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 pro