriptional and phenotypic analysis of ftsE and ftsX of Neisseria gonorrhoeae; Bernatchez S et al.; Although ftsE and ftsX are not universally present in bacteria, they are present in various Neisseria species as determined by Southern hybridization . The ftsE and ftsX genes of Neisseria gonorrhoeae (Ng) CH811 were cloned, sequenced and were shown to be co-transcribed from two promoters (P(E)1 and P(E)2) which were identified upstream of ftsE(Ng) by primer extension . Sequence analysis of FtsE(Ng) and alignment with other FtsE indicated that it contained the conserved motifs of ABC domains while sequence alignment of FtsX(Ng) with other published FtsX sequences predicted that they all contain four transmembrane segments and a conserved motif (Leu-hydrophobic aa-Gly-Ala/Gly) which may prove to be important for FtsX function . The viability of ftsE(Ng) and ftsX(Ng) mutants that were constructed by insertional inactivation indicated that these genes are not essential . The role of FtsE and FtsX is controversial . Analysis of ftsE(Ng) and ftsX(Ng) mutants by transmission electron microscopy showed that both exhibited morphological abnormalities indicative of defective division sites and in some cases aberrant condensation of DNA.

Ann N Y Acad Sci, 2000, 900, 309 - 15
Conservative management of PID; Georgilis K; The goals in the management of pelvic inflammatory disease (PID) are not only treatment of the infection and prevention of immediate complications, but also prevention of its long-term consequences . There are criteria for hospitalization, but patients who do not meet them can be safely treated as outpatients . A variety of sexually transmitted and other microorganisms can cause this infection, but the most important are Chlamydia trachomatis and Neisseria gonorrhoeae . Regimens with activity against gonococci, chlamydiae, streptococci, gram-negative bacteria, and anaerobes should be administered . Several such antimicrobial regimens have shown very good clinical and microbiologic efficacy . However, their efficacy in preventing long-term complications, such as infertility, has not been established . Close follow-up is an important part of management . Evaluation of male sexual partners is imperative to prevent reinfection . Better diagnostic techniques and treatment modalities for PID must be developed to prevent its long-term consequences.

Trop Med Int Health, 2000 Apr, 5(4), 280 - 7
Microheterogeneity of serogroup A (subgroup III) Neisseria meningitidis during an outbreak in northern Ghana; Gagneux S et al.; During a meningitis outbreak in the eastern subdistrict of the Kassena-Nankana District of the Upper East Region of Ghana, we analysed cerebrospinal fluid from suspected meningitis cases for the most common causative organisms . In 50 of 92 samples analysed, serogroup A Neisseria meningitidis were detected . The ages of serogroup A N . meningitidis patients ranged from 4 months to 64 years . The case fatality ratio was 20% . Coma or stupor on presentation worsened the prognosis . All serogroup A N . meningitidis isolates recovered revealed the A: 4: P1.9, 20 phenotype characteristic for the subgroup III clonal grouping . No evidence for resistance to penicillin G, chloramphenicol, cefotaxime, ciprofloxacin, rifampicin or tetracycline was found . All strains were resistant to sulphadiazine . Restriction analysis patterns of opa, iga and ingA genes were characteristic for the majority of N . meningitidis serogroup A subgroup III bacteria isolated in Africa after the 1987 epidemic in Mecca . Differences in pulsed-field gel electrophoresis patterns of NheI and SpeI digested DNA revealed microheterogeneity among the Ghanaian isolates.

J Biol Chem, 2000 Aug 4, 275(31), 23569 - 76
Two glycosylase/abasic lyases from Neisseria mucosa that initiate DNA repair at sites of UV-induced photoproducts; Nyaga SG et al.; Diverse organisms ranging from Escherichia coli to humans contain a variety of DNA repair proteins that function in the removal of damage caused by shortwave UV light . This study reports the identification, purification, and biochemical characterization of two DNA glycosylases with associated abasic lyase activity from Neisseria mucosa . These enzymes, pyrimidine dimer glycosylase I and II (Nmu-pdg I and Nmu-pdg II), were purified 30,000- and 10,000-fold, respectively . SDS-polyacrylamide gel electrophoresis analysis indicated that Nmu-pdg I is approximately 30 kDa, whereas Nmu-pdg II is approximately 19 kDa . The N-terminal amino acid sequence of Nmu-pdg II exhibits 64 and 66% identity with E . coli and Hemophilus parainfluenzae endonuclease III, respectively . Both Nmu-pdg I and Nmu-pdg II were found to have broad substrate specificities, as evidenced by their ability to incise DNA containing many types of UV and some types of oxidative damage . Consistent with other glycosylase/abasic lyases, the existence of a covalent enzyme-DNA complex could be demonstrated for both Nmu-pdg I and II when reactions were carried out in the presence of sodium borohydride . These data indicate the involvement of an amino group in the catalytic reaction mechanism of both enzymes.

Trop Med Int Health, 2000 Apr, 5(4), 280 - 7
Microheterogeneity of serogroup A (subgroup III) neisseria meningitidis during an outbreak in northern ghana
Gagneux S.
During a meningitis outbreak in the eastern subdistrict of the Kassena-Nankana District of the Upper East Region of Ghana, we analysed cerebrospinal fluid from suspected meningitis cases for the most common causative organisms . In 50 of 92 samples analysed, serogroup A Neisseria meningitidis were detected . The ages of serogroup A N . meningitidis patients ranged from 4 months to 64 years . The case fatality ratio was 20% . Coma or stupor on presentation worsened the prognosis . All serogroup A N . meningitidis isolates recovered revealed the A: 4: P1.9, 20 phenotype characteristic for the subgroup III clonal grouping . No evidence for resistance to penicillin G, chloramphenicol, cefotaxime, ciprofloxacin, rifampicin or tetracycline was found . All strains were resistant to sulphadiazine . Restriction analysis patterns of opa, iga and ingA genes were characteristic for the majority of N . meningitidis serogroup A subgroup III bacteria isolated in Africa after the 1987 epidemic in Mecca . Differences in pulsed-field gel electrophoresis patterns of NheI and SpeI digested DNA revealed microheterogeneity among the Ghanaian isolates.

Subcell Biochem, 2000, 33, 61 - 96
Host cell invasion by pathogenic Neisseriae; Dehio C et al.; As outlined in this review, various experimental techniques have been employed in an attempt to understand neisserial pathogenesis . In vitro genetic analysis has been used to study the genetic basis for the structural variability of cell surface components . Transformed or primary epithelial cell cultures have provided the simplest model to analyze bacterial adherence and invasion, while the infection of polarized epithelial monolayers, fallopian tube and nasopharyngeal organ cultures, and ureteral tissue have each been used to more closely represent the events which occur in vivo . Finally, the in vivo infection of human volunteers with N . gonorrhoeae has provided a powerful means to confirm and expand the results obtained in vitro . By these various approaches, a number of neisserial adhesins (i.e . pilli, Opa, Opc and P36) and additional putative virulence determinants which affect bacterial adherence and invasion into host cells (i.e . LOS, capsule, PorB) have been identified . Clearly, neisserial surface variation serves as an adaptive mechanism which can modulate tissue tropism, immune evasion and survival in the changing host environment . Important progress has been made in recent years with respect to the host cellular receptors and subsequent signal transduction processes which are involved in neisserial adherence, invasion and transcytosis . This has led to the identification of (i) CD46 as a receptor for pilus which allows adherence to epithelial and endothelial cells, (ii) HSPGs, in cooperation with vitronectin and fibronectin, as receptors for a particular subset of Opa proteins and Opc, which may both mediate invasion into most epithelial and endothelial cells, and (iii) CD66 as the receptors for most Opa variants, potentially being involved in cellular interactions including adherence, invasion and transcytosis with epithelial, endothelial and phagocytic cells . As most of these data have been obtained using transformed cell lines growing in vitro, attempts must be made to translate these basic observations into a more natural situation . It can be expected that the successful ongoing integration of laboratory findings from the various infection models with human volunteer studies will further increase our understanding of the biology of neisserial infection . Perhaps the most difficult but also most rewarding challenge for the future will be to use volunteer studies to identify and understand the role of host factors which are important for the infectious process . Hopefully, insights gained from each of these studies will reveal new and useful strategies for the preventive and/or therapeutic intervention into infection and disease by these fascinating microbes.

Drugs, 2000 Apr, 59(4), 801 - 13
Concurrent gonococcal and chlamydial infection: how best to treat; Robinson AJ et al.; Clinicians treating concurrent gonococcal and chlamydial infections have a variety of drugs to choose from . Neisseria gonorrhoeae is adept at developing resistance and the choice of antibiotic must be dictated to some extent by the patterns of resistance in the locality of the clinician . In contrast, resistance of Chlamydia trachomatis to some classes of drugs has been shown in vitro but does not appear to be clinically important at present . The success of treatment depends on patient compliance with the drug administration schedule . With these organisms, which can be carried asymptomatically, many patients are unlikely to comply with courses of antibiotics . Although single-dose therapy with azithromycin is available and established for chlamydial genital infection, it is more expensive and difficult to justify in a cash limited Healthcare system, and its efficacy for treating concurrent gonococcal infection requires further study . In patients where compliance is likely to be of concern, its use may be justified . Another major deterrent for completing antibiotic courses is the adverse effect profile . Most of the available drugs cause only minor adverse effects, in particular gastrointestinal . Ofloxacin has a better profile than doxycycline but is considerably more expensive . Newer fluoroquinolones, found to be effective in vitro, are being assessed in clinical studies . However, more evidence is required before recommending these over the tried and tested therapies.

FEMS Microbiol Lett, 2000 May 15, 186(2), 197 - 201
The genome of Neisseria gonorrhoeae retains the remnants of a two-component regulatory system that once controlled piliation; Carrick CS et al.; An intact activator-binding site upstream of the sigma(54) promoter of the pilin-encoding pilE gene of Neisseria gonorrhoeae suggests gonococci produce a protein capable of binding this sequence . We cloned a chimeric gene, rsp, that has sequence similarity to both the pilS and pilR genes of Pseudomonas aeruginosa encoding a two-component regulatory system that controls piliation . This gene is transcribed in N . gonorrhoeae and indirect evidence suggests that Rsp binds to the activator-binding site of the pilE gene . Despite this, mutation of rsp has no effect on piliation in N . gonorrhoeae, suggesting that the remnants of this regulatory system have persisted in the genome, despite the loss of its original function.

Cochrane Database Syst Rev . 2000;(2):CD000098.
Interventions for treating gonorrhoea in pregnancy; Brocklehurst P; BACKGROUND: Neisseria gonorrhoeae can be transmitted from the mother's genital tract to the newborn during birth and can cause gonococcal opthalmia neonatorum . OBJECTIVES: The objective of this review was to assess the effects of antibiotic regimens in the treatment of genital infection with gonorrhoea during pregnancy with respect to neonatal and maternal morbidity . SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched . Date of last search: October 1998 . SELECTION CRITERIA: Randomised trials of one regimen of antibiotic versus another in pregnant women with culture confirmed genital gonococcal infection . DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer . MAIN RESULTS: Two trials involving 329 women were included . Amoxycillin with probenicid or spectinomycin or ceftriaxone have a similar effect on microbiological cure, defined by negative gonococcal culture . REVIEWER'S CONCLUSIONS: Any of the antibiotic regimens tested in these trials appear to be effective for the treatment of gonorrhoea in pregnancy in terms of their effect on microbiological cure . For women who are allergic to penicillin, this review provides reassurance that treatment with ceftriaxone or spectinomycin appears to be at least as equally effective in producing microbiological cure.

FEBS Lett, 2000 Apr 28, 472(2-3), 287 - 92
IgA1 protease from Neisseria gonorrhoeae inhibits TNFalpha-mediated apoptosis of human monocytic cells; Beck SC et al.; The modulation of programmed cell death is a common theme in the patho-physiology of inflammation and infectious disease . The synthesis and secretion of an IgA1 protease is strictly associated with virulence of the Neisseria species . Here, we report on the inhibition of tumor necrosis factor alpha (TNFalpha)-mediated apoptosis of the human myelo-monocytic cell line U937 by highly purified IgA1 protease . Apoptosis was verified by the cell surface exposure of phosphatidyl serine and by terminal transferase mediated end-labeling of fragmented DNA . Interestingly, IgA1 protease specifically cleaved the TNF receptor II (TNF-RII) on the surface of intact cells whereas TNF-RI was not affected by the enzyme . Therefore, inhibition of TNFalpha-mediated apoptosis might be correlated to specific cleavage of the TNF-RII by neisserial IgA1 protease.

Microbiology, 2000 Apr, 146 ( Pt 4), 967 - 79
Genetic characterization of pilin glycosylation in Neisseria meningitidis; Power PM et al.; Pili of Neisseria meningitidis are a key virulence factor, being the major adhesin of this capsulate organism and contributing to specificity for the human host . Pili are post-translationally modified by addition of an O-linked trisaccharide, Gal(beta1-4)Gal(alpha1-3)2,4-diacetimido-2,4,6-trideoxyhexose++ + . In a previous study the authors identified and characterized a gene, pglA, encoding a galactosyltransferase involved in pilin glycosylation . In this study a set of random genomic sequences from N . meningitidis strain MC58 was used to search for further genes involved in pilin glycosylation . Initially, an open reading frame was identified, and designated pglD (pilin glycosylation gene D), which was homologous to genes involved in polysaccharide biosynthesis . The region adjacent to this gene was cloned and nucleotide sequence analysis revealed two further genes, pglB and pglC, which were also homologous with genes involved in polysaccharide biosynthesis . Insertional mutations were constructed in pglB, pglC and pglD in N . meningitidis C311#3, a strain with well-defined LPS and pilin-linked glycan structures, to determine whether these genes had a role in the biosynthesis of either of these molecules . Analysis of these mutants revealed that there was no alteration in the phenotype of LPS in any of the mutant strains as judged by SDS-PAGE gel migration . In contrast, increased gel migration of the pilin subunit molecules of pglB, pglC and pglD mutants by Western analysis was observed . Pilin from each of the pglB, pglC and pglD mutants did not react with a terminal-galactose-specific stain, confirming that the gel migration differences were due to the alteration or absence of the pilin-linked trisaccharide structure in these mutants . In addition, antisera specific for the C311#3 trisaccharide failed to react with pilin from the pglB, pglC, pglD and galE mutants . Analysis of nucleotide sequence homologies has suggested specific roles for pglB, pglC and pglD in the biosynthesis of the 2,4-diacetimido-2,4,6-trideoxyhexose structure.

Sex Transm Dis, 2000 Apr, 27(4), 236 - 40
Comparative prevalence of infection with Trichomonas vaginalis among men attending a sexually transmitted diseases clinic; Joyner JL et al.; OBJECTIVE: Although established in women as a common cause of vaginal discharge, the prevalence of Trichomonas vaginalis (TV) in men compared with other classic urethral pathogens has not been well characterized . To assess this issue, the authors compared the prevalence of Neisseria gonorrhoeae (GC), Chlamydia trachomatis (CT), and TV in consecutive men attending a sexually transmitted diseases (STD) clinic . METHODS: From June 1, 1998 to July 27, 1998, 454 consecutive men presenting to the Denver Metro Health Clinic with a new problem were tested for GC by urethral swab culture, for CT by polymerase chain reaction of urine, and for TV by urine sediment culture . RESULTS: GC, CT, and TV were detected in 23 (5.1%), 34 (7.5%), and 13 (2.8%) of men, respectively . There were significant differences by age for both CT (11.3% in men younger than 30 years versus 3.3% in men 30 years and older, P < 0.05) and TV (0.8% in men younger than 30 years versus 5.1% in men 30 years and older, P < 0.05) . In 50 men 30 years or older with symptoms of urethral discharge, TV prevalence (12.0%) rivalled that of GC (12.0%) and CT (14.0%) . In 45 men 30 years and older with nongonococcal urethritis, the prevalence of TV and CT were each 13.3% . Multivariate logistical regression analysis showed the presence of discharge and nongonococcal urethritis in men 30 years and older to be an independent predictor of TV . CONCLUSIONS: TV is common in men attending sexually transmitted disease clinics, especially in those 30 years or older, in whom it may account for as much urethritis as GC or CT . These findings suggest that in older men with nongonococcal urethritis, diagnostic evaluation, empiric treatment, and partner management should include the possibility of TV infection.

FEMS Microbiol Lett, 2000 May 1, 186(1), 103 - 8
Characterisation of the activator effect of glycogen on amylosucrase from Neisseria polysaccharea; Potocki de Montalk G et al.; Amylosucrase produces an insoluble alpha-1,4-linked glucan from sucrose, releasing fructose . In addition to polymerisation, in the presence of sucrose as sole substrate, amylosucrase catalyses sucrose hydrolysis and oligosaccharide synthesis in significant proportions . The effects of both glycogen acceptor and sucrose concentrations on the reactions catalysed by the highly purified amylosucrase from Neisseria polysaccharea were investigated . Sucrose hydrolysis decreased strongly with the increase of the concentration of glycogen, as did oligosaccharide synthesis, by glucose transfer onto glucose and fructose . The glucosyl units consumed were then preferentially used for elongation of glycogen chains . The study of the kinetic behaviour of amylosucrase revealed a strong, sucrose concentration dependent activator effect of glycogen . This activation was decreased at high sucrose concentration . The optimal sucrose concentrations increased with glycogen concentration, suggesting competition between sucrose and glycogen, and the presence of a second non-catalytic acceptor binding site which could bind various acceptors (glucose, maltose, glycogen) and also sucrose.

Int J STD AIDS, 2000 Apr, 11(4), 235 - 40
Can a two-glass urine test or leucocyte esterase test of first-void urine improve syndromic management of male urethritis in southern Thailand?
Chandeying V, Skov S, Tabrizi SN, Kemapunmanus M, Garland S.
The goal of this study was to determine whether a urine two-glass test or a leucocyte esterase (LE) test of first-void urine (FVU) improve the sensitivity or specificity of the World Health Organization (WHO) algorithm for the syndromic management of men with urethritis in southern Thailand . A secondary aim was to determine whether infection with Trichomonas vaginalis was sufficiently common to include treatment for it in a syndromic management protocol . One hundred and twenty-nine men with symptoms of urethritis seen at 2 STD clinics in Songkla Province, Thailand were enrolled . Symptoms and signs of each man were recorded and a urethral swab collected for microscopy and culture for Neisseria gonorrhoeae . A two-glass urine test and an LE test of an FVU specimen were performed . The FVU was tested by polymerase chain reaction (PCR) for N . gonorrhoeae, Chlamydia trachomatis and T . vaginalis . Dysuria was a symptom in 78% of men . A urethral discharge was a symptom in 68% but was evident on examination in 95% of the men . The prevalences of infection were 32.6% for N . gonorrhoeae, 23.3% for C . trachomatis, 1.6% for T . vaginalis and 51.9% for any infection . The sensitivities and specificities of urethral discharge on examination, two-glass test and LE test of FVU as indicators of infection with either or both of N . gonorrhoeae or C . trachomatis were 97% and 8%; 57% and 83%; and 59% and 78% respectively . Combinations of urethral discharge on examination and one of the other indicators were more specific but much less sensitive than the presence of discharge alone . Culture for N . gonorrhoeae was found to be only 43% sensitive compared with an expanded gold standard involving a PCR test . Our analysis demonstrates that neither the two-glass test nor the LE test of FVU were useful in improving on the WHO algorithm for management of men with urethritis . T . vaginalis was not common enough to include in a first-line syndromic management protocol for male urethritis . We recommend that, in southern Thailand, men with symptoms of urethritis in whom a urethral discharge is present on examination be offered immediate treatment for both N . gonorrhoeae and C . trachomatis as per the WHO algorithm.

Antimicrob Agents Chemother, 2000 May, 44(5), 1365 - 6
Spectinomycin resistance in Neisseria spp . due to mutations in 16S rRNA; Galimand M et al.; Spectinomycin resistance in clinical isolates of Neisseria meningitidis and Neisseria gonorrhoeae was found to be due to mutations G1064C and C1192U (Escherichia coli numbering) in 16S rRNA genes, respectively.

Clin Infect Dis, 2000 Apr, 30(4), 736 - 41 Epub 2000 Apr 03.
Features of urethritis in a cohort of male soldiers; McKee KT Jr et al.; Of 400 cases of urethritis in male soldiers enrolled in a behavioral intervention project, the etiology of 69% was defined at study enrollment, as well as the etiology of 72% of 25 repeated episodes involving 21 men during the first 78 days of active follow-up (5% of the cohort) . Chlamydia trachomatis (36%), Neisseria gonorrhoeae (34%), and Ureaplasma urealyticum (19%) were the most common causes of infection identified at enrollment and during subsequent visits (44%, 28%, and 12%, respectively) . By univariate analysis, patients with repeated infection ("repeaters") were significantly more likely to report a history of sexually transmitted disease (STD; relative risk {RR}, 3) and sex with sex workers (RR, 4) than were nonrepeaters . By multivariate analysis, only STD history was significant (RR, 2.8) . Characteristics of repeaters in this cohort suggest that specific patterns may be used to establish screening "profiles" of potential repeaters, by which such individuals might be targeted for aggressive intervention at the time of the initial diagnosis.

Gene, 2000 Apr 4, 246(1-2), 59 - 68
Genetic organization and heterogeneity of the iceA locus of Helicobacter pylori; Figueiredo C et al.; The genetic organization and sequence heterogeneity of the iceA locus of Helicobacter pylori was studied, and the existence of two distinct gene families, iceA1 and iceA2, at this locus was confirmed . iceA1 has significant sequence homology to nlaIIIR, encoding an endonuclease in Neisseria lactamica, but the similarity at the protein level is limited, due to frameshift mutations of iceA1 in most H . pylori strains . In only five of the 19 iceA1 strains studied, a full-length open reading frame (ORF), capable of encoding a 228aa protein, with 52% homology to NlaIII was observed . The region upstream of iceA2 is highly variable in length, containing up to 15 copies of 8bp tandem repeats . iceA2 can encode proteins of 24, 59, 94, or 129 amino acids, consisting of 14 and 10aa domains, conserved in all iceA2 strains, flanking 0, 1, 2, or 3 copies of a 35aa cassette . This 35aa cassette consists of domains of 13, 16 and 6aa, respectively . The 13aa and 6aa domains are highly conserved, but the 16aa domain exists in two variants . In total, five distinct iceA2 subtypes were defined . Database searches did not reveal any homologous sequences . Recombinant IceA1 and IceA2 proteins were expressed in Escherichia coli, confirming the predicted ORFs . Genotype-specific PCR primers permitted iceA genotyping in 318 (99 . 1%) of a worldwide collection of 321 H . pylori strains . The conserved sizes of the amplification products confirmed the worldwide distribution of discrete variants of iceA1 and iceA2.

FEBS Lett, 2000 Apr 14, 471(2-3), 219 - 23
Amylosucrase from Neisseria polysaccharea: novel catalytic properties; Potocki de Montalk G et al.; Amylosucrase is a glucosyltransferase that synthesises an insoluble alpha-glucan from sucrose . The catalytic properties of the highly purified amylosucrase from Neisseria polysaccharea were characterised . Contrary to previously published results, it was demonstrated that in the presence of sucrose alone, several reactions are catalysed, in addition to polymer synthesis: sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis - these two sucrose isomers being obtained by glucosyl transfer onto fructose . The effect of initial sucrose concentration on initial activity demonstrated a non-Michaelian profile never previously described.

East Afr Med J, 1999 Jun, 76(6), 330 - 4
Susceptibility pattern of Neisseria gonorrhoeae to antimicrobial agents in Dar es Salaam; Mbwana J et al.; OBJECTIVE: To determine the susceptibility pattern of local strains of Neisseria gonorrhoeae from Dar es salaam, Tanzania to locally used antibiotics . METHOD: Out of 429 Neisseria gonorrhoeae strains isolated between 1993 and 1995, one hundred and ninety nine were recovered and tested . Minimum inhibitory concentrations (MIC) of penicillin, doxycycline, erythromycin, cefuroxime and ciprofloxacin were determined by the E-test method while that of spectinomycin was measured by the agar dilution method . Penicillinase producing N . gonorrhoeae were identified by the chromogenic cephalosporin method . RESULTS: Of the 199 strains tested 128 (64%) were found to be penicillinase producing Neisseria gonorrhoeae (PPNG) . Only 19 (10%) were penicillin sensitive while all penicillin resistant strains were found to be PPNG . One hundred and seventy five (88%), 11(5%) and 13 (7%) of the tested isolates were resistant, less susceptible and fully susceptible to doxycycline respectively . Resistance to cotrimoxazole, cefuroxime and ciprofloxacin was 36 (18%), 11 (6%), and 3 (2%) respectively . The trend of antibiotic susceptibility rates over the three year period of study showed a significant increase in the proportion of susceptible strains to cotrimoxazole . All of the 75 strains tested against spectinomycin were susceptible . There was a statistically significant difference between the susceptibility patterns of non-PPNG and PPNG . Non-PPNG isolates were more susceptible to doxycycline (chi 2 = 78.2, df 2, p = < 0.0001) . CONCLUSION: These findings have shown that spectinomycin, ciprofloxacin and cefuroxime could continue to be used to treat gonorrhoea in our settings . Continuous surveillance of susceptibility to the commonly used antibiotics is important in order to detect emergence of resistance early and control the possible wide spread of resistant strainsPIP: This article presents a study on the susceptibility pattern of local strains of Neisseria gonorrheae (NG) to antimicrobial agents (penicillin, doxycyline, erythromycin, cefuroxime, ciprofloxacin, cotrimoxazole, and spectinomycin) in Dar Es Salaam, Tanzania . Out of the 429 isolated strains of NG in 1993-95, 199 were included in the study . Susceptibility patterns to the six antimicrobials was determined through the E-test method, a test that measures their minimum inhibitory concentrations (MICs) . On the other hand, the spectinomycin MIC was determined through the antibiotic agar dilution method . Results revealed the following patterns of susceptibility of isolates: spectinomycin (100%), ciprofloxacin (97%), Cefuroxime (89%), erythromycin (57%), cotrimoxazole (40%), doxycycline (7%), and penicillin (10%) . It was also noted that NG strains are highly resistant to penicillin (64%) and doxycycline (88%) . The study concludes that three drugs--spectinomycin, ciprofloxacin, and spectinomycin--could be effective in treating gonorrhea . However, a continued surveillance of common antibiotics against gonococcus is necessary for the early detection and control of strain resistance .

Br J Gen Pract, 2000 Feb, 50(451), 129 - 30
Survey of the use of selective culture for Neisseria gonorrhoeae in specimens from the female genital tract sent by general practitioners to a microbiology laboratory; Gillespie T et al.; A retrospective survey of the number of cultures found to be positive for Neisseria gonorrhoeae in genital specimens from female patients sent by general practitioners (GPs) over a three-year period was carried out . The organism could be detected in only four specimens out of over 28,000 specimens sent . Specifically, additional selective culture for N gonorrhoeae had been carried out in 8529 of these specimens . An estimate of the cost savings achievable if this laboratory was no longer to culture routinely for N gonorrhoeae was made . GPs should be aware of their local laboratory's normal practice when processing such specimen and should request specific culture if appropriate . The low number of specimens from which N gonorrhoeae could be cultured might suggest that GPs are referring 'at-risk' patients to genitourinary medicine clinics already.

J Clin Microbiol, 2000 Apr, 38(4), 1638 - 40
Long-term persistence of a discotheque-associated invasive Neisseria meningitidis group C strain as proven by pulsed-field gel electrophoresis and porA gene sequencing; Riesbeck K et al.; A cluster of a Neisseria meningitidis serogroup C strain causing invasive disease was investigated . Five out of seven cases were associated with a particular discotheque . The strains were indistinguishable, as revealed by pulsed-field gel electrophoresis and sequencing of variable regions of the porA gene, but caused strikingly different clinical presentations during 5 months.

J Med Virol, 2000 May, 61(1), 107 - 10
Hydrogels containing monocaprin prevent intravaginal and intracutaneous infections with HSV-2 in mice: impact on the search for vaginal microbicides; Neyts J et al.; Hydrogel formulations containing the 1-monoglyceride of capric acid (monocaprin) possess potent in vitro microbicidal activity against HIV and HSV, Chlamydia trachomatis and Neisseria gonorrhoeae . These formulations were studied to determine whether they prevent intracutaneous and intravaginal infections of mice with HSV-2, a virus that is in vitro as sensitive to the virucidal action of the compound as is HIV . In mice intravaginal infection with HSV-2 and the associated mortality was prevented completely when the infection was carried out in the presence of a 20 mM monocaprin containing gel formulation . Similarly, virtually complete protection of lesion development and associated mortality was observed when mice were infected intracutaneously with HSV-2 in the presence of gels containing 10 or 20 mM monocaprin . No irritation or toxicity was observed following application of the gel to the skin or the vaginal mucosa . Hydrogel formulations of monocaprin could thus be pursued as vaginal microbicides for the prevention of sexual transmission of HSV, HIV and other infectious pathogens .

APMIS, 2000 Feb, 108(2), 107 - 12
Lack of endotoxin tolerance with respect to TNF alpha production in the subarachnoid space; Steinshamn S et al.; To study endotoxin tolerance in the subarachnoid space 0.1 mg of endotoxin derived from Neisseria meningitidis was injected intracisternally into rabbits on 2 consecutive days . On day 1 the maximum peak level of TNF alpha was 7 ng/ml 2 h after injection, whereas on day 2 the highest levels were 3.6 ng/ml and 3.7 ng/ml, respectively, 1 and 2 h after injection . Pretreatment with intravenous endotoxin 5 or 21 h before consecutive intracisternal endotoxin did not affect the cerebrospinal fluid (CSF) levels of TNF alpha . In contrast, there was a marked endotoxin tolerance with respect to TNF alpha in the systemic circulation . Cells appearing in the CSF 5, 12 and 20 h after intracisternal injection of endotoxin were harvested, cultured, and then stimulated with 0.1 mg/ml of endotoxin . In 10 experiments a marked TNF alpha production in the range 10-70 ng/ml was detected in the supernatants, whereas unstimulated cells did not produce TNF alpha . We conclude that tolerance to endotoxin does not develop in the subarachnoid space as evaluated by the present experimental design . The pattern of TNF alpha production and endotoxin tolerance is distinctly different in the subarachnoid space and systemic circulation.

Commun Dis Intell, 2000 Jan 20, 24(1), 1 - 4
Surveillance of antibiotic resistance in Neisseria gonorrhoeae in the WHO Western Pacific Region, 1998 . The WHO Western Pacific Gonococcal Antimicrobial Surveillance Programme; Tapsall JW; Effective treatment of gonorrhoea in the World Health Organization's Western Pacific Region is hampered by the emergence and spread of antibiotic resistant strains of Neisseria gonorrhoeae . A programme of surveillance of gonococcal susceptibility to antibiotics (GASP) continued in the region in 1998 . A high proportion of isolates in many participating countries was resistant to quinolones and penicillins, continuing trends observed by this programme since 1992 . Resistance to the later generation cephalosporins and to spectinomycin was absent or infrequent . Options for effective treatment of gonorrhoea in the region have been severely compromised by antibiotic resistance.

Diagn Microbiol Infect Dis, 2000 Mar, 36(3), 163 - 8
Evaluation of four commercial transport media for the survival of Neisseria gonorrhoeae; Arbique JC et al.; We evaluated four commercial transport systems with a standardized inoculum of clinical isolates of N . gonorrhoeae (NG), and assessed survival after holding for up to 48 hours at both ambient and refrigeration temperatures . Suspensions of clinical isolates of NG were standardized and adsorbed onto four transport swab types: Culturette EZ (Becton Dickinson {BD}, Cockeysville, MD, USA); Cultureswab (Difco Laboratories, Detroit, MI, USA); Venturi Transystem (Copan Italia, Bovezzo, Italy); and a recently modified Starswab (Starplex Scientific, Etobicoke, ON) . Swabs were plated to chocolate agar at 0, 6, 24, and 48 hours, and colonies counted . Each swab type was tested in quadruplicate with each NG strain for all time and temperature variables . There was a marked reduction in NG CFUs after only 6 hours incubation with each of the swabs tested . Survival was best using Venturi Transystem and Cultureswab transports (colony counts were reduced to 15.3% and 13.0%, respectively, at 6 hours) when compared with the Culturette EZ and Starswab (colony counts were reduced to 2.2% and 4.3%, respectively, at 6 hours) . After the 24-hour holding period, 94% of the cultures from the Venturi Transystem were positive, 82% from the Cultureswab, 24% from the Starswab; and 17% from the Culturette EZ . After 48 hours, recovery dropped to 72%, 43%, 14%, and 0.04%, respectively . All of the systems tested had at least an 80% decrease in recovered colonies after only 6 hours . Further studies are required to determine how poor transport conditions influence the number of positive cultures and what the public health implications are . Of the swabs tested, Cultureswab and Venturi Transystem were most acceptable.

Akush Ginekol (Sofiia), 1999, 38(2), 23 - 5
{The etiology of infectious cervicitis in women}; Borisov I et al.; Infectious cervicitis is a common disease in women of reproductive age . A prospective study was carried out including 70 women with clinical manifestations of cervicitis during gynecological and colposcopic examination . In 19 women (27%), Chlamydia trachomatis was demonstrated by direct immunofluorescence in cervical smears . Neisseria gonorrhoeae was isolated in 2 women and 1 had a concomitant infection with Neisseria gonorrhoeae and Chlamydia trachomatis . Mycoplasma hominis and Ureaplasma urealyticum were isolated in 7 patients and 4 had cytological smears consistent with Human Papilloma Virus infection . In the rest 47 patients no etiological agent was found . The results of the present study indicate that the most common etiological cause for infectious cervicitis can be Chlamydia trachomatis and an examination for this organism should me mandatory in women with infectious cervicitis for guiding the correct diagnosis and treatment.

Biochim Biophys Acta, 2000 Apr 5, 1464(2), 284 - 98
Biochemical and biophysical characterization of in vitro folded outer membrane porin PorA of Neisseria meningitidis; Jansen C et al.; Two subtypes of the outer membrane porin PorA of Neisseria meningitidis, P1.6 and P1.7,16, were folded in vitro after overexpression in, and isolation from Escherichia coli . The PorA porins could be folded efficiently by quick dilution in an appropriate buffer containing the detergent n-dodecyl-N, N-dimethyl-1-ammonio-3-propanesulphonate . Although the two PorA porins are highly homologous, they required different acidities for optimal folding, that is, a pH above the pI was needed for efficient folding . Furthermore, whereas trimers of PorA P1.7,16 were almost completely stable in 2% sodium dodecyl sulphate (SDS), those of P1.6 dissociated in the presence of SDS . The higher electrophoretic mobility of the in vitro folded porins could be explained by the stable association of the RmpM protein to the porins in vivo . This association of RmpM contributes to the stability of the porins . The P1.6 pores were moderately cation-selective and displayed a single-channel conductance of 2.8 nS in 1 M KCl . The PorA P1.6 pores, but not the PorA P1.7,16 pores, showed an unusual non-linear dependence of the single-channel conductance on the salt concentration of the subphase . We hypothesize that a cluster of three negatively charged residues in L5 of P1.6 is responsible for the higher conductance at low salt concentrations.

J Exp Med, 2000 Mar 20, 191(6), 949 - 60
Role of lipooligosaccharide in Opa-independent invasion of Neisseria gonorrhoeae into human epithelial cells; Song W et al.; Lipooligosaccharide (LOS) has been implicated in the adhesion and invasion of host epithelial cells . We examined the adhesive and invasive abilities of isogenic gonococcal opacity-associated outer membrane protein-negative, pilus-positive (Opa-Pil+) Neisseria gonorrhoeae strains expressing genetically defined LOS . Strain F62 (Opa-Pil+), expressing the lacto-N-neotetraose and the galNac-lacto-N-neotetraose LOS, and its isogenic derivative that expressed only the lacto-N-neotetraose LOS (F62 Delta lgtD), adhered to, and invaded, to the same extent the human cervical epidermoid carcinoma cell line, ME180 . While the adhesive abilities of Opa-Pil+ isogenic strains that express LOS molecules lacking the lacto-N-neotetraose structure were similar to that seen for F62, their invasive abilities were much lower than the strains expressing lacto-N-neotetraose . Fluorescence microscopy studies showed that the adherence of F62, but not the strains lacking lacto-N-neotetraose, induced the rearrangement of actin filaments under the adherent sites . Electron microscopy studies demonstrated that F62, but not the strains lacking lacto-N-neotetraose, formed extensive and intimate associations with epithelial cell membranes . Thus, in the absence of detectable Opa protein, the lacto-N-neotetraose LOS promotes gonococcal invasion into ME180 cells . The data also suggest that LOS is involved in the mobilization of actin filaments in host cells, and in the formation of a direct interaction between the bacterial outer membrane and the plasma membrane of ME180 cells.

Int J STD AIDS, 2000 Mar, 11(3), 187 - 90
Symptomatic and asymptomatic chlamydial non-gonococcal urethritis in Jamaica: the potential for HIV transmission; Dowe G et al.; To determine the contribution of Chlamydia trachomatis to non-gonococcal urethritis (NGU) in men attending sexually transmitted disease (STD) clinics in Jamaica we studied men with NGU (n=339), and control groups including asymptomatic men who were STD contacts (n=61), asymptomatic men who were not STD contacts (n=32) and men with gonococcal urethritis (GU) (n=61) . Urethral specimens were examined for C . trachomatis and Neisseria gonorrhoeae . Serological tests for syphilis (STS) and HIV-1 infection were also performed . C . trachomatis accounted for 63% of cases of NGU but high prevalences were also found in asymptomatic STD contacts (59%), asymptomatic STD non-contacts (78%) and men with GU (48%) . The prevalence of C . trachomatis in men with GU differed significantly from that in men with NGU and asymptomatic STD non-contacts (P<0.05) . C . trachomatis infection in men with NGU was associated with multiple sex partners (71% vs 58%; chi2=4.78; odds ratio (OR)=1.76; P<0.05) and previous history of gonococcal infection (83% vs 42%; chi2=59.8; OR=6.8; P<0.0001) . Concomitant infection with HIV-1 occurred in 5.2% of cases of NGU and 50% and 90%, respectively, of the HIV-positive men had chlamydia or reactive STS . As a cost effective strategy in the control of STD and HIV we recommend presumptive treatment for C . trachomatis in men seeking STD treatment in Jamaica.

Sex Transm Dis, 2000 Mar, 27(3), 146 - 51
Urine screening for gonococcal and chlamydial infections at community-based organizations in a high-morbidity area; Jones CA et al.; BACKGROUND: Increasing availability of urine testing for Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT) offers expanded opportunities to collaborate with community-based organizations (CBOs) to screen high-risk populations for sexually transmitted diseases (STDs) . GOAL: To determine the prevalence and correlates of genital tract gonococcal and chlamydial infection among CBO clients, and to assess the feasibility of implementing widespread community-based STD screening programs . DESIGN: Free, voluntary, confidential first-catch urine screening was conducted at 20 CBOs serving disadvantaged populations in St . Louis, MO . Brief demographic, behavioral, and sexual contact data were obtained from all participants . Urine samples were tested by ligase chain reaction (LCR) . Persons testing positive were promptly notified and directed to seek treatment . RESULTS: A GC and/or CT infection was identified in 24 of 277 persons (8.7%) screened; 2 persons were infected with GC only, 17 with CT only, and 5 with GC and CT . Treatment was documented for 22 persons (91.7%) testing positive . The highest rates of infection were found at shelters (12.3%) and residence facilities (11.1%) . Costs of screening were $38 per sample collected and $453 per case identified . CONCLUSION: Community-based urine testing successfully identified GC and CT infections, and was well accepted by community members and CBOs . Community-based screening can significantly impact STD epidemiology by facilitating early detection, treatment, and interruption of transmission.

Sex Transm Dis, 2000 Mar, 27(3), 133 - 7
Evaluation of patient-administered tampon specimens for Chlamydia trachomatis and Neisseria gonorrhoeae; Tabrizi SN et al.; BACKGROUND: The patient-administered tampon specimen has proven to be an easy and sensitive method for the diagnosis of genital Chlamydia trachomatis and Neisseria gonorrhoeae infections in women by polymerase chain reaction (PCR) . This method avoids the need for endocervical sampling and stringent criteria for transport . GOAL: To evaluate two commercial amplification systems for the detection of C trachomatis and N gonorrhoeae from tampon specimens . STUDY DESIGN: A group of 400 positive and negative tampon specimens tested by an in-house PCR method were selected from a pool of more than 2,000 previously collected tampons . Overall, 93 C trachomatis-positive and 77 N gonorrhoeae-positive specimens were evaluated . Each specimen was tested by Roche Cobas Amplicor and Abbott LCx (LCR), and results were compared to the in-house PCR method . RESULTS: Detection of C trachomatis by both assays was not significantly different from the in-house PCR assay . Fewer tampons were positive for N gonorrhoeae by LCR than either the in-house assay (P = 0.0001) or by Roche Amplicor (P = 0.01) . However, tampon specimens tested by Roche Amplicor required DNA extraction to achieve comparative sensitivity . CONCLUSION: Both commercial assays can be applied to tampon-collected specimens for automated detection of sexually transmitted diseases . The detection of C trachomatis was similar to the in-house PCR test for both assays (P = 0.73, 0.68) . Detection of N gonorrhoeae resulted in fewer positive tampon specimens when tested by ligase chain reaction than both Roche Amplicor and in-house PCR.

Curr Opin Urol, 1999 Jan, 9(1), 65 - 70
Antimicrobial resistance in Neisseria gonorrhoeae; Fox KK et al.; Despite a declining incidence of gonorrhea in much of the industrialized world, antimicrobial resistance continues to present a challenge to the treatment of gonococcal infections . Although no clinically significant resistance to the broad-spectrum cephalosporins has been identified, fluoroquinolone-resistant gonococcal strains are now prevalent in Australia and much of the Far East . Ongoing surveillance for antimicrobial resistance is crucial to obtain the information needed to choose effective empiric therapies for gonorrhea.

Prim, Care Update Ob Gyns . 2000 Mar 1, 7(2), 49 - 54
Bacterial meningitis; Bartfield AA; With nearly 8,000 cases in the United States per year, and 2,000 deaths annually, bacterial meningitis continues to be a significant source of morbidity and mortality . The principal pathogens are Neisseria meningitidis, Streptococcus pneumoniae, group B streptococci, and Hemophilus influenzae . In immunocompromised patients, Listeria monocytogenes is also an important pathogen . Rapid identification and evaluation of the patient with bacterial meningitis and prompt initiation of antibiotics are the cornerstones of therapy . Except in the rare patient with papilledema, focal neurologic symptoms, or a seizure, a lumbar puncture should be performed without delay, and antibiotic therapy should be administered promptly . Patients without a readily identifiable source of infection should be treated empirically with intravenous ceftriaxone . Ampicillin should also be administered in populations at increased risk for L . monocytogenes . The risk of meningitis in some populations can be reduced by administration of vaccines against selected pathogens such as N . meningitidis, S . pneumoniae, and H . influenzae.

J Immunol Methods, 2000 Apr 3, 237(1-2), 73 - 84
Enzyme linked immunosorbent assay (ELISA) for the detection of serum antibodies to the inner core lipopolysaccharide of Neisseria meningitidis group B; Plested JS et al.; We have developed a solid-phase ELISA to study the human immune response to inner core lipopolysaccharide (LPS) of Neisseria meningitidis (Nm) using structurally defined glycolipids from a genetically defined mutant (galE) of a serogroup B Nm strain . Previous studies had demonstrated that a galE (inner core) LPS epitope is conserved in approximately 70% Nm strains and was accessible to antibody in fully encapsulated wild-type Nm strains . A murine monoclonal antibody, MAb B5, raised to a galE mutant of serogroup B Nm strain, immunotype L3 (B.15.P1.7,16) was used to determine the specificity of the inner core LPS ELISA by inhibition studies using purified galE LPS and human sera . The intra-assay coefficient of variation (CV) was 5-6% and inter-assay CV was 19-22% . Using this ELISA, significant differences in the geometric mean titres (GMTs) of naturally occurring serum antibodies (specific to inner core LPS) between healthy adults (18-65 years, N=54) and healthy infants (3-4 months, N=144) of both IgG and IgM classes were found (P<0.0001) . GMTs were expressed in galE arbitrary units (AU/ml) (95% confidence intervals): IgG antibodies in adults 5.7 (5 . 0,6.9) and in infants 1.1 (1.0,1.3); IgM antibodies in adults 7.7 (5 . 7,10.4), and in infants 0.85 (0.7,1.1) . In age-matched children aged 26-113 months a difference (P=0.04) in specific IgG was found in healthy infants and infants in the acute phase of invasive Nm disease (GMT (95%CI) in AU/ml: in healthy infants 7.7 (5.3,11.0), in acute phase infants 4.2 (2.5,7.2) . However, there was no difference in specific IgM (P=0.98) between these groups healthy infants 4.7 (3 . 1,7.0), acute phase 4.6 (2.9, 7.4) . In eleven children (5-181 months) there were differences in the GMTs of specific IgG and IgM (P=0.02, P=0.008 respectively) between paired acute and convalescent sera (GMT) (95%CI) in AU/ml: IgG acute 1.95 (0.98, 3.8), convalescent 5.2 (2.2,12.4); IgM acute 1.78 (1.05,3.0), convalescent 4.38 (2.6,7.3) . We conclude that ELISA is a specific, sensitive and reproducible method for the detection of antibodies to inner core LPS of Nm and that an epitope defined by MAb B5 can be immunogenic in infants and adults . These findings are relevant to the potential candidacy of inner core LPS as a vaccine.

Mol Biol Evol, 2000 Mar, 17(3), 423 - 36
Population genetics of the porB gene of Neisseria gonorrhoeae: different dynamics in different homology groups; Posada D et al.; The porB locus codes for the major outer membrane protein of Neisseria gonorrhoeae . Alleles of this locus have been assigned to two homology groups based on close sequence and immunological relationships and are designated as either PIA or PIB . Several population parameters were estimated and compared among these two groups using a data set of 22 PIA sequences and 91 PIB sequences obtained from diverse geographic localities and from time periods spanning approximately 50 years . Recombination appears to be extensive in the porB gene . While the recombination rates are similar for the PIA and PIB sequences, the relative contribution of recombination to genetic diversity is higher for the PIA sequences . Alleles belonging to the PIB group show greater genetic diversity than do those in the PIA group . Although phylogenetic analysis did not reveal temporal or geographic clustering of sequences, estimates of gene flow and the fixation index suggested that PIB sequences exhibit population substructure based on geographic locality . Selection acts in these homology groups in a different way . While positive Darwinian selection is the dominant force driving the evolution of the PIA sequences, purifying selection operates also on the PIB sequences . These differences may be attributable to the greater propensity of PIA strains, as compared with PIB strains, to cause disseminated gonococcal infection, which would expose the former to intense selection pressure from the host immune system . The molecular evolution of Neisseria gonorrhoeae seems to be driven by the simultaneous action of selection and recombination, but under different rates and selection pressures for the PIA and PIB homology groups.

FEMS Microbiol Lett, 2000 Mar 15, 184(2), 161 - 4
The siaA gene involved in capsule polysaccharide biosynthesis of Neisseria meningitidis B codes for N-acylglucosamine-6-phosphate 2-epimerase activity; Petersen M et al.; The capsule polysaccharide of Neisseria meningitidis serogroup B is composed of a homopolymer of alpha-2-->8 linked N-acetyl-neuraminic acid (sialic acid) . The enzymes required for sialic acid biosynthesis and polymerization are encoded in region A of the capsule gene complex . We here describe the enzymatic activity of the siaA gene product as determined by biochemical analysis . siaA was overexpressed in Escherichia coli and the SiaA protein was purified to homogeneity . Enzymatic assays revealed that SiaA did not accept N-acetyl-glucosamine as substrate, but only N-acetyl-glucosamine-6-phosphate (EC 5.1.3.9) . SiaA catalyzes the isomerization of N-acetyl-glucosamine-6-phosphate to form N-acetyl-mannosamine-6-phosphate . This reaction represents the first step in capsule biosynthesis of N . meningitidis B.

Science, 2000 Mar 10, 287(5459), 1809 - 15
Complete genome sequence of Neisseria meningitidis serogroup B strain MC58; Tettelin H et al.; The 2,272,351-base pair genome of Neisseria meningitidis strain MC58 (serogroup B), a causative agent of meningitis and septicemia, contains 2158 predicted coding regions, 1158 (53.7%) of which were assigned a biological role . Three major islands of horizontal DNA transfer were identified; two of these contain genes encoding proteins involved in pathogenicity, and the third island contains coding sequences only for hypothetical proteins . Insights into the commensal and virulence behavior of N . meningitidis can be gleaned from the genome, in which sequences for structural proteins of the pilus are clustered and several coding regions unique to serogroup B capsular polysaccharide synthesis can be identified . Finally, N . meningitidis contains more genes that undergo phase variation than any pathogen studied to date, a mechanism that controls their expression and contributes to the evasion of the host immune system.

Vet Microbiol, 2000 Mar 1, 72(1-2), 153 - 61
Survival of avian strains of Pasteurella multocida in chicken serum; Diallo IS et al.; The ability of bacteria to survive in serum is considered a likely virulence determinant in diseases where the infective bacteria become septicaemic . Optimal conditions were established to test the survival of Pasteurella multocida in chicken serum . Serum was used at 90%, the inoculum was 10(3)-10(4)cfu in phosphate buffered saline pH 7.4 . Survival was measured after incubation for 2-4 h; if survival was <50% the strain was considered serum susceptible . Susceptible strains were either killed or their growth was inhibited . Some resistant strains not only survived but grew rapidly in unheated serum . Thirty-five strains, all originally isolated from clinical fowl cholera, were tested; eight were susceptible, of which three were killed and five inhibited, and the remainder (27) were resistant . Ten serum-resistant P . multocida serogroup A strains were grown in hyaluronidase to remove the capsule and survival in chicken serum was re-tested . Three strains became susceptible, while seven strains remained resistant . Three serum susceptible strains were then tested in the presence of cytidine monophosphate-N-acetylneuraminic acid (CMP-NANA) . This substance is present in the human serum, and is known to mask the effect of complement on Neisseria gonorrhoeae rendering susceptible strains resistant . Two of the three serum susceptible strains became resistant in the presence of CMP-NANA . Serum susceptibility/resistance was more complex than that of Escherichia coli, and the role of resistance to avian complement in the pathogenesis of fowl cholera remains to be determined.

EMBO J, 2000 Mar 1, 19(5), 1098 - 107
Charged tmRNA but not tmRNA-mediated proteolysis is essential for Neisseria gonorrhoeae viability; Huang C et al.; tmRNA, through its tRNA and mRNA properties, adds short peptide tags to abnormal proteins, targeting these proteins for proteolytic degradation . Although the conservation of tmRNA throughout the bacterial kingdom suggests that it must provide a strong selective advantage, it has not been shown to be essential for any bacterium . We report that tmRNA is essential in Neisseria gonorrhoeae . Although tagging per se appears to be required for gonococcal viability, tagging for proteolysis does not . This suggests that the essential roles of tmRNA in N.gonorrhoeae may include resolving stalled translation complexes and/or preventing depletion of free ribosomes . Although derivatives of N.gonorrhoeae expressing Escherichia coli tmRNA as their sole tmRNA were isolated, they appear to form colonies only after acquiring an extragenic suppressor(s).

J Clin Microbiol, 2000 Mar, 38(3), 1244 - 6
Etiology of sexually transmitted infections among street-based female sex workers in Dhaka, Bangladesh; Rahman M et al.; An etiological study of sexually transmitted infections (STIs) was conducted among female sex workers (FSWs) in Dhaka, Bangladesh . Endocervical swab and blood samples from 269 street-based FSWs were examined for Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis as well as for antibodies to Treponema pallidum and herpes simplex virus 2 (HSV-2) . Sociodemographic data and data regarding behavior were also collected . A total of 226 of the 269 FSWs (84%) were positive for the STI pathogens studied . Among the 269 FSWs, 35.5% were positive for N . gonorrhoeae, 25% were positive for C . trachomatis, 45.5% were positive for T . vaginalis, 32.6% were seropositive for T . pallidum, 62.5% were seropositive for HSV-2, and 51% had infections with two or more pathogens.

Int J STD AIDS, 2000 Feb, 11(2), 115 - 8
Trend of antimicrobial resistance in Neisseria gonorrhoeae at New Delhi, India; Ray K et al.; We aim to monitor the trends of antimicrobial resistance in Neisseria gonorrhoeae and to compare the results of antimicrobial sensitivity by disc diffusion and minimum inhibitory concentration (MIC) . Two hundred and eleven confirmed strains of N . gonorrhoeae were subjected to antimicrobial sensitivity testing by disc diffusion using penicillin, tetracycline, ciprofloxacin and ceftriaxone from 1995 to June 1999 . Penicillinase-producing Neisseria gonorrhoeae (PPNG) were detected by lodometric method . Minimum inhibitory concentration was determined by E test . A low level of penicillin resistance and PPNG detected in 1996 was maintained over the years . Significant increasing trend of tetracycline and ciprofloxacin resistance with high MIC i.e . 2-96 microg/ml and 1-32 microg/ml respectively were found . Ceftriaxone was found to be the drug of choice, being 100% sensitive . Comparison of resistance pattern by the 2 tests showed satisfactory agreement . Emergence of penicillin, quinolone and tetracyline resistance in N . gonorrhoeae isolates from a major STD centre at New Delhi indicates the need for increased awareness, prudent use of antimicrobials, and evaluation of new antimicrobials for the treatment of gonorrhoeaPIP: This study determined the prevalence of antimicrobial resistance by Neisseria gonorrhoeae, including penicillinase-producing gonorrhea (PPNG) strains, and monitored the trends . It further compared the results of antimicrobial sensitivity by disc diffusion and minimum inhibitory concentration (MIC) . A total of 211 confirmed gonorrhea strains were subjected to antimicrobial sensitivity testing by disc diffusion using penicillin, tetracycline, ciprofloxacin, and ceftriaxone from 1995 to June 1999 . PPNG strains were detected by lodometric method, and an E test method of 55 strains isolated in 1997-98 determined MIC . Statistical analysis of the results indicates that a low level of penicillin resistance and PPNG detected in 1996 was maintained over the years . In addition, a significant increasing trend of tetracycline and ciprofloxacin resistance with high MIC was found . Ceftriaxone, being 100% sensitive, was found to be the drug of choice . Moreover, comparison of resistance pattern by the two tests showed satisfactory agreement . Findings indicate the need for increased awareness, prudent use of antimicrobials, and evaluation of new antimicrobials for the treatment of gonorrhea .

Southeast Asian J Trop Med Public Health, 1999 Mar, 30(1), 52 - 7
The prevalence of Chlamydia trachomatis infection in rural Thai women; Thongkrajai P et al.; A cross sectional study was designed to investigate the prevalence of Chlamydia trachomatis among different groups of rural women in the northeast Thailand . The presence of chlamydial antigens in endocervical swabs was detected by ELISA . The prevalences of Chlamydia trachomatis were 6.8% (31/485), 5.2% (24/466) and 6.7% (12/179) in women attending antenatal, postpartum and family planning clinics respectively . The average prevalences of C . trachomatis among hospital-based and community-based women were 6.1% (67/1,103) and 3.6% (15/411) respectively . In addition, the prevalences of some pathogens including Candida albicans, Trichomonas vaginalis, Treponema pallidum and Neisseria gonorrhoea among hospital-based and community-based women were 14.2, 2.8, 0.7, 0.2 and 10.9, 5.1, 2.7, 0.0% respectively . It was concluded that C . trachomatis was a problem of woman's reproductive health.

Mol Pathol, 1999 Aug, 52(4), 220 - 30
Cell adhesion molecules in the pathogenesis of and host defence against microbial infection; Kerr JR; Eukaryotic cell adhesion molecules (CAMs) are used by various cells and extracellular molecules in host defence against infection . They are involved in many processes including recognition by circulating phagocytes of a site of inflammation, transmigration through the endothelial barrier, diapedesis through basement membrane and extracellular matrix, and release of effector mechanisms at the infected site . CAMs involved in leucocyte-endothelial cell interaction include the selectins, integrins, and members of the immunoglobulin superfamily . However, CAMs are also used by various microorganisms (protozoa, fungi, bacteria, and viruses) during their pathogenesis . For example, bacteria that utilise CAMs include Mycobacterium tuberculosis, Listeria monocytogenes, Yersinia spp, enteropathogenic Escherichia coli, Shigella spp, Neisseria spp, Bordetella spp, and Borrelia burgdorferi . In addition, CAMs are involved in the pathogenetic effects of the RTX toxins of Pasteurella haemolytica, Actinobacillus actinomycetemcomitans, and the superantigen exotoxins of Staphylococcus aureus and Streptococcus pyogenes . A recurrent and topical theme of potential importance within the bacterial group is the intimate relation between CAMs, bacterial protein receptors, and type III secretion systems . For example, the IpaBCD protein complex is secreted by the type III system of Shigella flexneri and interacts with alpha 5 beta 1 integrin on the eukaryotic cell surface, followed by Rho mediated internalisation; this illustrates the relevance of cellular microbiology . CAMs might prove to be novel therapeutic targets . Comparative genomics has provided the knowledge of shared virulence determinants among diverse bacterial genera, and will continue to deepen our understanding of microbial pathogenesis, particularly in the context of the interaction of prokaryotic and eukaryotic molecules.

Infect Immun, 2000 Mar, 68(3), 1696 - 9
Macropinocytosis as a mechanism of entry into primary human urethral epithelial cells by Neisseria gonorrhoeae; Zenni MK et al.; Gonococcal entry into primary human urethral epithelial cells (HUEC) can occur by macropinocytosis . Scanning and transmission electron microscopy revealed lamellipodia surrounding gonococci, and confocal laser scanning microscopy analysis showed organisms colocalized with M(r) 70,000 fluorescein isothiocyanate-labeled dextran within the cells . Phosphoinositide 3-kinase inhibitors and an actin polymerization inhibitor prevented macropinocytic entry of gonococci into HUEC.

Infect Immun, 2000 Mar, 68(3), 1164 - 70
Actinobacillus pleuropneumoniae iron transport: a set of exbBD genes is transcriptionally linked to the tbpB gene and required for utilization of transferrin-bound iron; Tonpitak W et al.; Upon iron restriction, Actinobacillus pleuropneumoniae has been shown to express the transferrin-binding proteins TbpB and TbpA, both of which have been implied to be important virulence factors . In order to identify additional iron-regulated proteins, we cloned and analyzed the region upstream of the transferrin-binding protein genes in an A . pleuropneumoniae serotype 7 strain . We located immediately upstream of the tbpB gene two open reading frames which were 43% homologous to the neisserial ExbBD protein genes . By raising specific antibodies, we showed that ExbB is expressed under iron-limiting growth conditions only, and RT-PCR analysis revealed that the exbBD genes and the tbpB gene are transcribed on a single polycistronic mRNA . By constructing an isogenic and nonpolar exbBD mutant, we showed that the exbBD genes are required by A . pleuropneumoniae for utilization of transferrin-bound iron . Using PCR and Western blotting, we showed that the genetic organization found in A . pleuropneumoniae serotype 7 is similar in all 12 A . pleuropneumoniae serotype reference strains.

Hunan Yi Ke Da Xue Xue Bao, 1998, 23(2), 169 - 71
{Infection of sexually transmitted diseases in female patients with inflammation of genitourinary tract by polymerase chain reaction}; Li D et al.; Using polymerase chain reaction (PCR), Neisseria gonorrhoeae (NG) from 5899 female out-patients with inflammation of genitourinary tract was detected . Among 18.57% positive cases, the effective rates of the gonorrhea (26.07%) and urethritis (23.73%) were higher than those of the Condyloma acuminatum (CA) (17.73%), vaginitis (17.70%), and cervicitis (11.54%)(P< 0.01), indicating that there were various degrees of NG infection in genitourinary tract diseases . Moreover, Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), NG, and human papilloma viruses (HPV) were also detected in 8329 patients with vaginitis . Among 20.01% positive cases, the positive rate of UU was 39.95%, CT 21.36%, NG 17.70%, and HPV 16.40% . Variance analysis showed that significant differences existed in the vaginitis caused by different pathogens ( P <0.01); namely, four kinds of pathogens were present in the patients with vaginitis.

Sex Transm Dis, 2000 Feb, 27(2), 115 - 8
Resistance of Neisseria gonorrhoeae epidemic strains to antibiotics: report of resistant isolates and surveillance in Zhanjiang, China: 1998 to 1999; Guoming L et al.; BACKGROUND: Antibiotics are widely used to treat gonorrhea . Changes in the susceptibility of Neisseria gonorrhoeae to these agents may influence their use . GOAL: To measure the antibiotic susceptibility of N gonorrhoeae epidemic strains in Zhanjiang (Guangdong) and to evaluate the prevalence of strains with reduced susceptibility . STUDY DESIGN: A total of 98 gonococcal isolates obtained from 1998 through 1999 in Zhanjiang were tested for antibiotic susceptibility based on the systemic identification . The inhibitory zone diameters (mm) and the MICs of penicillin, tetracycline, spectinomycin, ceftriaxone, and ciprofloxacin were determined using disk-diffusion and agar-dilution methods, respectively . The susceptibilities of these isolates were defined using criteria of the National Committee for Clinical Laboratory Standards . RESULTS: The percentages of gonococci-resistant strains to penicillin, tetracycline, spectinomycin, ceftriaxone, and ciprofloxacin were 32.65%, 69.39%, 8.16%, 13.27%, 82.65% by disk-diffusion method and 23.91%, 49.46 %, 11.11%, 16.48%, 59.34% by agar-dilution method, respectively . CONCLUSIONS: The resistant strains of contemporary gonococci in Zhanjiang were serious, especially for ciprofloxacin resistance . Continued active surveillance is needed to detect and control the spread of ceftriaxone-resistant and spectinomycin-resistant N gonorrhoeae.

J Infect Dis, 2000 Feb, 181(2), 770 - 3
Poly(sodium 4-styrene sulfonate): an effective candidate topical antimicrobial for the prevention of sexually transmitted diseases; Herold BC et al.; Presently marketed vaginal barrier agents are cytotoxic and damage the vaginal epithelium and natural vaginal flora with frequent use . Novel noncytotoxic agents are needed to protect women from sexually transmitted diseases . One candidate compound is a high-molecular-mass form of soluble poly(sodium 4-styrene sulfonate) (T-PSS) . The antimicrobial activity of T-PSS was evaluated in primary culture systems and in a genital herpes murine model . Results obtained indicate that T-PSS is highly effective against herpes simplex viruses, Neisseria gonorrhoeae, and Chlamydia trachomatis in vitro . A 5% T-PSS gel protected 15 of 16 mice from vaginal herpes, compared with 2 of 16 mice treated with a placebo gel . Moreover, T-PSS exhibited little or no cytotoxicity and has an excellent selectivity index . T-PSS is an excellent candidate topical antimicrobial that blocks adherence of herpes simplex virus at low concentrations, inactivates virus at higher concentrations, and exhibits a broad spectrum of antimicrobial activity.

Int J STD AIDS, 2000 Jan, 11(1), 45 - 8
Disease prevalence in women attending the STD clinic in Mumbai (formerly Bombay), India; Divekar AA et al.; Our objectives were to determine the prevalence of Neisseria gonorrhoeae and its association with other STD causing organisms . Three hundred and thirty-six consecutive women (female sex workers (FSWs) and married contacts), attending a sexually transmitted disease (STD) clinic in Mumbai, were screened for N . gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis . Per speculum examination was performed and clinical signs were recorded . Symptoms perceived by the women were also recorded . The mean age for married contacts, FSWs and gonorrhoea-positive women was 27.9, 29.7 and 27.5, respectively . 9.7% of the women were positive for N . gonorrhoeae, 23.2% were chlamydia-positive and 5.9% had trichomoniasis . N . gonorrhoeae was isolated more frequently from FSWs as compared to the married contacts . The prevalence of HIV was significantly higher among women with multiple sex partners (FSWs) (P<0.001) . Gonococcal infection is significantly associated with the presence of HIV . A significant association between sexual habits and prevalence of gonorrhoea, trichomoniasis and HIV was observed . The prevalence of gonorrhoea over 1988 to 1996 remained approximately the samePIP: This clinical trial was conducted to determine the prevalence of Neisseria gonorrhea and its correlation with other sexually transmitted disease (STD) causing organisms . A total of 336 consecutive women (female sex workers (FSWs) and married contacts) attending an STD clinic in Mumbai, India, were screened for N . gonorrhea, Chlamydia trachomatis, and Trichomonas vaginalis . Speculum examinations were performed, the vulva and external genitalia were assessed for ulcers and warts, and clinical signs were recorded . The symptoms reported by the women were also documented . The mean age of married contacts was 27.9 years, while that of FSWs was 29.7 years . The mean age of gonorrhea-, chlamydia-, and trichomonas-positive women was 27.3, 28.0, and 28.8 years, respectively . This study showed that 9.7% of the women were gonorrhea-positive, 23.3% were chlamydia-positive, and 5.9% were trichomonas-positive . N . gonorrhea was more prevalent among FSWs than married contacts . Moreover, HIV was more prevalent among FSWs (P 0.001) . Gonococcal infection is significantly associated with HIV . An important correlation between sex behavior and the prevalence of gonorrhea, trichomoniasis, and HIV was found . Between 1988 and 1996 the prevalence of gonorrhea remained approximately the same .

Acta Crystallogr D Biol Crystallogr, 2000 Feb, 56 ( Pt 2), 203 - 5
Crystallization and preliminary X-ray studies of recombinant amylosucrase from Neisseria polysaccharea; Skov LK et al.; Recombinant amylosucrase from Neisseria polysaccharea was crystallized by the vapour-diffusion procedure in the presence of polyethylene glycol 6000 . The crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 95.7, b = 117.2, c = 62.1 A, and diffract to 1.6 A resolution . A p-chloromercuribenzene sulfonate (pcmbs) derivative has been identified and a selenomethionine-substituted protein has been produced and crystallized.

Folia Microbiol (Praha), 1999, 44(3), 333 - 7
Gram staining with an automatic machine; Felek S et al.; This study was undertaken to develop a new Gram-staining machine controlled by a micro-controller and to investigate the quality of slides that were stained in the machine . The machine was designed and produced by the authors . It uses standard 220 V AC . Staining, washing, and drying periods are controlled by a timer built in the micro-controller . A software was made that contains a certain algorithm and time intervals for the staining mode . One-hundred and forty smears were prepared from Escherichia coli, Staphylococcus aureus, Neisseria sp., blood culture, trypticase soy broth, direct pus and sputum smears for comparison studies . Half of the slides in each group were stained with the machine, the other half by hand and then examined by four different microbiologists . Machine-stained slides had a higher clarity and less debris than the hand-stained slides (p < 0.05) . In hand-stained slides, some Gram-positive organisms showed poor Gram-positive staining features (p < 0.05) . In conclusion, we suggest that Gram staining with the automatic machine increases the staining quality and helps to decrease the work load in a busy diagnostic laboratory.

Trends Biochem Sci, 2000 Feb, 25(2), 70 - 4
Anticodon nucleases; Kaufmann G; A tRNALys-specific anticodon nuclease is kept in a latent form in a rare Escherichia coli strain, complexed with a DNA restriction enzyme . A phage T4 inhibitor of DNA restriction activates anticodon nuclease, but other T4 proteins restore tRNALys . Detection of a homologous system in Neisseria and a different anticodon nuclease in colicin E5 suggest ubiquity and diversity of such tRNA toxins . Analysis of these systems could reveal novel RNA recognition and cleavage mechanisms.

Sante, 1999 Sep-Oct, 9(5), 332 - 4
{Frequency and susceptibility to antibiotics of Neisseria gonorrhoeae at Lomé}; Dagnra AY et al.; A cross-sectional study was carried out by the bacteriology laboratory of the Lome-Tokoin University Hospital, to determine the frequency of gonorrhea in patients providing vaginal or urethral samples, and to investigate the susceptibility of N . gonorrhoeae to antibiotics . The samples were treated according to classic bacteriological methods and the disk diffusion tests were used to investigate antibiotic susceptibility . The samples were taken from 420 patients, 246 of whom were male (26.86% inverted question markNdT: d'ou vient ce chiffre??, 58.33% si 420 patients, 61.19% si 402 patients inverted question mark) and 156 of whom were female (37.14% inverted question markNdT: plutot 38.8% si 402 patients inverted question mark) inverted question markNdT: et le reste?? Il me semble que c'est plutot 402 patients en total (= 246 + 156) . Sinon il y a 18 patients qui ne sont ni male ni femelle?! inverted question mark . N . gonorrhoeae was isolated from 88 patients, giving a prevalence of 20.95% inverted question markNdT: ou 21.89% si 402 patients inverted question mark . We isolated strains of the bacterium from 76 men (25.78% inverted question markNdT: incorrect, 30 . 89% inverted question mark and 12 women (7.69%) . Ciprofloxacin, ofloxacin and netilmicin were the most effective antibiotics . The cyclins, cotrimoxazole and penicillin were less effective.

J Immunol, 2000 Feb 15, 164(4), 1839 - 46
Membrane cofactor protein (MCP; CD46): isoform-specific tyrosine phosphorylation; Wang G et al.; Membrane cofactor protein (MCP; CD46) is a widely expressed type 1 transmembrane glycoprotein that inhibits complement activation on host cells . It also is a receptor for several pathogens including measles virus, Streptococcus pyogenes, Neisseria gonorrhea, and Neisseria meningitidis . That MCP may have signaling capability was suggested by its microbial interactions . That is, binding of MCP on human monocytes by measles virus hemagglutinin or cross-linking by an anti-MCP Ab resulted in IL-12 down-regulation, while binding to MCP by Neisseria on epithelial cells produced a calcium flux . Through alternative splicing, MCP is expressed on most cells with two distinct cytoplasmic tails of 16 (CYT-1) or 23 (CYT-2) amino acids . These play pivotal roles in intracellular precursor processing and basolateral localization . We investigated the putative signal transduction pathway mediated by MCP and demonstrate that CYT-2, but not CYT-1, is phosphorylated on tyrosine . We examined MCP tail peptides and performed Ab cross-linking experiments on several human cell lines and MCP isoform transfectants . We found an MCP peptide of CYT-2 was phosphorylated by a src kinase system . Western blots of the cells lines demonstrated that cells bearing CYT-2 were also phosphorylated on tyrosine . Additionally, we provide genetic and biochemical evidence that the src family of kinases is responsible for the latter phosphorylation events . In particular, the src kinase, Lck, is required for phosphorylation of MCP in the Jurkat T cell line . Taken together, these studies suggest a src family-dependent pathway for signaling through MCP.

J Clin Microbiol, 2000 Feb, 38(2), 521 - 5
Antimicrobial resistance of Neisseria gonorrhoeae and high prevalence of ciprofloxacin-resistant isolates in Japan, 1993 to 1998; Tanaka M et al.; To assess the antimicrobial resistance of Neisseria gonorrhoeae isolated from 1993 through 1998 in Japan, susceptibility testing was conducted on 502 isolates . Selected isolates were characterized by auxotype and analysis for mutations within the quinolone resistance-determining region (QRDR) in the gyrA and parC genes, which confer fluoroquinolone resistance on the organism . Plasmid-mediated penicillin resistance (penicillinase-producing N . gonorrhoeae) decreased significantly from 1993-1994 (7.9%) to 1997-1998 (2.0%) . Chromosomally mediated penicillin resistance decreased from 1993-1994 (12.6%) to 1995-1996 (1.9%) and then increased in 1997-1998 (10.7%) . Chromosomally mediated tetracycline resistance decreased from 1993-1994 (3.3%) to 1997-1998 (2.0%), and no plasmid-mediated high-level tetracycline resistance was found . Isolates with ciprofloxacin resistance (MIC >/= 1 microg/ml) increased significantly from 1993-1994 (6.6%) to 1997-1998 (24.4%) . The proline-requiring isolates were less susceptible to ciprofloxacin than the prototrophic or arginine-requiring isolates . Ciprofloxacin-resistant isolates contained three or four amino acid substitutions within the QRDR in the GyrA and ParC proteins.

Genetics, 2000 Feb, 154(2), 523 - 32
A homologue of the recombination-dependent growth gene, rdgC, is involved in gonococcal pilin antigenic variation; Mehr IJ et al.; Neisseria gonorrhoeae pilin undergoes high-frequency changes in primary amino acid sequence that aid in the avoidance of the host immune response and alter pilus expression . The pilin amino acid changes reflect nucleotide changes in the expressed gene, pilE, which result from nonreciprocal recombination reactions with numerous silent loci, pilS . A series of mini-transposon insertions affecting pilin antigenic variation were localized to three genes in one region of the Gc chromosome . Mutational analysis with complementation showed that a Gc gene with sequence similarity to the Escherichia coli rdgC gene is involved in pilus-dependent colony phase variation and in pilin antigenic variation . Furthermore, we show that the Gc rdgC homologue is transcriptionally linked in an operon with a gene encoding a predicted GTPase . The inability to disrupt expression of this gene suggests it is an essential gene (engA, essential neisserial GTPase) . While some of the transposon mutations in rdgC and insertions in the 5'-untranslated portion of engA showed a growth defect, all transposon insertions investigated conferred an aberrant cellular morphology . Complementation analysis showed that the growth deficiencies are due to the interruption of RdgC expression and not that of EngA . The requirement of RdgC for efficient pilin variation suggests a role for this protein in specialized DNA recombination reactions.

Sex Transm Dis, 2000 Jan, 27(1), 1 - 8
Evidence of declining STD prevalence in a South African mining community following a core-group intervention; Steen R et al.; OBJECTIVES: To reduce the prevalence of curable sexually transmitted diseases (STDs) in a South African mining community through provision of STD treatment services, including periodic presumptive treatment and prevention education to a core group of high-risk women living in areas around the mines . METHODS: Women at high risk for STDs attended a mobile clinic monthly for examination and counseling, and were treated presumptively for bacterial STDs with a directly observed 1-g dose of azithromycin . Gonococcal and chlamydial infection rates were measured by urine ligase chain reaction, and genital ulcers were assessed by clinical examination . Changes in STD prevalence among local miners were assessed through comparison of prevalence in two cross-sectional samples of miners taken 9 months apart, and through routine disease surveillance at mine health facilities . RESULTS: During the first 9 months of the intervention, 407 women used the services . Baseline prevalence of Neisseria gonorrhoeae and/or Chlamydia trachomatis in women was 24.9%; 9.7% of these women had clinical evidence of genital ulcer disease (GUD) . The proportion of women with incident gonococcal or chlamydial infections at the first monthly return visit (69% follow-up rate) was 12.3%, and genital ulcers were found in 4.4% of these women . In the miner population, the prevalence of N gonorrhoeae and/or C trachomatis was 10.9% at baseline and 6.2% at the 9-month follow-up examination (P<0.001) . The prevalence of GUD by clinical examination was 5.8% at baseline and 1.3% at follow-up examination (P< 0.001) . Rates of symptomatic STDs seen at mine health facilities decreased among miners in the intervention area compared with miners living farther from the site and with less exposure to the project . DISCUSSION: Provision of STD treatment services to a core group of high-risk women may significantly reduce their burden of disease, and may contribute to a reduction in community STD prevalence . In the absence of sensitive and affordable screening tests for STDs in women, periodic presumptive treatment coupled with prevention education is a feasible approach to providing STD services in this populationPIP: This intervention-linked study was conducted to reduce the prevalence of curable sexually transmitted diseases (STDs) in a South African mining community through provision of STD treatment services, including periodic presumptive treatment and prevention education to a core group of high-risk women living in areas around the mines . In this article, the impact of such an intervention is assessed on the women as well as the male migrant community in the intervention area . During the 9 months of the intervention, 407 women used the services . Baseline prevalence of gonococcal or chlamydial infections in women was 24.9%; 9.7% of these women had clinical evidence of genital ulcer disease (GUD) . At the first monthly return, baseline for gonococcal or chlamydial infected women was 12.3%, and genital ulcers were found in 4.4% of the women . In the miner population, the overall result was similar to the result observed in the women: a decrease in rate of baseline prevalence of gonococcal or chlamydial infections and GUD was noted . In addition, rates of symptomatic STDs seen at mine health facilities decreased more among miners living closer within the intervention area than among those living farther away . In conclusion, provision of STD treatment services to a core group of high-risk women may significantly reduce their burden of disease, and may contribute to a reduction in community STD prevalence .

Proteins, 1999 Dec 1, 37(4), 717 - 28
Structural characterization of two tandemly arranged DNA methyltransferase genes from Neisseria gonorrhoeae MS11: N4-cytosine specific M.NgoMXV and nonfunctional 5-cytosine-type M.NgoMorf2P; Radlinska M et al.; Two adjacent genes encoding DNA methyltransferases (MTases) of Neisseria gonorrhoeae MS11, an active N4-cytosine specific M . NgoMXV and an inactive 5-cytosine type M . NgoMorf2P, were cloned into Escherichia coli and sequenced . We analyzed the deduced amino acid sequence of both gene products and localized conserved regions characteristic for DNA MTases . Structure prediction, threading-derived alignments, and comparison with the common fold for DNA MTases allowed for construction of super-secondary and tertiary models for M.NgoMorf2P and M.NgoMXV, respectively . These models helped in identification of amino acids and structural elements essential for function of both enzymes . The implications of this putative structural model on the catalytic mechanism of M.NgoMXV and its possible relation to the common ancestor of modern DNA amino-MTases are also discussed.

Rheumatol Int, 1999, 19(1-2), 1 - 5
Infective arthritis in adults--experience at a teaching hospital in Kuwait; Kaushik P et al.; Infective arthritis (InfectA) has a variable geographical pattern . A 'Medline' search did not yield any earlier series on epidemiological pattern of InfectA from Kuwait or the Gulf region . Therefore, an observational prospective study of sequential cases seen over a period of 4 years was carried out . Of a total of 2021 patients seen during this period, 36 (1.8%) were diagnosed to have InfectA . There were 6 (16.7%) cases of neisserial and 30 (83.3%) of non-neisserial InfectA . Predisposing factors were seen in 17 (47.2%), all with non-neisserial InfectA . Microbiological diagnosis was proven in 15 (41.7%) cases, and included Neisseria gonorrhoeae in 5, Brucella spp . in 4, Staphylococcus aureus in 3, and Neisseria meningitidis, Salmonella spp . and Mycobacterium tuberculosis in 1 each . In the remaining 21 (58.3%) cases the diagnosis was made on clinical grounds and confirmed on therapeutic response . The most common presentation of neisserial InfectA was acute polyarthritis while that of non-neisserial InfectA was acute monoarthritis . All patients recovered with little disability and no mortality.

FEMS Microbiol Lett, 2000 Feb 1, 183(1), 49 - 53
Association of iron-regulated outer membrane proteins of Neisseria meningitidis with the RmpM (class 4) protein; Prinz T et al.; The RmpM (class 4) protein of Neisseria meningitidis has previously been shown to be associated with the outer membrane porins . In the present study, we demonstrate that this protein forms complexes with the lactoferrin receptor LbpA, the transferrin receptor TbpA and the siderophore receptor FrpB as well . This complexation apparently resulted in a stabilization of oligomeric forms of these iron-regulated proteins . In vitro experiments further revealed a reduced ability to acquire iron from human lactoferrin in the rmpM mutant . Furthermore, all TonB-dependent receptors investigated here appeared to exist as oligomers (probably dimers), suggesting that this is a general feature of this class of proteins.

Arch Microbiol, 2000 Jan, 173(1), 10 - 20
Characterization of the ftsZ cell division gene of Neisseria gonorrhoeae: expression in Escherichia coli and N . gonorrhoeae; Salimnia H et al.; We cloned the cell division gene ftsZ of the gram-negative coccus Neisseria gonorrhoeae (Ng) strain CH811, characterized it genetically and phenotypically, and studied its localization in N . gonorrhoeae and Escherichia coli (Ec) . The 1,179-bp ORF of ftsZ(Ng) encodes a protein with a predicted molecular mass of 41.5 kDa . Protein sequence alignments indicate that FtsZ(Ng) is similar to other FtsZ proteins and contains the conserved GTP binding motif . FtsZ homologues were identified in several N . gonorrhoeae strains and in Neisseria lactamica, Neisseria sicca, Neisseria polysaccharae and Neisseria cinerea either by Western blot or by PCR-Southern blot analysis . Attempts to inactivate the ftsZ(Ng) on the chromosome failed, indicating that it is essential for gonococcal growth . FtsZ(Ng) was synthesized in an in vitro transcription/translation system and was shown to be 43 kDa, the same size as in Western blots . Expression of the ftsZ(Ng) gene from nongonococcal promoters resulted in a filamentous phenotype in E . coli . Under controlled expression, the FtsZ(Ng)-GFP fusion protein localized at the mid-cell division site in E . coli . E . coli expressing high levels of the FtsZ(Ng)-GFP fusion protein formed filaments and exhibited different fluorescent structures including helices, spiral tubules extending from pole to pole, and regularly spaced dots or bands that did not localize at the middle of the cell . Expression of the FtsZ(Ng)-GFP fusion protein in N . gonorrhoeae resulted in abnormal cell division as shown by electron microscopy . FtsZ(Ng)-GFP fusions were also expressed in a gonococcal background using a unique shuttle vector.

Obstet Gynecol Surv, 2000 Jan, 55(1), 51 - 7
Epidemiologic issues of sexually transmitted diseases in sexual assault victims; Reynolds MW et al.; The objective of this study was to investigate the epidemiologic aspects of sexually transmitted diseases (STDs) in victims of sexual assault and to discuss the methodological issues in determining risk of STD acquisition . We performed a comprehensive review of the medical literature to determine the prevalence of STDs in victims of sexual assault . A MEDLINE search and a search of bibliographies of published manuscripts was performed to discover relevant articles published in the English language . Studies were included in our review if they provided estimates of the prevalence of infection with Neisseria gonorrhoeae, Chlamydia trachomatis, Treponema pallidum, Trichomonas vaginalis, or human papillomavirus (HPV) . The main outcome measure was prevalence rates of STDs in victims of sexual assault . It is difficult to determine the rate of newly acquired STDs from sexual assault . In the populations studied the prevalence of STDs can be summarized as follows: N . gonorrhoeae 0.0 to 26.3 percent; C . trachomatis 3.9 to 17 percent; T . pallidum 0.0 to 5.6 percent; T . vaginalis 0.0 to 19.0 percent; and HPV 0.6 to 2.3 percent . We conclude that prevalence estimates vary widely depending on the population studied and known risk factors for STDs . Given the limited follow-up rates in this population, preventive treatment for STDs in addition to emergency contraception should be offered in most instances . TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians . LEARNING OBJECTIVES: After completion of this article, the reader will be able to estimate the prevalence of STD transmission from an act of sexual assault, to describe the various types of STDs that a sexual assault victim is at risk for, and to explain the role of preventive therapy in this setting.

Obstet Gynecol Surv, 2000 Jan, 55(1), 41 - 50
Pelvic inflammatory disease after tubal sterilization: a review; Levgur M et al.; Exceptions ot the common notion that tubal occlusion protects from recurrent pelvic inflammatory disease (PID) do exist . Since 1975, 71 cases of salpingitis and 38 tubo-ovarian abscesses (TOA) in sterilized women have been published . The majority of cases of salpingitis after previous tubal occlusion (SPOT) developed more than a year after either laparoscopic or laparotomy sterilization procedures . For tubo-ovarian abscess after previous tubal occlusion (TOAPOT), this time interval ranged from several weeks to almost two decades . Most cases of salpingitis showed inflammation of both tubal segments . When only one segment was involved, it was generally the proximal segment . The appearance of the TOAPOT at the time of surgery was typical to TOA . The symptoms of salpingitis were not different from symptoms in any other case of PID, and those associated with TOAPOT were typical of TOA . Laboratory findings included leucocytosis and growth of Neisseria gonorrohoea and Chlamydia trachomatis from the cervix, the infected tube, and the peritoneal fluid . Pus cultures obtained from cases of TOAPOT grew mixed or single organisms . Detailed histopathologic studies in tubal specimens after the failure of an occlusion procedure are available from cases with no infection . They have demonstrated distortion, loss of musculature, and loss of lumen configuration, all of which may have been the result of compromised blood supply to the tube . These findings may be extrapolated to cases of SPOT and TOAPOT, assuming similar changes may be present before the development of infection . The mechanisms by which infection may develop in previously occluded tubes are divided into three groups: The first group consists of situations where there is persistence of free passage between the proximal and distal portions of the tube . These include toboperitoneal fistula, spontaneous anastomosis at the occlusion site, recanalization of the occluded site, incomplete tubal occlusion due to a faulty surgical technique or rupture of the weakened tubal wall . The second group consists of infections initiated by the surgical procedure itself, such as introduction of pathogens at surgery, exacerbation of chronic PID, and ascending infection secondary to surgical manipulation . In the third group, the infection is initiated systemically by hematogeneous spread, lymphatic spread, or change in immunologic status.

Infect Immun, 2000 Feb, 68(2), 906 - 11
Effects of the immunoglobulin A1 protease on Neisseria gonorrhoeae trafficking across polarized T84 epithelial monolayers; Hopper S et al.; We previously demonstrated that the Neisseria IgA1 protease cleaves LAMP1 (lysosome-associated membrane protein 1), a major integral membrane glycoprotein of lysosomes, thereby accelerating its degradation rate in infected A431 human epidermoid carcinoma cells and resulting in the alteration of lysosomes in these cells . In this study, we determined whether the IgA1 protease also affects the trafficking of Neisseria gonorrhoeae across polarized T84 epithelial monolayers . We report that N . gonorrhoeae infection of T84 monolayers, grown on a solid substrate or polarized on semiporous membranes, also results in IgA1 protease-mediated reduction of LAMP1 . We demonstrate that iga mutants in two genetic backgrounds exited polarized T84 monolayers in fewer numbers than the corresponding wild-type strains . Finally, we present evidence that these mutants have a statistically significant and reproducible defect in their ability to traverse T84 monolayers . These results add to our previous data by showing that the IgA1 protease alters lysosomal content in polarized as well as unpolarized cells and by demonstrating a role for the protease in the traversal of epithelial barriers by N . gonorrhoeae.

Infect Immun, 2000 Feb, 68(2), 896 - 905
Isolation of Neisseria gonorrhoeae mutants that show enhanced trafficking across polarized T84 epithelial monolayers; Hopper S et al.; Initiation of a gonococcal infection involves attachment of Neisseria gonorrhoeae to the plasma membrane of an epithelial cell in the mucosal epithelium and its internalization, transepithelial trafficking, and exocytosis from the basal membrane . Piliation and expression of certain Opa proteins and the immunoglobulin A1 protease influence the transcytosis process . We are interested in identifying other genetic determinants of N . gonorrhoeae that play a role in transcellular trafficking . Using polarized T84 monolayers as a model epithelial barrier, we have assayed an N . gonorrhoeae FA1090 minitransposon (mTn) mutant bank for isolates that traverse the monolayer more quickly than the isogenic wild-type (WT) strain . From an initial screen, we isolated four mutants, defining three genetic loci, that traverse monolayers significantly more quickly than their WT parent strain . These mutants adhere to and invade cells normally and do not affect the integrity of the monolayer barrier . Backcrosses of the mutations into the WT FA1090 strain yielded mutants with a similar fast-trafficking phenotype . In two mutants, the mTns had inserted 370 bp apart into the same locus, which we have named fit, for fast intracellular trafficker . Backcrosses of one of these mutants into the MS11A genetic background also yielded a fast-trafficking mutant . The fit locus contains two overlapping open reading frames, fitA and fitB, whose deduced amino acid sequences have predicted molecular weights of 8.6 and 15.3, respectively . Neither protein contains a signal sequence . FitA has a potential helix-turn-helix motif, while the deduced sequence of FitB offers no clues to its function . fitA or fitB homologues are present in the genomes of Pseudomonas syringae and Rhizobium meliloti, but not Neisseria meningitidis . Replication of the MS11A fitA mutant in A431 and T84 cells is significantly accelerated compared to that of the isogenic WT strain . In contrast, growth of this mutant in liquid media is normal . Taken together, these results strongly suggest that traversal of N . gonorrhoeae across an epithelial barrier is linked to intracellular bacterial growth.

Mol Microbiol, 2000 Jan, 35(1), 32 - 43
Interaction of primary human endometrial cells with Neisseria gonorrhoeae expressing green fluorescent protein; Christodoulides M et al.; Infection of the endometrium by Neisseria gonorrhoeae is a pivotal stage in the development of pelvic inflammatory disease in women . An ex vivo model of cultures of primary human endometrial cells was developed to study gonococcal-host cell interactions . To facilitate these studies, gonococci were transformed with a hybrid shuttle vector containing the gfp gene from Aequoria victoria, encoding the green fluorescent protein (GFP), to produce intrinsically fluorescent bacteria . The model demonstrated that both pili and Opa proteins were important for both mediating gonococcal interactions with endometrial cells and inducing the secretion of pro-inflammatory cytokines and chemokines . Pil+ gonococci showed high levels of adherence and invasion, regardless of Opa expression, which was associated with increased secretion of IL-8 chemokine and reduced secretion of IL-6 cytokine . Gonococcal challenge also caused increased secretion of TNF-alpha cytokine, but this did not correlate with expression of pili or Opa, suggesting that release of components from non-adherent bacteria may be involved in TNF-alpha induction . Thus, the use of cultured primary endometrial cells, together with gonococci expressing green fluorescent protein, has the potential to extend significantly our knowledge, at the molecular level, of the role of this important human pathogen in the immunobiology of pelvic inflammatory disease.

J Bacteriol, 2000 Jan, 182(2), 439 - 47
Use of heme compounds as iron sources by pathogenic neisseriae requires the product of the hemO gene; Zhu W et al.; Heme compounds are an important source of iron for neisseriae . We have identified a neisserial gene, hemO, that is essential for heme, hemoglobin (Hb), and haptoglobin-Hb utilization . The hemO gene is located 178 bp upstream of the hmbR Hb receptor gene in Neisseria meningitidis isolates . The product of the hemO gene is homologous to enzymes that degrade heme; 21% of its amino acid residues are identical, and 44% are similar, to those of the human heme oxygenase-1 . DNA sequences homologous to hemO were ubiquitous in commensal and pathogenic neisseriae . HemO genetic knockout strains of Neisseria gonorrhoeae and N . meningitidis were unable to use any heme source, while the assimilation of transferrin-iron and iron-citrate complexes was unaffected . A phenotypic characterization of a conditional hemO mutant, constructed by inserting an isopropyl-beta-D-thiogalactopyranoside (IPTG)-regulated promoter upstream of the ribosomal binding site of hemO, confirmed the indispensability of the HemO protein in heme utilization . The expression of HemO also protected N . meningitidis cells against heme toxicity . hemO mutants were still able to transport heme into the cell, since both heme and Hb could complement an N . meningitidis hemA hemO double mutant for growth . The expression of the HmbR receptor was reduced significantly by the inactivation of the hemO gene, suggesting that hemO and hmbR are transcriptionally linked . The expression of the unlinked Hb receptor, HpuAB, was not altered . Comparison of the polypeptide patterns of the wild type and the hemO mutant led to detection of six protein spots with an altered expression pattern, suggesting a more general role of HemO in the regulation of gene expression in Neisseriae.

Microbiology, 1999 Dec, 145 ( Pt 12), 3529 - 37
Representational difference analysis of cDNA for the detection of differential gene expression in bacteria: development using a model of iron-regulated gene expression in Neisseria meningitidis; Bowler LD et al.; Representational difference analysis of cDNA (cDNA RDA) provides a powerful technique for the identification of specific differences between two mRNA populations . The method has previously been used to analyse differential gene expression in eukaryotes, but until now has not been successfully applied to prokaryotes . A strain of Neisseria meningitidis with a deletion of the iron-regulated lactoferrin-binding protein A (IbpA) gene, grown under iron-replete conditions, and the isogenic parent strain, grown under iron limitation, were used as a model for developing cDNA RDA for use with bacteria . In this system, the technique should specifically detect the differential expression of the IbpA gene in the parent strain, along with other genes whose expression is switched on (or up-regulated) under iron-deficient conditions . Since cDNA RDA requires high-quality, representative mRNA, a variety of methods for the isolation of RNA were evaluated . A triisopropylnaphthalene sulphonic acid/ p-aminosalicylic acid-based technique was found to give the best results . cDNA was prepared from total RNA isolated from the two N . meningitidis strains and subjected to an adapted cDNA RDA procedure . The method resulted in the amplification of five major PCR products, which included fragments of the IbpA gene and the iron-regulated RTX-like toxin gene (frpC), thus validating the technique for use with bacteria.

Sex Transm Infect, 1999 Oct, 75(5), 327 - 31
Peripheral blood T cell proliferative response to chlamydial organisms in gonococcal and non-gonococcal urethritis and presumed pelvic inflammatory disease; Shahmanesh M et al.; OBJECTIVE: To study peripheral blood mononuclear cell (PBMC) proliferative response to Chlamydia trachomatis elementary bodies in (a) controls, (b) various stages of gonococcal (c) and non-gonococcal urethritis, and (d) women with a clinical diagnosis of pelvic inflammatory disease (PID) . METHODS: We categorised 102 men presenting to a GUM clinic with urethritis by organisms (C trachomatis (CT) or Neisseria gonorrhoeae (NG) (both by culture), and whether it was their first (urethritis naive) or subsequent (urethritis experienced) attack . 23 women presenting to the clinic with a clinical diagnosis of PID were also investigated . We measured PBMC proliferative responses to C trachomatis (DK20--an oculogenital strain, serovar E), lysate of McCoy cells (used to propagate chlamydiae), and the recall antigen PPD . Controls were 37 men and women without present or past history of urethritis or chlamydial infection . Results were expressed as the ratio of the stimulation index (SI) obtained with DK20 compared with McCoy cells (DK index), and the ratio of the SI obtained with DK20 compared with PPD (PPD index) . RESULTS: The median SI to DK20 in the urethritis was 12.7 which was significantly higher than the controls (7.6, p < 0.003) . The median SI to the recall antigen PPD was similar in the urethritis patients (17.4) and the controls (22.4) . All urethritis patient subgroups had a significantly higher DK index and PPD index than the controls . There was no difference in the PPD and DK index between urethritis naive and urethritis experienced patients and between the culture positive and culture negative urethritis subgroups . In PID patients only the PPD index was significantly higher than the controls . CONCLUSION: Men presenting with urethritis and women presenting with PID both have significantly greater peripheral blood mononuclear cell proliferative responses to the DK20 strain of C trachomatis than controls . A similar T cell proliferative response pattern in urethritis naive patients with either gonococcal or non-gonococcal urethritis could be because low sensitivity of CT culture failed to diagnose some cases of C trachomatis . However, it may also signify earlier exposure of the patients to chlamydial antigens (for example, C pneumoniae), cross reacting antigens such as heat shock proteins from other microbial species, or a "bystander" activation of chlamydia specific memory T cells trafficking through mucosal lymphoid tissue during urethritis . These results suggest evidence of T cell mediated response to C trachomatis cannot be used as a diagnostic tool.

Sex Transm Infect, 1999 Oct, 75(5), 312 - 6
Risk factors for the acquisition of genital warts: are condoms protective?
Wen LM, Estcourt CS, Simpson JM, Mindel A.
OBJECTIVES: To characterise risk factors for the acquisition of genital warts and specifically to determine whether condoms confer protection from infection . METHODS: A retrospective case-control study comparing demographic, behavioural, and sexual factors in men and women with and without newly diagnosed genital warts, who attended Sydney Sexual Health Centre (SSHC), an inner city public sexual health centre, in 1996 . Data were extracted from the SSHC database . Crude odds ratios (OR) were calculated to compare cases and controls and significant factors were then controlled for using multivariate logistic regression to obtain adjusted odds ratios (ORs) . RESULTS: 977 patients with warts and 977 controls matched by sex and date of attendance were included . In both sexes, univariate analysis revealed that younger age, more lifetime sexual partners, failure to use condoms, greater cigarette smoking and alcohol consumption were associated with warts, and there was a negative association with previous infection with Chlamydia trachomatis, Neisseria gonorrhoeae, hepatitis B, and genital herpes . In males, on multivariate analysis, factors which remained significant were younger age, more lifetime sexual partners; failure to use condoms, greater cigarette smoking, and previous chlamydia . In women, factors which remained significant were younger age, more lifetime sexual partners, condom use, marital status, and previous infections with Chlamydia trachomatis and herpes . CONCLUSIONS: Independent risk factors for genital warts include younger age, greater number of lifetime sexual partners, and smoking . Consistent condom use significantly reduces the risk of acquiring genital warts.

J Bacteriol, 2000 Jan, 182(1), 171 - 8
Opa expression correlates with elevated transformation rates in Neisseria gonorrhoeae; Hill SA; Neisseria gonorrhoeae is naturally competent for DNA transformation . Under most conditions encountered in vivo, gonococci express one or more opacity (Opa) proteins on their surfaces . Recently, it was shown that DNA preferentially binds to the surfaces of Opa-expressing organisms compared to those of isogenic Opa-negative strains, presumably due to the numerous cationic residues in the predicted surface-exposed loops of the Opa protein . This study examined whether Opa-DNA interactions actually influence DNA transformation of the gonococcus . The data show that Opa-expressing gonococci are more efficient recipients of DNA for transformation and are more susceptible to exogenous DNase I treatment at early stages during the DNA transformation process than non-Opa expressors . Furthermore, inhibition of the transformation process was demonstrable for Opa(+) populations when either nonspecific DNA or the polyanion heparin was used . Overall, the data suggest that Opa expression, with its presumptive positive surface charge contribution, promotes DNA transformation by causing a more prolonged sequestration of donor DNA at the cell surface, which translates into more efficient transformation over time.

Plasmid, 2000 Jan, 43(1), 24 - 34
Sequence analysis of the family of penicillinase-producing plasmids of Neisseria gonorrhoeae; Pagotto F et al.; The exact nature of the sequence differences between the medically important family of gonococcal penicillinase-producing plasmids has been ascertained . The entire DNA sequence of the Asia-type plasmid, pJD4, demonstrated that it is 7426 bp and contains two direct repeats (DR30) that are implicated in the formation of deletion variant plasmids, such as the Africa-type plasmid . We have identified putative DnaA and IHF binding sites, various open reading frames that are thought to specify functional proteins, and some important DNA sequences involved with conjugative transfer of gonococcal beta-lactamase plasmids . The deletion in the Africa-type plasmid is 1827 bp and one of the DR30 repeats is also missing . The deletion in the Rio-type plasmid and several Toronto-type plasmids was determined to be 2273 bp and the sequence spanning the deletion was identical irrespective of geographic or temporal origin . The &Ncirc;imes-type plasmid is an Africa-type plasmid and also contains an IS5 insertion sequence . Since IS5 has not been identified in gonococcal isolates, we suggest that this sequence may have been inserted after the original gonococcal plasmid was transformed into Escherichia coli . The New Zealand plasmid is an Asia-type plasmid that contains an endogenous tandem duplication of 1883 bp and the direct DR2 is implicated in this duplication . The nature of the defined truncation of Tn2 present in the various plasmids is also discussed .

An Med Interna, 1999 Sep, 16(9), 451 - 6
{A management analysis of bacterial meningitis in a hospital emergency service: the delay from the start of treatment and related factors}; Morano Amado LE et al.; OBJECTIVE: To establish the time elapsed from the patient arrival to the emergency room to the beginning of antibiotic therapy . To identify etiologic factors for treatment delay . METHODS: 73 patients diagnosed of bacterial meningitis in the emergency room and admitted to the hospital were studied . Patient characteristics as well as meningitis predisposing factors, symptoms, physical examination, laboratory data, radiological studies and previous ambulatory treatment, were recorded retrospectively . Arrival time, time expended at diagnostic procedures and time of administration of the first antibiotic dose, as well as the administration place were registered . Patients clinical evolution, and factors influencing the delay of antibiotic administration were analyzed . RESULTS: Median age was 17 years . Patient care was evenly distributed along the day, 80% had a light base risk, 29% had at least a risk factor for meningitis, 22% received antibiotic previously . Clinical presentation was classic in more than 71% of patients . Blood cultures were positive in 41%, and CSF cultures were positive in 63%, 43% of cases were related to Neisseria meningitidis, 20% Streptococcus pneumoniae and unknown bacteria in 31.5% . Computerized Tomography (CT) was performed in 9 cases . Median time from the arrival to the Emergency Room until antibiotic administration was 5 hours and 25 minutes: When antibiotics were given before Lumbar Puncture (LP), it was 2 hours and 50 minutes, 5 hours 20 minutes when therapy was started after LP, and 7 hours and 22 minutes when CT was performed before LP . The only factor showing a statistically significant relation with the time to antibiotic administration was the patient being sent by the primary care physician to the hospital with a presumptive diagnosis of bacterial meningitis (1 hour 20 minutes vs . 5 hours 51 minutes) . CONCLUSION: Only a small part of bacterial meningitis cases start antibiotic treatment in the first 30 minutes . Delay is high and it increases when certain diagnostic tests are performed . Information received from the primary care physician, has the highest influence on the beginning of treatment.

J Biol Chem, 1999 Dec 31, 274(53), 37717 - 22
Glycosyl fluorides can function as substrates for nucleotide phosphosugar-dependent glycosyltransferases; Lougheed B et al.; alpha-Galactosyl fluoride is shown to function as a substrate, in place of uridine-5'-diphosphogalactose, for the alpha-galactosyltransferase from Neisseria meningitidis . The reaction only occurs in the presence of catalytic quantities of uridine 5'-diphosphate . In the presence of galactosyl acceptors, the expected oligosaccharide product is formed in essentially quantitative yields, reaction having been performed on multi-milligram scales . In the absence of a suitable acceptor, the enzyme synthesizes uridine-5'-diphosphogalactose, as demonstrated through a coupled assay in which uridine-5'-diphosphogalactose is converted to uridine-5'-diphosphoglucuronic acid with conversion of NAD to NADH . These glycosyl fluoride substrates therefore offer the potential of an inexpensive alternative donor substrate in the synthesis of oligosaccharides as well a means of generating steady state concentrations of nucleotide diphosphate sugars for in situ use by other enzymes . Further, they should prove valuable as mechanistic probes.

Microbes Infect, 1999 May, 1(6), 455 - 64
The epidemiology of Neisseria gonorrhoeae in Europe; van Duynhoven YT; This review addresses the occurrence, the trends, and the risk groups of Neisseria gonorrhoeae in Europe over the past decade . A decline has been observed in most of Europe since the 1980s, except for an increase in the Baltic countries (early 1990s) and an increase among men who have sex with men (between 1989 and 1991), and heterosexuals in some countries (between 1995 and 1997) . Despite the overall fall in the incidence of gonorrhoea, plasmid-mediated resistance to penicillin and tetracycline increased in Europe . More recently, resistance to fluoroquinolones has been documented, mainly imported from Southeast Asia . Until now, no resistance to third-generation cephalosporins has been observed.

Trends Microbiol, 1999 Dec, 7(12), 482 - 7
Multilocus sequence typing; Enright MC et al.; Multilocus sequence typing (MLST) provides a new approach to molecular epidemiology that can identify and track the global spread of virulent or antibiotic-resistant isolates of bacterial pathogens using the Internet . MLST databases, together with interrogation software, are available for Neisseria meningitidis and Streptococcus pneumoniae and databases for Streptococcus pyogenes and Staphylococcus aureus will be released shortly.

Infect Immun, 2000 Jan, 68(1), 239 - 46
Murine immune response to Neisseria meningitidis group C capsular polysaccharide: analysis of monoclonal antibodies generated in response to a thymus-independent antigen and a thymus-dependent toxoid conjugate vaccine; Garcia-Ojeda PA et al.; Antibody (Ab) responses to polysaccharides (PSs) such as Neisseria meningitidis group C PS (MCPS) are characterized as being thymus independent (TI) and are restricted with regard to clonotype and isotype expression . PS conjugated to proteins, e.g., MCPS coupled to tetanus toxoid (MCPS-TT), elicits a thymus-dependent (TD) response . In order to understand the influence of the form of a vaccine (TI versus TD) on the Ab repertoire, we generated monoclonal antibody (MAb) panels from mice immunized and boosted with MCPS or MCPS-TT in different ways . The panels of MAbs were examined for isotype, fine specificity, affinity, and V(H) gene family usage . The use of MCPS-TT resulted in a shift in the isotype from immunoglobulin M (IgM) and IgG3 elicited in response to the MCPS to primarily IgG1 . This isotype shift was accompanied by a change in the fine specificity of the response to the conjugate compared to that of PS . New fine specificities and increased affinity were observed in response to the TD antigen (Ag) . Dot blot and Northern analyses of MCPS MAbs revealed that V(H) gene family usage is dominated by V(H)J558, used by 23 of 39 MAbs . V(H)3609 was seen in three MAbs of restricted fine specificity . V(H)Q52, V(H)7183, and V(H)VGAM3-8 were seen in more than one MAb across these panels, while V(H)10 and V(H)X24 were detected only once in response to the TI-2 Ag . All MAbs in the panels utilized kappa light chains, and all functional J(kappa) genes were expressed.

Antimicrob Agents Chemother, 2000 Jan, 44(1), 210 - 2
Complete sequence of a beta-lactamase-encoding plasmid in Neisseria meningitidis; Backman A et al.; Identical beta-lactamase-encoding (TEM-1) plasmids were found in two different clinical Neisseria meningitidis strains . They were completely sequenced (5,597 bp) and designated pAB6 . The plasmid is almost identical to Neisseria gonorrhoeae plasmid pJD5 (5,599 kb) and may have been picked up from a gonococcus in vivo.

Antimicrob Agents Chemother, 2000 Jan, 44(1), 192 - 5
Susceptibilities of Neisseria gonorrhoeae isolates containing amino acid substitutions in GyrA, with or without substitutions in ParC, to newer fluoroquinolones and other antibiotics; Tanaka M et al.; We examined the antimicrobial susceptibilities of 85 Neisseria gonorrhoeae isolates, classified according to the presence of amino acid substitutions in the GyrA and ParC proteins, to 12 fluoroquinolones and 7 other antibiotics . Sitafloxacin and HSR-903 showed excellent activity against N . gonorrhoeae, including strains with both GyrA and ParC substitutions . Among the strains with various GyrA substitutions, strains with a serine-91-to-phenylalanine mutation required the highest MICs of all of the fluoroquinolones tested and were cross-resistant to structurally unrelated beta-lactams.

Am J Obstet Gynecol, 1999 Dec, 181(6), 1374 - 81
Effects of human immunodeficiency virus 1 infection on microbial origins of pelvic inflammatory disease and on efficacy of ambulatory oral therapy; Bukusi EA et al.; OBJECTIVE: This study was undertaken to determine the effects of human immunodeficiency virus 1 infection on the clinical presentation, severity, causal organisms, and response to ambulatory therapy of pelvic inflammatory disease . STUDY DESIGN: Women 18 to 40 years old with lower abdominal pain for <1 month were recruited . Participants underwent a standardized questionnaire, physical examination, screening for human immunodeficiency virus 1 and other sexually transmitted infections, and endometrial biopsy to detect plasma cell endometritis . Reevaluations were performed at 1 and 4 weeks to assess response to therapy . RESULTS: Among 162 women with adequate endometrial biopsy specimens 63 (39%) had histologically confirmed endometritis . Endometritis was more frequent among women who were seropositive for human immunodeficiency virus 1 than among women who were seronegative (odds ratio, 3.0; 95% confidence interval, 1.5-5.9) . Infections with either Neisseria gonorrhoeae or Chlamydia trachomatis, or both, were least common and bacterial vaginosis was most common among human immunodeficiency virus 1-infected women with CD4 T-lymphocyte counts <400 cells/microL (P < . 04, P <.03, respectively) . After oral antibiotic therapy, similar proportions of both women who were seropositive and women who were seronegative for human immuno-deficiency virus 1 had a >/=75% reduction in clinical severity score (81% vs 86%) . CONCLUSION: Outpatient treatment of pelvic inflammatory disease was successful regardless of human immunodeficiency virus 1 serostatus.

Glycoconj J, 1999 Mar, 16(3), 205 - 12
The living factory: in vivo production of N-acetyllactosamine containing carbohydrates in E . coli; Bettler E et al.; Scientific and commercial interest in oligosaccharides is increasing, but their availability is limited as production relies on chemical or chemo-enzymatic synthesis . In search for a more economical, alternative procedure, we have investigated the possibility of producing specific oligosaccharides in E . coli that express the appropriate glycosyltransferases . The Azorhizobium chitin pentaose synthase NodC (a beta(1,4)GlcNAc-oligosaccharide synthase), and the Neisseria beta(1,4)galactosyltransferase LgtB, were co-expressed in E . coli . The major oligosaccharide isolated from the recombinant strain, was subjected to LC-MS, FAB-MS and NMR analysis, and identified as betaGal(1,4){betaGlcNAc(1,4)}4GlcNAc . High cell density culture yielded more than 1.0 gr of the hexasaccharide per liter of culture . The compound was found to be an acceptor in vitro for betaGal(1,4)GlcNAc alpha(1,3)galactosyltransferase, which suggests that the expression of additional glycosyltransferases in E . coli will allow the production of more complex oligosaccharides.

Mol Diagn, 1999 Sep, 4(3), 233 - 9
Development of a simplex polymerase chain reaction-based assay for the detection of Neisseria meningitidis; Glustein JZ et al.; BACKGROUND: This study was undertaken to develop a sensitive and specific polymerase chain reaction (PCR)-based assay for Neisseria meningitidis and Neisseria gonorrhoeae that could ultimately be incorporated into a multiplex assay designed to screen cerebrospinal fluid (CSF) for a panel of pyogenic bacterial species associated with bacterial meningitis . METHODS: N.meningitidis-specific primers were designed from porA gene sequences with the aid of a commercial software program and were used to develop and validate a clinical assay using a liquid hybridization-gel retardation detection system . The analytic sensitivity of the assay was determined using CSF spiked with N . meningitidis and comparing the PCR-based results with culture . RESULTS: Analytic sensitivity experiments showed the assay's limit of detection to be 100 fg purified input target DNA and 0.0125 colony-forming units of N . meningitidis spiked into CSF . Specificity experiments showed the assay could detect all strains of N . meningitidis and N . gonorrhoeae tested, but did not support amplification of the commensal neisserial species or a panel of other human bacterial pathogens . CONCLUSIONS: This PCR-based assay for pathogenic neisserial species is sensitive and specific and suitable for incorporation into a multiplex assay for the clinical differentiation of aseptic and septic meningitis.

J Biol Chem, 1999 Dec 17, 274(51), 36550 - 8
Structural and immunochemical characterization of a Neisseria gonorrhoeae epitope defined by a monoclonal antibody 2C7; the antibody recognizes a conserved epitope on specific lipo-oligosaccharides in spite of the presence of human carbohydrate epitopes; Yamasaki R et al.; Lipo-oligosaccharides (LOS) produced by Neisseria gonorrhoeae are important antigenic and immunogenic components of the outer membrane complex . Previously, we showed that murine monoclonal antibody (mAb) 2C7 did not cross-react with human glycosphingolipids but identified the LOS epitope that is widely expressed in vivo and in vitro (Gulati, S., McQuillen, D . P., Mandrell, R . E., Jani, D . B., and Rice, P . A . (1996) J . Infect . Dis . 174, 1223-1237) . In the present study, we analyzed the structure of gonococcal strain WG LOS containing the 2C7 epitope and investigated the structural requirements for expression of the epitope . We determined that the WG LOS components are Hep{1}-elongated forms of 15253 LOS that have a lactose on both Hep{1} and Hep{2} (Yamasaki, R., Kerwood, D . E., Schneider, H., Quinn, K . P., Griffiss, J . M., and Mandrell, R . E . (1994) J . Biol . Chem . 269, 30345-30351) . In addition, we found that expression of the 2C7 epitope within the LOS is blocked when the Hep{2}-lactose is elongated . Based on the structural data of these LOS and the results obtained from immunochemical analyses, we conclude the following: 1) mAb 2C7 requires both the 15253 OS minimum structure and the N-linked fatty acids in the lipoidal moiety for expression of the epitope; 2) mAb 2C7 binds to the LOS that elongates the lactose on Hep{1} of the 15253 OS, but not the one on Hep{2}; and 3) the 2C7 epitope is expressed on gonococcal LOS despite the presence of human carbohydrate epitopes such as a lactosamine or its N-acetylgalactosaminylated (globo) form . Our study shows that the conserved epitope defined by mAb 2C7 could potentially be used as a safe site for the development of a vaccine candidate.

Acta Paediatr, 1999 Nov, 88(11), 1216 - 22
Bacterial flora of the lower respiratory tract in children with bronchial asthma; Fayon M et al.; The aims of this retrospective study were to (i) determine the risk of contamination of lower respiratory tract samples obtained during fiberoscopy in children; (ii) determine the incidence and profile of the bacterial flora of the lower respiratory tract in a selected group of asthmatic children at high risk for bacterial infection; and (iii) identify risk markers for such findings . In 29 asthmatic children, comparison of bacterial cultures of specimens obtained from the upper and, lower respiratory tracts showed that contamination was a possibility in only 3.4% (1/29) of cases . The results from bacterial samples obtained via flexible bronchoscopy in a further 273 consecutively investigated physician-diagnosed asthmatic children were analysed . Patients were selected for bronchoscopy if they had severe chronic asthma or in order to exclude other diseases able to provoke wheezing . Their mean (SD) and median ages were 32.2 (38.3) and 17.5 mo, respectively . The incidence of positive bacterial cultures was 12.1% (33/273 patients) . Bacterial flora included H . influenzae (39.5%, 15/38), B . catarrhalis (23.7%, 9/38), Neisseria species (7.9%, 3/38), M . pneumoniae (7.9%, 3/38), P . non-aeruginosa (5.3%, 2/38) and P . aeruginosa (2.6%, 1/38) . No clinical or radiological markers were significantly associated with lower respiratory tract bacterial infection . Large quantities of bacteria were present in the lower respiratory tracts of a substantial number of children (1/8) in this selected group of asthmatics . For the moment, however, the clinical implications of this finding remain unclear.

Rev Epidemiol Sante Publique, 1999 Oct, 47(5), 397 - 402
{Incidence and management of male urethritis in the district of Tunis}; Ben Hamida A et al.; BACKGROUND: The aim of the study was to evaluate the incidence of male urethritis and the relative frequency of the different etiological agents in order to adapt standard case management . METHODS: The incidence of urethritis has been estimated with a postal study made on a sample of druggists (1/10) of the area . The total observation period was four weeks . The relative frequency of the etiological agents and the positive predictive value (PPV) of the therapeutic approach based on antibiotic treatment of gonorrhoea and Chlamydia trachomatis was achieved on 92 cases of male urethritis attending general physicians in two polyclinics . RESULTS: The annual incidence of male urethritis was estimated at 680 per 100,000 . The relative frequency of etiological agents was as 34.7% for Neisseria gonorrhoea, 7.6% for Chlamydia trachomatis and was found at 3.3% for Trichomonas vaginalis; the PPV was only 43% because of the high proportion of negative results reported by the laboratory . CONCLUSION: Gonococcal urethritis incidence tends to decrease regarding non gonococcal urethritis and the therapeutic approach appears to be warranted.

Biosci Biotechnol Biochem, 1999 Oct, 63(10), 1708 - 13
Purification and characterization of an esterase involved in cellulose acetate degradation by Neisseria sicca SB; Moriyoshi K et al.; An esterase catalyzing the hydrolysis of acetyl ester moieties in cellulose acetate was purified 1,110-fold to electrophoretic homogeneity from the culture supernatant of Neisseria sicca SB, which can assimilate cellulose acetate as the sole carbon and energy source . The purified enzyme was a monomeric protein with a molecular mass of 40 kDa and the isoelectric point was 5.3 . The pH and temperature optima of the enzyme were 8.0-8.5 and 45 degrees C . The enzyme catalyzed the hydrolysis of acetyl saccharides, p-nitrophenyl esters of short-chain fatty acids, and was slightly active toward aliphatic and aromatic esters . The K(m) and Vmax for cellulose acetate (degree of substitution, 0.88) and p-nitrophenyl acetate were 0.0162% (716 microM as acetyl content in the polymer) and 36.0 microM, and 66.8 and 39.1 mumol/min/mg, respectively . The enzyme was strongly inhibited by phenylmethylsulfonyl fluoride and diisopropyl fluorophosphate, which indicated that the enzyme was a serine esterase.

J Immunol, 1999 Dec 15, 163(12), 6732 - 40
NADPH oxidase activation and assembly during phagocytosis; DeLeo FR et al.; Generation of superoxide (O2-) by the NADPH-dependent oxidase of polymorphonuclear leukocytes is an essential component of the innate immune response to invading microorganisms . To examine NADPH oxidase function during phagocytosis, we evaluated its activation and assembly following ingestion of serum-opsonized Neisseria meningitidis, serogroup B (NMB), and compared it with that elicited by serum-opsonized zymosan (OPZ) . Opsonized N . meningitidis- and OPZ-dependent generation of reactive oxygen species by polymorphonuclear leukocytes peaked early and then terminated . Phosphorylation of p47phox coincided with peak generation of reactive oxygen species by either stimulus, consistent with a role for p47phox phosphorylation during NADPH oxidase activation, and correlated with phagosomal colocalization of flavocytochrome b558 (flavocytochrome b) and p47phox and p67phox (p47/67phox) . Termination of respiratory burst activity did not reflect dephosphorylation of plasma membrane- and/or phagosome-associated p47phox; in contrast, the specific activity of phosphorylated p47phox at the phagosomal membrane increased . Most significantly, termination of oxidase activity paralleled the loss of p47/67phox from both NMB and OPZ phagosomes despite the continued presence of flavocytochrome b . These data suggest that 1) the onset of respiratory burst activity during phagocytosis is linked to the phosphorylation of p47phox and its translocation to the phagosome; and 2) termination of oxidase activity correlates with loss of p47/67phox from flavocytochrome b-enriched phagosomes and additional phosphorylation of membrane-associated p47phox.

Pediatrics . 1999 Dec;104(6):e72.
Neisseria gonorrhea infections in girls younger than 12 years of age evaluated for vaginitis; Shapiro RA et al.; OBJECTIVE: This study examined the prevalence of gonorrhea in girls <12 years of age who presented with vaginitis and in whom sexual abuse was not suspected . DESIGN: A prospective, consecutive patient series was performed in a pediatric emergency department with 90 000 visits per year and in 2 affiliated pediatric continuity clinics . All girls (Tanner I or II) between the ages of 12 months and 12 years, presenting with a chief complaint of vaginal discharge, burning, pain, or itching, were enrolled (n = 93) . Patients were excluded (n = 6) if there was a history of sexual abuse . The presence or absence of vaginal discharge, vaginal erythema, or trauma was recorded . Physicians were instructed to collect cultures for Neisseria gonorrhea (GC), Chlamydia trachomatis, and bacteria/yeast . Wet prep, urinalysis, urine culture, serum rapid plasma reagin, and fungal culture were obtained at the physician's discretion . RESULTS: Of the girls, 43 had a vaginal discharge on examination . Of these girls, 4 (9%) had GC, 9 (26%) had group A, B, or F streptococcus and 1 had Staphylococcus aureus . Of the girls, 44 had no discharge on examination . In this group, 3 had streptococcus infection and 2 had Candida albicans . Both children with C albicans had been treated recently with systemic antibiotics . Those girls with a vaginal discharge on examination had a microbial etiology significantly more often than did those without discharge . All of the girls with infection were Tanner I on genital examination . CONCLUSIONS: The prevalence of unsuspected GC infection was high and emphasizes the importance of culturing Tanner I girls for GC when they have a vaginal discharge along with routine bacterial cultures . Testing and/or treating for C albicans should be considered when there has been recent antibiotic use . Girls with vaginal complaints but without vaginal discharge may have a bacterial infection, but such diagnoses occur less frequently than with girls who have a discharge.

Int J STD AIDS, 1999 Oct, 10(10), 646 - 51
Subtyping of high-level plasmid-mediated tetracycline resistant Neisseria gonorrhoeae isolated in Scotland between 1992 and 1998; Beattie T et al.; Tetracycline resistant Neisseria gonorrhoeae (TRNG) contain a 25.2 MDa TetM plasmid encoding a 68 KDa cytoplasmic protein which confers high-level tetracycline resistance . The aim of this study was to subtype all TRNG isolated in Scotland between 1992 and 1998 . Subtyping was performed by a polymerase chain reaction (PCR) assay which characterizes the TetM plasmid as either the Dutch variant (443 base pair product) or the American variant (777 base pair product) . Of the 78 TRNG isolates, 35 were the American variant and 43 were the Dutch variant . TRNG were distributed amongst 30 serovar/auxotype classes, the most common being 1A6/NR (11.5%), 1A6/P (14.1%) and 1B4/NR (14.1%) . The country where infection was acquired was known for 36 of the 46 TRNG strains isolated between 1996 and 1998 . All infections acquired in Asia and South America were the Dutch variant whereas all infections acquired in Africa were the American variant . A penicillinase plasmid was present in 66% (23/35) of the American variant TRNG compared with 51% (22/43) of the Dutch variant: the 3.2 MDa penicillinase plasmid was found in 87% of the American variant TRNG whereas the 4.4 MDa penicillinase plasmid was found in 68% of the Dutch variant TRNG . We conclude that subtyping of TRNG by PCR is a useful tool in studying the epidemiology of gonococcal infection due to plasmid-mediated resistant isolates.

Lancet, 1999 Nov 20, 354(9192), 1776 - 81
Reproductive-tract infections in women in low-income, low-prevalence situations: assessment of syndromic management in Matlab, Bangladesh; Hawkes S et al.; BACKGROUND: In the control of reproductive-tract infections, including sexually transmitted infections (STIs), in low-income and middle-income countries, WHO recommends syndromic management for individuals with symptoms . This intervention was initially developed in areas where prevalence of such infections is high . We investigated the clinical effectiveness and cost of this approach among a group of women with a low prevalence of infection . METHODS: During a 5-month period, we investigated all women complaining of abnormal vaginal discharge and seeking care at maternal and child health/family-planning centres in Matlab, Bangladesh, for the presence of laboratory-diagnosed reproductive-tract infections and STIs . Syndromic diagnoses made by trained health-care workers were compared with laboratory diagnosis of infection . We then calculated the costs of treating women by means of the recommended WHO algorithm and an adapted algorithm incorporating use of a speculum and simple diagnostic tests . FINDINGS: The prevalence of endogenous infections among 320 women seen was 30% . Cervical infections (Neisseria gonorrhoeae and Chlamydia trachomatis) were found in only three women . The WHO algorithm had a high sensitivity (100%) but a low specificity (zero for bacterial vaginosis, candida, and Trichomonas vaginalis) . The speculum-based algorithm had a low sensitivity (between zero and 59%) but a higher specificity (79-97%) . Between 36% and 87% of costs would have been spent on uninfected women . INTERPRETATION: The high rate of overtreatment in the population studied carries both financial and social costs--the latter in potentially exposing women misdiagnosed as having an STI to threats of domestic disruption or even violence . We make recommendations for management programmes in areas of low STI prevalence and low incomePIP: This paper presents a study on the syndromic management of reproductive tract infections among women in low-income and low-prevalence situations . Women complaining of abnormal vaginal discharge and seeking care at health centers in Matlab, Bangladesh, were examined for the presence of laboratory-diagnosed reproductive tract infections and sexually transmitted infections . In the results, 30% of 320 women were diagnosed as having endogenous infections . Overall result of the study revealed a low prevalence of sexually transmitted infections among these women . The WHO algorithm had 100% sensitivity but a low specificity, while the speculum-based algorithm had a low sensitivity (0-59%) but a higher specificity (79-97%) . Cost analysis indicated that 87% of expenditure was wasted on overtreatment under the WHO algorithm, while only 36% of expenditure was wasted on overtreatment using the speculum-based algorithm . In conclusion, the development of simple, affordable and effective diagnostic tests should be prioritized by policymakers and public health specialists to ensure the provision of adequate services among the higher risk groups in society .

Mol Cell Biol Res Commun, 1999 Jun, 1(3), 167 - 75
Variability in the 28-kDa surface antigen protein multigene locus of isolates of the emerging disease agent Ehrlichia chaffeensis suggests that it plays a role in immune evasion; Reddy GR et al.; Infections caused by rickettsial pathogens persist in vertebrate hosts for long periods of time, despite the active host immune response . We recently described the multigene locus encoding 28 kDa surface antigen proteins (28 kDa SAPs) for two closely related rickettsials, Ehrlichia chaffeensis and Ehrlichia canis (Reddy, G . R., et al . (1998) Biochem . Biophys . Res . Commun . 247, 636-643), that share extensive structural homology to antigenic variant surface protein genes of Neisseria and Borrelia species . In this study, we describe motifs for recombinase binding sites and a high frequency of repeat elements in the cloned 28 kDa SAP genes . The locus for two newly established E . chaffeensis isolates also was characterized, and immunological specificity of the 28 kDa SAPs was investigated . The study identified variant forms of the 28 kDa SAPs and extensive restriction fragment length polymorphisms among isolates . The molecular data suggest that the locus is influenced by short-term evolutionary changes such as genetic recombinations leading to the generation of antigenic variants . Antigenic variation could contribute to the mechanism of immune evasion and the emergence of new diseases.

Gene, 1999 Nov 15, 240(1), 175 - 82
Cell to cell transmission of donor DNA overcomes differential incorporation of non-homologous and homologous markers in Neisseria gonorrhoeae; Hill SA; The neisseriae are naturally competent for DNA transformation . This genetic study examines whether the modification status of chromosomal donor DNA affects transformation of Neisseria gonorrhoeae to drug resistance . When a single modification system was inactivated, unmodified chromosomal donor DNA was not restricted when used to transform the cognate restriction+ host, irrespective of whether the donor DNA carried a point mutation (homologous marker) or a drug-resistance gene cassette (non-homologous marker) . These observations contrasted transformations performed with unmodified plasmid donor DNAs, where the incoming DNA was excluded . However, during the study, it became apparent that certain strains of gonococci showed differential incorporation of non-homologous markers when compared with the incorporation of the homologous marker, even when the donor DNAs were prepared from parental strains . Differential incorporation of markers could be rescued either through cell to cell transmission of donor DNA, or by performing in vitro transformations with donor DNA preparations that were obtained from spent culture supernatants . Overall, the data indicate that, in addition to the exclusion of foreign DNA through the requirement for a genus-specific uptake sequence, gonococci appear capable of excluding DNA on the basis of homology.

Int J STD AIDS, 1999 Nov, 10(11), 741 - 3
An outreach programme for sexually transmitted infection screening in street sex workers using self-administered samples; Morton AN et al.; Street sex workers represent an at-risk group of individuals who find it difficult to access mainstream health services . This was a cross-sectional study of street sex workers in Melbourne, Australia using a self-administered method to detect chlamydial, gonorrhoea and trichomonas infections . Of the 81 individuals approached, 63 (78%) (95% CI: 67-86%) agreed to participate . Overall, 87% of the participants obtained their results . Of the 63 participants, 53 (84%) had a past history of injecting drug use (95% CI: 73-92%), and 21 (33%) had a history of a sexually transmitted infection (STI) (95% CI: 22.0-46.3%) . Neisseria gonorrhoeae was identified in 7 (11%) participants, Trichomonas vaginalis in 7 (11%), Chlamydia trachomatis in 1 (1.6%) . None of the 19 (30%) participants who had been screened for an STI in the preceding 3 months were infected . Our results demonstrated that this method of testing for STIs was acceptable to the street sex workers, and demonstrated a disturbingly high proportion with infections.

J Clin Pathol, 1999 Jun, 52(6), 411 - 4
Evaluation of an in-house polymerase chain reaction for detection of Neisseria gonorrhoeae in urogenital samples; Roymans R et al.; AIM: To develop and evaluate a one day in-house polymerase chain reaction (PCR) assay for the detection of Neisseria gonorrhoeae DNA in urogenital samples . METHODS: 429 urogenital specimens were tested for the presence of N gonorrhoeae by in-house PCR and by Gen-Probe . The PCR assay amplifies target sequences within the N gonorrhoeae cppB gene on the 4.2 kb cryptic plasmid, after which amplicons are detected by a streptavidinbiotin based enzyme immunoassay using an internal probe . Discordant specimens were further evaluated by repeating the PCR and the Gen-Probe assay, and by an additional PCR using another set of 16S primers followed by radioactive detection of amplicons on a Southern blot . RESULTS: Of the 429 samples tested, 15 were found positive by in-house PCR, eight of which were confirmed by Gen-Probe . Of the seven discrepant samples, five were confirmed by 16S PCR and are also considered true positive . The remaining two samples were positive in the in-house PCR only, and are considered false positive . After resolution of discrepant samples, the sensitivities of the N gonorrhoeae assays were 100% and 61.5% for the in-house PCR and Gen-Probe, respectively, while specificities were comparable at 99.5% and 100% . CONCLUSIONS: The in-house PCR for the detection of N gonorrhoeae DNA is at least comparable to Gen-Probe in performance . An extended evaluation period should elucidate if the additional five GO-PCR positive specimens, confirmed by 16S PCR, are caused by persistence of DNA or whether the method is indeed more sensitive.

J Immunol, 1999 Nov 15, 163(10), 5566 - 73
An immunodominant conserved region within the variable domain of VlsE, the variable surface antigen of Borrelia burgdorferi; Liang FT et al.; Antigenic variation is an effective strategy evolved by pathogenic microbes to avoid immune destruction . Variable Ags such as the variable major protein of Borrelia hermsii, the variant surface glycoprotein of African trypanosomes, and the pilin of Neisseria gonorrhoeae include an immunodominant variable domain and one or more invariable domains that are not antigenic . Short, nonantigenic, invariable regions also may be present within the variable domain . VlsE (variable major protein-like sequence, expressed), the variable surface Ag of Borrelia burgdorferi, the Lyme disease spirochete, also contains both variable and invariable domains . In addition, interspersed within the VlsE variable domain there are six invariable regions (IR1-6) that together amount to half of this portion's primary structure . We show here that these IRs are conserved among strains and genospecies of the B . burgdorferi sensu lato complex . Surprisingly, unlike the invariable regions of variable major protein, variant surface glycoprotein, and pilin, which are not antigenic in natural infections, the most conserved of the IRs, IR6, is immunodominant in Lyme disease patients and in monkeys infected with B . burgdorferi . IR6 is exposed on the surface of VlsE, as assessed by immunoprecipitation experiments, but is inaccessible to Ab on the spirochete's outer membrane, as demonstrated by immunofluorescence and in vitro killing assays . VlsE thus significantly departs from the antigenic variation paradigm, whereby immunodominance is only manifest in variable portions . We submit that IR6 may act as a decoy epitope(s) and contribute to divert the Ab response from other, perhaps protective regions of VlsE.

Mol Microbiol, 1999 Nov, 34(3), 538 - 51
Critical determinants of host receptor targeting by Neisseria meningitidis and Neisseria gonorrhoeae: identification of Opa adhesiotopes on the N-domain of CD66 molecules; Virji M et al.; The human pathogens Neisseria meningitidis and Neisseria gonorrhoeae express a family of variable outer membrane opacity-associated (Opa) proteins that recognize multiple human cell surface receptors . Most Opa proteins target the highly conserved N-terminal domain of the CD66 family of adhesion molecules, although a few also interact with heparan sulphate proteoglycans . In this study, we observed that at least two Opa proteins of a N . meningitidis strain C751 have the dual capacity to interact with both receptors . In addition, all three Opa proteins of C751 bind equally well to HeLa cells transfected with cDNA encoding the carcinoembryonic antigen {CEA (CD66e)} subgroup of the CD66 family, but show distinct tropism for CGM1- (CD66d) and NCA (CD66c)-expressing cells . Because the C751 Opa proteins make up distinct structures via the surface-exposed hypervariable domains (HV-1 and HV-2), these combinations appear to be involved in tropism for the distinct CD66 subgroups . To define the determinants of receptor recognition, we used mutant proteins of biliary glycoprotein {BGP (CD66a)} carrying substitutions at several predicted exposed sites in the N-domain and compared their interactions with several Opa proteins of both N . meningitidis and N . gonorrhoeae . The observations applied to the molecular model of the BGP N-domain that we constructed show that the binding of all Opa proteins tested occurs at the non-glycosylated (CFG) face of the molecule and, in general, appears to require Tyr-34 and Ile-91 . Further, efficient interaction of distinct Opa proteins depends on different non-adjacent amino acids . In the three-dimensional model, these residues lie in close proximity to Tyr-34 and Ile-91 at the CFG face, making continuous binding domains (adhesiotopes) . The epitope of the monoclonal antibody YTH71.3 that inhibits Opa/CD66 interactions was also identified within the Opa adhesiotopes on the N-domain . These studies define the molecular basis that directs the Opa specificity for the CD66 family and the rationale for tropism of the Opa proteins for the CD66 subgroups.

Obstet Gynecol, 1999 Nov, 94(5 Pt 1), 653 - 8
Seroprevalence of human papillomavirus type 16 in pregnant women; Hagensee ME et al.; OBJECTIVE: To determine the seroprevalence of and risk factors for human papillomavirus (HPV) type 16 capsid antibodies in a large cohort of pregnant women . METHODS: Antibodies against in vitro produced HPV-16 capsids were measured in stored sera from 2597 pregnant women enrolled from 1984 through 1989 in the Vaginal Infection and Prematurity Study, New Orleans site . RESULTS: Women in this study were primarily black (83.4%) with a mean age of 23.4 years (standard deviation {SD}, 5.1), mean number of sexual partners in lifetime was 3.3 (SD, 6.6), and the mean age at sexual debut was 16.7 years (SD, 2.2) . Overall, 28.0% (n = 727) of these women were positive for HPV-16 capsid antibodies . In bivariate analysis, the presence of antibodies against HPV-16 was correlated with numerous demographic characteristics as well as history of various sexually transmitted diseases . However, neither current cervical or vaginal infection nor adverse obstetric outcome was associated with increased detection of HPV-16 antibodies . In multivariate logistic regression analysis, factors predictive of HPV-16 seropositivity were: more than five lifetime sexual partners (odds ratio {OR}, 1.80; 95% confidence interval {CI}, 1.28, 2.52), 6 or more years of sexual activity (OR, 1.84; 95% CI, 1.22, 2.78), level of education (OR, 1.26; 95% CI, 1.03, 1.55), and history of Neisseria gonorrhoeae infection (OR, 1.53; 95% CI, 1.20, 1.96) . CONCLUSION: HPV-16 seropositivity correlates with measures of sexual activity, confirming its role as a sexually transmitted disease, and its prevalence is similar to that in nonpregnant populations . HPV-16 seropositivity does not predict an adverse obstetric outcome.

Rapid Commun Mass Spectrom, 1999, 13(22), 2252 - 9
252Cf-plasma desorption mass spectrometry of unmodified lipid A: fragmentation patterns and localization of fatty acids; Karibian D et al.; The fragmentation patterns of synthetic Escherichia coli-type lipid A in plasma desorption mass spectrometry (PDMS) in both negative- and positive-ion modes were determined . Negative-ion spectra gave signals for the main diphosphorylated (intact) molecular species in their native proportions . Intact and alkaline-treated lipid A in this mode gave, for the glucosamine I moiety, easily identified signals that have not been previously reported in PDMS . These spectra gave enough information to localize the fatty acids . The procedure was verified with relatively homogeneous lipids A prepared from Salmonella minnesota R595 and Neisseria meningitidis lipopolysaccharides, and then applied to the previously unstudied Yersinia entercolitica O:11,24 lipid A to obtain the localization of its fatty acids . The possibility of obtaining this much information from two negative-ion spectra was attributed to the method, described earlier, of preparing the samples . In the positive-ion mode, about half of the E . coli ions containing diglucosamine appeared as monodephosphorylated species and/or as Na adducts . The intact glucosamine II moiety and its fragment ions gave signals none of which were Na adducts . With lipids A prepared from S . minnesota, N . meningitidis, and Y . enterocolitica, similar fragmentation patterns were observed . For lipid A structure determination, the positive-ion mode could play a confirmatory role . The above results and some of those reported by others were compared .

Antimicrob Agents Chemother, 1999 Nov, 43(11), 2790 - 2
In vitro susceptibilities of Neisseria gonorrhoeae to fatty acids and monoglycerides; Bergsson G et al.; The susceptibility of Neisseria gonorrhoeae to several medium-chain fatty acids and their 1-monoglycerides was tested at a short inactivation time of 1 min . The results indicate that monocaprin, a monoglyceride of capric acid (10 carbon atoms, no double bonds), causes the fastest and most effective killing of all strains of N . gonorrhoeae tested.

Microbiology, 1999 Oct, 145 ( Pt 10), 2967 - 75
Neisseria gonorrhoeae bacterioferritin: structural heterogeneity, involvement in iron storage and protection against oxidative stress; Chen CY et al.; The iron-storage protein bacterioferritin (Bfr) from Neisseria gonorrhoeae strain F62 was identified in cell-free extracts and subsequently purified by column chromatography . Gonococcal Bfr had an estimated molecular mass of 400 kDa by gel filtration; however, analysis by SDS-PAGE revealed that it was composed of 18 kDa (BfrA) and 22 kDa (BfrB) subunits . DNA encoding BfrB was amplified by PCR using degenerate primers derived from the N-terminal amino acid sequence of BfrB and from a C-terminal amino acid sequence of Escherichia coli Bfr . The DNA sequence of bfrA was subsequently obtained by genome walking using single-specific-primer PCR . The two Bfr genes were located in tandem with an intervening gap of 27 bp . A potential Fur-binding sequence (12 of 19 bp identical to the consensus neisserial fur sequence) was located within the 5' flanking region of bfrA in front of a putative -35 hexamer . The homology between the DNA sequences of bfrA and bfrB was 55.7%; the deduced amino acid sequences of BfrA (154 residues) and BfrB (157 residues) showed 39.7% identity, and showed 41.3% and 56.1% identity, respectively, to E . coli Bfr . Expression of recombinant BfrA and BfrB in E . coli strain DH5alpha was detected on Western blots probed with polyclonal anti-E . coli Bfr antiserum . Most Bfrs are homopolymers with identical subunits; however, the evidence presented here suggests that gonococcal Bfr was composed of two similar but not identical subunits, both of which appear to be required for the formation of a functional Bfr . A Bfr-deficient mutant was constructed by inserting the omega fragment into the BfrB gene . The growth of the BfrB-deficient mutant in complex medium was reduced under iron-limited conditions . The BfrB-deficient mutant was also more sensitive to killing by H2O2 and paraquat than the isogenic parent strain . These results demonstrate that gonococcal Bfr plays an important role in iron storage and protection from iron-mediated oxidative stress.

J Med Virol, 1999 Dec, 59(4), 469 - 73
Cervical shedding of cytomegalovirus in human immunodeficiency virus type 1-infected women; Mostad SB et al.; Cervical shedding of cytomegalovirus (CMV) is important in transmission of CMV to exposed sexual partners and neonates . We evaluated prevalence and correlates of CMV DNA shedding in cervical secretions from a large cohort of HIV-1-seropositive women . Using polymerase chain reaction (PCR) assays, CMV DNA was detected in 183 (59%) cervical swab samples from 311 women . Cervical shedding of CMV DNA was significantly associated with shedding of HIV-1 DNA (odds ratio 1.8; 95% confidence interval 1.1-2.8) . CMV shedding was also more frequent in women with Neisseria gonorrhoeae and Trichomonas vaginalis infections, but these associations were not statistically significant . Cervical shedding of CMV in HIV-1-infected women is very frequent and may reflect higher risk of transmission to sexual partners and neonates than previously appreciated .

Sex Transm Dis, 1999 Oct, 26(9), 531 - 6
Double-blind comparison of trovafloxacin and doxycycline in the treatment of uncomplicated Chlamydial urethritis and cervicitis . Trovafloxacin Chlamydial Urethritis/Cervicitis Study Group; McCormack WM et al.; BACKGROUND: Chlamydia trachomatis is among the most common sexually transmitted bacteria worldwide . With excellent activity against C . trachomatis and Neisseria gonorrhoeae and prolonged elimination half-life allowing once-daily dosage, the fluoroquinolone trovafloxacin has potential advantages in the treatment of uncomplicated chlamydial infection . GOAL OF THIS STUDY: This study compared the efficacy of trovafloxacin with that of doxycycline for the treatment of uncomplicated chlamydial infection . STUDY DESIGN: In a double-blind, multicenter trial, trovafloxacin 200 mg was administered once daily for 5 days and doxycycline 100 mg was administered twice daily for 7 days to patients with uncomplicated chlamydial urethritis or cervicitis . Follow-up visits were conducted 10, 21, and 35 days after enrollment . RESULTS: Of the 970 patients (403 men, 567 women) observed, 511 were microbiologically evaluable and 360 were clinically evaluable . C . trachomatis eradication rates in the trovafloxacin and doxycycline groups were equivalent in women (95% and 97%, respectively), but not in men (89% and 99%) . Similarly, rates of clinical success (cure plus improvement) demonstrated equivalence of trovafloxacin and doxycycline in women (96% and 94%), but not in men (94% and 100%) . The most frequent treatment-related adverse events were dizziness, nausea, and headache in patients given trovafloxacin, and nausea, vomiting, and headache in patients given doxycycline . Treatment-related discontinuations were comparable between the drug groups . CONCLUSION: Trovafloxacin given once daily for 5 days was clinically and bacteriologically equivalent to doxycycline given twice daily for 7 days in women with uncomplicated chlamydial cervicitis . This equivalence was not demonstrated in men with uncomplicated chlamydial urethritis.

J AAPOS . 1999 Oct;3(5):316.
An uncommon cause of ophthalmia neonatorum: Neisseria meningitidis; Lehman SS; Ophthalmia neonatorum is defined as conjunctivitis appearing during the first month of life . The differential diagnosis includes chemical, bacterial, viral, and other pathogens, including Neisseria gonorrhoeae, herpes simplex, and Chlamydia trachomatis . Neisseria meningitidis is not commonly specifically included in the differential.

Infect Immun, 1999 Nov, 67(11), 6119 - 29
Identification of regions of the chromosome of Neisseria meningitidis and Neisseria gonorrhoeae which are specific to the pathogenic Neisseria species; Perrin A et al.; Neisseria meningitidis and Neisseria gonorrhoeae give rise to dramatically different diseases . Their interactions with the host, however, do share common characteristics: they are both human pathogens which do not survive in the environment and which colonize and invade mucosa at their port of entry . It is therefore likely that they have common properties that might not be found in nonpathogenic bacteria belonging to the same genetically related group, such as Neisseria lactamica . Their common properties may be determined by chromosomal regions found only in the pathogenic Neisseria species . To address this issue, we used a previously described technique (C . R . Tinsley and X . Nassif, Proc . Natl . Acad . Sci . USA 93:11109-11114, 1996) to identify sequences of DNA specific for pathogenic neisseriae and not found in N . lactamica . Sequences present in N . lactamica were physically subtracted from the N . meningitidis Z2491 sequence and also from the N . gonorrhoeae FA1090 sequence . The clones obtained from each subtraction were tested by Southern blotting for their reactivity with the three species, and only those which reacted with both N . meningitidis and N . gonorrhoeae (i.e., not specific to either one of the pathogens) were further investigated . In a first step, these clones were mapped onto the chromosomes of both N . meningitidis and N . gonorrhoeae . The majority of the clones were arranged in clusters extending up to 10 kb, suggesting the presence of chromosomal regions common to N . meningitidis and N . gonorrhoeae which distinguish these pathogens from the commensal N . lactamica . The sequences surrounding these clones were determined from the N . meningitidis genome-sequencing project . Several clones corresponded to previously described factors required for colonization and survival at the port of entry, such as immunoglobulin A protease and PilC . Others were homologous to virulence-associated proteins in other bacteria, demonstrating that the subtractive clones are capable of pinpointing chromosomal regions shared by N . meningitidis and N . gonorrhoeae which are involved in common aspects of the host interaction of both pathogens.

Infect Immun, 1999 Nov, 67(11), 5699 - 708
Experimental gonococcal genital tract infection and opacity protein expression in estradiol-treated mice; Jerse AE; The development of effective prophylactic agents against gonorrhea and the study of adaptation by Neisseria gonorrhoeae to the urogenital mucosa are hindered by the lack of a well-established animal model of gonococcal genital tract infection . Here, a murine model of long-term gonococcal genital tract infection is described . Female BALB/c mice were treated with 17-beta-estradiol and inoculated intravaginally with wild-type gonococcal strain FA1090 or MS11 . N . gonorrhoeae was recovered from vaginal swabs for an average of 12 to 13 days following inoculation with 10(6) CFU of either strain . Inflammation occurred in over 80% of infected mice, and diplococci were associated with epithelial cells and neutrophils in stained vaginal smears . Ascended infection occurred in 17 to 20% of mice inoculated with strain FA1090 . An outbred mouse strain (SLC:ddY) previously reported to be naturally susceptible to N . gonorrhoeae was also tested; however, as with BALB/c mice, estradiol was required for prolonged infection . Although piliation was not maintained during experimental murine infection, 46 to 100% of vaginal isolates from four of eight BALB/c mice and three of four SLC:ddY mice expressed one or more opacity (Opa) proteins within 4 days after inoculation with an Opa-negative variant of strain FA1090 . The observed selection for and/or induction of gonococcal Opa protein expression during murine infection appears to parallel events that occur during experimental urethritis in volunteers.

Pediatr Infect Dis J, 1999 Oct, 18(10 Suppl), S42 - 9
Bacterial and viral etiology of severe infection in children less than three months old in the highlands of Papua New Guinea; Lehmann D et al.; OBJECTIVE: Determine the bacterial and viral etiology of severe infection in young Papua New Guinean infants as part of a multicenter study in four developing countries aimed at improving case management guidelines . METHODS: Between March, 1991, and April, 1993, children aged <3 months were recruited at the outpatient department of Goroka Base Hospital, Papua New Guinea (PNG) . Children with pre-defined inclusion criteria were enrolled, a history was taken and clinical examination was performed . Blood and urine were collected from children with signs suggestive of severe disease together with eye, umbilical and pernasal swabs as appropriate . Nasopharyngeal aspirates (NPAs) were collected from children with and without signs of severe disease for identification of viruses and Chlamydia trachomatis by direct fluorescent antibody staining . RESULTS: 3280 infants were triaged and 2168 enrolled, among whom 968 had signs suggestive of severe disease . Group A Streptococcus (Streptococcus pyogenes) and Staphylococcus aureus were the most important bacterial pathogens isolated from children < 1 month old with severe infections, and Streptococcus pneumoniae, S . pyogenes and Staphylococcus aureus were most important in older children . Of 292 eye swabs 19 (7%) grew Neisseria gonorrhoeae . Of 116 umbilical swabs 51 (44%) grew S . pyogenes and 45 (39%) grew Staphylococcus aureus . Respiratory syncytial virus was the most important viral cause of acute lower respiratory infection . CONCLUSIONS: S . pyogenes, S . pneumoniae and Staphylococcus aureus are important causes of severe infection in young children in the PNG highlands . It is necessary to improve access to clean water, promote hand-washing in the hospital and at home and investigate further the use of maternal immunization for the prevention of severe disease in young infants.

Int J Antimicrob Agents, 1999 Aug, 12 Suppl 2, S45 - 58
Fusidic acid in vitro activity; Collignon P et al.; Fusidic acid is a narrow spectrum agent that acts to inhibit protein synthesis by inhibition of elongation factor G at the level of the ribosome . Because of high protein binding susceptibility testing in vitro is affected by the presence of blood or serum . In addition, there is a modest inoculum effect in vitro . A breakpoint of 1 or 2 mg/l is most widely used for defining resistance to systemic treatment with fusidic acid . Fusidic acid activity is principally directed at staphylococci, both Staphylococcus aureus and coagulate-negative species which are highly susceptible . It is also active against Gram-positive anaerobic activity, and shows in vitro activity against Neisseria spp., Bordetella pertussis and Moraxella catarrhalis . It has no activity against other aerobic Gram-negative species . Modest activity (MICs just above breakpoint values) is seen with Streptococcus and Enterococcus spp . as well as Gram-negative anaerobic bacteria . Fusidic acid is defined as bacteriostatic . For staphylococci MBC values are generally 8--32-fold that of the MIC . Interaction studies with other antibiotics give varying results depending on methodology . However, interaction with beta-lactams is generally indifferent, as it is with rifampicin, while aminoglycosides and macrolides appear to be synergistic and fluoroquinolones antagonistic . Fusidic acid appears to inhibit the function of neutrophils and T-lymphocytes at clinically achieved concentrations.

J Mol Neurosci, 1999 Apr, 12(2), 81 - 7
Intrathecal synthesis of immunoglobulins in Neisseria meningitidis and echovirus 6 meningoencephalitis; Dorta Contreras AJ; Infectious diseases of the central nervous system (CNS) constitute the main cause of death amidst the neurological diseases in pediatric patients in Cuba . This factor, among others, made us study the immune response in children with such disorders . Cerebrospinal fluid (CSF) and serum samples were collected from 69 patients: 26 with Neisseria meningitidis meningoencephalitis, and 43 with echovirus 6 . IgA, IgM, IgG, and albumin were measured by immunodiffusion in serum and CSF . Local synthesis of immunoglobulins was calculated using Reiber's formula . The detection of intrathecal synthesis of IgG represents an important event in the majority of cases . Detection depends on the acuity of the process, the nature of the biologic agent and its virulence, and the immunological status of the patient . Local IgG synthesis is a very frequent finding in Neisseria meningoencephalitis and echovirus 6 meningoencephalitis . Intrathecally synthesized IgA in patients suffering from N . meningitidis meningoencephalitis is an important indication.

Clin Infect Dis, 1999 Nov, 29(5), 1145 - 50
Prevalence of lower genital tract infections among human immunodeficiency virus (HIV)-seropositive and high-risk HIV-seronegative women . HIV Epidemiology Research Study Group; Cu-Uvin S et al.; This study was undertaken to assess whether the prevalence of lower genital tract infections among human immunodeficiency virus (HIV)-seropositive women was higher than among high-risk HIV-seronegative women at their baseline visit for the HIV Epidemiology Research Study . Results were available for 851 HIV-seropositive and 434 HIV-seronegative women . Human papilloma virus (HPV) infection was more prevalent among HIV-seropositive women (64% vs . 28%) . Bacterial vaginosis was common (35% vs . 33%), followed by trichomoniasis (12% vs . 10%), syphilis (8% vs . 6%), Chlamydia trachomatis infection (4% vs . 5%), candidal vaginitis (3% vs . 2%), and Neisseria gonorrhoeae infection (0.8% vs . 0.3%) . Alcohol use (odds ratio {OR}, 1.8; 95% confidence interval {CI}, 1 . 3-2.4) and smoking (OR, 1.8; 95% CI, 1.3-2.5) were associated with bacterial vaginosis . Bacterial vaginosis (OR, 2.3; 95% CI, 1.5-3.4), trichomoniasis (OR, 2.3; 95% CI, 1.1-4.7), and syphilis (OR, 3.1; 95% CI, 1.3-7.4) were found to be more prevalent among black women . Our study showed no statistically significant difference in the prevalence of lower genital tract infections except for HPV between HIV-infected and demographically and behaviorally similar HIV-uninfected high-risk women.

J Clin Microbiol, 1999 Nov, 37(11), 3668 - 71
Ability of the digene hybrid capture II test to identify Chlamydia trachomatis and Neisseria gonorrhoeae in cervical specimens; Schachter J et al.; The Digene Hybrid Capture II (HCII CT/GC) test is a combination test designed to detect Chlamydia trachomatis and Neisseria gonorrhoeae in a single specimen . It is a nucleic acid hybridization test which uses signal amplification to increase sensitivity . We compared its performance to that of culture on cervical specimens from 1,370 women . Direct fluorescent-antibody assay was used to resolve discrepant results for C . trachomatis . Samples were collected with a proprietary cervical brush or with endocervical swabs . The HCII CT/GC test proved to be sensitive and specific in detecting these organisms . Compared to N . gonorrhoeae culture, it had a sensitivity of 93% (87/94) and a specificity of 98.5% (1,244/1,263) . Compared to C . trachomatis culture, the sensitivity was 97.7% (129/132) and specificity was 98.2% (1,216/1,238) . Testing of some specimens with discrepant results by PCR suggested that the test would actually prove to be even more specific if it were compared to a nucleic acid amplification test (NAAT) . The sensitivity of C . trachomatis culture was somewhat less, at 88.6% (117/132) . The endocervical brush appeared to be better than Dacron swabs for collecting specimens . The HCII CT/GC test offers an attractive format that allows simultaneous detection of C . trachomatis and N . gonorrhoeae with a single specimen . An initial positive result is followed by repeat tests with probes to identify chlamydiae or gonococci . This test is more sensitive than C . trachomatis culture and is at least as sensitive as culture for gonococci . It deserves further evaluation and comparison with NAATs and may well offer an attractive alternative for diagnosis and screening of these infections.

J Clin Microbiol, 1999 Nov, 37(11), 3583 - 5
Comparison of direct inoculation and Copan transport systems for isolation of Neisseria gonorrhoeae from endocervical specimens; Olsen CC et al.; Two commercial swab transport systems, Copan Amies gel agar with and without charcoal (Copan Diagnostics, Corona, Calif.), were compared to direct inoculation onto modified Thayer-Martin medium for detection of Neisseria gonorrhoeae in 1,490 endocervical specimens obtained from women attending a sexually transmitted disease clinic . Copan swabs were held in the transport system for 24 h at room temperature prior to inoculation onto modified Thayer-Martin medium . All cultures were incubated at 35 degrees C in 5% CO(2), and bacteria were identified on the basis of Gram stain, oxidase, and biochemical reactions . Copan Amies gel agar transport system without charcoal detected 77 of 81 (95%) direct inoculation culture-positive specimens, and Copan Amies gel agar transport system with charcoal detected 53 of 56 (95%) directly inoculated culture-positive specimens . Copan Amies gel agar without charcoal inoculated after 6 h supported growth of 56 (98%) positive cultures out of only 55 directly inoculated culture-positive specimens . This study demonstrates that Copan swabs represent a reasonable alternative, providing convenience, low cost, and ease of use while still maintaining a satisfactory recovery rate of N . gonorrhoeae from clinical specimens, if specimens can be inoculated onto selective media within a relatively short time period not involving overnight shipment.

DNA Seq, 1998, 9(5-6), 341 - 5
Nucleotide sequence of a three gene cluster in Neisseria Gonorrhoeae encoding ribosomal proteins S6, S18, and L9; Ropp PA et al.; A cluster of three genes, rpsF, rpsR, and rpII, encoding the ribosomal proteins S6, S18, and L9, respectively, were cloned and sequenced from Neisseria gonorrhoeae . The order of the genes within the cluster was established as rpsF-rpsR-rpII . Within this cluster an additional open reading frame of unknown identity spanning 108 bp was found between rpsF and rpsR . The putative amino acid sequences deduced from all three genes show a high degree of homology to other bacterial ribosomal proteins.

J Exp Med, 1999 Oct 18, 190(8), 1049 - 58
Immunoglobulin A1 protease, an exoenzyme of pathogenic Neisseriae, is a potent inducer of proinflammatory cytokines; Lorenzen DR et al.; A characteristic of human pathogenic Neisseriae is the production and secretion of an immunoglobulin (Ig)A1-specific serine protease (IgA1 protease) that cleaves preferentially human IgA1 and other target proteins . Here we show a novel function for native IgA1 protease, i.e., the induction of proinflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, and IL-8 from peripheral blood mononuclear cells . The capacity of IgA1 protease to elicit such cytokine responses in monocytes was enhanced in the presence of T lymphocytes . IgA1 protease did not induce the regulatory cytokine IL-10, which was, however, found in response to lipopolysaccharide and phytohemagglutinin . The immunomodulatory effects caused by IgA1 protease require a native form of the enzyme, and denaturation abolished cytokine induction . However, the proteolytic activity is not required for the cytokine induction by IgA1 protease . Our results indicate that IgA1 protease exhibits important immunostimulatory properties and may contribute substantially to the pathogenesis of neisserial infections by inducing large amounts of TNF-alpha and other proinflammatory cytokines . In particular, IgA1 protease may represent a key virulence determinant of bacterial meningitis.

Glycobiology, 1999 Oct, 9(10), 1061 - 71
High-level expression of the Neisseria meningitidis lgtA gene in Escherichia coli and characterization of the encoded N-acetylglucosaminyltransferase as a useful catalyst in the synthesis of GlcNAc beta 1-->3Gal and GalNAc beta 1-->3Gal linkages; Blixt O et al.; We have expressed the Neisseria meningitidis lgtA gene at a high level in Escherichia coli . The encoded beta-N-acetylglucosaminyltransferase, referred to as LgtA, which in the bacterium is involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosaccharide, was obtained in an enzymatically highly active form . This glycosyltransferase appeared to be unusual in that it displays a broad acceptor specificity toward both alpha- and beta-galactosides, whether structurally related to N- or O-protein-, or lipid-linked oligosaccharides . Product analysis by one- and two-dimensional 400 MHz 1H- and 13C-NMR spectroscopy reveals that LgtA catalyzes the introduction of GlcNAc from UDP-GlcNAc in a beta 1-->3-linkage to accepting Gal residues . The enzyme can thus be characterized as a UDP-GlcNAc:Gal alpha/beta-R beta 3-N-acetylglucosaminyltransferase . Although lactose is a highly preferred acceptor substrate the recombinant enzyme also acts efficiently on monomeric and dimeric N-acetyllactosamine revealing its potential value in the synthesis of polylactosaminoglycan structures in enzyme assisted procedures . Furthermore, LgtA shows a high donor promiscuity toward UDP-GalNAc, but not toward other UDP-sugars, and can catalyze the introduction of GalNAc in beta 1-->3-linkage to alpha- or beta-Gal in the acceptor structures at moderate rates . LgtA therefore shows promise to be a useful catalyst in the preparative synthesis of both GlcNAc beta 1-->3Gal and GalNAc beta 1-->3Gal linkages.

J Pharm Sci, 1999 Oct, 88(10), 1011 - 5
Development and evaluation of microbicidal hydrogels containing monoglyceride as the active ingredient; Kristmundsdottir T et al.; A number of medium-chain saturated and long-chain unsaturated fatty acids and their monoglycerides were tested against herpes simplex virus (HSV-1) to determine which lipids were most active during a short incubation time . The aim was to find which lipid would be preferable as the active ingredient in a virucidal hydrogel formulation for the purpose of preventing transmission of pathogens to mucosal membranes, particularly sexually transmitted viruses, such as herpes simplex virus and human immunodeficiency virus (HIV), and bacteria, such as Chlamydia trachomatis and Neisseria gonorrheae . The main strategy was that the formulations would be fast-acting, killing large numbers of virus or bacteria on contact in a short time, preferably causing at least a 10000-fold reduction in virus/bacteria titer in 1-5 min . Monocaprin, the 1-monoglyceride of capric acid, and lauric acid were found to be most active of all the lipids tested, causing a greater than 100000-fold reduction in virus titer in 1 min at a concentration of 20 mM . When tested at a concentration of 10 mM for 1 min, monocaprin was still fully active whereas lauric acid had no or negligible activity . It was concluded that monocaprin was most suitable as the active ingredient in a fast-acting virucidal gel formulation, and several hydrogel formulations containing monocaprin were tested . Formulations where the monoglyceride was dissolved in glycofurol were found to be active against HSV-1 . The hydrogel formulations containing 20 mM monocaprin were highly virucidal in vitro and caused a greater than 100000-fold (HSV-1) inactivation of virus in human semen in 1 min . Formulations in dilution 1:10 were cytotoxic in monolayers of CV-1 cells, but they were 10-100 fold less cytotoxic than a commercial product which contains 2% nonoxynol-9.

Antonie Van Leeuwenhoek, 1999 May, 75(4), 369 - 79
Regulation of the lipopolysaccharide-specific sialyltransferase activity of gonococci by the growth state of the bacteria, but not by carbon source, catabolite repression or oxygen supply; Regan T et al.; The enzyme sialyltransferase (STase) of Neisseria gonorrhoeae is a major pathogenicitiy determinant . Using a refined method for assaying the STase activity, the Km for CMP-NANA was shown to be 14 +/- 2 microM, higher than that reported previously . Rates of sialylation by Nonidet extracts, prepared under conditions that optimise solubilisation of the membrane-bound enzyme, were 6 to 20 nmol of NANA transferred from CMP-14C-NANA onto isolated lipopolysaccharide/min./mg of extracted protein, far higher than the previously reported rates of less than 1 nmol of NANA transferred/min./mg of extracted protein . Gonococci grew more slowly with lactate or pyruvate than with glucose as the carbon source . Although growth with a mixture of limiting concentrations of both glucose and lactate was biphasic, diauxic growth was also found in the control culture supplied with glucose alone . The growth rate in the presence of lactate alone was slower than with glucose . The growth rate increased slightly relative to the glucose culture when both substrates were available; lactate was consumed more rapidly than glucose . Higher STase activities were found in bacteria harvested in the exponential than in the stationary phase of aerobic growth: the activity in aerated cultures was higher than those of oxygen-limited or anaerobic cultures . Similar STase activities were found in bacteria that had been grown with glucose, lactate or pyruvate as the carbon and energy source . Sialyltransferase synthesis is essentially constitutive: it is not regulated by glucose repression or by induction by lactate or anaerobiosis.

Mol Microbiol, 1999 Sep, 33(6), 1232 - 43
Probing secretion and translocation of a beta-autotransporter using a reporter single-chain Fv as a cognate passenger domain; Veiga E et al.; The mechanism of protein secretion mediated by the beta-domain of the Neisseria gonorrhoeae IgA protease, a paradigm of a family of secreted polypeptides of Gram-negative bacteria called autotransporters, has been examined using a single-chain antibody (scFv) as a reporter passenger domain to monitor the translocation process . Fusion of a scFv to the beta-module of the IgA protease allowed us to investigate the passage of the chimeric protein through the periplasm, its insertion into the outer membrane and the movement of the N-terminal moiety towards the cell surface . As the binding activity of the scFv to its target antigen is entirely dependent on the formation of disulphide bonds, the relationship between secretion, folding and formation of S-S bridges could be analysed in detail . In contrast to the current notion that only an unfolded N-passenger domain can be translocated through the beta-domain, our results show that the scFv is able to pass through the outer membrane, albeit at a threefold reduced level, in an active conformation with its disulphide bonds preformed in the periplasm through the action of the DsbA product . These data call for a re-evaluation of the prevailing model for secretion of the N-domain of autotransporters.

Antimicrob Agents Chemother, 1999 Oct, 43(10), 2468 - 72
Decreased azithromycin susceptibility of Neisseria gonorrhoeae due to mtrR mutations; Zarantonelli L et al.; Single-dose azithromycin therapy has recently been used in Uruguay for the treatment of uncomplicated gonococcal infections . As part of an active surveillance study to monitor the emergence of antibiotic resistance in gonococcal isolates, we examined the levels of azithromycin susceptibility in 51 consecutive isolates obtained from males with uncomplicated gonococcal urethritis . Isolates with decreased susceptibility to azithromycin (MICs, 0.25 to 0.5 microg/ml) were common, and these isolates often displayed cross-resistance to hydrophobic antimicrobial agents (erythromycin and Triton X-100) . Resistance to erythromycin and Triton X-100 is frequently due to overexpression of the mtrCDE-encoded efflux pump mediated by mutations in the mtrR gene, which encodes a transcriptional repressor that modulates expression of the mtrCDE operon . Accordingly, we questioned whether clinical isolates that express decreased azithromycin susceptibility harbor mtrR mutations . Promoter mutations that would decrease the level of expression of mtrR as well as a missense mutation at codon 45 in the mtrR-coding region that would result in a radical amino acid replacement within the DNA-binding motif of MtrR were found in these strains . When these mutations were transferred into azithromycin-susceptible strain FA19 by transformation, the susceptibility of gonococci to azithromycin was decreased by nearly 10-fold . The mtrCDE-encoded efflux pump system was responsible for this property since insertional inactivation of the mtrC gene resulted in enhanced susceptibility of gonococci to azithromycin . We conclude that the mtrCDE-encoded efflux pump can recognize azithromycin and that the emergence of gonococcal strains with decreased susceptibility to azithromycin can, in part, be explained by mtrR mutations.

Wiad Lek, 1999, 52(5-6), 242 - 5
{The analysis of the incidence of meningitis in children}; Buszman Z et al.; The aim of the study is to present the results of epidemiological investigation in 142 infants and young children who suffered from meningitidis in the years 1987-1995 . The most frequent cause of the disease was bacterial infection . The bacterial factors have been identified in 49% of those cases . Neisseria meningitidis was found to be most common bacteria that was responsible for the disease.

J Bacteriol, 1999 Oct, 181(19), 6133 - 41
Insertion mutations in pilE differentially alter gonococcal pilin antigenic variation; Howell-Adams B et al.; Pilus antigenic variation in Neisseria gonorrhoeae occurs by the high-frequency, unidirectional transfer of DNA sequences from one of several silent pilin loci (pilS) into the expressed pilin gene (pilE), resulting in a change in the primary pilin protein sequence . Previously, we investigated the effects of large or small heterologous insertions in conserved and variable portions of a pilS copy on antigenic variation . We observed differential effects on pilin recombination by the various insertions, and the severity of the defect correlated with the disruption or displacement of a conserved pilin DNA sequence called cys2 . In this study, we show that disruption or displacement of the pilE cys2 sequence by the same insertions or a deletion also affects pilin recombination . However, in contrast to the insertions in pilS, the analogous insertions in pilE impaired, but did not block, recombination of the flanking pilin sequences . These results, the change in the spectrum of donor silent copies used during variation, and our previous results with pilS mutations show that the donor pilS and recipient pilE play different roles in antigenic variation . We conclude that when high-frequency recombination mechanisms are blocked, alternative mechanisms are operative.

J Bacteriol, 1999 Oct, 181(19), 6063 - 72
Use of heme-protein complexes by the Yersinia enterocolitica HemR receptor: histidine residues are essential for receptor function; Bracken CS et al.; The abilities of two bacterial active heme transporters, HmbR of Neisseria meningitidis and HemR of Yersinia enterocolitica, to use different heme sources were compared . While HmbR-expressing cells used only hemoglobin (Hb) and heme, HemR-expressing bacteria were able to grow on Hb, heme, myoglobin, hemopexin, catalase, human and bovine serum albumin-heme, and haptoglobin-hemoglobin complexes as sources of iron . Expression of functional HemR allowed Escherichia coli cells to respond to heme-containing peptides, microperoxidases MP-8, MP-9, and MP-11, suggesting the ability of HemR to transport heme covalently linked to other molecules . Comparison of HemR with other heme receptors identified several highly conserved histidine residues as well as two conserved amino acid motifs, the FRAP and NPNL boxes . A site-directed mutagenesis approach was used to investigate the roles of His128, His192, His352, and His461 residues in HemR function . The HemR receptor with histidine changed to lysine at position 128 (HemR(H128K)), HemR(H461L), HemR(H461A), and HemR(H128A,H461A) mutant receptors were unable to use Hb, human serum albumin-heme, and myoglobin as sources of porphyrin and iron . Utilization of free heme was also severely affected, with some residual heme uptake in cells expressing HemR(H128K), HemR(H461A), and HemR(H461L) . Conversely, the HemR(H192T), HemR(H352A), HemR(H352K), and HemR(H192T,H352K) mutant receptors were fully functional . All mutant HemR proteins were expressed in the outer membrane at levels similar to that of the wild-type HemR receptor . Nonfunctional HemRs were able to bind heme- and Hb-agarose . A hypothetical model of the HemR function in which two conserved histidine residues, His128 and His461, participate in the transport of heme through the receptor pore is postulated.

Commun Dis Intell . 1999 Aug 5;23(8):221.
Neisseria canis infection: a case report; Safton S et al.; The third case report, which is the first in Australia, of human infection with Neisseria canis is documented . This is the first case report in which the pathogenicity of this organism for humans is unequivocally demonstrated.

Infect Immun, 1999 Oct, 67(10), 5490 - 4
Tyrosine phosphatase SHP-1 is involved in CD66-mediated phagocytosis of Opa52-expressing Neisseria gonorrhoeae; Hauck CR et al.; Opa proteins of Neisseria gonorrhoeae bind to CD66 receptors on human phagocytes, thereby inducing efficient uptake of the bacteria in the absence of opsonins . The interaction of Opa proteins and CD66 receptors leads to activation of Src family tyrosine kinases, a process that is of critical importance for the efficient, CD66-mediated internalization . Here we show that during Opa-mediated stimulation of CD66 the activity of the host cell tyrosine phosphatase SHP-1 is strongly downregulated, concomitant with increases in the tyrosine phosphorylation of several cellular proteins . Since the SHP-1 tyrosine phosphorylation level itself is influenced by Opa-induced events, this phosphatase comprises an important regulatory checkpoint of the pathogen-triggered signaling cascade in human phagocytes.

Infect Immun, 1999 Oct, 67(10), 5417 - 26
Conservation and accessibility of an inner core lipopolysaccharide epitope of Neisseria meningitidis; Plested JS et al.; We investigated the conservation and antibody accessibility of inner core epitopes of Neisseria meningitidis lipopolysaccharide (LPS) because of their potential as vaccine candidates . An immunoglobulin G3 murine monoclonal antibody (MAb), designated MAb B5, was obtained by immunizing mice with a galE mutant of N . meningitidis H44/76 (B . 15.P1.7,16 immunotype L3) . We have shown that MAb B5 can bind to the core LPS of wild-type encapsulated MC58 (B.15.P1.7,16 immunotype L3) organisms in vitro and ex vivo . An inner core structure recognized by MAb B5 is conserved and accessible in 26 of 34 (76%) of group B and 78 of 112 (70%) of groups A, C, W, X, Y, and Z strains . N . meningitidis strains which possess this epitope are immunotypes in which phosphoethanolamine (PEtn) is linked to the 3-position of the beta-chain heptose (HepII) of the inner core . In contrast, N . meningitidis strains lacking reactivity with MAb B5 have an alternative core structure in which PEtn is linked to an exocyclic position (i.e., position 6 or 7) of HepII (immunotypes L2, L4, and L6) or is absent (immunotype L5) . We conclude that MAb B5 defines one or more of the major inner core glycoforms of N . meningitidis LPS . These findings support the possibility that immunogens capable of eliciting functional antibodies specific to inner core structures could be the basis of a vaccine against invasive infections caused by N . meningitidis.

Commun Dis Public Health, 1999 Sep, 2(3), 198 - 202
Epidemiology and treatment outcome of infection with antibiotic resistant strains of Neisseria gonorrhoeae in Scotland; Young H et al.; Antibiotic resistance in Neisseria gonorrhoeae occurs worldwide and may limit the efficacy of treatment . This study reports the epidemiology and treatment outcome for 125 episodes of infection with antibiotic resistant gonococci diagnosed in Scotland in 1996 . Infections with penicillinase producing N . gonorrhoeae (PPNG) and/or high level plasmid mediated tetracycline resistant N . gonorrhoeae (TRNG) and isolates showing reduced susceptibility to ciprofloxacin were more likely to belong to an unusual serovar and have been acquired abroad by heterosexual intercourse than isolates showing chromosomal resistance to penicillin or tetracycline, which were more likely to belong to a common serovar and to have been acquired in the United Kingdom by homosexual intercourse . Among the 88 episodes of infection whose outcome was known initial treatment with an antibiotic to which the isolate was resistant failed in nine out of 16 infections and in three out of the 72 infections treated with an antibiotic to which the organism was sensitive . In the case of high level plasmid mediated resistance, treatment with an inappropriate antibiotic always resulted in failure . Continuous monitoring of treatment outcome is essential to guide clinicians in prescribing the most appropriate antibiotic for individual patients.

Sex Transm Dis, 1999 Sep, 26(8), 476 - 82
Predicting Neisseria gonorrhoeae and Chlamydia trachomatis infection using risk scores, physical examination, microscopy, and leukocyte esterase urine dipsticks among asymptomatic women attending a family planning clinic in Kenya; Tyndall MW et al.; BACKGROUND: Sexually transmitted infections (STIs) continue to exert a tremendous health burden on women in developing countries . Poor socioeconomic status, inadequate knowledge, lack of diagnostic facilities, and shortages of effective treatment all contribute to the high incidence of STIs . The use of clinical algorithms for the detection and management of STIs has gained widespread acceptance in settings where there are limited resources . Evaluation of these algorithms have been few, especially in women who are not recognized as members of high-risk groups . OBJECTIVES: To develop a simple scoring system based on historical and demographic data, physical findings, microscopy, and leukocyte esterase (LE) urine dipsticks to predict cervical gonococcal and chlamydial infection among asymptomatic women . METHODS: One thousand and forty-eight women attending an urban family planning clinic in Nairobi were randomly selected to participate . After the identification of factors that were associated with infection, we assigned one point each for: age 25 or younger, single status, two or more sex partners in the past year, cervical discharge, cervical swab leukocytes, and a positive LE urine dipstick . Identification of any one of these six factors gave a sensitivity of 85% and a specificity of 30% for the detection of cervical infections . A positive LE urine dipstick had a sensitivity of 63 % and a specificity of 47% when used alone and did not contribute to the identification of infection if a physical examination was performed . The application of existing clinical algorithms to this population performed poorly . CONCLUSIONS: The use of risk scores, physical examination, microscopy, and the urine LE dipstick, used alone or in combination, as predictors of gonococcal or chlamydial cervical infection was of limited utility in low-risk, asymptomatic women . Accurate diagnostic testing is necessary to optimize treatmentPIP: This cross sectional study presents a risk scoring system that would identify women at highest risk for sexually transmitted infections (STIs) . 1058 randomly selected women participated in the study in Nairobi, Kenya; of these, 1048 participants were included in the analysis . The study was conducted from May 1994 to July 1995 at a clinic sponsored by the Family Planning Association of Kenya . Information pertaining to the demographic, behavioral and social characteristics of the participants was gathered . In addition, a clinical algorithm, which includes physical examination, microscopy, and leukocyte esterase (LE) urine dipsticks, was employed to detect gonorrhea and chlamydia infections among asymptomatic women . The results revealed that the prevalence of STIs, including HIV-1, was high among women attending this urban family planning clinic . Standard demographic, behavioral, and clinical characteristics were only weakly associated with infection, resulting in poor sensitivity and specificity calculations in the risk scores . Detection of cervical infections gave a sensitivity of 85% and a specificity of 30% . A positive LE urine dipstick had a sensitivity of 63% and a specificity of 47% . Although the addition of physical examination and LE dipstick to the work-up improved the sensitivity of case detection, it did not improve the overall validity of the scoring system .

Int J Antimicrob Agents, 1999 Aug, 12(4), 325 - 32
Alterations within the quinolone resistance-determining regions of GyrA and ParC of Neisseria gonorrhoeae isolated in the Far East and the United States; Trees DL et al.; The genetic mutations within the quinolone resistance-determining regions (QRDRs) of gyrA and parC of 234 Neisseria gonorrhoeae strains isolated in the Far East and the United States, which exhibited either clinically significant ciprofloxacin resistance (CipR) or intermediate ciprofloxacin resistance (CipI) were characterized . A number of GyrA/ParC amino acid alteration patterns were identified, the most prevalent alteration pattern among CipR isolates being GyrA-91,95/ParC-Asp-86- > Asn (91,95/Asp-86- > Asn) . Isolates containing 91,95/Asp-86- > Asn belonged to a number of A/S classes, penicillin/tetracycline resistance phenotypes, and plasmid profiles . These results strongly suggest that the continuing emergence of ciprofloxacin-resistant gonococci is not due to the spread of a single or a few strains but to numerous factors such as 'spread of existing strains, importation of new strains and, possibly, de novo development of ciprofloxacin resistance in previously susceptible strains.

Int J STD AIDS, 1999 Sep, 10(9), 609 - 14
Sexually transmitted infections in young pregnant women in Bangui, Central African Republic; Blankhart D et al.; In early 1996, 481 women visiting the antenatal services of the 3 major governmental health centres in the capital city of the Central African Republic (CAR) were included in the study . All study participants underwent the health centre's routine gynaecological examination, including laboratory diagnosis of trichomoniasis, candidiasis, gonorrhoea, syphilis and bacterial vaginosis . Cervical secretions and blood samples from study participants were sent to the National STD Reference Centre for diagnosis of Chlamydia trachomatis, Neisseria gonorrhoeae, Candida albicans, Treponema pallidum, and HIV . Overall, 34% of the study women were diagnosed with at least one sexually transmitted infection (STI) (3.1% N . gonorrhoeae, 6.2% C . trachomatis, 9.9% T . vaginalis, 6.7% T . pallidum, 12.2% HIV-1) . In addition, 29.1% of women were diagnosed with bacterial vaginosis and 46.6% with candidiasis . Only a small proportion of these women had sought treatment during the weeks before, despite the recognition of genital symptoms . Self-reported and health worker-recognized symptoms, signs and laboratory results exhibited only low sensitivities, specificities, and positive predictive values in the diagnosis of STIs . These findings confirm the high vulnerability of young African women to STIs and emphasize the need for specific control interventions which should include affordable and user-friendly services . Moreover, these results call for more effective quality control in case of laboratory-based STI control strategies and question the validity of syndromic STI management strategies in women attending antenatal care services in AfricaPIP: This study reports on the prevalence of sexually transmitted infections (STIs) among pregnant women in Bangui, Central African Republic . A total of 481 pregnant women visiting the antenatal services of the 3 major governmental health centers were included in the study . All these women were interviewed and underwent gynecological examination, including laboratory diagnosis of trichomoniasis, candidiasis, gonorrhea, syphilis and bacterial vaginosis . The results revealed that 34% of the women were diagnosed with at least one STI . Gonorrhea accounted for 3.1%, chlamydiasis for 6.2%, trichomoniasis for 9.9%, syphilis for 6.7%, and HIV-1 for 12.2% . In addition, 29.1% of women were diagnosed with bacterial vaginosis and 46.6% with candidiasis . The majority of the women diagnosed with an STI reported specific clinical symptoms during the week before they entered into the study, but only a minority of them actively sought treatment, and only half of those consulted the formal health sector . Self-reported and health worker-recognized symptoms, signs and laboratory results manifested only low sensitivities, specificities, and positive predictive values in STI diagnosis . These results confirmed the high susceptibility of young African women to STIs . Therefore, specific control interventions should be carried out which incorporate affordable and user-friendly services .

J Bacteriol, 1999 Sep, 181(18), 5591 - 9
Genetic diversity and mosaicism at the por locus of Neisseria gonorrhoeae; Fudyk TC et al.; The por genes of the predominant serovars of Neisseria gonorrhoeae circulating in a high-frequency transmitter core group located in Nairobi, Kenya, were examined for nucleotide sequence polymorphism . The level of por gene diversity did not differ significantly between core group-derived gonococcal strains and gonococcal strains originating elsewhere . However, por mosaicism appeared to be more frequent among core group-derived strains, suggesting that recombination of different por sequences may be a important strategy by which N . gonorrhoeae generates por gene diversity within core group populations . Despite extensive sequence variability, por expressed by gonococcal isolates of different geographic origin exhibited conserved patterns of nucleotide change, suggesting that diversity among por alleles may also be finite.

Commun Dis Intell, 1999 Jul 8, 23(7), 193 - 7
Annual report of the Australian Gonococcal Surveillance Programme, 1998; Emergency department characteristics of male sexual assault; Department of Emergency Medicine, Saint Vincent's Hospital, New York, NY 10011, USAOBJECTIVE: To characterize the nature, frequency, and treatment of male sexual assault encountered by physicians in an ED . A minor objective compared the lengths of time the victim knew the assailant between males and females to determine whether there were differences between male and female victims . METHODS: Retrospective review over four years of all male patients with a diagnosis of sexual assault presenting to an urban academic ED in New York City . Demographics, types of injury, assailant/victim information, and treatment of the assault were obtained . RESULTS: Twenty-seven male sexual assaults (approximately 12% of all sexual assaults) were documented during this time period . Forcible rectal, oral, or both rectal and oral intercourse occurred 14, 4, and 9 times, respectively . Documented physical trauma occurred in nine of 27 visits, with two admissions for head trauma . Five additional patients complained of rectal pain with no independent objective evidence of trauma . Prophylactic treatment with antibiotics for the prevention of Neisseria gonorrhoeae and Chlamydia trachomatis was offered in 26 of 27 episodes and prophylactic HIV protection was given to only two of 21 eligible patients . Documentation of HIV status and HIV counseling occurred only in ten and five visits, respectively . The male-to-female odds ratios for whether the assailant was unknown, known less than 24 hours, or known more than 24 hours were 0.43 (95% CI = 0.15 to 1.26), 2.48 (95% CI = 0.94 to 6.53), and 0.92 (95% CI = 0.31 to 2.71), respectively . CONCLUSIONS: The percentage of total sexual assaults occurring in males who present to an ED is now more than 10% of all sexual assaults seen in this urban ED . These assaults are associated with a high proportion of patients with documented trauma . Although the treatment of traditional sexually transmitted diseases appears to be covered well, the need for HIV documentation, counseling, and possible HIV prophylaxis should be addressed more aggressively . In comparing the lengths of the relationships between the victim and the assailant, it appears twice as likely that the female victim will not know her assailant compared with the male, and twice as likely that the male victim will know his assailant less than 24 hours compared with the female.

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