Neisseria

J Am Chem Soc, 2002 Dec 18, 124(50), 14879 - 92
Oxidation of heme to beta- and delta-biliverdin by Pseudomonas aeruginosa heme oxygenase as a consequence of an unusual seating of the heme; Caignan GA et al.; The origin of the unusual regioselectivity of heme oxygenation, i.e . the oxidation of heme to delta-biliverdin (70%) and beta-biliverdin (30%), that is exhibited by heme oxygenase from Pseudomonas aeruginosa (pa-HO) has been studied by (1)H NMR, (13)C NMR, and resonance Raman spectroscopies . Whereas resonance Raman indicates that the heme-iron ligation in pa-HO is homologous to that observed in previously studied alpha-hydroxylating heme oxygenases, the NMR spectroscopic studies suggest that the heme in this enzyme is seated in a manner that is distinct from that observed for all other alpha-hydroxylating heme oxygenase enzymes for which a structure is known . In pa-HO, the heme is rotated in-plane approximately 110 degrees, so the delta-meso-carbon of the major orientational isomer is located within the HO-fold in the place where the alpha-hydroxylating enzymes typically place the alpha-meso-carbon . The unusual heme seating displayed by pa-HO places the heme propionates so that these groups point in the direction of the solvent-exposed heme edge and appears to originate in large part from the absence of stabilizing interactions between the polypeptide and the heme propionates, which are typically found in alpha-hydroxylating heme oxygenase enzymes . These interactions typically involve Lys-16 and Tyr-112, in Neisseriae meningitidis HO, and Lys-16 and Tyr-134, in human and rat HO-1 . The corresponding residues in pa-HO are Asn-19 and Phe-117, respectively . In agreement with this hypothesis, we found that the Asn-19 Lys/Phe-117 Tyr double mutant of pa-HO exists as a mixture of molecules exhibiting two distinct heme seatings; one seating is identical to that exhibited by wild-type pa-HO, whereas the alternative seating is very similar to that typical of alpha-hydroxylating heme oxygenase enzymes and is related to the wild-type seating by approximately 110 degrees in-plane rotation of the heme . Furthermore, each of these heme seatings in the pa-HO double mutant gives rise to a subset of two heme isomeric orientations that are related to each other by 180 degrees rotation about the alpha-gamma-meso-axis . The coexistence of these molecules in solution, in the proportions suggested by the corresponding area under the peaks in the (1)H NMR spectrum, explains the unusual regioselectivity of heme oxygenation observed with the double mutant, which we found produces alpha- (55%), delta- (35%), and beta-biliverdin (10%) . Alpha-biliverdin is obtained by oxidation of the heme seated similar to that of alpha-hydroxylating enzymes, whereas beta- and delta-biliverdin are formed from the oxidation of heme seated as in wild-type pa-HO.

J Cancer Res Clin Oncol, 2002 Dec, 128(12), 669 - 77 Epub 2002 Nov 14.
Role of cell surface GM3 ganglioside and sialic acid in the antitumor activity of a GM3-based vaccine in the murine B16 melanoma model; Gabri MR et al.; PURPOSE: To examine the role of GM3 monosialoganglioside and sialic acid in the antitumor activity of a vaccine based on GM3, hydrophobically conjugated with the outer-membrane-protein complex from Neisseria meningitidis (GM3/VSSP) . METHODS: In order to evaluate the relationship between antitumor activity and the presence of GM3 on the surface of tumor cells, we used two murine tumor cell lines with different ganglioside expression . Syngeneic mice were immunized with four i.m . doses of GM3/VSSP (120 micro g) at 14-day intervals and challenged subcutaneously with tumor cells . RESULTS: B16 melanoma cells showed GM3 on cell surface and GM3-dependent in vitro growth . As expected, preimmunization with the vaccine significantly inhibited tumor formation and prolonged survival in mice challenged with B16 cells . In contrast, no antitumor effect was observed in mice challenged with GM3-negative F3II mammary carcinoma cells . The reactivity of sera from immunized mice against B16 cells was confirmed by flow cytometry and immunoperoxidase staining . Depletion of sialic acid residues from the cell surface completely abolished antibody response against melanoma cells . CONCLUSIONS: These results indicate that the antitumor activity of GM3/VSSP is associated with GM3 expression on tumor cell surface and demonstrate a major role of sialic acid in the humoral response of vaccinated mice.

Sex Transm Infect, 2002 Dec, 78(6), 440 - 4
Mutation patterns in gyrA and parC genes of ciprofloxacin resistant isolates of Neisseria gonorrhoeae from India; Chaudhry U et al.; AIM: To analyse mutations in the gyrA and parC genes leading to possible increase in ciprofloxacin resistance (high MIC values for ciprofloxacin) in clinical isolates of Neisseria gonorrhoeae in Delhi, India . METHOD: MIC of ciprofloxacin for 63 clinical isolates of N gonorrhoeae were examined by the Etest method . Subsequently, gyrA and parC genes of these isolates were amplified and sequenced for possible mutations . RESULTS: Out of the 63 clinical isolates tested, only five (8%) isolates were found to be susceptible to ciprofloxacin (MIC <0.06 micro g/ml) . DNA sequence analysis of the gyrA and the parC genes of all these isolates (n = 63) revealed that all isolates which were not susceptible to ciprofloxacin (n=58) had mutation(s) in gyrA and parC genes . 12 isolates (19%) exhibited high resistance with an MIC for ciprofloxacin of 32 micro g/ml . Two out of these 12 isolates (UD62 and UD63), harboured triple mutations (Ser-91 to Phe, Asp-95 to Asn and Val-120 to Leu) in the gyrA gene . The third mutation of Val-120 to Leu, lies downstream of the quinolone resistance determining region (QRDR) of the gyrA and has not been described before in gonococcus . In addition, both these isolates had a Phe-100 to Tyr substitution in the parC, a hitherto unknown mutation . CONCLUSIONS: Emergence of ciprofloxacin resistance with high levels of MIC values (up to 32 micro g/ml) in India is alarming . Double and triple mutations in gyrA alone or together in gyrA and parC could be responsible for such a high resistance.

Rev Saude Publica, 2002 Oct, 36(5), 545 - 52 Epub 2002 Dec 02.
{Prevalence of associated microorganisms in genital discharge, Argentina}; Di Bartolomeo S et al.; OBJECTIVE: There was a significant increase in the number of women demanding gynecological care in public hospital of the Great Buenos Aires, Argentina, between 1997 and 1998 . It was necessary to update the prevalence of associated microorganisms in order to review the laboratory support and adjust prevention and control guidelines . METHODS: Samples from vaginal and endocervical discharge, from total cases: 84 adolescents (15 to 19 years) and 784 adults (20-60 years) attended in 1997-1998, were studied . Neisseria gonorrhoeae, Streptococcus agalactiae, Trichomonas vaginalis, Candida spp, and bacterial vaginosis, were diagnosed applying direct detection methods and specific culture isolation . Chlamydia trachomatis (antigen detection), Ureaplasma urealyticum and Mycoplasma hominis (culture) were also studied in part of the population . RESULTS: Patient care increased steadily from 1997 and there was an increase of 2.1 times from the first semester to the last one in 1998 . Bacterial vaginosis was the most prevalent disease in the adult group, with 23.8%; followed by Candida spp 17.8%; S . agalactiae 5.6%; T.vaginalis 2.4% . In 50.3% of total adult cases neither bacterial vaginosis or presence of any sought microorganisms, was detected . In the adolescents group the most frequent detection was Candida spp with a 29.7%; bacterial vaginosis in 17.8%; followed by S . agalactiae 3.6%, T.vaginalis 2.4% . Also in this group on an important number of cases, 46.4%, none bacterial vaginosis or the presence of the sought microorganism were found . In some of the adult group, C . trachomatis (7/400) 1.76%, U . urealyticum (209/340) 61.4% and M . hominis (45/272) 16.5% were detected . CONCLUSIONS: Bacterial vaginosis and Candida spp prevalence is important in both groups . The absence of N . gonorrhoeae and lower prevalence of T . vaginalis and C . trachomatis is remarkable . A high prevalence of U . urealyticum and M . hominis were also detected, but the actual pathogenic role in adult women is still under discussion . The significant increase of attendance in the public system is due to social changes in the population and not due to changes in any particular pathology . Laboratory support has to be adjusted to the most cost-effective procedures to solve the most prevalent problems.

Sex Transm Dis, 2002 Dec, 29(12), 840 - 6
Sexually transmitted diseases in Manitoba: evaluation of physician treatment practices, STD drug utilization, and compliance with screening and treatment guidelines; Moses S et al.; BACKGROUND AND GOAL: There is little information in Manitoba on the utilization of drugs for sexually transmitted disease (STD) treatment and the extent to which physicians comply with STD screening and treatment guidelines . This study was undertaken to provide such information to inform policy and program development . METHODS: Physicians providing STD care were asked to complete a simple record for each new STD index client or contact seen . This information was subsequently linked with data from provincial diagnostic and treatment databases . RESULTS: Between October 1997 and September 1998, there were 2535 reports of STD treatments . Only about 25% of drugs provided by the provincial health department to physicians for STD treatment could be accounted for on the basis of the treatment reports received . Seventy-four percent of all treatments provided were presumptive ones for urethral or cervical infection or pelvic inflammatory disease (PID), and 14.4% of these were associated with subsequent positive tests for Chlamydia trachomatis or Neisseria gonorrhoeae . About three quarters of presumptive treatments were in compliance with provincial guidelines, as were most etiology-based treatments for chlamydial infection and 72% of etiology-based treatments for gonorrhea . Noncompliance with guidelines was commonly due to presumptive treatment that covered only C trachomatis, nonrecommended treatment for N gonorrhoeae, and incorrect treatment of PID . Only about 25% of women and 4% of men aged 15 to 24 years who visited a physician in 1997 were tested for C trachomatis . CONCLUSION: Increased educational efforts are required to improve physician compliance with STD screening and treatment guidelines, as well as ensure the appropriate use of STD drugs provided . Opportunities are being missed for screening for C trachomatis among young people, the majority of whom are seen regularly by a physician.

Sex Transm Dis, 2002 Dec, 29(12), 828 - 33
Prevalence of gonococcal and chlamydial infections and sexual risk behavior among youth in Addis Ababa, Ethiopia; Taffa N et al.; BACKGROUND: No community-based study on the magnitude of sexually transmitted diseases (STDs) has ever been conducted among young people in Ethiopia . GOAL: To assess the magnitude of Chlamydia trachomatis and Neisseria gonorrhoeae infections and status of sexual risk behavior among youths (15-24 years old) in Addis Ababa, Ethiopia . STUDY DESIGN: Youths in or out of school residing in two (of the six) administrative zones in Addis Ababa served as the study population . Participants filled out a self-administered questionnaire related to sexuality and its sociocultural determinants . First-void urine (FVU) was analyzed for gonorrhea and chlamydial infection by polymerase chain reaction (PCR) . RESULTS: A total of 561 youths took part in the study . Urine PCR was performed for 522 of them . Nine subjects (1.7%) were found to have and N gonorrhoeae and C trachomatis infections . There were five cases (1.0%) involving each agent . Double infection was noted in one female subject . All but one of the infections were detected among the out-of-school youths (chi-square = 4.5; < 0.05) . None of these subjects complained of symptoms suggestive of an active STD . One-third (188/561) reported having had sexual intercourse . The prevalence among sexually active youths was thus 4.8% (9/188) for both infections combined (2.7% for each agent) . While 7/52 (13.5%) of the sexually active females were found to also have STDs, only 2/136 (1.5%) of the males had an STD (chi-square = 8.0; < 0.01) . Report of sexual activity was significantly associated with being male, an age of >/=20 years, out-of-school status, and report of alcohol/khat (amphetamine-like substance) consumption . Females reported less condom use, whether they were in or out of school and independent of age . CONCLUSIONS: Out-of-school youths, especially females, took more sexual risk and were exceedingly susceptible to STDs . This calls for alternative group-targeted strategies for sex education, disease prevention, and STD screening and management.

Sex Transm Dis, 2002 Dec, 29(12), 792 - 7
Detection of a novel Tet M determinant in tetracycline-resistant Neisseria gonorrhoeae from Uruguay, 1996-1999; Marquez CM et al.; BACKGROUND: Determination of the diversity within the tet(M) sequence from N gonorrhoeae is a useful epidemiologic tool for monitoring the movement or importation of strains within a geographic region . Only two distinct tet(M) genes in clinical gonococcal isolates have been described up to now: the Dutch and the American types . GOAL: The study involved surveillance of the tet(M) gene types in high-level-tetracycline-resistant gonococcal isolates from Uruguay during the period 1996 to 1999 . STUDY DESIGN: Among 181 gonococcal isolates, those showing MICs >/=16 microg/ml to tetracycline were analyzed for detection and characterization of the tet(M) gene by a polymerase chain reaction (PCR) and further HpaII restriction fragment polymorphism methods, respectively . The plasmid content and antibiogram were determined . RESULTS: Twenty-two of 181 isolates (12%) exhibited high levels of resistance to tetracycline (MICs >/=16 microg/ml) and harbored a putative 25.2-Mda plasmid that contained the tet(M) gene . A high percentage of isolates (95%; 21/22) presented the Dutch type tet(M) gene . One isolate from 1999 revealed a new restriction pattern . Such a pattern had been previously noted in 1991 . This new restriction pattern has not been described previously as occurring in isolates of N gonorrhoeae . The tet(M) amplimer sequence showed 100% identity with a previously described tet(M)-carrying plasmid from N meningitidis . CONCLUSION: A new HpaII restriction pattern of the tet(M) gene is present in low frequency . The tet(M) sequence was different from the gonococcal tet(M) sequences already known and not typable with the use of a differential PCR assay . Accordingly, with the genetic diversity already present within the tet(M) sequence of N gonorrhoeae isolates, we should be aware of the sensitivity of the PCR assays in use for tetracycline-resistant N gonorrhoeae detection.

Sex Transm Dis, 2002 Dec, 29(12), 780 - 8
Characterization of ciprofloxacin resistance in Neisseria gonorrhoeae isolates in Canada; Ng LK et al.; BACKGROUND: Ciprofloxacin (500 mg orally, single dose) is one of the recommended therapies for gonorrhea in Canada . In Canada, the first ciprofloxacin-resistant (CipR) Neisseria gonorrhoeae strain was isolated in 1993 . Antimicrobial susceptibilities of N gonorrhoeae isolates were monitored as part of a national surveillance program to ensure efficacy of antimicrobial therapies . GOAL: The goal was to determine the characteristics of ciprofloxacin resistance in Canadian gonococcal isolates . STUDY DESIGN: Susceptibility testing was performed on gonococcal strains from different provinces in Canada to determine the prevalence of CipR strains and their distribution . The CipR strains were further differentiated according to auxotype (A), serotype (S), plasmid profile (P), and pulsed-field gel electrophoresis (PFGE) profile . DNA sequencing and DNA microarray technology were used to determine mutations in gyrA and parC . RESULTS: In Canada, between 1997 and 1999, 4.8% of resistant strains (130 of 2687 antibiotic-resistant N gonorrhoeae isolates) were CipR (MICs of 1-32 microg/l) and belonged to 48 A/S/P classes . Sixty-eight of the strains that were not differentiated by A/S/P were subtyped into 47 classes with PFGE . DNA sequencing and DNA microarray showed that the most common mutations had amino acid substitutions of Ser-->Phe at codon 91 and Asp-->Gly at codon 95 of the gyrA and Ser-->Arg at codon 87 of parC . CONCLUSION: The CipR strains isolated in Canada are phenotypically and genotypically diverse, indicating that they were imported from overseas and not endemic in Canada . Mutations in gyrA and parC previously only identified by DNA sequencing were successfully identified with DNA microarray technology . DNA microarray technology could be an alternative tool for identifying point mutations in resistance genes or other epidemiologic markers when clinical laboratories replace culture methods with rapid and automated molecular methods for diagnosis.

Sex Transm Dis, 2002 Dec, 29(12), 775 - 9
High prevalence of Neisseria gonorrhoeae and multiple sexually transmitted diseases among rural women in the Eastern Highlands Province of Papua New Guinea, detected by polymerase chain reaction; Mgone CS et al.; BACKGROUND: In a previous community-based study among rural women in the Eastern Highlands Province (EHP) of Papua New Guinea we determined that the prevalences of Trichomonas vaginalis infection, Chlamydia trachomatis infection, and syphilis were 46%, 26%, and 4%, respectively . Surprisingly, however, the prevalence of Neisseria gonorrhoeae infection was only 1%, which we considered low in consideration of the high prevalence of other sexually transmitted diseases (STDs) . The aim of the current study was to reexamine samples that were collected in that survey and retest them with use of polymerase chain reaction (PCR) . STUDY DESIGN: Using a cluster-sampling method, we surveyed 201 women aged 15 to 45 years in a population of approximately 19,000 people . In addition, 243 other women living in the same area who wished to be screened for STDs were included in the study . METHODS: Endocervical samples that were stored frozen at -80 degrees C were retested with multiplex PCR (M-PCR) for the detection of both N gonorrhoeae and C trachomatis and with a separate PCR for the detection of T vaginalis . RESULTS: A total of 373 samples that were still available were analyzed . The prevalences of T vaginalis, C trachomatis, and N gonorrhoeae infections were 42.6%, 26.5%, and 18.2%, respectively; 59.8% of the women had at least one STD, while 21.7% had mixed infections, 5.9% of them with all three pathogens . CONCLUSIONS: STDs are very common among rural women in the EHP of Papua New Guinea and often present as multiple infections.

Arch Inst Pasteur Madagascar, 2000, 66(1-2), 46 - 9
{Management of sexually transmitted diseases by the syndrome approach and voluntary HIV screening in a specialized dispensary in Antananarivo (Madagascar)}; Gleize L et al.; In 1994, Medecins du Monde opened a free health centre specialized in STD/AIDS in an ill-favored district of Antananarivo, the Malagasy capital of Madagascar . Besides the medical treatment of Sexually Transmitted Diseases (STD) and AIDS, the centre is responsible for the Information, Education and Communication activities (IEC) within and without the centre towards the residents of the 67 hectares district and the high-risk populations (prostitutes, truck-drivers and transvestites) . The project aimed at both preventing the spreading of the VIH infection and reducing the incidence of STD . As the Ministry of Health directed, a syndromic method was applied since 1997 regarding STD . Results for 1998 showed the predominance of the association Neisseria gonorrhae-Chlamydiae among the consultants of both sexes . Negative results from 1,218 HIV serological tests carried out seemed confirm the low prevalence of the HIV infection in Madagascar . Yet, the percentage of positive syphilis serology among the tested consultants was lower than that mentioned in previous surveys . Finally, it appears that the syndromic method is of high interest for the countries with limited laboratory capacities.

Trop Med Int Health, 2002 Dec, 7(12), 1053 - 63
A community randomized controlled trial to investigate impact of improved STD management and behavioural interventions on HIV incidence in rural Masaka, Uganda: trial design, methods and baseline findings; Kamali A et al.; OBJECTIVE: To describe study design, methods and baseline findings of a behavioural intervention alone and in combination with improved management of sexually transmitted diseases (STDs) aimed at reducing HIV incidence and other STDs . DESIGN: A three-arm community randomized controlled trial (RCT) of 18 rural communities (approximately 96 000 adults) in SW Uganda . A standardized behavioural intervention was implemented in 12 communities (arms A and B) through community-based education, meetings and information leaflets . Six of these communities in addition received improved STD management through government and private health units (arm B) . Arm C communities received routine government health services . Impact assessment was through three questionnaire and serological surveys of 750-1000 adults in each community at 18-24-month intervals . The primary outcome measure was HIV incidence and secondary measures were syphilis and herpes simplex virus type 2 incidence, prevalence of Neisseria gonorrhoea and Chlamydia trachomatis and sexual behaviour changes . RESULTS: Approximately 15 000 adults (72% of eligible population) were enrolled at baseline . HIV baseline prevalence rates were 9-10% in all arms and demographic and behavioural characteristics and STD prevalence were also similar . In intervention communities, there were 391 995 attendance at 81 502 activities (6.1 per target adult), 164 063 leaflets distributed (2.6 per person) and 1 586 270 condoms (16.5 condoms per adult) . In the STD communities a total of 12 239 STD cases (65% women) were seen over a 5-year period (7.7 per 100 adults/year) . CONCLUSION: This is the first community RCT of its type with a behavioural component . There is fair baseline comparability between study arms and process data suggest that interventions were adequately implemented.

J Fam Plann Reprod Health Care, 2001 Apr, 27(2), 81 - 4
Genital infection and termination of pregnancy: Are patients still at risk?
Smith CD, Carlin EM, Heason J, Liu DT, Jushuf IA, Hammond RH.
OBJECTIVES: To identify the treatment patterns and early complication rates in women identified with a genital infection prior to a termination of pregnancy (TOP) and to re-assess our earlier work on contact attendance and treatment . The aim was to refine current prophylactic measures to provide the most efficient and effective way of reducing post-termination complications . METHOD: Six-month retrospective analysis of all women with genital Chlamydia trachomatis or Neisseria gonorrhoeae identified prior to TOP . Data on the timing of positive results and treatment in relation to the TOP were recorded . Clinical signs of pelvic infection and the success of contact tracing and treatment were noted . RESULTS: Forty women were identified with infection, 31 (78%) proceeded to TOP; only five were treated adequately in the peri-TOP period . Twenty-six (84%) of the women undergoing TOP attended the genitourinary medicine (GUM) department after the TOP and received antimicrobial therapy where necessary . In total, four women (10%) were untreated . Approximately two thirds of results were reported prior to, or on the day of, the TOP . The majority of TOPs (71%) were carried out at 5 days or less from the initial assessment . Clinical signs of post-TOP pelvic inflammation developed in seven (28%) women with chlamydial infection . Contact tracing was successful in 24 (69%) of the 35 women who attended the GUM department . CONCLUSION: The debate about the best option for the most effective management of women undergoing TOP continues . The options with respect to C . trachomatis include universal prophylaxis, screening and treating as necessary, or a 'belt and braces' approach of screening all and treating all . Fundamental is the continuation of active contact tracing and GUM remains integral to this.

J Clin Microbiol, 2002 Dec, 40(12), 4567 - 70
Typing of Neisseria gonorrhoeae reveals rapid reinfection in rural South Africa; Moodley P et al.; A recent study afforded us the opportunity to collect pre- and post-treatment isolates of Neisseria gonorrhoeae from women who supposedly failed to eradicate the organism when tested 8 to 10 days following treatment with a single, directly observed 250-mg dose of ciprofloxacin . In an attempt to differentiate true treatment failure from reinfection, we determined the ciprofloxacin MICs and performed auxotyping, serotyping, and opa typing of the pre- and post-treatment isolates . Paired isolates of N . gonorrhoeae were obtained from seven different women, despite susceptibility of the initial isolates to ciprofloxacin . Six of seven patients were infected with gonococcal isolates that differed significantly from their primary isolate . These most probably represent reinfection with a different strain, which could originate from the same partner infected with multiple strains or reinfected with a new strain or from a different partner . The susceptibility to ciprofloxacin of all isolates makes the possibility of multiple strains in the patient unlikely . The diversity of the isolates within the pairs therefore suggests rapid reinfection within the partnerships.

J Clin Microbiol, 2002 Dec, 40(12), 4520 - 7
Sequence diversity of Neisseria meningitidis 16S rRNA genes and use of 16S rRNA gene sequencing as a molecular subtyping tool; Sacchi CT et al.; We investigated the diversity of the primary sequences of 16S rRNA genes among Neisseria meningitidis strains (Men) and evaluated the use of this approach as a molecular subtyping tool . We aligned and compared a 1,417-bp fragment of the 16S rRNA gene from 264 Men strains of serogroups A, B, C, and Y (MenA, MenB, MenC, and MenY, respectively) isolated throughout the world over a 30-year period . Thirty-one positions of difference were found among 49 16S types: differences between types ranged from 1 to 14 positions (0.07 to 0.95%) . 16S types and serogroups were highly associated; only 3 out 49 16S types were shared by two or more serogroups . We have identified 16S types that are exclusively associated with strains of certain hypervirulent clones: 16S type 5 with MenA subgroup III, 16S type 4 with the MenB electrophoretic type 5 (ET-5) complex, and 16S types 12 and 13 with MenC of the ET-37 complex . For MenC strains, 16S sequencing provided the highest sensitivity and specificity and the best overall association with the outbreak-related versus sporadic isolates when compared with pulsed-field gel electrophoresis, multilocus enzyme electrophoresis, and multilocus sequence typing . We demonstrated for the first time an unexpected diversity among 16S rRNA genes of Men strains, identified 16S types associated with well-defined hypervirulent clones, and showed the potential of this approach to rapidly identify virulent strains associated with outbreaks and/or an increased incidence of sporadic disease.

Mol Microbiol, 2002 Dec, 46(5), 1441 - 50
An inhibitor of DNA binding and uptake events dictates the proficiency of genetic transformation in Neisseria gonorrhoeae: mechanism of action and links to Type IV pilus expression; Aas FE et al.; Although natural genetic transformation is a widely disseminated form of genetic exchange in prokaryotic species, the proficiencies with which DNA recognition, uptake and processing occur in nature vary greatly . However, the molecular factors and interactions underlying intra- and interspecies diversity in levels of competence for natural genetic transformation are poorly understood . In Neisseria gonorrhoeae, the Gram-negative aetiologic agent of gonorrhoea, DNA binding and uptake involve components required for Type IV pilus (Tfp) biogenesis as well as those which are structurally related to Tfp biogenesis components but dispensable for organelle expression . We demonstrate here that the gonococcal PilV protein, structurally related to Tfp pilin subunits, is an intrinsic inhibitor of natural genetic transformation which acts ultimately by reducing the levels of sequence-specific DNA uptake into the cell . Specifically, we show that DNA uptake is enhanced in strains bearing pilV mutations and reduced in strains overexpressing PilV . Furthermore, we show that PilV exerts its effect by acting as an antagonist of ComP, a positive effector of sequence-specific DNA binding . As it prevents the accumulation of ComP at a site where it can be purified by shear extraction of intact cells, the data are most consistent with PilV either obstructing ComP trafficking or altering ComP stability . In addition, we report that ComP and PilV play overlapping and partially redundant roles in Tfp biogenesis and document other genetic interactions between comP and pilV together with the pilE and pilT genes required for the expression of retractile Tfp . Together, the results reveal a novel mechanism by which the levels of competence are governed in prokaryotic species and suggest unique ways by which competence might be modulated.

J Infect Dis, 2002 Dec 1, 186(11), 1669 - 72 Epub 2002 Nov 06.
Cervical shedding of human T cell lymphotropic virus type I is associated with cervicitis; Zunt JR et al.; Human T cell lymphotropic virus type I (HTLV-I) is sexually transmitted . The purpose of this study was to determine the prevalence and risk factors for cervical shedding of HTLV-I DNA among Peruvian sex workers . HTLV tax DNA was detected in cervical specimens from 43 (68%) of 63 HTLV-I-infected sex workers and in samples obtained during 113 (52%) of 216 clinic visits between 1993 and 1997 . Detection of HTLV DNA was associated with the presence of > or =30 polymorphonuclear cells (PMNs) within cervical mucus per 100x microscopic field (odds ratio {OR}, 4.3, 95% confidence interval {CI}, 1.8-10.1) and with the presence of cervical secretions (OR, 2.0; 95% CI 1.2-3.4) . Hormonal contraceptive use (OR 1.7; 95% CI, 0.8-3.6) and concomitant cervical infection by Chlamydia trachomatis (OR, 1.5; 95% CI, 0.3-4.3) or Neisseria gonorrhoeae (OR, 1.1; 95% CI, 0.6-3.7) were not significantly associated with HTLV-I shedding . Our results suggest that cervicitis may increase cervical HTLV-I shedding and the sexual transmission of this virus.

Proc Natl Acad Sci U S A, 2002 Dec 10, 99(25), 16012 - 7 Epub 2002 Nov 22.
Single pilus motor forces exceed 100 pN; Maier B et al.; Force production by type IV pilus retraction is critical for infectivity of Neisseria gonorrhoeae and DNA transfer . We investigated the roles of pilus number and the retraction motor, PilT, in force generation in vivo at the single-molecule level and found that individual retraction events are generated by a single pilus fiber, and only one PilT complex powers retraction . Retraction velocity is constant at low forces but decreases at forces greater than 40 pN, giving a remarkably high average stall force of 110 +/- 30 pN . Further insights into the molecular mechanism of force generation are gained from the effect of ATP-depletion, which reduces the rate of retraction but not the stall force . Energetic considerations suggest that more than one ATP is involved in the removal of a single pilin subunit from a pilus . The results are most consistent with a model in which the ATPase PilT forms an oligomer that disassembles the pilus by a cooperative conformational change.

Biochem Soc Trans, 2002 Nov, 30(Pt 6), 978 - 82
Structural and functional studies of complement inhibitor C4b-binding protein; Blom AM; C4b-binding protein (C4BP) is a potent inhibitor of the classical pathway of the complement system . This large plasma glycoprotein consists of seven identical alpha-chains and a unique beta-chain held together by disulphide bridges . Both types of subunits are composed almost exclusively of complement control protein domains (CCPs) . Using homology-based computer modelling and mutagenesis of recombinant proteins we have localized binding sites for several ligands of C4BP: complement factor C4b, heparin and vitamin K-dependent anticoagulant protein S (PS) . We found that C4b requires CCP1-3 of the alpha-chain for binding . The interaction is ionic in nature and mediated by a cluster of positively charged amino acids present on the interface between CCP1 and CCP2 of the alpha-chain . Loss of C4b-binding resulted in a loss of all inhibitory functions of C4BP within the classical pathway of complement . Binding of heparin required CCPs 1-3 of the alpha-chain, with CCP2 being the most important, as well as the cluster of positively charged amino acids involved in binding of C4b . The interaction between C4BP and PS is of very high affinity and conveyed by a cluster of surface exposed hydrophobic amino acids localized on CCP1 of the beta-chain . Furthermore, C4BP is captured on the surface of several pathogens, which may contribute to their serum resistance and pathogenicity . We have localized interaction of C4BP with Neisseria gonorrhoeae, Bordetella pertussis, Streptococcus pyogenes and Escherichia coli to various regions of the alpha-chain.

Sex Transm Dis, 2002 Nov, 29(11), 668 - 73
Multiclonal increase in ciprofloxacin-resistant Neisseria gonorrhoeae, Thailand, 1998-1999; Trees DL et al.; BACKGROUND: isolates exhibiting clinically significant resistance to fluoroquinolones have been isolated most frequently in Asian and western Pacific countries, including Thailand . In Bangkok, Thailand, ciprofloxacin has been used to treat gonorrhea since 1987 . GOAL: Our goal was to determine the prevalence of isolates of that exhibit resistance to ciprofloxacin in Bangkok and to characterize these strains with regard to ciprofloxacin MICs, auxotype/serovar (A/S) classification, A and C mutations responsible for ciprofloxacin resistance, and outer membrane lipoprotein (Lip) subtype analysis . STUDY DESIGN: MICs of gonococcal isolates from consecutive patients attending the Bangrak Hospital STD Clinic in Bangkok were determined by agar dilution . A/S class was determined by established procedures . Mutations within A and C were determined by DNA sequencing . Lip subtypes were determined by PCR and DNA sequencing . RESULTS: In 1998 and 1999, 115 of 168 isolated strains of exhibited decreased susceptibility or resistance to ciprofloxacin, and three cases of possible ciprofloxacin treatment failure were identified . Ciprofloxacin-resistant (CipR) strains increased from 13.8% (8/58) in 1998 to 25.4% (28/110) in 1999 ( = 0.08) . Ciprofloxacin MICs of CipR isolates ranged from 1.0 microg/ml to 32.0 microg/ml . CipR strains belonged to a number of A/S classes and Lip subtypes . Different CipR strains contained alterations at both amino acid 91 and amino acid 95 of A and also contained an amino acid alteration in C . These alterations are known to be involved in gonococcal resistance to ciprofloxacin . CONCLUSIONS: The prevalence of CipR strains of isolated in Bangkok increased substantially in the 1990s . Characterization of the CipR isolates revealed a number of different strain subtypes, indicating that CipR isolates in Bangkok are not from a single clonal source and therefore result from multiple cases of importation or local emergence . Because of the high level of CipR isolates at Bangrak Hospital, in 2000 the Thai Ministry of Public Health issued recommendations against the use of fluoroquinolones for the treatment of gonococcal infection in Thailand.

Sex Transm Dis, 2002 Nov, 29(11), 647 - 54
Evaluation of self-collected samples in contrast to practitioner-collected samples for detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis by polymerase chain reaction among women living in remote areas; Knox J et al.; BACKGROUND: Self-collected samples have been shown to be an acceptable and sensitive method for the detection by polymerase chain reaction (PCR) of sexually transmitted infections (STIs) among women.GOAL The goal of the study was to compare self-collected sampling methods to conventional practitioner endocervical sampling for the PCR detection of Chlamydia trachomatis and Neisseria gonorrhoeae to compare two self-collected sampling methods for the detection of T vaginalis by PCR . STUDY DESIGN: Women (n = 318) from urban and remote areas of central Australia participated in the study when attending their health clinic for a check-up . They each provided a FVU sample, self-collected vaginal swab specimen, and tampon specimen . This was followed by a clinical examination by a practitioner, with collection of endocervical and high vaginal swabs for testing by conventional microscopy and culture for N gonorrhoeae and T vaginalis, respectively . The FVU, self-collected vaginal swab, tampon, and endocervical swab specimens were tested by Roche Cobas Amplicor for C trachomatis and N gonorrhoeae . The self-collected vaginal swab and tampon specimens were also tested by an in-house PCR method for the detection of T vaginalis . RESULTS: In toto, C trachomatis was detected by PCR in 11.5%, N gonorrhoeae in 11.8%, and T vaginalis in 24.6% . Molecular diagnostics for N gonorrhoeae and T vaginalis were significantly more sensitive than traditional assays with microscopy and culture . For the detection of C trachomatis by PCR, tampons were the most sensitive (100.0%) and urine the least sensitive (72.7%) specimens ( = 0.01) . For the detection of by PCR, the self-collected tampon was the most sensitive specimen, followed by the endocervical swab, self-collected swab, and urine specimen, with sensitivities of 97.2%, 92.6%, 71.9%, and 31.2%, respectively . For detection of N gonorrhoeae, statistically significant differences were detected for urine versus tampon ( < 0.0001), endocervical swab ( < 0.001), and self-collected swab ( = 0.01) and for self-collected swab versus tampon ( = 0.01) . Subsequent data collection showed that sensitivity of urine PCR for detection of N gonorrhoeae improved with freezing of urine specimens and shorter transport time . Tampons were also more sensitive than self-collected swabs for detection of T vaginalis (sensitivity of 100% versus 87.7%) . CONCLUSION: Self-collected specimens offer women in remote communities an acceptable and sensitive alternative method of testing for STIs . The low sensitivity of N gonorrhoeae PCR of urine specimens may reflect poor transport and storage conditions, which we have shown can be improved by freezing urine specimens and reducing transport delays.

Antimicrob Agents Chemother, 2002 Dec, 46(12), 3744 - 9
Mosaic-like structure of penicillin-binding protein 2 Gene (penA) in clinical isolates of Neisseria gonorrhoeae with reduced susceptibility to cefixime; Ameyama S et al.; Neisseria gonorrhoeae strains with reduced susceptibility to cefixime (MICs, 0.25 to 0.5 micro g/ml) were isolated from male urethritis patients in Tokyo, Japan, in 2000 and 2001 . The resistance to cephems including cefixime and penicillin was transferred to a susceptible recipient, N . gonorrhoeae ATCC 19424, by transformation of the penicillin-binding protein 2 gene (penA) that had been amplified by PCR from a strain with reduced susceptibility to cefixime (MIC, 0.5 micro g/ml) . The sequences of penA in the strains with reduced susceptibilities to cefixime were different from those of other susceptible isolates and did not correspond to the reported N . gonorrhoeae penA gene sequences . Some regions in the transpeptidase-encoding domain in this penA gene were similar to those in the penA genes of Neisseria perflava (N . sicca), Neisseria cinerea, Neisseria flavescens, and Neisseria meningitidis . These results showed that a mosaic-like structure in the penA gene conferred reductions in the levels of susceptibility of N . gonorrhoeae to cephems and penicillin in a manner similar to that found for N . meningitidis and Streptococcus pneumoniae.

BMC Microbiol . 2002 Nov 18;2(1):34.
Sulphonamide resistant commensal Neisseria with alterations in the dihydropteroate synthase can be isolated from carriers not exposed to sulphonamides; Qvarnstrom Y et al.; BACKGROUND: Development of sulphonamide resistance in Neisseria meningitidis has been suggested to involve horizontal DNA-transfer from a commensal Neisseria species . In this study, we isolated commensal Neisseria from throat specimens and examined the isolates with respect to sulphonamide resistance . RESULTS: Three resistant clones were identified and the resistance phenotype could be explained by amino acid variations in their dihydropteroate synthase, the target molecule for sulphonamides . Some of these variations occurred in positions corresponding to previously detected variations in resistant N . meningitidis . CONCLUSIONS: Sulphonamide resistant commensal Neisseria were isolated from an environment not exposed to sulphonamides, suggesting that resistant Neisseria has become a natural part of the commensal throat flora.

Indian Pediatr, 2002 Oct, 39(10), 914 - 21
Diagnosis and outcome of acute bacterial meningitis in early childhood; Chinchankar N et al.; OBJECTIVE: To estimate frequency of acute bacterial meningitis (ABM) in early childhood in hospital admissions, to describe clinical and diagnostic features, and to analyze mortality, complications and long term sequelae . DESIGN: Prospective study . SETTING: Pediatric wards and Rehabilitation Center of KEM Hospital, Pune . METHOD: Study subjects between the ages of 1 months to 5 years with ABM were recruited . Clinical details were recorded . CSF was analysed by routine biochemical methods, antigen detection tests (Latex agglutination LAT) and microbiological studies on special media . Management was as per standard protocols . Survivors were followed up long term with neurodevelopmental studies and rehabilitation programmes . RESULTS: In a study period of 2 years, 54 children (1.5% of all admissions) satisfied the criteria of ABM in early childhood; 78% were below one year and 52% were under the age of six months . Chief presentation was high fever, refusal of feeds, altered sensorium and seizures . Meningeal signs were present in only 26% . CSF C-reactive protein was positive in 41%, gram stain was positive in 67% LAT in 78% and cultures grew causative organisms in 50% of the cases . The final etiological diagnosis (as per LAT and/or cultures) were Streptococcus pneumoniae 39% Hemophilus influenzae type b 26% and others in 35% The others included one case of Neisseria meningitidis and 10 who were LAT negative and culture sterile . 39% patients developed acute neurological complications during the hospital course . 31% children with ABM died in hospital or at home soon after discharge . Six were lost to follow up . Of the 31 children, available for long term follow up (1-3 years), 14 (45%) had no sequelae . The remaining had significant neurodevelopmental handicaps ranging from isolated hearing loss to severe mental retardation with multiple disabilities . CONCLUSION: ABM in early childhood has a considerable mortality, morbidity and serious long term sequelae . Neurodevelopmental follow up and therapy should begin early . Etiological diagnosis can be enhanced by LAT and good culture media . H . influenzae b and S . pneumoniae account for more than 60% of ABM in early childhood.

J Bacteriol, 2002 Dec, 184(23), 6410 - 6
Biochemical properties of Neisseria gonorrhoeae LgtE; Piekarowicz A et al.; A fragment of chromosomal DNA encoding the lgtE gene of Neisseria gonorrhoeae strain F62 was amplified by PCR and cloned into the expression vector pET15b . Functional LgtE was purified and its biochemical properties were determined . The purified enzyme was maximally active in buffer containing manganese; minimal activity was obtained in buffer containing other divalent cations . LgtE was only able to mediate the addition of UDP-galactose into neisserial lipooligosaccharides (LOSs) . We used a variety of genetically defined and chemically verified LOS structures to determine acceptor specificity . LgtE was able to mediate the addition of galactose into a variety of LOS structures, indicating the this enzyme possesses broad acceptor specificity . Furthermore, it was able to add multiple galactose residues onto LOS . We also determined that this enzyme was capable of adding galactose onto both the alpha and beta chains of neisserial LOS.

FEMS Immunol Med Microbiol, 2002 Nov 15, 34(3), 193 - 200
Immunologic and genetic characterization of lipooligosaccharide variants in a Neisseria meningitidis serogroup C strain; Zhu P et al.; Neisseria meningitidis shows great variation in expression of structurally different lipooligosaccharides (LOS) on its cell surface . To better understand the LOS diversity that may occur within an individual strain, a group C wild-type strain, BB305-Tr4, and two stable isogenic LOS variants, Tr5 and Tr7, were selected for this study . SDS-PAGE analysis showed a size reduction of Tr5 and Tr7 LOS compared to that of Tr4 . Immunoblotting showed that parental Tr4 LOS reacted with L1, L2 and L3,7 antibodies, variant Tr5 LOS with L1 and L6 antibodies, while Tr7 LOS was non-typeable . Genetic analysis showed that the gene organization at the lgt-1 locus in the three strains was lgtZ,C,A,B,H4 in Tr4, lgtZ,C,A,H4 in Tr5 and lgtZ,C,A,H9 in Tr7 . The genetic differences in the three strains were consistent with their phenotypic changes . Sequence comparison revealed two independent recombination events . The first was the recombination of repeated DNA fragments in the flanking regions to delete lgtB in Tr5 . The second was the recombination of a fragment of two genes, lgtB and lgtH4, to create an inactive lgtH9 allele with a mosaic structure in Tr7 . These findings suggest that besides phase variation, homologous recombination can contribute to the genetic diversity of the lgt locus and to the generation of LOS variation in N . meningitidis.

MMWR Recomm Rep, 2002 Oct 18, 51(RR-15), 1 - 38; quiz CE1-4
Screening tests to detect Chlamydia trachomatis and Neisseria gonorrhoeae infections--2002; Johnson RE et al.; Since publication of CDC's 1993 guidelines (CDC, Recommendations for the prevention and management of Chlamydia trachomatis infections, 1993 . MMWR 1993;42{No . RR-12}:1-39), nucleic acid amplification tests (NAATs) have been introduced as critical new tools to diagnose and treat C . trachomatis and Neisseria gonorrhoeae infections . NAATs for C . trachomatis are substantially more sensitive than previous tests . When using a NAAT, any sacrifice in performance when urine is substituted for a traditional swab specimen is limited, thus reducing dependence on invasive procedures and expanding the venues where specimens can be obtained . NAATs can also detect both C . trachomatis and N . gonorrhoeae organisms in the same specimen . However, NAATs are usually more expensive than previous tests, making test performance from an economic perspective a key consideration . This report updates the 1993 guidelines for selecting laboratory tests for C . trachomatis with an emphasis on screening men and women in the United States . (In this report, screening refers to testing persons in the absence of symptoms or signs indicating C . trachomatis or N . gonorrhoeae infection.) In addition, these guidelines consider tests from an economic perspective and expand the previous guidelines to address detection of N . gonorrhoeae as well as C . trachomatis infections . Because of the increased cost of NAATs, certain laboratories are modifying manufacturers' procedures to improve test sensitivity without incurring the full cost associated with screening with a NAAT . Such approaches addressed in these guidelines are pooling of specimens before testing with a NAAT and additional testing of specimens whose non-NAAT test result is within a gray zone . This report also addresses the need for additional testing after a positive screening test to improve the specificity of a final diagnosis . To prepare these guidelines, CDC staff identified pertinent concerns, compiled the related literature published during 1990 or later, prepared tables of evidence, and drafted recommendations . Consultants, selected for their expertise or disciplinary and organizational affiliations, reviewed the draft recommendations . These final guidelines are the recommendations of CDC staff who considered contributions from scientific consultants . These guidelines are intended for laboratorians, clinicians, and managers who must choose among the multiple available tests, establish standard operating procedures for collecting and processing specimens, interpret test results for laboratory reporting, and counsel and treat patients.

Pediatrics . 2002 Nov;110(5):e57.
Gang involvement and the health of African American female adolescents; Wingood GM et al.; OBJECTIVE: To examine the association between gang involvement and female adolescents' health . METHODS: African American adolescent females (N = 522) completed a survey that assessed their history of gang involvement and health behaviors and provided specimens that were analyzed for marijuana use and sexually transmitted diseases . RESULTS: In logistic regression analyses, adolescents with a history of gang involvement were more likely to have been expelled from school (odds ratio {OR}: 3.6), be a binge drinker (OR: 3.3), have a positive toxicologic test for marijuana (OR: 2.6), have been in 3 or more fights in the past 6 months (OR: 3.8), have a nonmonogamous partner (OR: 2.4), and test positive for Trichomonas vaginalis (OR: 2.2) and Neisseria gonorrhoeae (OR: 3.6) . CONCLUSION: This study extends the current research on risk behaviors associated with gang involvement to include biological markers for substance use and sexual health outcomes, namely, marijuana use and sexually transmitted diseases.

Mol Microbiol, 2002 Nov, 46(3), 749 - 60
Competence for natural transformation in Neisseria gonorrhoeae: components of DNA binding and uptake linked to type IV pilus expression; Aas FE et al.; The mechanisms by which DNA is taken up into the bacterial cell during natural genetic transformation are poorly understood . Although related components essential to the uptake of DNA during transformation have been defined in Gram-negative species, it remains unclear whether DNA binding and uptake are dissociable events . Therefore, DNA uptake has been the earliest definable step in any Gram-negative transformation pathway . In the human pathogen Neisseria gonorrhoeae, sequence-specific DNA uptake requires an intact type IV pili (Tfp) biogenesis machinery along with three molecules that are dispensable for Tfp expression: ComP (a pilin subunit-like molecule), PilT (a cytoplasmic protein involved in pilus retraction) and ComE (a periplasmic protein with intrinsic DNA-binding activity) . By conditionally altering the levels of ComP and PilT expression, we show here that DNA binding and uptake are resolvable events . Consequently, we are able to demonstrate that PilT is largely dispensable for functional DNA binding and, therefore, contributes specifically to uptake . Furthermore, sequence specificity in this system is imposed at the level of DNA binding, a process that is influenced by both ComP and PilE . However, sequence-specific DNA binding is not attributable to an intrinsic property of the Tfp subunit protein . Finally, we demonstrate the existence of a robust, non-specific DNA-binding activity associated with the expression of both Tfp and PilT, which is unrelated to transformation but obscures the observation of specific binding events.

Clin Infect Dis, 2002 Nov 15, 35(10), 1167 - 73 Epub 2002 Oct 21.
Mycoplasma genitalium infections in asymptomatic men and men with urethritis attending a sexually transmitted diseases clinic in New Orleans; Mena L et al.; We report the results of a study of Mycoplasma genitalium (detected with a modified polymerase chain reaction {PCR} assay) in men with urethritis and in asymptomatic control subjects at a sexually transmitted diseases clinic in New Orleans . Data for 97 men with urethritis and 184 asymptomatic men were available for analysis . M . genitalium infection rates in symptomatic and asymptomatic men who were negative for Chlamydia trachomatis and Neisseria gonorrhoeae were 25% and 7%, respectively (P=.006) . M . genitalium coinfection rates among men with chlamydial and gonococcal urethritis were 35% and 14%, respectively . Men with M . genitalium urethritis resembled those with C . trachomatis in that both groups were younger and more likely to experience milder urethral symptoms . Among men with urethritis, the sensitivities of PCR of urine and swab specimens for the detection of M . genitalium were 87% and 91%, respectively . M . genitalium is associated with nongonococcal urethritis in this population.

Sex Transm Infect, 2002 Oct, 78(5), 365 - 8
Post-treatment sexual and prevention behaviours of adolescents with sexually transmitted infections; Fortenberry JD et al.; OBJECTIVE: To evaluate sexual behaviour (including abstinence), sex partner change, and condom use during the 3 month period following treatment for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, or non-gonococcal urethritis . METHODS: 251 14-21 year old participants (83% female; 83% African-American) diagnosed with gonorrhoea, chlamydia, trichomonas, or non-gonococcal urethritis or sexual contacts of infected partners . Participants were clients of a public sexually transmitted diseases clinic or primary care adolescent clinics . Data were collected by structured interview at treatment, 1 month post-treatment, and 3 months post-treatment . At each visit, participants were asked about coital frequency and condom use for each recent partner . At 1 month, participants were asked when coitus occurred following treatment . At each follow up visit, sex partners were compared to partners named at treatment and classified as "same partner(s)," "new partner(s)," or both "same and new partner(s)." RESULTS: Post-treatment abstinence was reported by 26% and 19% for the 1 month and 3 month visits, respectively . Abstinence was associated with greater likelihood of infection at enrolment although abstainers reported fewer lifetime STI and fewer lifetime sex partners . A substantial proportion of participants reported additional sexual contact with a previous partner . The average proportion of condom protected coital events increased from about 45% at enrolment to 64% at 1 month and 58% at 3 months (p<0.05) . Higher levels were sustained for the 3 months following treatment . CONCLUSIONS: Many adolescents adopt, at least temporarily, risk reduction behaviours such as abstinence or increased condom use . Sexual re-exposure to potentially untreated previous partners may increase risk of subsequent reinfection.

Sex Transm Infect, 2002 Oct, 78(5), 357 - 9
Reaching homeless youths for Chlamydia trachomatis and Neisseria gonorrhoeae screening in Denver, Colorado; Van Leeuwen JM et al.; BACKGROUND AND RATIONALE: The availability of urine based testing for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) provides a unique opportunity to screen for these pathogens outside traditional clinical settings . The researchers investigated the feasibility of street based CT/GC screening in the context of an outreach programme among street/homeless adolescents in Denver . OBJECTIVES: To describe the integration of urine based CT/GC screening into an existing outreach programme among street/homeless youths and the yield of CT/GC testing in this setting . METHODS: The Denver Department of Public Health (DPH) collaborated with outreach staff from Urban Peak (a community based organisation serving street/homeless youths in Denver) to offer urine based CT/GC testing to males and females in street settings . Tests were conducted on the street in areas where street/homeless youths congregate . RESULTS: Urban Peak outreach staff were trained by DPH staff to conduct CT testing, process urine specimens, and provide test results to participating youths . DPH remained responsible for treatment of people with CT or GC infection as well as CT/GC case reporting . CT testing started in January 2000; GC testing was added in July 2000 . Throughout April 2002 a total of 414 CT and 302 GC tests were conducted, respectively 11.6% and 2.7% of which were positive . Among first testers, 13.0% were positive for CT and 3.7% for GC . CONCLUSIONS: CT/GC urine testing can be incorporated into existing outreach programmes without considerable extra effort . Overall CT rates were high and suggest the need for ongoing screening in this manner . LEARNING OBJECTIVE: To understand the role of CT/GC screening in the context of services provided to street/homeless youths in outreach settings.

Mol Microbiol, 2002 Oct, 46(2), 493 - 504
Conservation of dynamic localization among MinD and MinE orthologues: oscillation of Neisseria gonorrhoeae proteins in Escherichia coli; Ramirez-Arcos S et al.; Min proteins are involved in the correct placement of division septa in many bacterial species . In Escherichia coli (Ec) cells, these proteins oscillate from pole to pole, ostensibly to prevent unwanted polar septation . Here, we show that Min proteins from the coccus Neisseria gonorrhoeae (Ng) also oscillate in E . coli . Green fluorescent protein (GFP) fusions to gonococcal MinD and MinE localized dynamically in different E . coli backgrounds . GFP-MinDNg moved from pole to pole in rod-shaped E . coli cells with a 70 +/- 25 s localization cycle when MinENg was expressed in cis . The oscillation time of GFP-MinDNg was reduced when wild-type MinENg was replaced with MinENg carrying a R30D mutation, but lengthened by 15 s when activated by MinEEc . Several mutations in the N-terminal domain of MinDNg, including K16Q and 4- and 19-amino acid truncations, prevented oscillation; these MinDNg mutants showed decreased or lost interaction with themselves and MinENg . Like MinEEc-GFP, MinENg-GFP formed MinE rings and oscillated in E . coli cells when MinDEc was expressed in cis . Finally, in round E . coli cells, GFP-MinDNg appeared to move in a plane parallel to completed septa . This pattern of movement is predicted to be similar in gonococcal cells, which also divide in alternating perpendicular planes.

West Afr J Med, 2002 Apr-Jun, 21(2), 153 - 6
HIV association with conventional STDS (sexual transmitted diseases) in Lagos State, Nigeria; Otuonye NM et al.; The study examined a possible association between HIV infection and conventional sexually transmitted diseases (STDS) in a population of 700 patients seen in some hospitals and clinics in Lagos State between November 1997 and December 1999 . The patients were drawn mainly from LUTH and Jolad hospitals in Lagos State . In these hospitals, patients who presented with symptoms of STDS were screened clinically and microbiologically for agents of STDS and HIV antibodies . Screening was carried out using conventional methods . A total of 150 (21.5%) were found positive for various STDS while 550 (78.5%) were negative Also, 109 (15.8%) were sero-positive for HIV while 591 (84.4%) were sero-negative . The frequency of STDS diagnosed were, Treponema pallidum, 38(25.3%), Neisseria gonorrhoea 3(2.0%), Chlamydia trachomatis 26(17.3), Hepatitis B virus 60(40.0%) Staphylococcus aureaus, 20 (13.3%) and Candida albicans 3(2.0%) . Data showed that Syphillis was the most prevalent STDS diagnosed while Calbicans and N . gonorrhoea are the least . Amongst the 150 (21.5%) patients positive with STDS, 82(54.65%) were found to be positive for HIV antibodies . The remaining 68(45.3%) patients were negative for HIV . The difference in sero-prevalence on the true group of patients rates was significant . The higher rate in the STDS patients strongly suggest some association between HIV infections andSTDS amongst the patients studied p = 0.05 . It was also recorded that HIV-1 infection is four times more prevalent than HIV-2 in these patients.

J Bacteriol, 2002 Nov, 184(22), 6163 - 73
Genome analysis and strain comparison of correia repeats and correia repeat-enclosed elements in pathogenic Neisseria; Liu SV et al.; Whole genome sequences of Neisseria meningitidis strains Z2491 and MC58 and Neisseria gonorrhoeae FA1090 were analyzed for Correia repeats (CR) and CR-enclosed elements (CREE) . A total of 533, 516, and 256 copies of CR and 270, 261, and 102 copies of CREE were found in these three genomes, respectively . The lengths of CREE range from 28 to 348 bp, and the lengths of multicopy CREE appear mainly in the ranges of 154 to 156 bp and 105 to 107 bp . The distribution of CREE lengths is similar between the two N . meningitidis genomes, with a greater number of 154- to 156-bp CREE (163 and 152 copies in N . meningitidis strain Z2491 and N . meningitidis strain MC58, respectively) than 105- to 107-bp CREE (72 and 77 copies) . In the N . gonorrhoeae strain FA1090 genome there are relatively more 105- to 107-bp CREE (51 copies) than 154- to 156-bp CREE (36 copies) . The genomic distribution of 107-bp CREE also shows similarity between the two N . meningitidis strains (15 copies share the same loci) and differences between N . meningitidis strains and N . gonorrhoeae FA1090 (only one copy is located in the same locus) . Detailed sequence analysis showed that both the terminal inverted repeats and the core regions of CREE are composed of distinct basic sequence blocks . Direct TA dinucleotide repeats exist at the termini of all CREE . A survey of DNA sequence upstream of the sialyltransferase gene, lst, in several Neisseria isolates showed that 5 N . meningitidis strains contain a 107-bp CREE in this region but 25 N . gonorrhoeae strains show an exact absence of a 105-bp sequence block (i.e., the 107-bp CREE without a 5' TA dinucleotide) in the same region . Whole-genome sequence analysis confirmed that this 105-bp indel exists in many homologous 107-bp CREE loci . Thus, we postulate that all CREE are made of target TA with indels of various lengths . Analysis of 107-bp CREE revealed that they exist predominantly in intergenic regions and are often near virulence, metabolic, and transporter genes . The abundance of CREE in Neisseria genomes suggests that they may have played a role in genome organization, function, and evolution . Their differential distribution in different pathogenic Neisseria strains may contribute to the distinct behaviors of each Neisseria species.

J Bacteriol, 2002 Nov, 184(22), 6138 - 45
Demonstration and characterization of a specific interaction between gonococcal transferrin binding protein A and TonB; Kenney CD et al.; Iron scavenging by Neisseria gonorrhoeae is accomplished by the expression of receptors that are specific for host iron-binding proteins, such as transferrin and lactoferrin . Efficient transferrin-iron acquisition is dependent on the combined action of two proteins, designated TbpA and TbpB . TbpA is a TonB-dependent outer membrane receptor, whereas TbpB is lipid modified and serves to increase the efficiency of transferrin-iron uptake . Both proteins, together or separately, can be isolated from the gonococcal outer membrane by using affinity chromatography techniques . In the present study, we identified an additional protein in transferrin-affinity preparations, which had an apparent molecular mass of 45 kDa . The ability to copurify this protein by transferrin affinity was dependent upon the presence of TbpA and not TbpB . The amino-terminal sequence of the 45-kDa protein was identical to the amino terminus of gonococcal TonB, indicating that TbpA stably interacted with TonB, without the addition of chemical cross-linkers . Using immunoprecipitation, we could recover TbpA-TonB complexes without the addition of transferrin, suggesting that ligand binding was not a necessary prerequisite for TonB interaction . In contrast, a characterized TonB box mutant of TbpA did not facilitate interaction between these two proteins such that complexes could be isolated . We generated an in-frame deletion of gonococcal TonB, which removed 35 amino acids, including a Neisseria-specific, glycine-rich domain . This mutant protein, like the parental TonB, energized TbpA to enable growth on transferrin . Consistent with the functionality of this deletion derivative, TbpA-TonB complexes could be recovered from this strain . The results of the present study thus begin to define the requirements for a functional interaction between gonococcal TbpA and TonB.

Acta Med Croatica, 2001, 55(4-5), 191 - 201
Epidemiologic characteristics of sexually transmitted infection/coinfection with Chlamydia trachomatis and Neisseria gonorrhoeae; Cerkez-Habek J et al.; Sexually transmitted infection/coinfection with Chlamydia trachomatis and Neisseria gonorrhoeae is responsible for pelvic inflammatory disease, ectopic pregnancy, infertility, neonatal conjunctivitis, and adhesive perihepatitis . The increase in the rate of chlamydial and gonococcal infection/coinfection has been related to promiscuity, inadequate use of mechanical contraception (condom), early sexual activity, and lack of proper sexual education . Therefore, the need of primary prevention and education of the young population, and introduction of amplification screening tests in the diagnosis of chlamydial infection at least before each intrauterine procedure, in women with mucopurulent discharge and in women below age 25 is emphasized.

Eur J Biochem, 2002 Nov, 269(21), 5215 - 23
Conformational analysis of opacity proteins from Neisseria meningitidis; de Jonge MI et al.; Opacity-associated (Opa) proteins are outer membrane proteins which play a critical role in the adhesion of pathogenic Neisseria spp . to epithelial and endothelial cells and polymorphonuclear neutrophils . The adherence is mainly mediated by the CD66-epitope-containing members of the carcinoembryonic-antigen family of human cell-adhesion molecules (CEACAM) . For the analysis of the specific interactions of individual Opa proteins with their receptors, pure protein is needed in its native conformation . In this study, we describe the isolation and structural analysis of opacity proteins OpaJ129 and OpaB128 derived from Neisseria meningitidis strain H44/76 . When the Opa proteins were produced with the phoE signal sequence in Escherichia coli, they were localized at the cell surface and the recombinant bacteria were found to specifically interact with CEACAM1 . For refolding and purification, the proteins were overproduced without their signal sequences in E . coli, resulting in its cytoplasmic accumulation in the form of inclusion bodies . After solubilization of the inclusion bodies in urea, the proteins could be folded efficiently in vitro, under alkaline conditions by dilution in ethanolamine and the detergent n-dodecyl-N,N-dimethyl-1-ammonio-3-propanesulfonate (SB12) . The structure of the refolded and purified proteins, determined by circular dichroism, indicated a high content of beta-sheet conformation, which is consistent with previously proposed topology models for Opa proteins . A clear difference was found between the binding of refolded vs . denatured OpaJ protein to the N-A1 domain of CEACAM1 . Almost no binding was found with the denatured Opa protein, showing that the Opa-receptor interaction is conformation-dependent.

Int J Technol Assess Health Care, 2002 Summer, 18(3), 485 - 96
Cost-utility analysis of patient care in children with meningeal signs; Oostenbrink R et al.; OBJECTIVES: We designed a model of diagnostic and therapeutic interventions applied in children with meningeal signs . Using this model, we determined in a cost-utility analysis the consequences for society of different diagnostic strategies in terms of quality-adjusted life-years (QALYs) and costs . METHODS: Data were used from 360 children (0.1-15 years) visiting the pediatric emergency department of the Sophia Children's Hospital Rotterdam, The Netherlands (1988-98) with meningeal signs . Model inputs included probabilities of meningitis and adverse outcome, QALYs for years lived with long-term sequelae, and costs of tests and treatments . Mean outcome measures were costs and effects of diagnostic and therapeutic interventions in children suspected of bacterial meningitis, key determinants of the model outcomes, and evaluation of alternative diagnostic strategies and two vaccination programs in an analysis . RESULTS: The population comprised 99 children with bacterial meningitis (adverse outcome in 10), 36 with serious other bacterial infections, and 225 with self-limiting diseases . Key determinants were the risk of bacterial meningitis or sequelae, costs of treatment, and long-term morbidity . Minimizing lumbar punctures and empirical treatments using a diagnostic decision rule, without missing a single case of meningitis, was a dominant strategy to actual practice . Vaccination strategies of Streptococcus pneumoniae and Neisseria meningitidis resulted in our model in incremental cost-utility ratios of 401,965 Euro dollar ({symbol: see text})/QALY and {symbol: see text}22,635/QALY, respectively . CONCLUSIONS: Costs of long-term morbidity of bacterial meningitis largely outweigh diagnostic and treatment costs . Modeling interventions in children at risk of bacterial meningitis should include long-term consequences in terms of costs and QALYs.

Cell Microbiol, 2002 Sep, 4(9), 585 - 98
The role of lipooligosaccharide in Neisseria gonorrhoeae pathogenesis of cervical epithelia: lipid A serves as a C3 acceptor molecule; Edwards JL et al.; The use of primary, human, ecto- and endocervical epithelial cell cultures has increased our understanding of the pathogenesis of gonococcal infection in women . Primary cervical epithelial cells express complement (C') receptor type 3 (CR3) and C' proteins required for alternative pathway (AP) activity . Gonococcus -induced membrane ruffling and cellular invasion of primary cervical epithelia is mediated by CR3 and requires co-operative CR3 binding by gonococcus-bound iC3b, porin and pilus . We have extended these studies to identify the site of C3 deposition upon gonococci within the cervical microenvironment . By immunoprecipitation and ELISA we demonstrate that covalent and non-covalent associations occurred between gonococcal LOS and C' protein C3 . Sialylation or LOS truncation did not alter the gonococcus-CR3 interaction . By Western blot analysis we observed comparable C3 opsonization patterns among a panel of LOS truncation mutants, sialylated wild-type gonococci, or wild-type bacteria that were not sialylated . Quantitative association/invasion assays performed in the presence or absence of LOS competimers support C3b deposition on the lipid A core structure . Our findings demonstrate a role for lipid A as a C3 acceptor site and suggest that multiple factors govern C3b deposition and its subsequent conversion to iC3b on the surface of the gonococcus within the cervical microenvironment.

Cell Microbiol, 2002 Sep, 4(9), 571 - 84
A co-operative interaction between Neisseria gonorrhoeae and complement receptor 3 mediates infection of primary cervical epithelial cells; Edwards JL et al.; Little is known about the pathogenesis of gonococcal infection within the lower female genital tract . We recently described the distribution of complement receptor 3 (CR3) on epithelia of the female genital tract . Our studies further indicate that CR3-mediated endocytosis serves as a primary mechanism by which N . gonorrhoeae elicits membrane ruffling and cellular invasion of primary, human, cervical epithelial cells . We have extended these studies to describe the nature of the gonococcus-CR3 interaction . Western Blot analysis demonstrated production of alternative pathway complement components by ecto- and endocervical cells which allows C3b deposition on gonococci and its rapid conversion to iC3b . Anti-iC3b and -factor I antibodies significantly inhibited adherence and invasion of primary cervical cells, suggesting that iC3b covalently bound to the gonococcus serves as a primary ligand for CR3 adherence . However, gonococcal porin and pili also bound to the I-domain of CR3 in a non-opsonic manner . Binding of porin and pili to CR3 were required for adherence to and invasion of cervical epithelia . Collectively, these data suggest that gonococcal adherence to CR3 occurs in a co-operative manner, which requires gonococcal iC3b-opsonization, porin and pilus . In conjunction, these molecules facilitate targeting to and successful infection of the cervical epithelium.

J Matern Fetal Neonatal Med, 2002 Jun, 11(6), 391 - 5
Screening positive urine pregnancy tests for sexually transmitted diseases expedites the treatment of infected adolescent gravidas; Stevens-Simon C et al.; OBJECTIVE: To test the utility of screening the urine samples used to diagnose pregnancies at urban teen clinics for Chlamydia trachomatis and Neisseria gonorrhoeae by polymerase chain reaction (PCR) . We hypothesized that urine screening would increase the proportion of teenagers treated for these two sexually transmitted diseases (STDs) before they initiated pregnancy-related care . DESIGN: A randomly selected subset of the urine samples used to diagnose 212 teen pregnancies were tested for C . trachomatis and N . gonorrhoeae by PCR . Endocervical testing was at the providers' discretion . Bivariate analyses were used to compare the teenagers randomized to the urine screening group (n = 102) and the non-screening group (n = 110) . RESULTS: Of the 102 urine PCR tests, 14 (13.7%) were positive . Endocervical swabs were obtained in 31 (14.6%) of the 212 teenagers and five (16.1%) were positive . Since pelvic examinations were performed so infrequently, the net endocervical swab detection rate was significantly lower than the urine-based detection rate (1.8% compared to 13.7%; p = 0.001) . Only one infected teenager was untreated when she initiated pregnancy-related care . Thus, the treatment rate was more than six times higher when urine samples were screened (12.7% compared to 1.8%; p = 0.003) . CONCLUSIONS: Screening the urine samples used to diagnose teen pregnancies for two common STDs is a simple, non-invasive procedure that is acceptable to providers and patients, and significantly increases the number of teenagers who are treated for genital infection before they initiate pregnancy-related care.

Minerva Pediatr, 2002 Dec, 54(6), 539 - 45
{Sexually transmitted infections}; De Toni T et al.; Sexually transmitted diseases (STD) are quite common and observed more frequently in teens . The adolescents represent a group at particular risk for STD due to biological, sociocultural and psychological factors . Undectected infections may lead to unwanted sequelae, including pelvic inflammatory disease, chronic abdominal pain, tubal scarring and increased risk of ectopic pregnancy . This paper deals with infections by Candida albicans, Chlamidia tracomatis, Neisseria gonorrheae, Gardnerella vaginalis, Treponema pallidum, Tricomonas vaginalis, Herpes simplex, Papilloma virus . In regard to gonorrhea, chlamydia, syphilis and papilloma virus, the expectation is that improved detection will decrease sequelae by early diagnosis and treatment . Prevention programs (information, use of hormonal contraception associated with condom use) and improved access to STD diagnosis and treatment services are useful to reduce the incidence of STD among young people.

J Am Assoc Gynecol Laparosc, 2002 Nov, 9(4), 503 - 7
Effects of laparoscopic lavage on adhesion formation and peritoneum in an animal model of pelvic inflammatory disease; Roberts LM et al.; STUDY OBJECTIVES: To develop an animal model of pelvic inflammatory disease (PID) that simulates intraluminal tubal damage and adhesion formation seen in human PID, and to evaluate peritoneal effects of laparoscopic lavage with 1% povidone-iodine, 0.5% povidone-iodine, and 0.05% chlorhexidine gluconate solutions . DESIGN: Prospective, randomized, controlled study (Canadian Task Force classification I) . SETTING: Animal research operating room facility . SUBJECTS: Forty-five 1-year-old Pasteurella-free New Zealand white rabbits . INTERVENTION: Laparoscopy with instillation of one of five lavage solutions after bacterial inoculation with Neisseria gonorrhoeae, Bacteroides fragilis, Escherichia coli, and Peptococcus niger . MEASUREMENTS AND MAIN RESULTS: Histologic evaluation revealed significantly more peritoneal inflammation after lavage with all three solutions than after no lavage or lavage with normal saline . Gross adhesion formation was visible in only one rabbit after lavage with normal saline . One animal having lavage with 1% povidone-iodine died immediately after injection for sedation before laparoscopic lavage, and was not included in the final results . CONCLUSION: Bacterial inoculation using an intrauterine insemination catheter did not produce clinical adhesions in 43 of 44 rabbits . Histologic evaluation revealed significantly more peritoneal inflammation in the lavage with povidone-iodine and chlorhexidine gluconate compared with no lavage or lavage with normal saline.

J Reprod Immunol, 2002 Oct-Nov, 57(1-2), 61 - 79
The innate and early immune response to pathogen challenge in the female genital tract and the pivotal role of epithelial cells; Quayle AJ; The female reproductive tract is immunologically unique in its requirement for tolerance to allogeneic sperm and, in the upper tract, to the conceptus . However, it must also be appropriately protected from, and respond to, a diverse array of sexually transmitted pathogens . Some of these infections can be lethal (e.g . Human Immunodeficiency Virus (HIV), Human Papilloma Virus (HPV)), and others (e.g . Chlamydia trachomatis and Neisseria gonorrhoeae) can have potentially devastating reproductive sequelae . Interactions between a host and a pathogen are complex, diverse and regulated, and are a function of the individual pathogen, and host immunity . Although there is undoubtedly commonality in the mucosal immune response, there is also evidence of a degree of site-specificity in immune mechanisms, dependent upon the function and anatomical location of an organ . In this article, we review the evidence on the pivotal role of epithelial cells in the innate and early immune response to pathogen challenge in female genital tract tissues, and examine the evidence that the 'sterile' upper and the 'non-sterile' lower female genital tract may maintain a different immunological surveillance milieu, and may also respond differentially to pathogen challenge . We also review the unique characteristics, and subsequent ramifications of the acute cervical immune response to C . trachomatis, and discuss how natural antimicrobial mediators of immunity may be utilized to decrease the spread of sexually transmitted infections.

Int J Antimicrob Agents, 2002 Oct, 20(4), 248 - 52
Lower dose of ciprofloxacin is adequate for the treatment of Neisseria gonorrhoeae in KwaZulu Natal, South Africa; Moodley P et al.; The response of male gonococcal urethritis to a single 250 mg dose of ciprofloxacin versus 500 mg was studied . Both regimens were given in combination with doxycycline in the context of the local syndromic management protocol . There was no significant difference in response between the regimens, inclusive/exclusive of tetracycline susceptible isolates . One patient in the 250 mg arm failed to respond clinically but was microbiologically cured and four patients in the 500 mg arm failed microbiologically but responded clinically . All four isolates had ciprofloxacin MICs </=0.007 mg/l.

Lancet, 2002 Sep 28, 360(9338), 971 - 7
Effectiveness of COL-1492, a nonoxynol-9 vaginal gel, on HIV-1 transmission in female sex workers: a randomised controlled trial; Van Damme L et al.; BACKGROUND: Nonoxynol-9 (rINN, nonoxinol-9) is an over-the-counter spermicide that has in-vitro anti-HIV-1 activity . Results of studies of its effectiveness in prevention of HIV-1 infection in women have been inconclusive . We aimed to assess effectiveness of this vaginal gel . METHODS: We did a randomised, placebo-controlled, triple-blinded, phase 2/3 trial with COL-1492, a nonoxynol-9 vaginal gel, in 892 female sex workers in four countries: Benin, Cote d'Ivoire, South Africa, and Thailand . 449 women were randomly allocated nonoxynol-9 and 443 placebo . Primary endpoint was incident HIV-1 infection . Secondary endpoints included Neisseria gonorrhoeae and Chlamydia trachomatis infections . Analysis was by intention to treat . FINDINGS: 765 women were included in the primary analysis . HIV-1 frequency in nonoxynol-9 users was 59 (16%) of 376 compared with 45 (12%) {corrected} of 389 in placebo users (402.5 vs 435.0 woman-years; hazard ratio adjusted for centre 1.5; 95% CI 1.0-2.2; p=0.047) . 239 (32%) women reported use of a mean of more than 3.5 applicators per working day, and in these women, risk of HIV-1 infection in nonoxynol-9 users was almost twice that in placebo users (hazard ratio 1.8; 95% CI 1.0-3.2) . 516 (68%) women used the gel less frequently than 3.5 times a day, and in these, risk did not differ between the two treatments . No significant effect of nonoxynol-9 on N gonorrhoeae (1.2; 0.9-1.6) or C trachomatis (1.2; 0.8-1.6) infections was reported . INTERPRETATION: This study did not show a protective effect of COL-1492 on HIV-1 transmission in high-risk women . Multiple use of nonoxynol-9 could cause toxic effects enhancing HIV-1 infection . This drug can no longer be deemed a potential HIV-1-prevention method . Assessment of other microbicides should continue.

Obstet Gynecol, 2002 Oct, 100(4), 808 - 12
Leukorrhea and bacterial vaginosis as in-office predictors of cervical infection in high-risk women; Hakakha MM et al.; OBJECTIVE: To evaluate 1) whether microscopic detection of leukorrhea or bacterial vaginosis identifies patients at high risk for cervical infection with Chlamydia trachomatis or Neisseria gonorrhoeae, and 2) if pregnancy alters the predictive value of these findings . METHODS: Wet-mount screening examination of vaginal discharge was performed on all new patients seen at two resident-staffed clinics serving primarily indigent women . Leukorrhea was defined as >10 white blood cells per high-power field on microscopic examination; Amsel criteria were used to determine the presence of bacterial vaginosis, with a positive clue cell test result defined as >20% of epithelial cells . The diagnoses of C trachomatis and N gonorrhoeae infection were established by deoxyribonucleic acid amplification tests . RESULTS: The study population consisted of 194 women, 118 (61%) of whom were pregnant . Overall, 11% of women had positive cultures for chlamydia or gonorrhea . Although both leukorrhea and clue cells were independently associated with positive cervical cultures, multivariate analysis found that clue cells did not contribute to the predictive value of leukorrhea alone among both pregnant (relative risk {RR} = 15.7) and nonpregnant (RR = 58.7) women . Negative predictive values for the screening test were comparably high (98-100%), independent of pregnancy status . CONCLUSION: Leukorrhea, in the presence or absence of bacterial vaginosis, was strongly associated with cervical infections with C trachomatis or N gonorrhoeae among both pregnant and nonpregnant patients . In settings where patient follow-up is uncertain, on-site screening tests identify women for whom empiric antibiotic therapy for sexually transmitted diseases may be appropriate.

Infect Immun, 2002 Nov, 70(11), 5965 - 71
Neisseria gonorrhoeae porin P1.B induces endosome exocytosis and a redistribution of Lamp1 to the plasma membrane; Ayala P et al.; The immunoglobulin A (IgA) protease secreted by pathogenic Neisseria spp . cleaves Lamp1, thereby altering lysosomes in a cell and promoting bacterial intracellular survival . We sought to determine how the IgA protease gains access to cellular Lamp1 in order to better understand the role of this cleavage event in bacterial infection . In a previous report, we demonstrated that the pilus-induced Ca(2+) transient triggers lysosome exocytosis in human epithelial cells . This, in turn, increases the level of Lamp1 at the plasma membrane, where it can be cleaved by IgA protease . Here, we show that porin also induces a Ca(2+) flux in epithelial cells . This transient is similar in nature to that observed in phagocytes exposed to porin . In contrast to the pilus-induced Ca(2+) transient, the porin-induced event does not trigger lysosome exocytosis . Instead, it stimulates exocytosis of early and late endosomes and increases Lamp1 on the cell surface . These results indicate that Neisseria pili and porin perturb Lamp1 trafficking in epithelial cells by triggering separate and distinct Ca(2+)-dependent exocytic events, bringing Lamp1 to the cell surface, where it can be cleaved by IgA protease.

Chest, 2002 Oct, 122(4), 1389 - 99
The significance of distal bronchial samples with commensals in ventilator-associated pneumonia: colonizer or pathogen?
Lambotte O, Timsit JF, Garrouste-Orgeas M, Misset B, Benali A, Carlet J.
STUDY OBJECTIVE: To investigate the role of oropharyngeal and cutaneous commensal microorganisms (OCCs) as a cause of ventilator-associated pneumonia (VAP) . DESIGN: Retrospective analysis of the medical and microbiological records . SETTING: One medical-surgical ICU . PATIENTS: All VAP episodes recorded during a 10-year period were reviewed . All patients with suspected VAP underwent bronchoscopy with protected-specimen brush (PSB) sampling and BAL before any change in antibiotic therapy was made . OCC-VAP was defined as VAP with significant growth in quantitative cultures (PSB yielded > or = 10(3) cfu/mL and/or BAL yielded > or = 10(4) cfu/mL) of OCCs only . Three experts reviewed the episodes . Exposed patients (ie, those with OCC-VAP) and unexposed patients (ie, patients without VAP) matched on condition severity at ICU admission and mechanical ventilation duration were compared . RESULTS: Twenty-nine episodes in 28 patients with > or = 10(4) cfu/mL OCCs in BAL fluid and/or > or = 10(3) cfu/mL OCCs in PSB specimens were found . All patients in these episodes had new radiologic lung infiltrates, with 26 episodes involving purulent tracheal aspirates, 23 episodes involving temperatures > or = 38.5 degrees C, and 18 episodes involving > or = 11,000 leukocytes/ microL . The main OCCs found were non-beta-hemolytic Streptococcus spp (n = 12), Neisseria spp (n = 7), and coagulase-negative Staphylococcus spp (n = 6) . Other possible reasons for fever and the presence of new chest infiltrates were found in 20 and 17 patients, respectively . Histologic evidence of pneumonia was found in 2 of the 10 patients who died . The three experts agreed on the diagnosis for 23 patients . In the OCC-VAP group only, the mean (+/- SD) logistic organ dysfunction (LOD) scores increased significantly (LOD score, 2 +/- 4; p = 0.008) during the 3 days before bronchoscopy, and ICU stay duration was longer than in the unexposed group . The exposed/unexposed study found no difference in mortality . CONCLUSION: OCCs may behave like classic nosocomial pathogens in critically ill patients.

J Biol Chem, 2002 Dec 6, 277(49), 47818 - 25 Epub 2002 Oct 07.
An essential role for albumin in the interaction of endotoxin with lipopolysaccharide-binding protein and sCD14 and resultant cell activation; Gioannini TL et al.; Experiments utilizing endotoxin aggregates, lipooligosaccharides (LOS) isolated from metabolically labeled Neisseria meningitidis serotype group B, demonstrate that albumin is an essential component of lipopolysaccharide binding protein- (LBP) and sCD14-dependent 1) disaggregation of LOS and 2) LOS activation of human umbilical vein endothelial cells (HUVEC) . Aggregates of LOS (LOS(agg)) with an apparent M(r) >or= 2 x 10(7) were isolated by gel sieving on Sephacryl HR S500 in buffered balanced salts solution plus albumin . Incubation of LOS(agg) with LBP and sCD14 promoted LOS(agg) disaggregation in an albumin-dependent fashion to complexes that contain LOS and sCD14, but no LBP, with an apparent M(r) approximately 60,000 (LOS:sCD14) as determined by Sephacryl S200 chromatography . Isolation by gel filtration of LOS(agg):protein aggregates formed by the interaction of LOS(agg) with either LBP or sCD14 alone revealed that the sequence of LOS-protein interactions as well as the step(s) at which albumin is necessary for the production of bioactive LOS:sCD14 were specific . Efficient generation of LOS:sCD14 required 1) interaction of LOS(agg) with LBP before interaction with CD14 and 2) the presence of albumin during the interaction of LBP with LOS(agg) . Activation of HUVEC by LOS(agg), as measured by IL-8 production, required both LBP and sCD14 and was thirty times more potent in the presence of albumin . In contrast, LOS:sCD14 did not require additional LBP, sCD14, or albumin to activate HUVEC but depended on the presence of albumin for optimal solubility/stability once formed . The albumin effect is apparently specific, because neither ovalbumin nor gelatin substituted for albumin in facilitating LBP:sCD14-dependent disaggregation of LOS(agg) or activation of endothelial cells . These results indicate that albumin is an essential facilitator of LBP/sCD14-induced LOS disaggregation that is required for activation of endothelial cells by LOS(agg).

J Biol Chem, 2003 Jan 24, 278(4), 2490 - 502 Epub 2002 Oct 07.
Synergistic anion and metal binding to the ferric ion-binding protein from Neisseria gonorrhoeae; Guo M et al.; The 34-kDa periplasmic iron-transport protein (FBP) from Neisseria gonorrhoeae (nFBP) contains Fe(III) and (hydrogen)phosphate (synergistic anion) . It has a characteristic ligand-to-metal charge-transfer absorption band at 481 nm . Phosphate can be displaced by (bi)carbonate to give Fe.CO(3).nFBP (lambda(max) 459 nm) . The local structures of native Fe-PO(4)-nFBP and Fe.CO(3).nFBP were determined by EXAFS at the FeK edge using full multiple scattering analysis . The EXAFS analysis reveals that both phosphate and carbonate ligands bind to FBP in monodentate mode in contrast to transferrins, which bind carbonate in bidentate mode . The EXAFS analysis also suggests an alternative to the crystallographically determined position of the Glu ligand, and this in turn suggests that an H-bonding network may help to stabilize monodentate binding of the synergistic anion . The anions oxalate, pyrophosphate, and nitrilotriacetate also appear to serve as synergistic anions but not sulfate or perchlorate . The oxidation of Fe(II) in the presence of nFBP led to a weak Fe(III).nFBP complex (lambda(max) 471 nm) . Iron and phosphate can be removed from FBP at low pH (pH 4.5) in the presence of a large excess of citrate . Apo-FBP is less soluble and less stable than Fe.nFBP and binds relatively weakly to Ga(III) and Bi(III) but not to Co(III) ions, all of which bind strongly to apo-human serum transferrin.

Ther Umsch, 2002 Sep, 59(9), 459 - 63
{Chlamydia and other sexually transmitted bacterial infections}; Clad A; Chlamydia trachomatis is the most common sexually transmitted bacterium worldwide . In Western Europe, the prevalence of gonorrhoea has decreased by more than 95% since the 1970ies; "tripper" and syphilis are essentially confined to high-risk groups while genital chlamydial infections affect people of all social classes, but information about chlamydia is still scarce in many European countries . Clinically genital chlamydial infections resemble gonorrhoea (dysuria, discharge, irregular bleeding, dyspareunia, perihepatitis) and may be mistaken for appendicitis . However, Chlamydia trachomatis persists longer and more often asymptomatic than Neisseria gonorrhoeae in the urogenital tract of men and women . About 20% of all chlamydia infected women suffer from partial or complete tubal occlusion . Chlamydia trachomatis is the leading cause of female infertility, but most of these women never experienced any clinical sign of pelvic inflammatory disease . Since particle concentrations are often very low in urine and cervical secretions only DNA-amplification tests, e.g . PCR or LCR, exhibit sufficient sensitivity for direct detection Chlamydia trachomatis . While Neisseria gonorrhoeae is eradicated by single-shot treatment with commonly used antibiotics like penicillins or cephalosporins Chlamydia trachomatis affords treatment for at least 10 days with doxycyline or macrolides . Partner treatment is essential to avoid reinfections . Condoms not only protect against HIV, but also against chlamydia, gonorrhoea and syphilis.

Cell Microbiol, 2002 Oct, 4(10), 649 - 62
Galectin-3 binds lactosaminylated lipooligosaccharides from Neisseria gonorrhoeae and is selectively expressed by mucosal epithelial cells that are infected; John CM et al.; Galectins are a family of beta-galactoside binding proteins that have been proposed as host receptors for bacteria because beta-galactoside carbohydrates are common in bacterial membrane glycolipid lipooligosaccharides (LOS) and lipopolysaccharides . We investigated the interaction of galectin-3 with gonococcal LOS that make lactosyl (Lc2 or Lac), paraglobosyl (nLc4; LNnT; lacto-N-neotetraose), gangliosyl (IV3GalNAcnLc4), and neolactohexaosyl (nLc6, lactonorhexaosyl) oligosaccharides . All but gangliosyl LOS terminate in beta-galactoside . Galectin-3 had the highest affinity for the nLc6 LOS, which is made by a strain that is highly infectious for the male urethra, but also bound nLc4 LOS and to a Lac LOS . The lacto-N-neotetraose tetrasaccharide was a more potent inhibitor of galectin-3 binding to LOS than either lactose or N-acetyllactosamine . The relative affinity of galectin-3 for gonococci mirrored its affinity for purified LOS . Western blot analysis revealed expression of galectin-3 by human endometrial adenocarcinoma and prostatic epithelial cells that can be invaded by gonococci . Immunohistochemistry of human fallopian tube epithelium showed localized expression of galectin-3 by non-ciliated cells, the specific cell gonococci invade in this tissue . We conclude that because of its location and affinity for gonococcal LOS galectin-3 could play a role in gonococcal infection.

Asian J Androl, 2002 Sep, 4(3), 179 - 82
Influence of several uropathogenic microorganisms on human sperm motility parameters in vitro; Liu JH et al.; AIM: The effects of certain uropathogenic microorganisms (Neisseria gonorrhoeae, Staphylococcus aureus, Staphylococcus epidermidis and Mycobacterium tuberculosis) on human sperm motility characteristics were studied in vitro . METHODS: In 10 healthy fertile men, ejaculates were aseptically obtained by masturbation and with a swim-up technique, a sperm suspension of high motility and purity was obtained . Several uropathogenic bacteria were obtained from outpatients with genitourinary tract infections . The sperm suspension was incubated with the pathogens at a bacteria: sperm ratio of 50:1 at 37deg . The sperm mobility parameters were estimated with a computer-assisted sperm analyzer (CASA) provided with a multiple-exposure photography system (Madi Corp., Zhejiang, China) . Measurements were carried out at 0, 2 and 4 hours of incubation . RESULTS: Staphylococcus aureus significantly decreased the sperm motility and viability, but Staphylococcus epidermidis, Mycobacterium tuberculosis and Neisseria gonorrhoeae did not . CONCLUSION: Staphylococcus aureus has an inhibitory effect on human sperm motility in vitro.

Clin Microbiol Rev, 2002 Oct, 15(4), 527 - 44
Acute septic arthritis; Shirtliff ME et al.; Acute septic arthritis may develop as a result of hematogenous seeding, direct introduction, or extension from a contiguous focus of infection . The pathogenesis of acute septic arthritis is multifactorial and depends on the interaction of the host immune response and the adherence factors, toxins, and immunoavoidance strategies of the invading pathogen . Neisseria gonorrhoeae and Staphylococcus aureus are used in discussing the host-pathogen interaction in the pathogenesis of acute septic arthritis . While diagnosis rests on isolation of the bacterial species from synovial fluid samples, patient history, clinical presentation, laboratory findings, and imaging studies are also important . Acute nongonococcal septic arthritis is a medical emergency that can lead to significant morbidity and mortality . Therefore, prompt recognition, rapid and aggressive antimicrobial therapy, and surgical treatment are critical to ensuring a good prognosis . Even with prompt diagnosis and treatment, high mortality and morbidity rates still occur . In contrast, gonococcal arthritis is often successfully treated with antimicrobial therapy alone and demonstrates a very low rate of complications and an excellent prognosis for full return of normal joint function . In the case of prosthetic joint infections, the hardware must be eventually removed by a two-stage revision in order to cure the infection.

J Clin Microbiol, 2002 Oct, 40(10), 3741 - 9
Molecular epidemiology of Neisseria gonorrhoeae: sequence analysis of the porB gene confirms presence of two circulating strains; Unemo M et al.; The phenotypic and genotypic characteristics of Neisseria gonorrhoeae strains fluctuate over time both locally and globally, and highly discriminative and precise characterization of the strains is essential . Conventional characterization of N . gonorrhoeae strains for epidemiological purposes is mostly based on phenotypic methods, which have some inherent limitations . In the present study sequence analysis of porB1b gene sequences was used for examination of the genetic relationships among N . gonorrhoeae strains . Substantial genetic heterogeneity was identified in the porB genes of serovar IB-2 isolates (8.1% of the nucleotide sites were polymorphic) and serovar IB-3 isolates (5.2% of the nucleotide sites were polymorphic) as well as between isolates of different serovars . The highest degree of diversity was identified in the gene segments encoding the surface-exposed loops of the mature PorB protein . Phylogenetic analysis of the porB1b gene sequences confirmed previous findings that have indicated the circulation of one N . gonorrhoeae strain each of serovar IB-2 and serovar IB-3 in the Swedish community . These strains caused the majority of the cases in two domestic core groups comprising homosexual men and young heterosexuals, respectively, and were also detected in other patients . The phylogenetic analyses of porB gene sequences in the present study showed congruence, but not complete identity, with previous results obtained by pulsed-field gel electrophoresis of the same isolates . In conclusion, porB gene sequencing can be used as a molecular epidemiological tool for examination of genetic relationships among emerging and circulating N . gonorrhoeae strains, as well as for confirmation or discrimination of clusters of gonorrhea cases.

J Clin Microbiol, 2002 Oct, 40(10), 3596 - 601
Duration of persistence of gonococcal DNA detected by ligase chain reaction in men and women following recommended therapy for uncomplicated gonorrhea; Bachmann LH et al.; Neisseria gonorrhoeae infection remains relatively common in the United States, representing a public health challenge . Ligase chain reaction (LCR) is both highly sensitive and specific for the detection of N . gonorrhoeae in urine and patient-obtained vaginal swab specimens . Because of the LCR test's exquisite sensitivity, it may potentially detect DNA from nonviable organisms following effective therapy, leading to false-positive test results and unnecessary additional treatment . The purpose of the present study was to determine the duration that gonococcal DNA is detectable by LCR following therapy for uncomplicated gonococcal infection . One hundred thirty men and women between the ages of 16 and 50 years presenting to a sexually transmitted disease clinic with urogenital gonorrhea were enrolled . After the standard history was taken and a genital examination was done, the patients were asked to submit either a urine specimen (men) or a urine specimen plus a self-obtained vaginal swab specimen (women) for N . gonorrhoeae testing by LCR at the initial visit and each day during the study period . At enrollment, patients were treated with single doses of ofloxacin, cefixime, or ceftriaxone . The median time to a negative urine LCR test result was 1 day for the men (mean, 1.6 +/- 0.14 days) and 2 days for the women (mean, 1.7 +/- 0.19 days) . Among the women the clearance time was significantly longer for vaginal specimens (mean, 2.8 +/- 0.30 days) than for urine specimens (mean, 1.7 +/- 0.11 days) . Irrespective of patient gender and specimen type, gonococcal DNA can be expected to be absent from urogenital specimens within 2 weeks following successful therapy.

J Clin Microbiol, 2002 Oct, 40(10), 3558 - 64
Evaluation of the Digene Hybrid Capture II Assay with the Rapid Capture System for detection of Chlamydia trachomatis and Neisseria gonorrhoeae; Van Der Pol B et al.; Screening for chlamydial and gonococcal infection has been strongly recommended for all sexually active women under the age of 26 . Advances in the ability to detect infection by nucleic acid detection techniques have improved access to screening methods in routine clinical practices . To meet the increasing demand for testing, a high-throughput system is desirable . We evaluated the performance of the Hybrid Capture 2 CT/GC (HC2) assay with the Digene Rapid Capture System (HC2-RCS) . The results of HC2-RCS for endocervical samples from 330 women were compared to those of culture and the COBAS Amplicor PCR . For detection of chlamydial infection, HC2-RCS had a sensitivity and a specificity similar to those of PCR (P > 0.5) and an improved sensitivity compared to that of culture (P = 0.007) . For identification of gonococcal infections, all assays performed similarly (P > 0.5) . The performance of HC2-RCS was also compared to that of the manual HC2 format (HC2-M) with these samples and with 911 endocervical samples collected previously . The performance of HC2-RCS was equivalent to that of HC2-M; the overall concordance rates for the detection of chlamydia and gonorrhea were 99.7% (kappa = 0.97) and 99.8% (kappa = 0.97), respectively . When the HC2 assay was performed with a semiautomated system application designed for high throughput, it demonstrated high sensitivity and a high specificity for detection of both Chlamydia trachomatis and Neisseria gonorrhoeae.

J Br Fer Soc, 1996, 1(1), 42 - 5
Immunopathogenesis of pelvic inflammatory disease and infertility -- what do we know and what shall we do?
Paavonen J.
PIP: The most significant complication of sexually transmitted diseases (STDs) in women is pelvic inflammatory disease (PID), which is responsible for considerable medical, social, and economic problems . Chlamydia trachomatis, Neisseria gonorrhoeae, or both cause PID in at least 50% of cases . Other microorganisms that are part of the abnormal vaginal flora also cause PID . Gonorrhea rates have quickly fallen in most developed countries, but chlamydia infection rates are still high in developed countries as well as in developing countries . The clinical signs and symptoms of PID have changed over time . More and more PID cases are classified as atypical or subclinical . Typical PID is rare . A strong association exists between chlamydia infection and tubal factor infertility or ectopic pregnancy in women with or without a history of PID . Health providers are concerned about the problem of unrecognized PID . Thus, recommendations for PID diagnosis have changed . A syndromic diagnosis of PID is advocated . The minimum criteria for syndromic diagnosis of PID include lower abdominal tenderness, bilateral adnexal tenderness, cervical motion tenderness, and no evidence of competing diagnosis (e.g., acute appendicitis) . Application of this improved approach will provide appropriate treatment earlier in the course of PID . PID-related morbidity (i.e., infertility and ectopic pregnancy) is a considerable public health problem worldwide . In order to prevent PID, clinicians and public health specialists need to understand the interactions of PID-causing microorganisms with the host immune system . By the time PID symptoms are detected, considerable tubal damage already exists limiting the effect of tertiary prevention of PID . Secondary prevention keeps lower genital tract infection from moving up to the upper genital tract . Health providers play a key role in secondary prevention by screening for STDs and in primary prevention by counseling patients about safer sex practices .

J Br Fer Soc, 1996, 1(1), 23 - 30
Chlamydia and its effect on reproduction; Westrom LV; PIP: Epididymitis in the male and salpingitis in the female may cause permanent functional damage to the reproductive tract, resulting in infertility . Agents with proven connection to post-infection infertility are Neisseria gonorrhoeae and Chlamydia trachomatis . In men, infertility after chlamydia-associated epididymitis is uncommon, whereas in women sequelae after salpingitis--including chlamydia-associated disease--are the most common cause of acquired infertility . In a prospectively followed cohort of women who all sought pregnancy after one episode of laparoscopically verified acute salpingitis, 79/1025 (7.8%) were infertile because of post-salpingitic tubal occlusion, compared with 4/448 control women (0.9%) . Important factors in post-salpingitic infertility were: number of episodes {relative risk (RR) after none, 1, 2, and 3 or more episodes were 1.0, 5.2, 11.3, and 19.8, respectively, amounting to a total infertility rate after salpingitis of 15%}; in women with only one episode, the severity of infection (mild, moderate, and severe; RR, 1.0, 1.8, and 5.6, respectively) and delayed care (less than 3 days and 3 or more days; RR, 1.0 and 3.0, respectively); use of contraceptives (non-contraceptors, pill users, IUD users, and 'other'; RR, 1.0, 0.3, 0.5, and 0.8, respectively); and non-chlamydial and chlamydial infection (RR, 1.0 and 1.7, respectively) . Observations in infertile women suggest that an equally large number of women may have post-infectious tubal infertility also after asymptomatic salpingitis; in the majority of cases with serologic evidence of a passed genital chlamydial infection . Recently, a hypothesis has been presented that antigen-antibody reactions to chlamydial heat-shock proteins might be an important factor for the morphological tissue damage and scarring leading to impaired fertility . Post-pelvic inflammatory disease associated with sexually transmitted disease infertility is acquired and, hence, preventable . author's modified

ORGYN, 1994, (3), 12 - 5
Infertility: a global perspective . The role of pelvic infection; Sciarra JJ; PIP: The results of an examination of 5800 couples in 33 World Health Organization centers in 25 countries have provided the most comprehensive information on infertility in both developed and developing countries . Prior infection and bilateral tubal blockages were frequently traced as the cause of infertility . In the African sample; up to 64% of female patients had prior infection as the cause of infertility . A gynecological history of sexually transmitted disease, pelvic inflammatory disease (PID) and infection-related pregnancy complications of normal childbirth and abortion were directly related to infertility . 49% of the African couples and 11-15% of other patients in other parts of the world had infectious tubal disease . Westrom's study found that after PID 6-60% of patients became infertile . The factors affecting the infertility were the number and severity of infections, and the age when initial infection occurred . Between 1960 and 1980, in Sweden and the US, infertility doubled . Cates in the US has estimated that after 3 PID infections more than 50% of patients will be infertile . Ectopic pregnancy is another outcome of pelvic infection and sexually transmitted disease . Common PID infections are caused by Chlamydia trachomatis and Neisseria gonorrhea . These infections can cause tubal obstruction or pelvic adhesions and destructive tubal damage . After an ectopic pregnancy, 50% of patients are infertile and 10% have subsequent ectopic pregnancies . Some infertility may be treated . Microsurgery can be performed for tubal disease . Hysteroscopy can be used to treat intrauterine adhesions (Asherman's syndrome) . Success has been better with hysteroscopy . Tubal microsurgery had led to conceptions in only 30-30% of patients, and a very high percentage of tubal pregnancies . The most successful results have come from tubal reanastomosis for reversal of tubal sterilization . The most promising results have come from "in vitro" fertilization and other methods of assisted reproduction . However, there must be a more simplified technology, improved results, and reduced costs before large numbers worldwide can benefit . The best approach is prevention and early detection of infection, and risk assessment . Culturally sensitive education programs will generally alert the population to precursor conditions of infertility .

Afr Health, 1993 Mar, 15(3), 15 - 7
Pelvic inflammatory disease; Brabin L; PIP: The real prevalence of pelvic inflammatory disease (PID) is unknown since many women are either asymptomatic or have atypical symptoms . It is often difficult to detect, manage, and prevent PID . Since PID has obstetric, gynecologic, and contraceptive-related causes, its prevalence is quite high . About 70% of PID hospital admissions in sub-Saharan Africa are a result of reproductive tract infections (RTIs) while this figure is 34% in Asia and 31% in developed countries . Only 10-20% of lower RTIs ascend into the upper genital tract and an even smaller percentage of women with PID develop chronic sequelae . Still, just 1 episode carries an increased risk of a tubal infertility, ectopic pregnancy, chronic pelvic pain, considerable pain during coitus, a new episode, and menstrual irregularities . Neisseria gonorrhoea and Chlamydia trachomatis are the most common causative organisms of PID . In Africa, the risk factors for PID are the same as they are for sexually transmitted diseases (STDs): multiple sex partners, young age at first intercourse, high frequency of coitus, and a high rate of acquiring new partners . The largest percentage of women with RTIs are monogamous women who are infected and constantly reinfected by their promiscuous husbands . The primary means to prevent PID are promotion of safer sexual behavior and condom usage . Secondary measures include accessible, acceptable, and effective STD services and education and counseling during case management . WHO suggests that STD treatment become part of the primary health care system . It has developed flow charts on syndromic diagnosis for urethral discharge in men and genital ulcer disease in women . Health workers should assume increased PID risk if the partner has had a history of urethral discharge and/or treatment for gonorrhea or nongonococcal urethritis . Partner notification is also needed for case management, but stigmatization in some countries poses a problem . WHO also recommends use of drugs which have a 95% STD cure rate .

J Obstet Gynaecol East Cent Africa, 1988, 7(2), 71 - 3
The prevalence of Neisseria gonorrhoea and Chlamydia trachomatis in intra-uterine contraceptive acceptors in Kenya; Sinei SK et al.; PIP: The 1725 women presenting at Kenyatta National Hospital in 1984-86 for IUD insertion were screened for cervical Chlamydia trachomatis and Neisseria gonorrhoea before the IUD insertion . 207 (12%) cases of chlamydia trachomatis and 61 (3.5%) cases of Neisseria gonorrhoea were detected . There was no association between the ages of the women and the prevalence of these 2 sexually transmitted pathogens; however, there was a significant relationship between the prevalence of N gonorrhoea and marital status . N gonorrhoea was detected in 6.2% of never-married and 5.2% of formerly married women compared with 2.3% of currently married subjects (p0.001) . Although there was no significant relationship between parity and the rate of isolation of the 2 pathogens, infection tended to be lower in women with 5 or more children . Educational attainment was significantly associated with N gonorrhoea infection: 5.1% in women who had 0-7 years of schooling compared with 3.0% in those with 8 or more years of education (p0.05) . 12 women with C trachomatis infection were also positive for N gonorrhoea . There was no significant relationship between C trachomatis infection and any of the demographic variables examined . Given the finding that the greatest risk of pelvic inflammatory disease occurs in the 1st month of IUD use, it can be speculated that pathogens are inserted into the uterine cavity at the time of IUD insertion . It is therefore recommended that clients--especially the unmarried, the formerly unmarried, and those with low levels of education--be screened and treated for N gonorrhoea and C trachomatis before an IUD is inserted .

IPPF Med Bull, 1987 Feb, 21(1), 6 - 8
Sterility in women in sub-Saharan Africa; Frank O; PIP: This article examines the problem of sterility in women in sub-Saharan Africa . The term sterility indicates a physiological inability to bear children: conception and early pregnancy may occur, but successful pregnancy does not . Primary infertility is the total absence of live-born children, whereas 2ndary infertility is the absence of additional children after a reasonably long period . Some naturally occurring level of primary infertility is expected in all populations of women, and various research had shown this to be 3%, on the basis of the proportion of women who have reached the end of their childbearing years childless . Although measurement of 2ndary fertility is more difficult, studies show that almost 3 times as many women suffer from 2ndary infertility as from primary infertility . Using the 3% standard of childlessness in women past childbearing, studies show that infertility varies by region in Africa, with higher levels of childlessness in Central Africa and parts of East Africa . Given the predominant natural fertility pattern of the region, the variation levels of infertility alone accounts for 60% of the variation in fertility levels . These high levels of infertility are acquired through infection, most particularly from sexual transmission of Neisseria gonorrhea and Chlamydia trachomatis . Recent findings reveal that more than 85% of infertile women have diagnosed reproductive pathology indicative of such genital infections . Together, gonorrhea and chlamydia challenge the medical resources of Africa, posing serious problems on the count of diagnosis, treatment, and their non-genital as well as 2ndary pathologies . Although the role of gonorrhea in infertility in Africa has long been recognized, the importance of C . trachomatis was not recognized until the early 1980s . The earlier absence of chlamydial control may account for the meager improvement in infertility by 1980, despite large scale penicillin campaigns . The resulting need for new and multivalent drug regimens and for good patient compliance (for chlamydia treatment) to make case-management feasible in the light of still severe constraints on diagnosis may well mean that notable improvements in infertility will still be hard to achieve for some time to come .

IPPF Med Bull, 1987 Feb, 21(1), 1 - 4
Statement on steroidal oral contraception; Fathalla M et al.; PIP: This report summarizes a meeting of the IPPF International Medical Advisory Panel (IMAP) held in November, 1986, at which information on steroidal oral contraception (OC), Acquired Immunodeficiency Syndrome (AIDS), and female sterility were discussed . Regarding the multiphasic OC now in use, the benefits to health and well-being outweigh the possible side-effects and infrequent complications . Use is associated with a lower incidence of pelvic inflammatory disease, 96-98% effective prevention of pregnancy, a protective effect against ovarian and endometrial cancer, and regulation of erratic menstrual cycles . Minor side effects include nausea, vomiting, dizziness, headache, fluid retention, and inter-menstrual spotting . Adverse effects are circulatory system disease, myocardial infarction, venous thromboembolism, elevated blood pressure, and liver disease . Data on possible carcinogenicity have been conflicting . For women over age 40 OCs should be prescribed with caution . IMAP also drew up recommendations to assist FPAs to play a more active role in controlling the spread of AIDS . An effective program of Information and Education is of primary importance, targeting family planning workers and clients, teachers, parents, and employers . Wide promotion of condom use is a priority . Studies in Africa have revealed a major epidemic of AIDS, with the major mode of transmission heterosexual . The only immediate practical step in prevention of spread is by changes in sexual behavior . The last topic discussed is that of sterility in African women . The naturally occurring level of infertility expected in all populations of women is 3%; high levels in Africa vary by region from 3-32% . These levels of sterility are acquired through infection with Neisseria gonorrheae and Chlamydia trachomatis . Silent infection of women with Chlamydia make treatment especially difficult .

Afr J Sex Transmi Dis, 1984 Jan-Mar, 1(1), 11 - 3
Thiamphenicol in treatment of genital gonorrhoeae in Central African Republic; Widy-wirski et al.; PIP: Gonorrhea treatment has become a problem in most parts of the world and valid recommendations regarding treatment are an important control tool . In Central African Republic genital gonorrhea is one of the most common communicable diseases . In 1981 Penicillinase Producing Neisseria Gonorrhea (PPNG) was 1st isolated and, in spite of control measures, its prevalence reached 12% by the end of 1982 . Effective and inexpensive treatment regimens are now needed as an alternative to the Procain Penicillin + Probenicid regimen currently in use . Since its introduction, many authors in industrialized countries have reported excellent results with Thiamphenicol (TAP) in the treatment of gonococal infections, particularly in the single dose oral treatment . Gonococcal infections caused by Penicillinase Producing Neisseria gonorrhea (PPNG) also responded favorably to the antibiotic . However, few careful studies were done in Africa . This study evaluates the efficacy of TAP compared with the standard regimen of Aqueous Procain Penicillin + Probenicid in the treatment of uncomplicated genital gonorrhea in men in Central African Republic . 380 such cases were used . Patients with coexisting genital ulcers and those treated with antimicrobics within the last 2 weeks were excluded . Gram stained smears were examined and a urethral swab was taken and cultured on a modified Thayer Martin medium . Blood specimens were collected for VDRL and TPHA tests . During a 3-month period 338 patients were treated with a positive pretreatment culture for N . gonorrhea . 242 (71.6%) presented at follow-up, with similar follow-up rates in both treatment groups (25 mg . Thiamphenicol and Aqueous Procain Penicillin G . + 1g . Probenicid) . All failures but 1 in the APPG treatment group were due to PPNG infections . All of those cases were treated with Thiamphenicol regimen, 4 were cured and 6 lost for follow-up . Of 12 PPNG infections treated with Thiamphenicol, only 1 failed treatment and was cured with spectinomycin . Only few and mild side effects (diarrhea or dizziness) resulted from treatment . Thiamphenicol in single oral regimen is a low cost, effective therapy for uncomplicated gonococcal infection in males in the Central African Republic and may become potentially one of the recommended regimens if the prevalence of PPNG continues to increase .

Contracept Fertil Sex (Paris), 1983 May, 11(5), 719 - 24
{Oral contraceptive use and prevalence of infection with Chlamydia trachomatis in women (author's transl)}; Kinghorn GR et al.; PIP: 1080 nonpregnant women ages 16-34 years, presenting for the 1st time at a clinic for sexually transmitted diseases (STD), were examined and screened for infection with Chlamydia trachomatis, Neisseria gonorrheae, Trichomonas vaginalis, and Candida species . The respective prevalence rates were 21.1%, 20.7%, 13.4%, and 27.8% . Isolation rates for Chlamydia trachomatis, either occurring alone or in association with other genital infections, were significantly greater in women using oral contraceptives (OCs) . This was not because OC users were more promiscuous . The findings strengthen the case for providing a routine chlamydial culture service for women attending STD clinics . They also indicate that the likelihood of chlamydial infection in women taking OCs is greater . (author's)

Malays J Reprod Health, 1993 Jun, 11(1), 1 - 7
STD and contraception; Chandran R; PIP: Pelvic inflammatory disease (PID) is caused by several pathogens . It is difficult to diagnose on clinical grounds alone . The best diagnostic tool for PID is laparoscopy . Women using contraceptives tend to be more sexually active and to have multiple sex partners than their counterparts and hence are at a greater risk of sexually transmitted diseases (STDs) . Oral contraceptives (OCs) appear to protect against PID but may increase the risk of lower genital tract chlamydial infection . Among OC users who do develop PID, it is less severe than that of non-users . Three biases identified in early studies may account for the increased risk of PID in IUD users: inappropriate control groups, overdiagnosis of PID among IUD users, and failure to analyze the PID risk for specific IUD types (e.g., Dalkon Shield) . IUD users in a monogamous relationship face a low risk of STDs and PID . Data indicate that insertion of the IUD is what affects the PID risk in IUD users, not the IUD itself . Male condoms have been used since the times of the ancient Egyptians and Romans to protect against pregnancy and STDs . The increase in STDs and the AIDS epidemic has increased use of latex condoms . Condoms do fail sometimes (e.g., breakage rate, 1-12%) . The female condom has been developed to give the female partner greater control . It covers the perineal area and slips out of the vagina, not off into the vagina . Diaphragms and cervical caps prevent cervical and upper genital tract infection but not vaginal and external genital infection . They do not protect against HIV infection . The contraceptive sponge forms a mechanical barrier to the cervix, absorbs some of the semen, and releases a spermicide (nonoxynol-9) during intercourse . Nonoxynol-9 inactivates spermatozoa and, in vitro, Trichomonas vaginalis, Neisseria gonorrhoea, Treponema pallidum, chlamydia, herpes, and HIV . It can be used in conjunction with any barrier method .

J Biochem Mol Biol, 2002 May 31, 35(3), 330 - 6
Expression and characterization of beta-1,4-galactosyltransferase from Neisseria meningitidis and Neisseria gonorrhoeae; Park JE et al.; The lgtB genes that encode beta-1,4-galactosyltransferases from Neisseria meningitidis ATCC 13102 and gonorrhoeae ATCC 31151 were isolated by a polymerase chain reaction using the pfu DNA polymerase . They were expressed under the control of lac and T7 promoters in Escherichia coli M15 and BL21 (DE3) . Although the genes were efficiently expressed in E . coli M15 at 37 degrees C (33 kDa), most of the beta-1,4-galactosyltransferases that were produced were insoluble and proteolysed into enzymatically inactive polypeptides that lacked C-terminal residues (29.5 kDa and 28 kDa) during the purification steps . When the temperature of the cell growth was lowered to 25 degrees C, however, the solubility of the beta-1,4-galactosyltransferases increased substantially . A stable N-terminal his-tagged recombinant enzyme preparation could be achieved with E . coli BL21 (DE3) that expressed lgtB . Therefore, the cloned beta-1,4-galactosyltransferases were expressed under the control of the T7 promoter in E . coli BL21 (DE3), mostly to the soluble form at 25 degrees C . The proteins were easily purified to homogeneity by column chromatography using Ni-NTA resin, and were found to be active . The galactosyltransferases exhibited pH optimum at 6.5-7.0, and had an essential requirement for the Mn(+2) ions for its action . The Mg(+2) and Ca(+2) ions showed about half of the galactosyltransferase activities with the Mn(+2) ion . In the presence of the Fe(+2) ion, partial activation was observed with the beta-1,4-galactosyltransferase from N . meningitidis (64% of the enzyme activity with the Mn(+2) ion), but not from N . gonorrhoeae . On the other hand, the N(+2), Zn(+2), and Cu(+2) ions could not activate the beta-1,4- galactosyltransferase activity . The inhibited enzyme activity with the Ni(+2) ion was partially recovered with the Mn(+2) ion, but in the presence of the Fe(+2), Zn(+2), and Cu(+2) ions, the Mn(+2) ion could not activate the enzyme activities . Also, the beta-1,4-galactosyltransferase activity was 1.5-fold stimulated with the non-ionic detergent Triton X-100 (0.1-5 percent).

J Bras Ginecol, 1987 Apr, 97(4), 183 - 7
{Sexually transmitted agents in gynecology: incidence and importance}; Tomioka ES et al.; PIP: The study comprised the analysis of 77 patients between May 1985 and May 1986 in the emergency clinic of the Gynecological Clinic of the Faculty of Medicine of the University of Sao Paulo . Three different groups constituted the cases: A) 15 women devoid of infectious symptoms, aged 18-52 with an average of 31.5 years; B) 25 patients who were carriers of mucous purulent cervicitis, aged 16-42 with an average age of 27.6 years; and C) 37 carriers of acute salpingitis, aged 16-48 with an average age of 25 years . All patients had been sexually active, and none of them had used any local or systemic anti-infection medication for 30 days preceding the study . Pap smears were carried out in all patients . Laparoscopy indicated light salpingitis in 17 patients, moderate in 12, and grave in 8 patients . Neisseria gonorrhea was isolated from the endocervix of 14 women (37.8%) with acute salpingitis, and from none of the other 2 groups . Chlamydia trachomatis was found in 1 (6.7%), 3 (12%), and 14 (37.8%) patients in the Groups A, B, and C . Ureaplasma urealyticum was identified in 3 (20%), 9 (36%), and 21 (56.8%) in the respective groups . Mycoplasma hominis was isolated in 1 (6.7%), 1 (4%), and 2 (5.4%) patients in the respective groups . Cervical-vaginal cytology in the different groups broke down in this fashion; in Group A, 4 were normal (26.7%) and 11 were infectious (64.7%); in Group B, 3 were normal (12%) and 21 were infectious (84%), while 1 (4%) was suspected of dysplasia; and in Group C, 2 were normal (5.4%) and 31 were infectious (83%), while 4 (10.8%) were suspected of dysplasia . It is well known that colpocytological alterations are intimately related to sexual activity . These alterations may also have to do with cancer of the cervix . In view of the fact that the analyzed groups had been promiscuous, a high percentage of colpocytological alterations could be expected . Indeed, clear increases of inflammatory alterations and cervical displasias were found in the cervicitis and salpingitis groups .

Afr Health . 1994 May;16(4):40.
STDs: prevention and treatment {letter}; Aina JB; PIP: AIDS is no myth . It is preventable but there is no cure . But how much are we doing in Africa about those sexually transmitted diseases which are both preventable and curable? I feed sad, really sad, about the cases of men and women who engage in unprotected (without condoms) extra-marital sex and then wait for 2 to 3 weeks before they have sex with their spouses in the belief that, if they have no problems in the meantime, they must be free of infection . They are unaware that it is possible to harbor Neisseria gonorrhoea and other STD organisms in the genital system without having symptoms . Studies have shown that 10% of men and 60% of women infected with N . gonorrhoea have no symptoms . Some who resume a sexual relationship with their spouse after the 2 to 3 week 'watchful period' pass on an infection and then blame their innocent spouse for the consequences} We should seek to improve the knowledge of the public and to disabuse their faith in the watchful period . Another area where action is required concerns herpes genitals . Many doctors are not familiar with atypical presentations of this infection and, consequently, many cases are missed . In view of the importance of early detection and early commencement of therapy with acyclovir (Zovirax), I call upon Wellcome--as manufacturers of Zovirax--to publish and distribute in Africa a chart illustrating the various presentations of the infection . This would help prevent doctors from making such patients go from antibiotic to antibiotic for many months without cure . (full text) .

Indian J Sex Transm Dis, 1989, 10(2), 45 - 7
Penicillinase-producing Neisseria gonorrhoeae infection in India; Mishra D et al.; PIP: In 1986-1987, health workers at the Banaras Hindu University Hospital in Varanasi, India took gonococcal smears from both men and women so laboratory personnel could test for detection of penicillinase-producing Neisseria gonorrhoea (PPNG) . None of the cultures tested positive for PPNG . They were detected in 3 cases in Varanasi in 1983, however . Nevertheless PPNG strains remained rare in Varanasi . The 1st reported case of PPNG occurred in Madras in 1981 . In 1989, only 52 cases of PPNG had been reported nationwide . The researchers suggested that such a low yield of PPNG isolates may be due to underreporting of STDs in general and inadequate laboratories and equipment to culture gonococci at most centers . Furthermore, many health practitioners treat urethritis patients without laboratory confirmation and switch antibiotics at the 1st possible sign of treatment failure . The researchers proposed educating practitioners about wise use of recommended effective STD therapies to not deplete the antimicrobial reserve . In addition, researchers should conduct studies to determine the exact prevalence of PPNG strains in an area . They should also conduct studies on treatment failure rates with the 1st line drug of choice to monitor drug efficacy . For example, cure rates 90% should be unacceptable for gonorrhea . They concluded by advocating the provision of a test for penicillinase production at all centers in India .

Malays J Reprod Health, 1989 Dec, 7(2), 92 - 8
Commentary: family planning and sex education in STD; Ang Eng Suan; PIP: The incidence of Sexually Transmitted Diseases (STD) has risen in the past years to epidemic proportions even though medical understanding of STDs has considerably improved . The most common forms of contraception (e.g . condoms, diaphragms, vaginal spermicides, intrauterine devices, and oral contraception pills) vary in how effective each is in preventing STDs . Condoms offer almost complete protection against many STDs, including, gonorrhoea, syphilis, herpes virus, trichomonas, candidiases, chlamydia, and AIDS . Vaginal spermicides have a range of effectiveness against Neisseria gonorrhoea, Treponema pallidum, Trichomonas vaginalis, Herpes vaginalis, and Candida albicans . There are conflicting reports on correlating usage of IUDs and a higher incidence of pelvic inflammatory disease . Oral contraceptive pills have a varied effect on STDs . Long-time users of the pill are 1/2 as likely to develop pelvic inflammatory disease but pill users are also more prone to yeast infections . This article offered suggestions on how to improve usage of contraception as a protection against STDs . The message that any sexually active individual with multiple partners or one who is unsure about their partner should protect against STDS needs to be more broadly disseminated by using mass media . Firstly, people need to be made more comfortable with the discussion of STDs for any message to be effective . Family planning services should expand their promotion of appropriate protective contraception for high risk groups .

J Bras Ginecol, 1987 Jul, 97(7), 315 - 6
{A comparative study of the incidence of Gardnerella vaginalis in users of IUD and oral contraceptives}; Aleixo Neto A et al.; PIP: Leukorrhea constitutes one of the most frequent complaints by women visiting out-patient gynecological clinics . The most common etiological agents are Gardnerella vaginalis, Trichomonas vaginalis, Neisseria gonorrhoea and Candida albicans . Some authors have been able to verify an increased presence of certain pathogenic germs in the vaginal flora for users of contraceptive methods, e.g., for users of IUD, Kivijarvi et al . demonstrated a significantly increased presence of Gardnerella vaginalis considered responsible for a large number of inflammatory diseases of the pelvis in woman . In order to establish the frequency of colpo-cervicitis in users of contraceptive methods a study was initiated of women attending the Family Planning Clinic of the Department of Medicine at UFMG during the period January-December 1986 . 305 cytological smears obtained from 111 users of oral hormonal contraceptives and from 194 users of IUD were studied by using the Papanicolaou staining method . A microscopic examination enabling the detection of cells that indicate the presence of Gardnerella vaginalis, trichomonas, Candida, papilloma, and dysplastic cells was performed . The results are listed in two tables . In 34.5% of cases a significant association was established between users of IUD and cells indicating the presence of Gardnerella vaginalis . For users of oral contraceptives that figure was lower, 28.8% . This circumstance has been attributed to probable epithelial lesions caused by incorrect insertion of the IUD and more abundant menstrual flux provoked b the IUD in combination with the hemophilic characteristics of Gardnerella vaginalis . A significant prevalence of papilloma and trichomonas was also noted for women using IUD as opposed to those oral contraceptives . The results demonstrate the necessity of frequent clinical attendance and laboratory tests for women using contraceptive methods, particularly the IUD .

Adv Contracept Deliv Syst, 1986 Jun, 2(2-3), 112 - 58
The rise and fall of IUDS's: banning of IUD's in USA . Alarming interactions between IUD's and sexually transmitted diseases (STD); Krzyzaniak LT et al.; PIP: This supplement presents an extensive review of patterns of IUD use in the US, unanswered controversies related to the role of the IUD in pelvic inflammatory disease (PID) and sexually transmitted diseases, and the ethical and mediciolegal aspects of IUD marketing . Also included is a section on an Adolescent Festival scheduled to be held in Turkey in June 1986 which will focus on beginning a dialog between adolescents and parents, educators, and physicians . The text is amply illustrated with figures and includes 10 tables . There has been a constant decline in the number of IUD users in the US, from 2.2 million in 1981 to 1.4 million in 1985 . Many women asked to have their IUDs removed after successive withdrawals of 1st-generation IUDs from the US market . It has been well established that users of copper IUDs are at higher risk for pelvic infection, especially nulligravidas under 25 years of age . The PID seen in IUD users is caused by microbiological contaminants such as Chlamydia trachomatis, ureaplasma urealyticum, and Neisseria gonorrhoeae . The constant presence of the IUD tail in the cervical canal and vagina is associated with transuterine migration of virus, bacteria, and fungus to the upper urogenital tract . In addition, the microbiologic contaminants in semen colonize on selected IUD parts . The lack of sophisticated evaluation of types of IUDs has been in part a reflection of insufficient testing on the part of IUD manufacturers who have a financial interest in study results . The ethical aspects of IUD use should be considered in relation to religion, ethnic groups, cultural patterns, government regulations, the role of gynecologists, family planning centers, marketing promotion, package inserts, and prevention of adolescent pregnancy . A series of IUD Documentation workshops are planned to assess the current status of this contraceptive method .

Med Mal Infect, 1983, 13 Spec No, 677 - 8
{Systematic detection of Neisseria gonorrhoeae in an abortion center}; Quentin R et al.; PIP: The existence of asymptomatic gonorrhea, the difficulties of clinical diagnosis, and the risk of uteroadnexial gonococcal infection following abortion prompted a campaign to diagnose N . gonorrhoeae among women seeking abortions at the Center for Pregnancy Interruption in Tours . 1446 women aged 14-46 years were examined between October 1981 and December 1982 . Women having positive cultures received antibiotic treatment for a week and the abortion was not performed until a negative culture was obtained . 15 women, or 1%, had positive cultures . 1 of the 15 positive cases, but only 45.8% of patients overall, were 26 years old or younger . 10 of the 15 cases but only 37% of all clients were nulliparous . 12 of the 15 cases but only 38.6% of patients overall were single, divorced, or separated . 1 of the 15 cases belonged to a "non-active" social category of unemployed, students, and women without professions, compared to 45.5% of all clients . 13 of the cases had no clinical signs of genital infection but 3 of their partners had urethritis . 2 cases had purulent leukorrhea . Leukorrhea was suspected in 8.6% of the women with negative cultures . This practice should be adopted in all abortion centers because of its simplicity and low cost .

J Bacteriol, 2002 Oct, 184(20), 5619 - 24
Overexpression of the MtrC-MtrD-MtrE efflux pump due to an mtrR mutation is required for chromosomally mediated penicillin resistance in Neisseria gonorrhoeae; Veal WL et al.; The importance of the mtrCDE-encoded efflux pump in conferring chromosomally mediated penicillin resistance on certain strains of Neisseria gonorrhoeae was determined by using genetic derivatives of penicillin-sensitive strain FA19 bearing defined mutations (mtrR, penA, and penB) donated by a clinical isolate (FA6140) expressing high-level resistance to penicillin and antimicrobial hydrophobic agents (HAs) . When introduced into strain FA19 by transformation, a single base pair deletion in the mtrR promoter sequence from strain FA6140 was sufficient to provide high-level resistance to HAs (e.g., erythromycin and Triton X-100) but only a twofold increase in resistance to penicillin . When subsequent mutations in penA and porIB were introduced from strain FA6140 into strain WV30 (FA19 mtrR) by transformation, resistance to penicillin increased incrementally up to a MIC of 1.0 micro g/ml . Insertional inactivation of the gene (mtrD) encoding the membrane transporter component of the Mtr efflux pump in these transformant strains and in strain FA6140 decreased the MIC of penicillin by 16-fold . Genetic analyses revealed that mtrR mutations, such as the single base pair deletion in its promoter, are needed for phenotypic expression of penicillin and tetracycline resistance afforded by the penB mutation . As penB represents amino acid substitutions within the third loop of the outer membrane PorIB protein that modulate entry of penicillin and tetracycline, the results presented herein suggest that PorIB and the MtrC-MtrD-MtrE efflux pump act synergistically to confer resistance to these antibiotics.

Biotechniques, 2002 Sep, 33(3), 526 - 31
Determination of methylation site of DNA-methyltransferase NlaX by a hybrid method; Kubareva EA et al.; Using a new method based on a combination of bisulfite reaction, the repair enzyme uracil-DNA glycosylase, and synthetic oligodeoxyribonucleotides, the methylation site of DNA-methyltransferase NlaX (M.NlaX) from Neisseria lactamica was established to be the inner cytosine in the double-stranded pentanucleotide recognition sequence 5'-CCNGG-3' (where N = any nucleoside) . 5-Methylcytosine (m5C) type modification by M-N1aX was confirmed by the use of oligonucleotide substrates that contain 5-fluoro-2'-deoxycytidine.

Sex Transm Infect, 2002 Jun, 78(3), 208 - 9
Epidemiology and natural history of ligase chain reaction detected chlamydial and gonococcal infections; Kissin DM et al.; OBJECTIVES: Ligase chain reaction (LCR) technology has dramatically increased the sensitivity of tests for sexually transmitted infections (STIs) . It is unknown whether low copy infections (LCR positive, culture negative) have any clinical consequences . We assessed the clinical significance of untreated low copy Chlamydia trachomatis and Neisseria gonorrhoeae infections in a cohort of sexually active women . METHODS: We studied a cohort of sexually active women followed at 6 month intervals for up to 3 years . Frozen urine specimens from 181 women with negative cultures for C . trachomatis and N . gonorrhoeae who were 'high risk' (defined as being less than 40 years old at baseline, and having either Trichomonas vaginalis at baseline or a history of more than one sexual partner during the 12 months before baseline) were tested for C . trachomatis and N . gonorrhoeae by LCR (Abbott Laboratories, Abbott Park, IL, USA) . The specimens from all visits for each person were pooled and LCR was performed on the pool . Laboratory results were linked to clinical information . We also tested all urine samples obtained from patients with a positive culture . RESULTS: 10 additional infections (nine C . trachomatis and one N . gonorrhoeae) were detected with LCR technique . None of the women with low copy infection had evidence of subsequent pelvic inflammatory disease or ectopic pregnancy . Pooling of urine samples resulted in a 47% decline in the number of tests performed . CONCLUSIONS: Additional STIs can be identified when using LCR . Pooling of urine specimens is a cost saving technique for C . trachomatis and N . gonorrhoeae testing.

Sex Transm Infect, 2002 Jun, 78(3), 190 - 3
Prevalence of chlamydia and gonorrhoea among a population of men who have sex with men; Cook RL et al.; OBJECTIVES: Few data are available on the prevalence of sexually transmitted diseases (STDs) in men who have sex with men (MSM), making it difficult to develop STD screening guidelines for this population . The objective of the study was to determine the prevalence of urethral infections caused by Chlamydia trachomatis and Neisseria gonorrhoeae within a large, community based population of MSM, and to assess the feasibility of rectal screening in this population . METHODS: This was a cross sectional study of 566 MSM, who were predominantly middle aged, white, asymptomatic, and engaged in sex with multiple partners . All provided a urine sample to screen for chlamydial and gonorrhoea infections using a PCR assay; rectal screening was performed on 48 participants . RESULTS: Urethral C . trachomatis infections were detected in 1/566 participants (prevalence 0.2%, 95% CI 0.004% to 1.0%), and rectal C . trachomatis infections were detected in 2/48 men (prevalence 4.2%, 95% CI 0.5% to 14.2%) . No gonorrhoea infections were detected, and none of the 117 HIV positive men had either infection . CONCLUSIONS: Chlamydial and gonorrhoea infections were uncommon in this sample of MSM, even among those with multiple sexual partners or HIV infection . These data call into question recommendations to screen all MSM based on their individual sexual behaviours or HIV . Additional data are needed on the prevalence of these infections in MSM from different settings.

Sex Transm Infect, 2002 Jun, 78(3), 187 - 9
The feasibility of integrated STI prevalence and behaviour surveys in developing countries; MacLachlan EW et al.; BACKGROUND: In countries where STI/HIV prevalence data and behavioural data are scarce UNAIDS second generation HIV surveillance guidelines recommend measuring STI/HIV prevalence and risk behaviours in vulnerable populations but do not recommend conducting these surveys concurrently because of concerns about participation rates, cost, and provision of services . OBJECTIVES: To assess the feasibility of conducting a national combined STD prevalence and behaviour survey in Mali among vulnerable populations with the intention of institutionalisation . METHODS: From March to June 2000 an integrated STI prevalence and behaviour survey was conducted using cluster sampling among five risk groups in four sites in Mali, west Africa . 2229 individuals in non-traditional settings such as taxi/bus stations, market areas, households, and brothels participated in any one or all components of the study: (1) behavioural questionnaire, (2) urine sample for Neisseria gonorrhoeae (GC)/Chlamydia trachomatis (CT) testing, (3) a fingerstick drop of blood for syphilis, and/or (4) HIV testing . RESULTS: High participation rates of 84%-100% were achieved despite specimen collection and HIV testing . Rates fell only slightly when participants were asked to provide biological samples and participants were more likely to provide urine than blood . Rates among the different groups for HIV and syphilis testing are similar and suggest that refusal was most probably because of a reluctance to give blood rather than because of HIV testing . The cost of the biological component added approximately $30 per participant . Included in the $30 are the costs of training, participant services, laboratory personnel and supplies, STI drugs, and STI testing costs . The total cost of the survey was $154,905 . Biomarkers aided in validation of answers to behavioural questions . Consenting individuals received HIV pretest and post-test counselling and referral to a trained health provider for treatment of STI and the provision of services provided the framework for interventions in the groups following the survey . CONCLUSION: This represents an effective methodology for collecting risk behaviour and STI/HIV prevalence information concurrently and should be considered by countries expanding STI/HIV surveillance as part of UNAIDS second generation HIV surveillance.

Infect Immun, 2002 Oct, 70(10), 5808 - 15
Immortalization of human urethral epithelial cells: a model for the study of the pathogenesis of and the inflammatory cytokine response to Neisseria gonorrhoeae infection; Harvey HA et al.; The primary human urethral epithelial cells developed by our laboratory have been immortalized by transduction with a retroviral vector expressing the human papillomavirus E6E7 oncogenes . Analysis of telomerase expression and comparison to that in primary cells revealed detectable levels in the transduced human urethral epithelial cells . Immortalized urethral cells could be passaged over 20 times . Immunofluorescence microscopy studies showed that the immortalized cells were phenotypically similar and responded to gonococcal infection similarly to primary cells . Specifically, positive cytokeratin staining showed that the immortalized cells are keratinocytes; cell surface levels of human asialoglycoprotein receptor increase following gonococcal infection, and, like the primary cells, the immortalized urethral epithelial cells are CD14 negative . Using enzyme-linked immunosorbent assay, we found that interleukin-6 (IL-6) and IL-8 levels in primary urethral epithelial cell supernatants increase after challenge with N . gonorrhoeae . Likewise, the immortalized urethral epithelial cells produced higher levels of IL-6 and IL-8 cytokines in response to gonococcal infection . Cells challenged with a gonococcal lipid A msbB mutant produced reduced IL-6 and IL-8 levels when compared to the parent strain . Additionally, these data suggest that the 1291 msbB lipooligosaccharide may suppress cytokine induction.

Obstet Gynecol, 2002 Sep, 100(3), 579 - 84
Predicting chlamydial and gonococcal cervical infection: implications for management of cervicitis; Marrazzo JM et al.; OBJECTIVE: To define utility of age and cervical findings in predicting infection with Chlamydia trachomatis and Neisseria gonorrhoeae among women universally tested for both infections, and to assess the independent contribution of Gram stain (GS) smear of endocervical secretions . METHODS: Visits by women to Seattle sexually transmitted diseases clinics from 1995 through 1999 were retrospectively reviewed . All women had endocervical GS and cultures for C trachomatis and N gonorrhoeae performed . Predictive values of age, cervical signs, and inflammation on GS (more than 30 polymorphonuclear leukocytes per 1000x field) were calculated . RESULTS: Among 6230 women, prevalence of C trachomatis and N gonorrhoeae was 6.9% and 2.1%, respectively; 520 women (8.3%) had either organism detected . Age, cervical signs (mucopus, induced bleeding), and inflammation on endocervical GS were independently associated with infection . However, the positive predictive value (PPV) of any cervical finding for infection was less than 19% in women 25 years and older . Inflammation on endocervical GS was the sole indicator of infection in 79 of 520 (15%) infections, but was insensitive in the absence of mucopurulent cervicitis (sensitivity, 26%; PPV, 21%) . CONCLUSION: Cervical signs suggesting chlamydial or gonococcal infection have higher positive predictive value (PPV) in younger women . The PPV of inflammation on endocervical GS is too low to recommend its use to direct empiric treatment in the absence of mucopurulent cervicitis, especially in women 25 years and older . Further, its low sensitivity in detecting infection in women without mucopurulent cervicitis does not justify routine use . Signs suggesting mucopurulent cervicitis should be interpreted in the context of age, and empiric treatment may not be indicated in women aged 25 years and older.

Obstet Gynecol, 2002 Sep, 100(3), 456 - 63
Lower genital tract infection and endometritis: insight into subclinical pelvic inflammatory disease; Wiesenfeld HC et al.; OBJECTIVE: To investigate the association between lower genital tract infections and subclinical PID . Fallopian tube damage is a common complication of acute symptomatic pelvic inflammatory disease (PID), yet most women with tubal factor infertility do not have a history of acute PID . Subclinical PID is believed to be an important cause of tubal factor infertility . METHODS: We conducted a cross-sectional study among women attending a sexually transmitted diseases or ambulatory gynecology clinic . A convenience sample of 556 women with bacterial vaginosis, gonorrhea, or chlamydia, or women at risk for gonorrhea or chlamydia were enrolled . Women diagnosed with acute PID were not eligible to participate . The main outcome was subclinical PID, as defined by the presence of histologic endometritis . RESULTS: Subclinical PID was more common in women with lower genital tract infection than in uninfected women . Subclinical PID was present in 27% of women with Chlamydia trachomatis (odds ratio 3.4; 95% confidence interval {CI} 1.8, 6.3) and in 26% of women infected with Neisseria gonorrhoeae (odds ratio 2.4; 95% CI 1.1, 5.1) . Among women with bacterial vaginosis, 15% had endometritis (odds ratio 2.7; 95% CI 1.02, 7.2) . CONCLUSION: Subclinical PID is common among women with lower genital tract infections . Additional prospective studies are necessary to determine the reproductive impact of these asymptomatic upper genital tract infections.

FEBS Lett, 2002 Sep 11, 527(1-3), 67 - 70
Maltooligosaccharide disproportionation reaction: an intrinsic property of amylosucrase from Neisseria polysaccharea; Albenne C et al.; Amylosucrase from Neisseria polysaccharea (AS) is a remarkable transglycosidase of family 13 of the glycoside hydrolases that catalyses the synthesis of an amylose-like polymer from sucrose and is always described as a sucrose-specific enzyme . Here, we demonstrate for the first time the ability of pure AS to catalyse the disproportionation of maltooligosaccharides by cleaving the alpha-1,4 linkage at the non-reducing end of a maltooligosaccharide donor and transferring the glucosyl unit to the non-reducing end of another maltooligosaccharide acceptor . Surprisingly, maltose, maltotriose and maltotetraose are very poor glucosyl donors whereas longer maltooligosaccharides are even more efficient glucosyl donors than sucrose . At least five glucose units are required for efficient transglucosylation, suggesting the existence of strong binding subsites, far from the sucrose binding site, at position +4 and above.

Commun Dis Intell, 2002, 26(2), 242 - 7
Annual report of the Australian Gonococcal Surveillance Programme, 2001; A cost-effectiveness evaluation of testing and treatment of Chlamydia trachomatis infection among asymptomatic women infected with Neisseria gonorrhoeae; Division of STD Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USABACKGROUND: Because patients infected with Neisseria gonorrhoeae are frequently coinfected with Chlamydia trachomatis, routine dual treatment of patients with N gonorrhoeae infection is frequently practiced and has long been recommended . GOAL: The goal of this study was to examine the cost-effectiveness of routine dual treatment of women with infection, with or without separate testing for C trachomatis, compared with an alternative of testing for both infections and restricting treatment for C trachomatis to women testing positive for C trachomatis . STUDY DESIGN: A decision analysis compared the cost-effectiveness of these options using cases of pelvic inflammatory disease prevented as the outcome . Parameter values were taken from the literature . RESULTS: Routine dual treatment is not an effective or cost-effective replacement for testing for C trachomatis, but it can increase the number of cases of C trachomatis treated when combined with testing . Dual treatment results in more overtreatment of infection C trachomatis than treatment based on test results . CONCLUSIONS: Testing for both infections is more cost-effective than routine presumptive treatment for C trachomatis . Providing both presumptive treatment and testing for C trachomatis can also be cost-effective in some settings.

Glycobiology, 2002 Sep, 12(9), 523 - 33
Neisseria gonorrhoeae strain PID2 simultaneously expresses six chemically related lipooligosaccharide structures; Tong Y et al.; Neisseria gonorrhoeae strain PID2 was isolated from a woman suffering from pelvic inflammatory disease . When LOS expressed by this strain is analyzed on SDS-PAGE gels, at least six different lipooligosaccharide (LOS) components are visualized . We characterized the LOSs made by this strain by exoglycosidase digestion, sugar composition analysis, mass spectrometry, and analysis of the genes needed for its synthesis . DNA sequence analysis showed that the lgt gene cluster in this strain has undergone a rearrangement and that it possesses two copies of lgtA, one copy of lgtB and lgtC, and a hybrid gene containing sequences from lgtB and lgtE . We determined that the hybrid lgtB/E gene retained the lgtE gene function . DNA sequence analysis of the gene organization suggested that an intramolecular recombination between lgtA and lgtD and lgtB and lgtE had occurred via homologous recombination between similar sequences . Our studies demonstrated that fluorophore-assisted carbohydrate electrophoresis can be utilized to rapidly determine the composition of LOS . By combining exoglycosidase digestion, in combination with mass spectrometry analysis and compositional analysis, the data indicate that all of the LOS components produced by PID2 extend off of the alpha chain . The longest alpha chain oligosaccharide structure is Gal-GlcNAc-Gal-GlcNAc-Gal-Glc-Heptose I, and the six LOS components are built up by sequentially adding sugars onto the first heptose . PID2 LOS is the first Neisserial LOS to be shown to be devoid of phosphoethanolamine modifications . Because PID2 can surface express its LOS, it indicates that the addition of phosphoethanolamine is not required for LOS surface expression.

MMWR Morb Mortal Wkly Rep . 2002 Aug 16;51(32):709.
Recall of LCx Neisseria gonorrhoeae assay and implications for laboratory testing for N . gonorrhoeae and Chlamydia trachomatis; Identification of Xenorhabdus nematophila genes required for mutualistic colonization of Steinernema carpocapsae nematodes; Department of Bacteriology, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USAOne stage in the symbiotic interaction between the bacterium Xenorhabdus nematophila and its nematode host, Steinernema carpocapsae, involves the species-specific colonization of the nematode intestinal vesicle by the bacterium . To characterize the bacterial molecular determinants that are essential for vesicle colonization, we adapted and applied a signature-tagged mutagenesis (STM) screen to this system . We identified 15 out of 3000 transposon mutants of X . nematophila with at least a 15-fold reduction in average vesicle colonization . These 15 mutants harbour disruptions in nine separate loci . Three of these loci have predicted open reading frames (ORFs) with similarity to genes (rpoS, rpoE, lrp) encoding regulatory proteins; two have predicted ORFs with similarity to genes (aroA, serC) encoding amino acid biosynthetic enzymes; one, designated nilB (nematode intestine localization), has an ORF with similarity to a gene encoding a putative outer membrane protein (OmpU) in Neisseria; and three, nilA, nilC and nilD, have no apparent homologues in the public database . nilA, nilB and nilC are linked on a single 4 kb locus . nilB and nilC are > 104-fold reduced in their ability to colonize the nematode vesicle and are predicted to encode membrane-localized proteins . The nilD locus contains an extensive repeat region and several small putative ORFs . Other than reduced colonization, the nilB, nilC and nilD mutants did not display alterations in any other phenotype tested, suggesting a specific role for these genes in allowing X . nematophila to associate with the nematode host.

Nucleic Acids Res, 2002 Sep 1, 30(17), 3839 - 47
Functional analysis of iceA1, a CATG-recognizing restriction endonuclease gene in Helicobacter pylori; Xu Q et al.; iceA1 in Helicobacter pylori is a homolog of nlaIIIR, which encodes the CATG-specific restriction endonuclease NlaIII in Neisseria lactamica . Analysis of iceA1 sequences from 49 H.pylori strains shows that a full-length NlaIII-like ORF is present in 10 strains, including CH4, but in other strains, including strain 60190, the ORFs are truncated due to a variety of mutations . Our goal was to determine whether iceA1 can encode a NlaIII-like endonuclease . Overexpression in Escherichia coli of iceA1 from CH4, but not from 60190, yielded NlaIII-like activity, indicating that the full-length iceA1 is a functional endonuclease gene . Repair of the iceA1 frameshift mutation in strain 60190 and its expression in E.coli yielded functional NlaIII-like activity . We conclude that iceA1 in CH4 is a functional restriction endonuclease gene, while iceA1 in 60190 is not, due to a frameshift mutation, but that its repair restores its restriction endonuclease activity.

Indian J Med Res, 2002 Mar, 115, 113 - 7
Rising quinolone resistance in Neisseria gonorrhoeae isolates from New Delhi; Bhalla P et al.; BACKGROUND & OBJECTIVES: Treatment for gonorrhoea with fluoroquinolones is recommended . However, reduced susceptibility and treatment failure with fluoroquinolones has recently been reported . We undertook to study the antibiotic susceptibility pattern and the incidence of quinolone resistance in 36 consecutive isolates of Neisseria gonorrhoeae from April to November 2000 . METHOD: Antibiotic susceptibility testing was performed by the Kirby Bauer disc diffusion technique and minimum inhibitory concentration (MIC) of ciprofloxacin was determined by the agar dilution method . Penicillinase producing N . gonorrhoeae (PPNG) were identified by using the nitrocefin disc method . RESULTS: Thirty six strains of N . gonorrhoeae obtained from 44 consecutive male patients (81.9%) were studied . By the disc diffusion method, only 3 (8.3%) of these isolates were found to be sensitive to ciprofloxacin . All isolates were sensitive to ceftriaxone while 23 (63.9%) were sensitive to tetracycline and 12 (33.3%) to penicillin . Four (11.1%) of the N . gonorrhoeae isolates were PPNG . Twenty seven (75%) isolates were found to be resistant to ciprofloxacin by MIC determination . INTERPRETATION & CONCLUSION: Incidence of ciprofloxacin resistance amongst N . gonorrhoeae isolates is on the rise in New Delhi . Periodic monitoring of antimicrobial susceptibility pattern of N . gonorrhoeae to antimicrobials other than quinolones is essential to prevent treatment failure in patients with gonorrhoea.

Ukr Biokhim Zh, 2002 Jan-Feb, 74(1), 49 - 53
{Generation of proton motive force (Delta mu H+) in Neisseria gonorrhoeae cells}; Skliar TV; The value of the proton motive force in the gonococci cells under incubation medium pH changing from 5 to 8 was equal to 183-192 mB . The membrane potential changed in the limits from 103 to 145 mB, while the hydrogen ions concentration gradient (delta pH) from 47 to 90 mB . The character of phenyldicarbaundecaborane absorption by the N . gonorrhoeae vesicules displays the presence of two membrane potential generators presence: respiratory chain and H(+)-ATPase . It is shown, that the inhibitors of the energy processes KCN, DCCD, CCF cause the suppression of proton motive force generators and membrane potential dissipation It is marked, that in the gonococci strains resistant to antibiotics the membrane potential is higher, than at the sensitive ones.

J Infect Dis, 2002 Sep 15, 186(6), 792 - 7 Epub 2002 Aug 28.
Association between elevated neutrophil defensin levels and endometritis; Wiesenfeld HC et al.; The role of host defenses in the pathogenesis of pelvic inflammatory disease (PID) remains largely uncharacterized . The antimicrobial peptides defensins are important components of innate host defense . To explore the relationship between neutrophil defensins and upper genital tract infection, 377 women who were at risk for PID were enrolled in a study . Women infected with Neisseria gonorrhoeae, Trichomonas vaginalis, or Chlamydia trachomatis had higher median levels of neutrophil defensins (human neutrophil peptides 1-3) in the vagina than did uninfected women . Neutrophil defensins were strongly associated with the presence of endometritis after the analysis was controlled for the presence of sexually transmitted diseases . Vaginal neutrophils were associated with endometritis only in the presence of elevated defensin levels, which highlights the importance of neutrophil activation, rather than the presence of neutrophils alone, in this inflammatory process . Neutrophil defensins appear to participate in the host defense in ascending pelvic infection and the pathogenesis of PID.

Acta Crystallogr D Biol Crystallogr, 2002 Sep, 58(Pt 9), 1467 - 9 Epub 2002 Aug 23.
Crystallization and preliminary X-ray diffraction studies of the peptide methionine sulfoxide reductase B domain of Neisseria meningitidis PILB; Kauffmann B et al.; Peptide methionine sulfoxide reductases (Msr) are ubiquitous enzymes that catalyse the reduction of free and protein-bound methionine sulfoxide back to methionine via sulfenic acid chemistry . Two classes of Msrs, MsrA and MsrB, have been described . The fact that the two Msrs display opposite stereoselectivities and have no sequence identity suggests that there is no structural similarity between the two classes . No three-dimensional structure of a MsrB is known . In the present report, the MsrB subdomain of Neisseria meningitidis PILB was used to grow orthorhombic crystals by the hanging-drop vapour-diffusion technique . The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 44.0, b = 118.6, c = 138.5 A . Crystals of selenomethionine-substituted MsrB were grown under the same conditions in order to use the MAD method for structure determination . Three diffraction data sets at 1.8 A resolution were collected . The positions of the Se atoms were determined and should result in a full structure determination.

Int J STD AIDS, 2002 Aug, 13(8), 568 - 72
Pelvic inflammatory disease in the People's Republic of China: aetiology and management; Hong S et al.; OBJECTIVES: To study the causes of pelvic inflammatory disease (PID) in Shenyang, Northeastern China, and to assess the efficacy of the syndromic management flowchart for PID recommended by World Health Organization (WHO) . METHODS: 200 outpatients with PID attending Gynaecologic Clinic of No . 1 and No . 2 hospital of China Medical University in Shenyang, and 155 control women without symptoms underwent pelvic examination, and were tested for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma hominis (MH), bacterial vaginosis (BV), Candida albicans (CA) and Trichomonas vaginalis (TV) . RESULTS: In PID patients, the prevalence of MH was 26%, of BV 26%, of CT 16%, of CA 11%, of TV 4% and of NG 2.5% . In the control population, prevalences were 5.2%, 8.4%, 0%, 5.2%, 0.7%, 0% respectively . MH, BV and CT were significantly more common in PID patients than in controls (P<0.01) . One hundred and thirty-seven of the 200 patients with PID (68.5%) returned for follow-up . Symptoms had resolved completely in 64 (47%), and had improved in 68 (50%) . There was no improvement in five (4%) . CONCLUSIONS: The management protocol for female lower abdominal pain recommended by WHO is effective in this setting.

Int J STD AIDS, 2002 Aug, 13(8), 564 - 7
High incidence of treatment failure of Neisseria gonorrhoeae isolates to ciprofloxacin in male gonococcal urethritis in Delhi; Chowdhry S et al.; Several treatment failures and widespread antimicrobial resistance to ciprofloxacin have been documented, subsequent to its initial recommendation in 1989 as a single dose alternative therapy for gonorrhoea . Still, it continues to be part of various treatment schedules in National STD control guidelines including India . This prompted us to study the current status of this drug in the treatment of gonorrhoea . Thirty-five male patients with gonococcal urethritis were included in the study . The susceptibility to penicillin, tetracycline, ciprofloxacin and ceftriaxone was determined by Kirby-Bauer disc diffusion method and minimum inhibitory concentration (MIC) of ciprofloxacin by agar plate dilution method . The clinical and bacteriological response was assessed on day 5 after treatment with single dose ciprofloxacin, 500 mg . The sensitivity pattern of Neisseria gonorrhoeae was observed to be: ceftriaxone 100%, azithromycin 100%, tetracycline 65.7%, penicillin 40% and ciprofloxacin 5.7% by disc diffusion method . The MIC for ciprofloxacin was below 0.06 microg/mL (sensitive) in one (2.5%) isolate only . On the fifth day a large number of treatment failures (88.5%) were seen with ciprofloxacin while none was noted one week after re-treatment with ceftriaxone . The location of endemic quinolone-resistant N . gonorrhoeae (QRNG) in New Delhi has increased alarmingly, resulting in an extremely high proportion of therapeutic failures, and thus requiring appropriate alterations in the presently recommended treatment regimens.

MMWR Recomm Rep, 2002 May 10, 51(RR-6), 1 - 78
Sexually transmitted diseases treatment guidelines 2002 . Centers for Disease Control and Prevention; Mutation in 23S rRNA associated with macrolide resistance in Neisseria gonorrhoeae; National Laboratory for Sexually Transmitted Diseases, National Microbiology Laboratory, Population and Public Health Branch, Health Canada, Winnipeg, Manitoba, Canada . Lai_King_Ng@hc-sc.gc.ca

Fifty-six azithromycin-resistant (MICs, 2.0 to 4.0 micro g/ml) Neisseria gonorrhoeae strains with cross-resistance to erythromycin (MICs, 2.0 to 64.0 micro g/ml), isolated in Canada between 1997 and 1999, were characterized, and their mechanisms of azithromycin resistance were determined . Most (58.9%) of them belonged to auxotype-serotype class NR/IB-03, with a 2.6-mDa plasmid . Based on resistance to crystal violet (MICs >or= 1 micro g/ml), 96.4% of these macrolide-resistant strains appeared to have increased efflux . Nine of the eleven strains selected for further characterization were found to have a promoter region mtrR mutation, a single-base-pair (A) deletion in the 13-bp inverted repeat, which is believed to cause overexpression of the mtrCDE-encoded efflux pump . The two remaining macrolide-resistant strains (erythromycin MIC, 64.0 micro g/ml; azithromycin MIC, 4.0 micro g/ml), which did not have the mutation in the mtrR promoter region, were found to have a C2611T mutation (Escherichia coli numbering) in the peptidyltransferase loop in domain V of the 23S rRNA alleles . Although mutations in domain V of 23S rRNA alleles had been reported in other bacteria, including E . coli, Streptococcus pneumoniae, and Helicobacter pylori, this is the first observation of these mutations associated with macrolide resistance in N . gonorrhoeae.

Antimicrob Agents Chemother, 2002 Sep, 46(9), 2811 - 20
Identification and analysis of amino acid mutations in porin IB that mediate intermediate-level resistance to penicillin and tetracycline in Neisseria gonorrhoeae; Olesky M et al.; PenB is the third resistance determinant in the stepwise acquisition of multiple resistance genes in chromosomally mediated resistant Neisseria gonorrhoeae (CMRNG) . Alterations in por(IB), one of two alleles at the por locus that encodes the outer membrane protein porin IB (PIB), were recently reported to be responsible for the increased resistance to penicillin and tetracycline conferred by penB, but the specific mutations conferring antibiotic resistance were not identified experimentally . To determine which amino acids in PIB confer increased resistance, we transformed a recipient strain with chimeras of the por(IB) genes from strains FA1090 and FA140 (penB2) . These studies revealed that two amino acid changes, G120D and A121D, were both necessary and sufficient to confer increased resistance to penicillin and tetracycline . Site-saturation and site-directed mutagenesis of Gly-120 and Ala-121 revealed that both a single mutation, G120K, and the double mutations G120R A121H and G120P A121P also conferred antibiotic resistance to the recipient strain . The identical mutations in PIA increased penicillin and tetracycline resistance either moderately or not at all . Analysis of por(IB) genes present in the GenBank database from 51 clinical isolates demonstrated that lysine and aspartate mutations at positions 120 and/or 121 also occur in nature . These studies demonstrate that charged amino acids at positions 120 and 121 in PIB are highly preferential for conferring resistance to penicillin and tetracycline in N . gonorrhoeae.

Sex Transm Infect, 2002 Aug, 78(4), 278 - 81
The diagnosis of chlamydia, gonorrhoea, and trichomonas infections by self obtained low vaginal swabs, in remote northern Australian clinical practice; Garrow SC et al.; OBJECTIVE: To examine the diagnostic performance of self obtained low vaginal swabs (SOLVS) and polymerase chain reaction (PCR) techniques in the diagnosis of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV) infection in a variety of clinical practice settings in remote north western Australia . DESIGN: A cross sectional field study of microbiological collection techniques in women undergoing gynaecological investigation in remote settings performed by a variety of practitioner types over 10 months . Participants and setting: 349 women from remote towns and communities in the Kimberley region of north west Western Australia having gynaecological examinations for clinical reasons, well women screening, antenatal screening, and sexual health examinations . RESULTS: The overall prevalence of infection in the study population based on any positive conventional sample was 9.2%, 7.6%, and 16.1% for CT, NG, and TV respectively . The detection rates for CT and NG by SOLVS were 89% and 96% respectively, compared with 79% and 91% for endocervical swabs and 79% and 83% for first void urine . SOLVS had a sensitivity of 93% for TV detection, equal to that of clinician obtained low vaginal swabs . None of these differences reached statistical significance . A combination of SOLVS and first void urine detected 96% of the CT cases, 100% of the NG cases, and 96% of TV cases . CONCLUSIONS: Self obtained low vaginal swabs are an acceptable, simple and sensitive diagnostic sample for the detection of CT, NG, and TV, and have particular applications in remote clinical practice and as a screening technique.

Sex Transm Infect, 2002 Aug, 78(4), 274 - 7
Coexistence of urethritis with genital ulcer disease in South Africa: influence on provision of syndromic management; Ballard RC et al.; OBJECTIVE: To assess whether syndromic management of genital ulcer disease was sound, if based on the premise that men with genital ulcers rarely have a concomitant urethral infection . METHODS: Specimens were taken in 1998 from 186 mine workers in Carletonville, South Africa, who were seen consecutively with genital ulcers . The specimens comprised a swab from the ulcer, a urethral swab for a Gram stained smear, and 10-15 ml of a first catch urine sample . The latter was tested by ligase chain reaction assays for Neisseria gonorrhoeae and Chlamydia trachomatis specific DNA sequences and by a polymerase chain reaction (PCR) assay for Mycoplasma genitalium . Ulcer inducing micro-organisms were detected either by a multiplex PCR assay, or in the case of lymphogranuloma venereum (LGV) serologically, and human immunodeficiency virus (HIV) infection was detected by an enzyme linked immunosorbent assay (ELISA) test . RESULTS: Most (54%) of the ulcers were chancroidal, 18% were herpetic (HSV type 2), 6.5% primary syphilitic, and 3.2% due to LGV . More than one micro-organism was detected in 9.1% of the ulcers and less than 10% were undiagnosed . Microscopic examination of the urethral smears showed that 99 (53%) of the men had urethritis, of whom 45 (45%) were infected with N gonorrhoeae . Of the 54 men (55%) who had non-gonococcal urethritis (NGU), 11 (19.6%) harboured C trachomatis or M genitalium . Almost two thirds (64.5%) of the men had HIV infection, but this did not seem to have influenced the aetiology of the ulcers . Nor was a particular ulcer associated with one type of urethritis more than the other . Neither C trachomatis nor M genitalium was associated significantly with non-gonococcal urethritis (NGU) in either HIV positive or HIV negative men . CONCLUSION: The combination of antibiotics used for the management of genital ulcer disease in men in this South African mining population needs to be widened to encompass frequently occurring concomitant gonococcal urethritis and NGU infections . This means treatment with long acting penicillin, combined with ciprofloxacin and azithromycin or erythromycin . A similar situation may exist in other geographical locations with a need to provide appropriate antimicrobial combinations depending on the patterns of infection detected.

J Prev Soc Med, 1999 Jun, 18(1), 46 - 51
Multiple drug resistant Neisseria gonorrhoeae among females of high risk behaviour in Dhaka city; Jahan Y et al.; PIP: This study was conducted to determine the antimicrobial resistance pattern of Neisseria gonorrhoeae species among females engaging in high-risk behaviors in Dhaka, Bangladesh . Endocervical swabs were collected from 110 prostitutes aged 13-40 years who were inmates at the Mirpur Government Rehabilitation Center . Of the 110 specimens tested, 22 (20%) were found positive for N . gonorrhea . A total of 8 antimicrobials were used for in-vitro antibiotic susceptibility testing . Out of the 22 positive strains, 8 resistance profiles were found . 12 (54.54%) of the strains were found resistant to penicillin, ampicillin, and cephalexin, which was the common resistance profile . None of the strains was found sensitive to all of the antimicrobials . All (100%) of the strains were resistant to cephalexin; 19 (86.36%) to penicillin and ampicillin; 7 (31.82%) to trimethoprim-sulphamethoxazole; 4 (18.18%) to tetracycline; 3 (13.64%) to erythromycin and norfloxacin; and only 1 (4.55%) to ciprofloxacin . These findings indicate that there is an alarming situation of multiple drug resistant N . gonorrhea in Dhaka City . This study focuses on the serious and regular monitoring of the problem, and recommends further study with a large number of samples countrywide .

Res Front Fertil Regul, 1984 May, 3(1), 1 - 16
The etiology of pelvic inflammatory disease; Keith L et al.; PIP: The etiology of pelvic inflammatory disease (PID) is speculated upon based on reported incidence and epidemiological studies . In Western society, the incidence of PID (annual) is 1% among women aged 15-34 years and 2% in the high risk group of women aged 15-24 years . The annual incidence in the US is higher, at least 2% among fecund sexually active women aged 13-44 years . The medical consequences of PID are infertility, ectopic pregnancy, and chronic pelvic pain . Causative agents include Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis and various other aerobic and anaerobic microorganisms; however, the natural genital flora of females is so varied that determining actual causative agents is difficult . some case-control studies have determined risk factors for PID; these include particularly current or prior use of IUD, prior pelvic surgery, sexual activity (including number of partners), race, and prior PID acute infection . PID is not a sexually transmitted disease, but rather is classified as sexually derived . Use of barrier methods and oral contraceptives protects against PID . IUD use greatly increases the risk of PID, probably because of the avenue the device provides for organisms to ascend from the lower to the upper genital tract . The role of males in PID etiology is currently the subject of much discussion . It is theorized that the mechanical action of penis insertion in intercourse helps to move causative agents to the upper genital region; also, semen may carry vaginal flora through the cervical opening into the uterus and tubes . Menstruation and PID are closely associated, perhaps because the cervix dilates during bleedings . Research areas include: determination of role of sexual activity (and number of partners) in PID etiology; evaluation of events of menstruation that are predisposing; evaluation of relationship between bacteriosperma and lower and upper genital infections; relationship of particular contraceptive methods to PID incidence; and breakdown of risk factors .

Br Med Bull, 2002, 62, 45 - 58
Bacterial pathogen genomics and vaccines; Moxon R et al.; Infectious diseases remain a major cause of deaths and disabilities in the world, the majority of which are caused by bacteria . Although immunisation is the most cost effective and efficient means to control microbial diseases, vaccines are not yet available to prevent many major bacterial infections . Examples include dysentery (shigellosis), gonorrhoea, trachoma, gastric ulcers and cancer (Helicobacter pylori) . Improved vaccines are needed to combat some diseases for which current vaccines are inadequate . Tuberculosis, for example, remains rampant throughout most countries in the world and represents a global emergency heightened by the pandemic of HIV . The availability of complete genome sequences has dramatically changed the opportunities for developing novel and improved vaccines and facilitated the efficiency and rapidity of their development . Complete genomic databases provide an inclusive catalogue of all potential candidate vaccines for any bacterial pathogen . In conjunction with adjunct technologies, including bioinformatics, random mutagenesis, microarrays, and proteomics, a systematic and comprehensive approach to identifying vaccine discovery can be undertaken . Genomics must be used in conjunction with population biology to ensure that the vaccine can target all pathogenic strains of a species . A proof in principle of the utility of genomics is provided by the recent exploitation of the complete genome sequence of Neisseria meningitidis group B.

Trans R Soc Trop Med Hyg, 2002 May-Jun, 96(3), 232 - 5
Behavioural characteristics, prevalence of Chlamydia trachomatis and antibiotic susceptibility of Neisseria gonorrhoeae in men with urethral discharge in Thyolo, Malawi; Zachariah R et al.; A study was carried out in 2000/2001 in a rural district of Malawi among men presenting with urethral discharge, in order to (a) describe their health-seeking and sexual behaviour, (b) determine the prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis, and (c) verify the antibiotic susceptibility of N . gonorrhoeae . A total of 114 patients were entered into the study; 61% reported having taken some form of medication before coming to the sexually transmitted infections clinic . The most frequent alternative source of care was traditional healers . Sixty-eight (60%) patients reported sexual encounters during the symptomatic period, the majority (84%) not using condoms . Using ligase chain reaction on urine, N . gonorrhoeae was detected in 91 (80%) and C . trachomatis in 2 (2%) urine specimens . Forty five of 47 N . gonorrhoeae isolates produced penicillinase, 89% showing multi-antimicrobial resistance . This study emphasizes the need to integrate alternative care providers and particularly traditional healers in control activities, and to encourage their role in promoting safer sexual behaviour . In patients presenting with urethral discharge in our rural setting, C . trachomatis was not found to be a major pathogen . Antimicrobial susceptibility surveillance of N . gonorrhoeae is essential in order to prevent treatment failures and control the spread of resistant strains.

Mol Genet Genomics, 2002 Jul, 267(5), 613 - 21 Epub 2002 Jun 07.
Reverse genetic approaches in plants and yeast suggest a role for novel, evolutionarily conserved, selenoprotein-related genes in oxidative stress defense; Rodrigo MJ et al.; Oxidation of methionine residues during periods of oxidative stress can lead to loss of protein function . Organisms have developed defense strategies to minimize such damage . The PilB protein, which is involved in pilus formation in the pathogen Neisseria gonorrhoeae, is composed of three functional protein domains (I-III) with putative roles in oxidative stress defense . These domains are evolutionarily conserved and homologs have been discovered in diverse prokaryotes and eukaryotes . Domain III shows similarities to selenoproteins which contain selenium instead of sulfur in a conserved cysteine residue . The substitution of selenium for sulfur alters the redox properties of such proteins . Knock-out mutants were used to elucidate the function of these novel selenoprotein-like domains in yeast and in Arabidopsis thaliana . We show that organisms with non-functional genes for selenoprotein-like polypeptides accumulate higher levels of oxidized methionine residues on exposure to oxidative stress . The behavior of the mutants suggests that these novel selenoprotein-like gene products are part of a ubiquitous detoxification system that interacts with other redox-related proteins such as the thioredoxin-related protein and methionine sulfoxide reductase which are encoded by domains I and II of PilB . These proteins may be encoded by one gene as in the case of several prokaryotes, or by separate genes as in the eukaryotes examined here.

Sex Transm Dis, 2002 Jul, 29(7), 387 - 90
Associations between biologically confirmed marijuana use and laboratory-confirmed sexually transmitted diseases among African American adolescent females; Liau A et al.; BACKGROUND: Numerous studies have examined the association between adolescents' marijuana use and their high-risk sexual behaviors and sexually transmitted diseases (STDs) . However, the validity of the findings is questionable because most of the studies relied on self-reporting for measurement of marijuana use and key outcome (i.e., STDs) . GOAL: The goal was to investigate associations between biologically confirmed marijuana use and laboratory-confirmed STDs and condom use . STUDY DESIGN: African American females adolescents (n = 522) completed a self-administered survey and face-to-face interview . The adolescents provided urine and vaginal swab specimens that were analyzed for marijuana metabolites and STDs, respectively . RESULTS: Among the study subjects, 5.4% tested positive for marijuana . These adolescents were more likely to test positive for Neisseria gonorrhoeae (adjusted odds ratio {AOR} = 3.4) and Chlamydia trachomatis (AOR = 3.9) . They were more likely to have never used condoms in the previous 30 days (AOR = 2.9) and to have not used condoms consistently in the previous 6 months (AOR = 3.6) . CONCLUSION: The findings represent unique biologic evidence that STDs and sexual risk behavior may co-occur with marijuana use . Interventions designed to reduce adolescents' risk of STDs and HIV infection should address marijuana use.

Sex Transm Dis, 2002 Jul, 29(7), 376 - 8
In vitro susceptibility of 120 strains of Neisseria gonorrhoeae isolated in Kyrghyzstan; Dorlencourt F et al.; BACKGROUND: The World Health Organization has established a worldwide program for gonococcal antimicrobial surveillance, but so far no data on gonococcal susceptibility in Central Asia are available . GOAL: The need for biological data on the susceptibility of Neisseria gonorrhoeae in Kyrghyzstan, to enable adaptation of the national treatment protocol for gonococcal infections, led Medecins Sans Frontieres and Epicentre to conduct a survey in collaboration with the Alfred Fournier Institute in Paris and the health authorities in Bishkek . STUDY DESIGN: In vitro susceptibility of N gonorrhoeae strains was determined with use of the reference agar-plate dilution technique . RESULTS: Results for 11 antibiotics tested on 120 strains of gonococci showed a low proportion (11.7%) of penicillinase-producing N gonorrhoeae and high proportions of intermediate or resistant strains to the majority of the antibiotics tested, including fluoroquinolones (>or=25% of strains resistant) . All the strains were susceptible to spectinomycin, and only two strains had decreased susceptibility to cefixime . CONCLUSION: The therapeutic choices available in Kyrghyzstan appear to be limited to cephalosporins and spectinomycin.

Theor Popul Biol, 2002 Jun, 61(4), 435 - 47
Microevolutionary genomics of bacteria; Jordan IK et al.; The availability of multiple complete genome sequences from the same species can facilitate attempts to systematically address basic questions in genome evolution . We refer to such efforts as "microevolutionary genomics" . We report the results of comparative analyses of complete intraspecific genome (and proteome) sequences from four bacterial species--Chlamydophila pneumoniae, Escherichia coli, Helicobacter pylori and Neisseria meningitidis . Comparisons of average synonymous (K(s)) and nonsynonymous (K(a)) substitution rates were used to assess the influence of various biological factors on the rate of protein evolution . For example, E . coli experiences the most intense purifying selection of the species analyzed, and this may be due to the relatively larger population size of this species . In addition, essential genes were shown to be more evolutionarily conserved than nonessential genes in E . coli and duplicated genes have higher rates of evolution than unique genes for all species studied except C . pneumoniae . Different functional categories of genes were shown to evolve at significantly different rates emphasizing the role of category-specific functional constraints in determining evolutionary rates . Finally, functionally characterized genes tend to be conserved between strains, while uncharacterized genes are over-represented among the unique, strain-specific genes . This suggests the possibility that nonessential genes are responsible for driving the evolutionary diversification between strains .

Niger Postgrad Med J, 2002 Jun, 9(2), 59 - 62
Penicillinase producing Neisseria gonnorhoeae: the review of the present situation in Ibadan, Nigeria; Bakare RA et al.; All the isolates of Neisseria gonorrhoeae from patients that attended special treatment Clinic, University College Hospital, Ibadan, Nigeria between 15th January 1997 and 15th December 1999 were studied so as to review the present prevalence rate of penicillinase producing Neisseria gonorrhoeae (PPNG) in Ibadan . Of the 214 patients that had gonococcal infections, 161 were male (75.2%) and 53 (24.8%) were female . Ninety-two (57.1%) of the male and 28(52.8%) of the female were aged between 20- 29 years while 19.9% of the male and 15.1 % of the female were in the age of 40 and above . The sex difference is not statistically significant (chi2=2.19, df=3, p=0.53) . The present study revealed that PPNG strains have increased dramatically to 98.6 per cent . This has posed a great threat to the usefulness of penicillin and ampicillin as the drugs of choice in gonococcal therapy in Nigeria.

Acad Emerg Med, 2002 Aug, 9(8), 781 - 5
Cervical infections in emergency department patients with vaginal bleeding; Hoffman L et al.; To the best of the authors' knowledge, no previous study has compared the prevalences of Chlamydia trachomatis (CT) or Neisseria gonorrhoeae (NG) cervical infection among pregnant and non-pregnant patients presenting to the emergency department (ED) with vaginal bleeding . OBJECTIVES: To determine the prevalence of these infections in a simultaneously assembled cohort, and to determine whether pregnancy is associated with altered rates of CT or NG cervical infection among patients with vaginal bleeding . METHODS: This was a prospective, seven-month, cross-sectional study of a convenience sample of patients (aged > or = 15 years) who presented to an urban, teaching ED with the chief complaint of vaginal bleeding . A urine pregnancy test was administered to each patient . Cervical swab specimens were analyzed for CT or NG via polymerase chain reaction technology (DNA probe) . The hospital laboratory's baseline 18% rate of positive tests was used for sample size planning . Chisquare analysis and Fisher's exact test, where appropriate, were used to compare pregnant and non-pregnant patient groups . RESULTS: Cervical infection rates did not differ between pregnant and non-pregnant patients . Testing for CT showed that 13 of 114 (11%) of the pregnant patients were positive, while 11 of 147 (8%) non-pregnant patients tested positive (chi(2) = 0.759, p = 0.384) . Testing for NG revealed that seven of 114 (6%) pregnant patients tested positive, while 16 of 147 (11%) non-pregnant patients tested positive (chi(2) = 1.256, p = 0.262) . Combined testing showed that 18 of 114 (16%) of the pregnant patients and 25 of 147 (17%) of the non-pregnant patients harbored a cervical infection with one or both pathogens (chi(2) = 0.009, p = 0.925) . CONCLUSIONS: The prevalence of CT or NG cervical infection in pregnant patients presenting to the ED with vaginal bleeding is 16% . Pregnant and non-pregnant patients with vaginal bleeding are at similar risks for having CT or NG cervical infection . Cervical swab specimens should be obtained in all patients with vaginal bleeding.

J Clin Microbiol, 2002 Aug, 40(8), 3035 - 7
Isolation from a healthy carrier and characterization of a Neisseria meningitidis strain that is deficient in gamma-Glutamyl aminopeptidase activity; Takahashi H et al.; gamma-Glutamyl aminopeptidase (GGT) activity is used as a specific marker for the identification of Neisseria meningitidis . We isolated from a healthy carrier and characterized an N . meningitidis isolate which lacked the activity due to the insertional mutation of the ggt gene, suggesting that naturally occurring N . meningitidis isolates do not always possess GGT activity.

Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1377 - 82
Phylogeny of the genus Simonsiella and other members of the Neisseriaceae; Hedlund BP et al.; 16S rDNA was sequenced from 16 strains of the oral commensal Simonsiella and was used to assess relationships between Simonsiella strains and other members of the Neisseriaceae . In all analyses, Simonsiella strains grouped according to established species designations and the mammalian hosts from which they were isolated . The commensals from cats and dogs formed a monophyletic group . The monophyly of the genus Simonsiella, however, could be neither supported nor rejected; deep nodes in the trees were unstable depending on the phylogenetic method or on the particular sequences used in the analysis . Instabilities may be attributable to frequent gene transfer between Neisseria or other members of the Neisseriaceae and Simonsiella.

Protein Expr Purif, 2002 Jul, 25(2), 353 - 61
Bacterial expression and characterization of mature apolipoprotein A-I; Panagotopulos SE et al.; Plasma levels of apolipoprotein A-I (apoA-I) are correlated with reduced incidence of heart disease due to the critical role of this protein in reverse cholesterol transport . Because of its diversity of function and poorly understood structure, much research has sought to understand how the structure of apoA-I facilitates its function . A popular approach has been the use of site-directed mutagenesis followed by structural and functional studies . There are a wide variety of expression systems available to produce these mutant proteins including eukaryotic cell lines and prokaryotic cells such as Escherichia coli . Expression in a bacterial system is generally favorable because it can produce large amounts of pure protein quickly and economically through the use of affinity tags on the expressed protein . Unfortunately, many of these systems are not ideal for the production of apolipoproteins because, in many cases, the proteolytic digestion required to remove the affinity tag also cleaves the target protein . Here we describe a method that produces large amounts of recombinant protein that is easily purified using a histidine (His) affinity tag that is cleaved with IgA protease from Neisseria gonorrhoeae . This enzyme does not cleave the wild type apoA-I sequence, leaving intact, mature apoA-I (containing a Thr-Pro- on the N-terminus) . We show that this recombinant protein is similar to wild type protein in structure and function using circular dichroism analysis, lipid clearance assays, recombinant particle formation and cholesterol efflux assays . This system is particularly useful for the bacterial production of apolipoproteins because of the extreme specificity of IgA protease for its target cleavage site.

Protein Expr Purif, 2002 Jul, 25(2), 237 - 40
High-level expression of recombinant Neisseria CMP-sialic acid synthetase in Escherichia coli; Karwaski MF et al.; The CMP-sialic acid synthetase (CMP-Neu5Ac, synthetase) is responsible for the synthesis of CMP-Neu5Ac, which is the donor used by sialyltransferases to attach sialic acid to acceptor hydroxyl groups in various polysaccharides, glycolipids, and glycoproteins . Since CMP-Neu5Ac is unstable and relatively expensive, the CMP-Neu5Ac synthetase is valuable for the preparative enzymatic synthesis of sialylated oligosaccharides . We made a construct to over-express the Neisseria meningitidis CMP-Neu5Ac synthetase in Escherichia coli . The recombinant enzyme was expressed at very high level (over 70,000 U/L) in a soluble form . It was purified by a sequence of anion-exchange chromatography and gel filtration with an overall yield of 23% (specific activity 220 U/mg) . The purified CMP-Neu5Ac synthetase was used in the gram-scale synthesis of CMP-Neu5Ac.

Vaccine, 2002 Jul 26, 20(23-24), 2964 - 71
Interspecific neisserial high molecular weight proteins able to induce natural immunity responses are strongly correlated with in vitro bactericidal activity; Sanchez S et al.; Human sera from healthy volunteers and patients convalescent from meningitis were used to search for widely cross-reactive antigens implicated in vivo protective responses . Using the type strain Neisseria meningitidis B16B6 and a wide variety of both N . meningitidis and N . lactamica strains, several cross-reactive antigens and bactericidal sera were found, although the cross-reactivity patterns did not correlate with bactericidal activity, a total correlation was found between bactericidal activity and reaction with one or two high molecular weight proteins (162 and 138 kDa), and a mouse serum against the purified proteins showed a high bactericidal activity . Our results suggest that the high molecular weight proteins found are immunogenic and cross-reactive, eliciting bactericidal responses during infection and as a result of natural immunity . These proteins should be taken into account in studies for future vaccine formulations.

Proc R Soc Lond B Biol Sci, 2001 Jan 7, 268(1462), 53 - 60
The probability of severe disease in zoonotic and commensal infections; Frank SA et al.; Cross-species transfers of pathogens (zoonoses) cause some of the most virulent diseases, including anthrax, hantavirus and Q fever . Zoonotic infections occur when a pathogen moves from its reservoir host species into a secondary host species . Similarly, commensal infections often have a primary reservoir location within their hosts' bodies from which they rarely cause disease symptoms, but commensals such as Neisseria meningitidis cause severe disease when they cross into a different body compartment from their normal location . Both zoonotic and commensal infections cause either mild symptoms or severe disease, but rarely intermediate symptoms . We develop a mathematical model for studying three factors that affect the probability of severe disease: the size of the inoculum, the route of inoculation and the frequency of naturally occurring infections that do not cause symptoms but do induce protective immunity (vaccinating inoculations) . With a single route of infection, increasing pathogen density causes inoculations to develop more often into disease rather than asymptomatic vaccinations that provide protective immunity . With two routes of infection, it may happen that a lower density of a pathogen or of a particular antigenic variant leads to a relatively higher frequency of disease-inducing versus vaccinating inoculations . This reversal occurs when one route of infection tends to vaccinate against relatively common pathogens but less often vaccinates against relatively rare pathogens, whereas the other route of infection is susceptible to disease-inducing inoculation even at relatively low pathogen density.

J Exp Med, 2002 Jul 15, 196(2), 147 - 62
Implications of phase variation of a gene (pgtA) encoding a pilin galactosyl transferase in gonococcal pathogenesis; Banerjee A et al.; The pilin glycoprotein (PilE) is the main building block of the pilus of Neisseria gonorrhoeae (gonococcus {GC}) . GC pilin is known to carry a disaccharide O-glycan, which has an alphaGal attached to the O-linked GlcNAc by a 1-3 glycosidic bond . In this report, we describe the cloning and characterization of the GC gene, pilus glycosyl transferase A (pgtA), which encodes the galactosyl transferase that catalyzes the synthesis of this Gal-GlcNAc bond of pilin glycan . A homopolymeric tract of Gs (poly-G) is present in the pgtA gene of many GC strains, and this pgtA with poly-G can undergo phase variation (Pv) . However, in many other GC, pgtA lacks the poly-G and is expressed constitutively without Pv . Furthermore, by screening a large number of clinical isolates, a significant correlation was observed between the presence of poly-G in pgtA and the dissemination of GC infection . Poly-G was found in pgtA in all (24 out of 24) of the isolates from patients with disseminated gonococcal infection (DGI) . In contrast, for the vast majority (20 out of 28) of GC isolated from uncomplicated gonorrhea (UG) patients, pgtA lacked the poly-G . These results indicate that Pv of pgtA is likely to be involved in the conversion of UG to DGI.

Am J Med Genet, 2002 Jun 1, 110(1), 25 - 9
C7 complement deficiency in an Israeli Arab village; Behar D et al.; Deficiencies of terminal complement components, particularly the latter ones, are often detected because of increased susceptibility to Neisserial infections . Herein we document the first report of C7 deficiency among a highly inbred Arab population living in the lower Galilee region of Israel . Both biochemical and molecular analysis were performed on samples from infected survivors and parents of children who succumbed to Neisserial infections in a 4-year period . Only the index case who suffered recurrent infections and a sibling who had not suffered an infection during the outbreak were found to be C7-deficient . The mutation was found to be the one previously described to be prevalent among Israeli Jews of Moroccan ancestry (mutation G1135C) . The implications of this finding are discussed in the context of family pedigree, the protective effect of complement deficiency, and the clinical outcome .

Minerva Ginecol, 2002 Aug, 54(4), 309 - 16
{Cervico-vaginal infections . Study of a population in the Turin area}; Latino MA et al.; BACKGROUND: Aim of this study is to determine the prevalence, in the Turin area, of the pathogens chiefly involved in the genesis of the most common infections of the female genitalia . METHODS: This is a retrospective study . During the period of time beginning from January 1, 1997 and ending December 31, 1999, we examined 10,249 women from 14 years to 54 years of age, who were seen at the out-patient diagnosis service of the Sant'Anna Hospital . The patients' cervical specimens were screened for common germs, Candida spp., Trichomonas vaginalis, Bact . Vaginosis, Chlamydia trachomatis, Microplasms, and Neisseria gonorrhea . The prevalence of each micro-organism was found . The obtained data were cross-referenced with the risk factors in the clinical history of each patient . The chi(2) test with a C.I . of 95% was used for the statistical evaluations . RESULTS: It is shown by a detailed analysis of the data in our possession that an anamnesis oriented mainly for the evaluation of the various risk factors would be more effective than one oriented for the presence of a subjective symptomatology, since many of these infections are either weakly symptomatic or totally asymptomatic (especially in the case of C . trachomatis), as has been many times underlined in the international literature . CONCLUSIONS: Greater attention should be given to the collection of anamnestic information in order to more precisely target for examination those subjects at greater risk of contracting a sexually transmitted infection.

FEMS Microbiol Lett, 2002 Jul 2, 212(2), 165 - 9
Phylogenetic analysis of the genus Aquaspirillum based on 16S rRNA gene sequences; Ding L et al.; Phylogenetic analysis of 15 species of the genus Aquaspirillum based on 16S rRNA gene (rDNA) sequences indicated that the genus Aquaspirillum is phylogenetically heterogeneous and the species could be divided into four groups as follows: Aquaspirillum serpens, the type species of this genus, A . dispar and A . putridiconchylium are situated in the family Neisseriaceae; members of the second group, A . gracile, A . delicatum, A . anulus, A . giesbergeri, A . sinuosum, A . metamorphum and A . psychrophilum, are included in the family Comamonadaceae; the two members of the third group, A . arcticum and A . autotrophicum, are included in the family Oxalobacteriaceae; and members of the fourth group, A . polymorphum, A . peregrinum, and A . itersonii, are included in the alpha-subdivision of Proteobacteria . Thus, phylogenetic studies indicated that all the species excepting A . serpens, the type species, should be transferred to distinct genera.

Eur J Clin Microbiol Infect Dis, 2002 Jun, 21(6), 483 - 5 Epub 2002 Jun 12.
Serogroups, serotypes, serosubtypes and antimicrobial susceptibility of Neisseria meningitidis isolates in Casablanca, Morocco; Zerouali K et al.; Since antigenic characterization and antibiotic susceptibility testing are useful for generating prophylactic recommendations and treatment guidelines, a total of 163 Neisseria meningitidis isolates obtained between January 1992 and September 2000 at the microbiology laboratory of the IbnRochd University Hospital of Casablanca, Morocco, were serogrouped, serotyped, serosubtyped and tested for their susceptibility to five antibiotics . Serogroup B was detected most frequently (75.5%), followed by serogroup A (13.5%) . The phenotype B:4:P1.15 represented 74.8% of all serogroup B isolates . Seven (4.3%) isolates demonstrated decreased susceptibility to penicillin G . All isolates tested were susceptible to cefotaxime, chloramphenicol and rifampin . All isolates were inhibited by spiramycin at a concentration of 0.4 mg/l.

J Infect Chemother, 2002 Jun, 8(2), 145 - 50
Rapid detection of quinolone resistance-associated gyrA mutations in Neisseria gonorrhoeae with a LightCycler; Li Z et al.; Afluorometric real-time polymerase chain-reaction (PCR)-hybridization system, the LightCycler was developed for the detection of Neisseria gonorrhoeae in clinical samples and the analysis of point mutations associated with quinolone resistance in the gyrAgene . This system allowed us to amplify the N . gonorrhoeae-specific gyrA gene from an amount of DNA corresponding to five genome copies per reaction . We were able to detect N . gonorrhoeae in either 55 control strains or 36 nonisolated clinical urethral swab specimens, and to analyze the presence of mutations in codons Ser-91 and Asp-95 of the gyrA gene within 1 h . The mutation status in the gyrA gene assessed by the LightCycler assay was completely in agreement with the results of our previous conventional sequencing analysis, and was associated with the susceptibility to ciprofloxacin.

J Mol Evol, 2002 Aug, 55(2), 232 - 45
Gene conversion and the evolution of euryarchaeal chaperonins: a maximum likelihood-based method for detecting conflicting phylogenetic signals; Archibald JM et al.; Recombination is well known as a complicating factor in the interpretation of molecular phylogenies . Here we describe a maximum likelihood sliding window method based on a likelihood ratio test for scanning DNA sequence alignments for regions of incongruent phylogenetic signals, such as those influenced by recombination . Using this method, we identify several instances of gene conversion between paralogous chaperonin genes in euryarchaeote Archaea, many of which are not detected by two other widely used methods . In the Thermococcus/Pyrococcus lineage, where a gene duplication producing a and b paralogues predates the divergence of Thermococcus strains KS-1 and KS-8, gene conversion has homogenized portions of the a and b genes in KS-8 since the divergence of these two strains . A region near the 3' end of the a and b paralogues in the methanogen Methanobacterium thermoautotrophicum also appears to have undergone gene conversion . We apply the method to two additional test data sets, the argF gene of Neisseria and a set of actin paralogues in maize, and show that it successfully identifies all the recombinant regions that were previously detected with other methods . Our approach is relatively insensitive to the presence of divergent sequences in the alignment, making it ideal for detecting recombination between both closely and distantly related genes.

Mil Med, 2002 Jun, 167(6), 496 - 500
Correlates of sexually transmitted diseases in a young male deployed military population; Shafer MA et al.; OBJECTIVE: To examine risk factors of sexually transmitted diseases (STDs) in a non-clinic-based sample of young male military personnel . METHODS: A total of 1,028 enlisted U.S . Marines completed a self-report survey querying sexual behaviors in the previous 3 months and provided urine specimens for STD screening by nucleic acid amplification testing for Chlamydia trachomatis and Neisseria gonorrhoeae . Risk for STDs was assessed using a composite Sexual Risk Index . Recent STD infection was defined as a positive result on urine-based screening for chlamydial and/or gonococcal infection and/or a history of STDs in the past 3 months . RESULTS: Recent STDs were identified in 76 men (7.4%): 42 (4.1%) were diagnosed with a chlamydial infection with no gonorrhea found, and 34 (3.3%) reported a history of STDs . Men with the highest score on the Sexual Risk Index were 2.6 times more likely to have a current STD compared with those at lowest risk (odds ratio = 2.6, confidence interval = 1.01, 6.2) . CONCLUSIONS: Use of a composite measure of risk can identify the most significant factors associated with recent STDs in young military men . Such an index may be useful to target STD screening for those at greatest risk for STDs.

Proc Natl Acad Sci U S A, 2002 Jul 23, 99(15), 10108 - 13 Epub 2002 Jul 02.
The outer membrane localization of the Neisseria gonorrhoeae MsrA/B is involved in survival against reactive oxygen species; Skaar EP et al.; The PilB protein of Neisseria gonorrhoeae has been reported to be involved in the regulation of pilin gene transcription, but it also possesses significant homology to the peptide methionine sulfoxide reductase family of enzymes, specifically MsrA and MsrB from Escherichia coli . MsrA and MsrB in E . coli are able to reduce methionine sulfoxide residues in proteins to methionines . In addition, the gonococcal PilB protein encodes for both MsrA and MsrB activity associated with the repair of oxidative damage to proteins . In this work, we demonstrate that the PilB protein of Neisseria gonorrhoeae is not involved in pilus expression . Additionally, we show that wild-type N . gonorrhoeae produces two forms of this polypeptide, one of which contains a signal sequence and is secreted from the bacterial cytoplasm to the outer membrane; the other lacks a signal sequence and is cytoplasmic . Furthermore, we show that the secreted form of the PilB protein is involved in survival in the presence of oxidative damage.

FEBS Lett, 2002 Jul 3, 522(1-3), 52 - 8
Transposon-like Correia elements: structure, distribution and genetic exchange between pathogenic Neisseria sp; Buisine N et al.; Correia elements are a prominent feature of all four Neisseria genome sequences . We report an in silico analysis of the structure and genomic distribution of these elements and some preliminary biochemical data . Correia elements fall into four major families, distinguished by a 50 bp internal deletion and five point mutations . The elements resemble a transposon with 25 bp inverted repeats and a TA duplication at the target site . Within the element there is a functional integration host factor binding site . The genomic distribution of Correia elements is essentially random except for some small Correia-less regions apparently acquired by horizontal transfer . Phylogenetic analysis suggests that their presence predates the divergence of Neisseria meningitidis and Neisseria gonorrhoeae.

Diagn Microbiol Infect Dis, 2002 Jun, 43(2), 119 - 21
Value of Etest penicillin V and penicillin G strips for penicillin susceptibility testing of Neisseria meningitidis; Daher O et al.; The NCCLS agar dilution method and Etest are currently accepted methods for susceptibility testing of Neisseria meningitidis to penicillin . We determined the MIC of penicillin V and penicillin G by both the agar dilution method and Etest using 43 strains of N . meningitidis . Although results for reference strains were within the accepted quality control range of penicillin MICs for both drugs, differences of two to three dilutions were seen between the two antibiotics with both methods . Penicillin V results cannot correctly predict the susceptibility to penicillin G for N . meningitidis if penicillin G breakpoints are used for penicillin V . However, adjusting the penicillin V breakpoints two dilutions higher (i.e., S < or = 0.25 and R > or = 8 microg/ml), concordance could be achieved for susceptibility categorization by the two compounds . An agreement of 98% +/- 1 dilution was obtained between Etest and the reference method when using penicillin G strips . We conclude that Etest with penicillin G strips is a convenient and reliable alternative method for determining the MICs of penicillin for N . meningitidis.

J Bacteriol, 2002 Jul, 184(14), 3965 - 74
The gonococcal fur regulon: identification of additional genes involved in major catabolic, recombination, and secretory pathways; Sebastian S et al.; In this study, we have characterized the in vitro binding of Neisseria gonorrhoeae Fur to several well-defined iron transport genes, as well as to additional genes involved in major catabolic, secretory, and recombination pathways of gonococci . The gonococcal Fur protein was recombinantly expressed in Escherichia coli HBMV119 . Fur was isolated from inclusion bodies and partially purified by ion-exchange chromatography . Gonococcal Fur was found to bind to the promoter/operator region of a gene encoding the previously identified Fur-regulated periplasmic binding protein (FbpA) in a metal ion-dependent fashion, demonstrating that purified Fur is functional . In silico analysis of the partially completed gonococcal genome (FA1090) identified Fur boxes in the promoters of several genes, including tonB, fur, recN, secY, sodB, hemO, hmbR, fumC, a hypothetical gene (Fe-S homolog), and the opa family of genes . By using purified gonococcal Fur, we demonstrate binding to the operator regions of tonB, fur, recN, secY, sodB, hemO, hmbR, fumC, the Fe-S homolog gene, and the opa gene family as determined by an electrophoretic mobility shift assay . While gonococcal Fur was demonstrated to bind to the promoter regions of all 11 opa genes (opaA through -K), we did not detect binding of purified E . coli Fur with 8 of the 11 opa members, indicating that target DNA sequence specificities between these two closely related proteins exist . Furthermore, we observed differences in the relative strengths of binding of gonococcal Fur for these different genes, which most likely reflect a difference in affinity between gonococcal Fur and its DNA targets . This is the first report that definitively demonstrates the binding of gonococcal Fur to its own promoter/operator region, as well as to the opa family of genes that encode surface proteins . Our results demonstrate that the gonococcal Fur protein binds to the regulatory regions of a broad array of genes and indicates that the gonococcal Fur regulon is larger than originally proposed.

Sex Transm Infect, 2002 Apr, 78(2), 106 - 9
Gonorrhoea in London: usefulness of first line therapies; Ison CA et al.; OBJECTIVES: To determine the true prevalence and patterns of resistance of Neisseria gonorrhoeae to antimicrobial agents used for therapy in London . METHODS: A longitudinal study of a representative sample of isolates of N gonorrhoeae linked to demographic data of infected patients was undertaken . Isolates were collected from consecutive patients attending genitourinary medicine clinics in the North and South Thames regions of London during 3 months of each of 4 years, 1997-2000 . Prevalence of plasmid mediated resistance to penicillin and chromosomally mediated resistance to penicillin and ciprofloxacin was determined by susceptibility testing . The association of antimicrobial resistance among gonococci with sexual orientation of the patient and country of acquisition of infection was determined . RESULTS: Numbers of gonococcal isolates collected over the same time period between 1997-2000 have increased by 74% . Plasmid mediated resistance to penicillin was low but has risen above 5% in 2000 . Chromosomal resistance to penicillin has fallen below the 5% level but total resistance to penicillin, plasmid and chromosomally mediated, was above 5% in all 4 years . The incidence of resistance and reduced susceptibility to the alternative therapeutic choice, ciprofloxacin, is low but numbers are increasing in each year . High level resistance, to both penicillin and ciprofloxacin, has generally been found among heterosexual men and is often acquired abroad . However, there is some evidence of a change to endemic spread in 2000 . CONCLUSIONS: This surveillance programme shows that the epidemiology of gonorrhoea in London is changing with increasing numbers and changing patterns of resistance . If gonorrhoea is to be controlled and targets set by the sexual health strategy are to be met, intervention with effective and appropriate antimicrobial agents is essential.

FEMS Microbiol Lett, 2002 Jun 4, 211(2), 203 - 6
In vitro anti-Neisseria gonorrhoeae activity of Terminalia macroptera leaves; Silva O et al.; We used the agar dilution method to evaluate the antibacterial effect of Terminalia macroptera leaf (Tml) extract against nine reference and clinical Neisseria gonorrhoeae strains, including penicillin- and tetracycline-resistant and -susceptible strains . Tml possesses anti-N . gonorrhoeae activity against all of the strains and the minimum inhibitory concentrations (MIC) were between 100 and 200 microg ml(-1) . We then used a liquid-liquid partition method to divide the Tml extract into five fractions and determined the anti-N . gonorrhoeae activity of each of the fractions . All of the fractions showed antibacterial activity . The most active one was identified as the diethyl ether fraction and had MIC values of between 25 and 50 microg ml(-1) against all of the strains.

Cochrane Database Syst Rev . 2002;(2):CD000098.
Antibiotics for gonorrhoea in pregnancy; Brocklehurst P; BACKGROUND: Neisseria gonorrhoeae can be transmitted from the mother's genital tract to the newborn during birth and can cause gonococcal ophthalmia neonatorum as well as systemic neonatal infection . It can also cause endometritis and pelvic sepsis in the mother . OBJECTIVES: The objective of this review was to assess the effects of antibiotic regimens in the treatment of genital infection with gonorrhoea during pregnancy with respect to neonatal and maternal morbidity . SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (November 2001) and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2001) were searched . SELECTION CRITERIA: Randomised trials of one regimen of antibiotic versus another in pregnant women with culture confirmed genital gonococcal infection . DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer . MAIN RESULTS: Two trials involving 346 women were included . The only outcome included in these trials was the incidence of 'cure' assessed by bacterial culture . Failure to achieve 'microbiological cure' was similar for each antibiotic regimen: Amoxicillin plus probenecid compared with spectinomycin (odds ratio (OR) 2.40, 95% confidence interval (CI) 0.71-8.12), amoxicillin plus probenecid compared with ceftriaxone (OR 2.40, 95% CI 0.71-8.12) and ceftriaxone compared with cefixime (OR 1.22, 95% CI 0.16-9.04) . Side effects were uncommon for all the tested regimens . REVIEWER'S CONCLUSIONS: The number of women included in each of the comparisons is small and therefore, although no differences were detected between the different antibiotic regimens, the trials were limited in their ability to detect important but modest differences . For women who are allergic to penicillin, this review provides some reassurance that treatment with ceftriaxone or spectinomycin appears to have similar effectiveness in producing microbiological cure.

J Infect, 2002 Feb, 44(2), 96 - 8
A case of lower respiratory tract infection caused by Neisseria weaveri and review of the literature; Panagea S et al.; Neisseria weaveri (formerly CDC {Centers for Disease Control and Prevention} group M-5 is part of the normal canine oral flora . Infections in humans are usually associated with dog bite wounds . Very rarely the organism has been isolated from sites other than wounds, or from deep seated infections . A 60-year-old man was admitted to our hospital because of an acute exacerbation of his bronchiectasis . Gram stain of bronchial washings and expectorated sputum showed numerous polymorphs and Gram-negative bacilli . Routine bacterial culture yielded a heavy pure growth of a Gram-negative rod-shaped organism that was strongly oxidase and catalase positive, indole negative, non-motile and did not ferment carbohydrates . The organism was identified as N . weaveri by using 16S rRNA sequencing . The patient was treated with a 3 weeks course of ofloxacin and had a good response . Sputum culture after treatment yielded normal respiratory flora only . To our knowledge, this is the first reported case of lower respiratory tract infection caused by N . weaveri .

Eur J Clin Microbiol Infect Dis, 2002 May, 21(5), 368 - 72 Epub 2002 May 17.
Increasing incidence of gonorrhea in Israel associated with countrywide dissemination of a ciprofloxacin-resistant strain; Yagupsky P et al.; The annual incidence of gonorrhea in Israel has sharply increased during the past 2 years . At the end of 1999, high-level ciprofloxacin-resistant strains of Neisseria gonorrhoeae (MIC90, > or =32 mcg/ml) also exhibiting decreased susceptibility to penicillin and tetracycline were isolated for the first time in southern Israel, as well as in other regions of the country . The incidence of male gonococcal urethritis in the south increased in a 1.5-year period from 3/100,000 to 12/100,000 ( P<0.05) in correlation with increased isolation of ciprofloxacin-resistant organisms . A marked increase in the incidence of gonorrhea was also encountered in Jerusalem, where ciprofloxacin resistance affected 54.5% of the isolates in 2000 . Pulsed-field gel electrophoresis typing of gonococci from different areas of Israel indicated that all of the ciprofloxacin-resistant isolates belonged to identical or related strains . Since fluoroquinolone-resistant gonococci may emerge and disseminate extensively over a short period of time, continuous surveillance of antibiotic susceptibility among gonococcal isolates should be performed to guide empiric therapy.

Scand J Infect Dis, 2002, 34(5), 379 - 82
Sequelae after bacterial meningitis in childhood; Oostenbrink R et al.; The neurological outcome of bacterial meningitis in children was evaluated retrospectively . Data were obtained from a large study on children aged between 1 month and 15 y who initially visited the emergency department of Sophia Children's Hospital, Rotterdam, The Netherlands with meningeal signs . This study presents data from 103 patients in whom bacterial meningitis was diagnosed . Neisseria meningitidis was the dominant pathogen of meningitis . We found a 2% case-fatality rate in children with bacterial meningitis and a 13% rate of sequelae among survivors: 7% hearing impairment and 7% neurological sequelae . Children with bacterial meningitis caused by Streptococcus pneumoniae and those with acute focal neurological symptoms tended to have the worst prognosis.

Am J Obstet Gynecol, 2002 Jun, 186(6), 1320 - 5
Postpartum Chlamydia trachomatis and Neisseria gonorrhoeae infections; Mahon BE et al.; OBJECTIVE: The purpose of this study was to investigate Chlamydia trachomatis and Neisseria gonorrhoeae infections that are diagnosed during the postpartum period (defined as the 3 months after delivery) . STUDY DESIGN: This was a retrospective cohort study of women delivered of infants at >or=28 weeks of gestation at an urban hospital from 1992 through 1998, including each woman's first delivery during this period . Postpartum C trachomatis and N gonorrhoeae testing were routine . Data were extracted from an electronic medical record system . RESULTS: Of 13,086 women with eligible deliveries, 7258 women (55.5%) underwent postpartum C trachomatis tests; 191 of those women (2.6%) had positive test results . Similarly, 7148 women (54.6%) underwent postpartum N gonorrhoeae tests; 95 of those women (1.3%) had positive test results . Most women with positive postpartum tests had prenatal testing (182/191 with C trachomatis; 94/95 with N gonorrhoeae ), and the last prenatal test results were usually negative (153/182 women with negative prenatal C trachomatis testing; 91/94 women with negative prenatal N gonorrhoeae testing) . CONCLUSION: Substantial rates of C trachomatis and N gonorrhoeae were found in postpartum women, most of whom had negative results on the last prenatal test.

Infect Immun, 2002 Jul, 70(7), 3891 - 903
Modification of type IV pilus-associated epithelial cell adherence and multicellular behavior by the PilU protein of Neisseria gonorrhoeae; Park HS et al.; Expression of type IV pili (Tfp) correlates with the ability of Neisseria gonorrhoeae to colonize the human host, as well as with adherence to human epithelial tissue, twitching motility, competence for natural transformation, and autoagglutination . N . gonorrhoeae PilF (required for Tfp biogenesis) and PilT (required for twitching motility and transformation) share significant identities with members of a family of putative ATPases involved in membrane trafficking of macromolecules . An open reading frame downstream of the pilT locus encoding a 408-amino-acid protein with 33% identity with the gonococcal PilT protein and 45% identity with the PilU protein in Pseudomonas aeruginosa was characterized, and the corresponding gene was designated pilU . Unlike N . gonorrhoeae pilT mutants, pilU mutants express twitching motility and are competent for DNA transformation . However, loss-of-function mutations in pilU increased bacterial adherence to ME-180 human epithelial cells eightfold and disrupted in vitro Tfp-associated autoagglutination . Comparative alignment of N . gonorrhoeae PilU with other members of the TrbB-like family of traffic ATPases revealed a conserved carboxy-terminal domain unique to family members which are not essential for Tfp biogenesis but which specifically modify Tfp-associated phenotypes . Studies of the pilT-pilU locus by using Northern blotting, transcriptional fusions, and reverse transcription-PCR showed that the two genes encoding closely related proteins with dissimilar effects on Tfp phenotypes are transcribed from a single promoter.

Infect Immun, 2002 Jul, 70(7), 3744 - 51
The Neisseria lipooligosaccharide-specific alpha-2,3-sialyltransferase is a surface-exposed outer membrane protein; Shell DM et al.; Neisseria gonorrhoeae and Neisseria meningitidis express an approximately 43-kDa alpha-2,3-sialyltransferase (Lst) that sialylates the surface lipooligosaccharide (LOS) by using exogenous (in all N . gonorrhoeae strains and some N . meningitidis serogroups) or endogenous (in other N . meningitidis serogroups) sources of 5'-cytidinemonophospho-N-acetylneuraminic acid (CMP-NANA) . Sialylation of LOS can protect N . gonorrhoeae and N . meningitidis from complement-mediated serum killing and from phagocytic killing by neutrophils . The precise subcellular location of Lst has not been determined . We confirm and extend previous studies by demonstrating that Lst is located in the outer membrane and is surface exposed in both N . gonorrhoeae and N . meningitidis . Western immunoblot analysis of subcellular fractions of N . gonorrhoeae strain F62 and N . meningitidis strain MC58 not subset 3 (an acapsulate serogroup B strain) performed with rabbit antiserum raised against recombinant Lst revealed an approximately 43-kDa protein exclusively in outer membrane preparations of both pathogens . Inner membrane, periplasmic, cytoplasmic, and culture supernatant fractions were devoid of Lst, as determined by Western blot analysis . Consistent with this finding, outer membrane fractions of N . gonorrhoeae were significantly enriched for sialyltransferase enzymatic activity . A trace of enzymatic activity was detected in inner membrane fractions, which may have represented Lst in transit to the outer membrane or may have represented inner membrane contamination of outer membrane preparations . Subcellular preparations of an isogenic lst insertion knockout mutant of N . gonorrhoeae F62 (strain ST01) expressed neither a 43-kDa immunoreactive protein nor sialyltransferase activity . Anti-Lst rabbit antiserum bound to whole cells of N . meningitidis MC58 not subset 3 and wild-type N . gonorrhoeae F62 but not to the Lst mutant ST01, indicating the surface exposure of the enzyme . Although the anti-Lst antiserum avidly bound enzymatically active, recombinant Lst, it inhibited Lst (sialyltransferase) activity by only about 50% at the highest concentration of antibody used . On the contrary, anti-Lst antiserum did not inhibit sialylation of whole N . gonorrhoeae cells in the presence of exogenous CMP-NANA, suggesting that the antibody did not bind to or could not access the enzyme active site on the surface of viable Neisseria cells . Taken together, these results indicate that Lst is an outer membrane, surface-exposed glycosyltransferase . To our knowledge, this is the first demonstration of the localization of a bacterial glycosyltransferase to the outer membrane of gram-negative bacteria.

Infect Immun, 2002 Jul, 70(7), 3707 - 13
Effect of O acetylation of Neisseria meningitidis serogroup A capsular polysaccharide on development of functional immune responses; Berry DS et al.; The importance of O-acetyl groups to the immunogenicity of Neisseria meningitidis serogroup A polysaccharide (PS) was examined in studies using human sera and mouse immunization . In 17 of 18 postimmunization human sera, inhibition enzyme-linked immunosorbent assay indicated that the majority of antibodies binding to serogroup A PS were specific for epitopes involving O-acetyl groups . Studies with mice also showed an essential role for O-acetyl groups, where serum bactericidal titers following immunization with de-O-acetylated (de-O-Ac) conjugate vaccine were at least 32-fold lower than those following immunization with O-Ac PS-conjugate vaccine and 4-fold lower than those following immunization with native capsular PS . Inhibition studies using native and de-O-Ac PS confirmed the specificity of murine antibodies to native PS . The dramatic reduction in immunogenicity associated with removal of O-acetyl groups indicates that O acetylation is essential to the immunogenic epitopes of serogroup A PS . Since levels of bactericidal antibodies are correlated with protection against disease, O-acetyl groups appear to be important in protection.

Infect Immun, 2002 Jul, 70(7), 3324 - 9
Protection of mice against brucellosis by intranasal immunization with Brucella melitensis lipopolysaccharide as a noncovalent complex with Neisseria meningitidis group B outer membrane protein; Bhattacharjee AK et al.; Intranasal immunization of mice with purified Brucella melitensis lipopolysaccharide (LPS) as a noncovalent complex with Neisseria meningitidis group B outer membrane protein (GBOMP) elicited a high-titer anti-LPS systemic antibody response and a significant mucosal antibody response . The anti-LPS immunoglobulin G (IgG) antibody was predominantly of the IgG1 subtype, although there was some response of the IgG2a, IgG2b, and IgG3 subtypes . The antibody titer remained high for 16 weeks postimmunization . Immunized mice and sham-immunized control mice were challenged intranasally with 10(4) CFU of virulent B . melitensis strain 16 M 4 weeks after the second dose of vaccine . The numbers of bacteria in lungs, livers, and spleens at 3 days, 9 days, and 8 weeks postchallenge were determined . Bacteria were found in lungs of all mice on day 3, but there was no disseminated infection of liver or spleen . By day 9, 40% of the mice had infected spleens and livers . At 8 weeks postchallenge, spleens of 25 of 62 immunized mice were infected, compared to 61 of 62 control mice (P < 0.0001) . The livers of 12 of 43 immunized mice were infected, compared to 22 of 36 control mice (P = 0.005) . In contrast, the lungs of 26 of 46 immunized mice were still infected, compared to 27 of 44 control mice . The numbers of bacterial CFU in lungs of immunized and control animals were identical . These studies show that intranasal immunization with B . melitensis LPS-GBOMP subunit vaccine significantly protects mice against intranasal challenge with virulent B . melitensis . Vaccination reduces bacterial dissemination to spleen and liver but has no effect on the course of lung infection.

Med J Aust, 2002 Jun 3, 176(11), 551 - 7
Infectious diseases . 6: Sexually transmitted infections: new diagnostic approaches and treatments; Bowden FJ et al.; 1 . Commercially available nucleic acid amplification assays (eg, polymerase or ligase chain reaction) are now the "gold standard" tests for genital chlamydial infection and also have a role in screening for gonococcal infection . 2 . Single-dose oral antibiotics are available for treatment of Chlamydia trachomatis, Neisseria gonorrhoeae and Trichomonas vaginalis infections . 3 . Strains of N . gonorrhoeae in urban Australia are often penicillin resistant, while strains from South East Asia and those in homosexually active men may show high-level resistance to quinolones . 4 . Imiquimod, a novel immune-response modifier, is now available for effective, safe, self-administered treatment of genital warts . 5 . The Pap smear remains the cornerstone of screening for precursor lesions of cervical cancer, but human papillomavirus genotyping may have a role in clinical decision-making for women with equivocal or early precancerous lesions . 6 . Treatment of primary genital herpes changes the clinical course, and long-term suppressive therapy is effective for those with multiple recurrences . New technologies have made diagnosis and screening easier for patients and clinicians

Microbiology, 2002 Jun, 148(Pt 6), 1833 - 44
Genetic diversity of three lgt loci for biosynthesis of lipooligosaccharide (LOS) in Neisseria species; Zhu P et al.; Lipooligosaccharide (LOS) is a major virulence factor of the pathogenic Neisseria . Nine lgt genes at three chromosomal loci (lgt-1, 2, 3) encoding the glycosyltransferases responsible for the biosynthesis of LOS oligosaccharide chains were examined in 26 Neisseria meningitidis, 51 Neisseria gonorrhoeae and 18 commensal Neisseria strains . DNA hybridization, PCR and nucleotide sequence data were compared to previously reported lgt genes . Analysis of the genetic organization of the lgt loci revealed that in N . meningitidis, the lgt-1 and lgt-3 loci were hypervariable genomic regions, whereas the lgt-2 locus was conserved . In N . gonorrhoeae, no variability in the composition or organization of the three lgt loci was observed . lgt genes were detected only in some commensal Neisseria species . The genetic organization of the lgt-1 locus was classified into eight types and the lgt-3 locus was classified into four types . Two types of arrangement at lgt-1 (II and IV) and one type of arrangement at lgt-3 (IV) were novel genetic organizations reported in this study . Based on the three lgt loci, 10 LOS genotypes of N . meningitidis were distinguished . Phylogenetic analysis revealed a gene cluster, lgtH, which separated from the homologous genes lgtB and lgtE . The lgtH and lgtE genes were mutually exclusive and were located at the same position in lgt-1 . The data demonstrated that pathogenic and commensal Neisseria share a common lgt gene pool and horizontal gene transfer appears to contribute to the genetic diversity of the lgt loci in Neisseria.

Microbiology, 2002 Jun, 148(Pt 6), 1821 - 31
Differential cross-complementation patterns of Escherichia coli and Neisseria gonorrhoeae RecA proteins; Stohl EA et al.; The Escherichia coli RecA protein is one of the best-studied enzymes, but less is understood about how RecA homologues of other species are similar to or different from the E . coli RecA . In the Gram-negative pathogen Neisseria gonorrhoeae (the gonococcus; Gc), the causative agent of gonorrhoea, RecA is involved in DNA transformation, pilin antigenic variation, and DNA repair . By expressing the recA genes from Gc and E . coli under control of lac regulatory sequences in E . coli, the authors have shown that the Gc RecA fully complements an E . coli recA mutant for homologous recombination, but only partially complements for survival to DNA damage . By expressing similar constructs in Gc, it was shown that the E . coli RecA complements for pilin antigenic variation, partially complements for DNA transformation, but does not complement for survival to DNA damage, suggesting that species-specific interactions are important for DNA repair, but not for homologous recombination . Co-expression of the E . coli recA and recX genes in Gc suggests that in this heterologous system RecX modulates RecA-mediated processes.

Ned Tijdschr Geneeskd, 2002 May 18, 146(20), 954 - 7
{Fitz-Hugh-Curtis syndrome: 2 patients with perihepatitis and sepsis}; Kaan JA et al.; Two women aged 53 and 22 years presented with abdominal pain and signs of sepsis with metabolic acidosis . The first patient had ecchymoses all over her body, the second patient had an enlarged left kidney with wedge-shaped hypo-intense areas on the CT scan . The clinical condition of both women deteriorated . On laparoscopy perihepatitis with fibrin wires ('violin strings') was seen, pathognomonic for Fitz-Hugh-Curtis syndrome . Upon appropriate antibiotic treatment, both patients fully recovered . Although it is common belief that Fitz-Hugh-Curtis syndrome is caused by local spread from the fallopian tubes into the peritoneal cavity, these presentations suggest a haematogenous spread of Neisseria gonorrhoeae as well as Chlamydia trachomatis in the first case, and C . trachomatis in the second case.

Ginecol Obstet Mex, 2002 Apr, 70, 203 - 9
{Cervicovaginal infection as a risk factor for premature labor}; Beltran Montoya J et al.; OBJECTIVE: To identify the possible association between cervicovaginal infections (CVI) and preterm delivery . DESIGN: Cohorts . REFERENCE FRAME: Instituto Nacional de Perinatologia, Hospital Central Militar and Hospital General Regional No . 1, IMSS, Culiacan, Sinaloa, Mexico . PATIENTS: Four hundred and sixty eight patients attending prenatal control and delivery care . INTERVENTIONS: Fresh smears, Gram stain, and cervicovaginal sample culture from samples obtained during the following gestational stages: First sample at 16-24 weeks, second sample at 25-32 weeks, and third sample at 33-42 weeks . The following microorganisms were studied: Candida albicans, Gardnerella vaginalis, Ureaplasma urealyticum, Streptococcus agalactiae, Mycoplasma hominis, Neisseria gonorrhoeae, Listeria monocytogenes, and Chlamydia trachomatis . In case of a positive culture, the specific treatment was indicated . MEASUREMENTS: Positive or negative culture for each of the studied pathogens, and the presence or absence of a preterm delivery for each of the patients included in the study . RESULTS: Three hundred and ninety eight were still present at the end of the study, of which 156 had a CVI and 242 had no CVI . No differences between both groups were observed concerning preterm delivery . Significant relative risks were: In the first stage, Ureaplasma urealyticum and Mycoplasma hominis with RR = 9.0 (6.81, 11.8); in the second stage, Ureaplasma urealyticum with RR = 6.2 (3.30, 11.7) and Escherichia coli with RR = 3.4 (1.33, 8.6); in the third stage, Ureaplasma urealyticum with RR = 9.19 (6.93, 12.1) . The logistic regression analysis identified Ureaplasma urealyticum during the second stage with OR = 16.6 (2.9, 93.7), statistically significant with p = 0.001 . The survival analysis showed differences between the two groups concerning pregnancy duration (p < 0.001) . CONCLUSIONS: There is a difference in the duration in pregnancy in patients with CVI and without CVI . Ureaplasma urealyticum is consistently associated with preterm delivery.

Arch Pathol Lab Med, 1996 Nov, 120(11), 1019 - 22
Effect of off-site transportation on detection of Neisseria gonorrhoeae in endocervical specimens; Iwen PC et al.; OBJECTIVES: To evaluate both the effect of off-site transportation on detection of Neisseria gonorrhoeae in cultured endocervical specimens and the impact of transportation on viability of N . gonorrhoeae by comparison of culture with a nucleic acid probe assay . DESIGN: Three endocervical swabs were randomly collected; one was tested on-site using a nucleic acid-based assay (PACE 2NG System, Gen-Probe, Inc, San Diego, Calif), one was tested off-site following inoculation to modified Thayer-Martin agar (Remel, Lenexa, Kan), and a third swab was tested on-site by culture isolation . A nucleic acid amplification assay of the original swab for PACE 2NG testing was used to resolve discrepancies . SETTING: The emergency department of a university medical center . PATIENTS: Four hundred two patients were evaluated . The test population consisted of both asymptomatic and symptomatic patients . MAIN OUTCOME MEASURE: Positivity for N . gonorrhoeae by one or more of the test procedures, with discrepancy analysis when warranted . RESULTS: Of 402 specimens evaluated, the sensitivities for on-site and off-site testing using culture isolation for N . gonorrhoeae were 88.9% and 77.8%, respectively, in a population prevalence of 6.7% . However, the sensitivity for on-site PACE 2NG testing for N . gonorrhoeae was 96.3% . CONCLUSIONS: A decrease in sensitivity between on-site and off-site culture was found, which suggested transportation may have an adverse effect on the detection of N gonorrhoeae . However, with the limited population and prevalence, the difference was not found to be statistically significant . Further studies indicated that the nucleic acid probe assay was significantly more sensitive (P = .05) when compared with off-site testing using a culture isolation method, demonstrating that viability is an important consideration . These results suggested that a molecular probe assay should be considered in testing specimens for N . gonorrhoeae, especially when the specimen is to be transported off-site.

Clin Microbiol Infect, 2002 Apr, 8(4), 222 - 8
Evaluation of the VITAL (bioMérieux) automated blood culture system using blind subculture; Gimenez M et al.; OBJECTIVE: This study was performed to determine the ability of the VITAL system to detect and allow recovery of microorganisms that are difficult to grow, such as Brucella spp., yeasts, or anaerobes, as well as to determine the need for blind subcultures after the incubation period . METHODS: A prospective evaluation of the system was performed, and 8247 blood culture bottles were processed . The standard was blind subculture from all the bottles after 5 days of incubation . RESULTS: There were 3.2% false-positive and 0.6% false-negative results (72% of clinical importance) . The system sensitivity for yeasts was 41% . The mean time for detection of Neisseria meningitidis was 31.9 +/- 2.8 h, for Brucella spp . 119.7 +/- 2 h, and for yeast 51.5 +/- 27.8 h . CONCLUSIONS: The VITAL system poses has serious difficulties in the detection of N . meningitidis, Brucella spp., yeast and methicillin- and aminoglycoside-resistant Staphylococcus aureus (MARSA) . The low system sensitivity for yeast detection makes the blind subculture necessary after the incubation period.

Helicobacter, 2002 Jun, 7(3), 163 - 9
Helicobacter pylori does not mediate the formation of carcinogenic N-nitrosamines; Vermeer IT et al.; BACKGROUND: Both N-nitroso compounds and colonization with Helicobacter pylori represent known risk-factors for the development of gastric cancer . Endogenous formation of N-nitroso compounds is thought to occur predominantly in acidic environments such as the stomach . At neutral pH, bacteria can catalyze the formation of N-nitroso compounds . Based on experiments with a noncarcinogenic N-nitroso compound as end product, and using only a single H . pylori strain, it was recently reported that H . pylori only displays a low nitrosation capacity . As H . pylori is a highly diverse bacterial species, it is reasonable to question the generality of this finding . In this study, several genetically distinct H . pylori strains are tested for their capacity to form carcinogenic N-nitrosamines . MATERIALS AND METHODS: Bacteria were grown in the presence of 0-1000 microM morpholine and nitrite (in a 1 : 1 molar ratio), at pH 7, 5 and 3 . RESULTS: Incubation of Neisseria cinerea (positive control) with 500 microM morpholine and 500 microM nitrite, resulted in a significant increase in formation of N-nitrosomorpholine, but there was no significant induction of N-nitrosomorpholine formation by any of the H . pylori strains, at any of the three pH conditions . CONCLUSION: H . pylori does not induce formation of the carcinogenic N-nitrosomorpholine in vitro . The previously reported weak nitrosation capacity of H . pylori is not sufficient to nitrosate the more difficulty nitrosatable morpholine . This probably also holds true for other secondary amines . These results imply that the increased incidence of gastric cancer formation that is associated with gastric colonization by H . pylori is unlikely to result from the direct induced formation of carcinogenic nitrosamines by H . pylori . However, this has to be further confirmed in in vivo studies.

Glycobiology, 2002 Apr, 12(4), 235 - 40
A new fermentation process allows large-scale production of human milk oligosaccharides by metabolically engineered bacteria; Priem B et al.; When fed to a beta-galactosidase-negative (lacZ(-)) Escherichia coli strain that was grown on an alternative carbon source (such as glycerol), lactose accumulated intracellularly on induction of the lactose permease . We showed that intracellular lactose was efficiently glycosylated when genes of glycosyltransferase that use lactose as acceptor were expressed . High-cell-density cultivation of lacZ(-) strains that overexpressed the beta 1,3 N acetyl glucosaminyltransferase lgtA gene of Neisseria meningitidis resulted in the synthesis of 6 g x L(-1) of the expected trisaccharide (GlcNAc beta 1-3Gal beta 1-4Glc) . When the beta 1,4 galactosyltransferase lgtB gene of N . meningitidis was coexpressed with lgtA, the trisaccharide was further converted to lacto-N-neotetraose (Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc) and lacto-N-neoheaxose with a yield higher than 5 g x L(-1) . In a similar way, the nanA(-) E . coli strain that was devoid of NeuAc aldolase activity accumulated NeuAc on induction of the NanT permease and the lacZ(-) nanA(-) strain that overexpressed the N . meningitidis genes of the alpha2,3 sialyltransferase and of the CMP-NeuAc synthase efficiently produced sialyllactose (NeuAc alpha 2-3Gal beta 1-4Glc) from exogenous NeuAc and lactose.

S Afr Med J, 2002 Mar, 92(3), 238 - 40
Single low-dose ceftriaxone for the treatment of gonococcal ophthalmia--implications for the national programme for the syndromic management of sexually transmitted diseases; Hoosen AA et al.; We prospectively analysed a total of 21 baby-mother pairs with culture-proven Neisseria gonorrhoeae treated with a single low dose of ceftriaxone, namely 62.5 mg for babies and 125 mg for mothers respectively . N . gonorrhoeae was eradicated from all babies' eyes with no residual damage, as well as from the mothers' cervixes . A single low dose of 62.5 mg ceftriaxone has emerged as the treatment of choice for gonococcal ophthalmia neonatorum because of its excellent activity against N . gonorrhocae, including penicillinase-producing strains.

Carbohydr Res, 2002 Jun 5, 337(11), 969 - 76
Efficient chemoenzymatic synthesis of globotriose and its derivatives with a recombinant alpha-(1-->4)-galactosyltransferase; Zhang J et al.; A truncated alpha-(1-->4)-galactosyltransferase (LgtC) gene from Neisseria meningitidis was cloned . The recombinant glycosyltransferase was expressed in Escherichia coli BL21 (DE3) strain with high specific activity (5 units/mg protein) . Its acceptor specificity was carefully characterized . Then the purified enzyme was utilized in highly efficient syntheses of globotriose and a variety of alpha-(1-->4)-galactosylated derivatives as potential antibacterial agents.

Obstet Gynecol, 2002 Apr, 99(4), 603 - 7
Combination of bacterial vaginosis and leukorrhea as a predictor of cervical chlamydial or gonococcal infection; Steinhandler L et al.; OBJECTIVE: To evaluate whether the combination of bacterial vaginosis and leukorrhea on microscopic evaluation of a saline wet preparation is associated with cervical infection with Chlamydia trachomatis (C . trachomatis) or Neisseria gonorrhea (N . gonorrhea) . METHODS: A cross-sectional study of 598 patients was performed . Nonpregnant patients undergoing a saline wet preparation and microbiologic testing for C . trachomatis and N . gonorrhea were eligible . Providers prospectively collected data from saline microscopic analysis . Bacterial vaginosis was documented based on Amsel's criteria, and ligase chain reaction testing of the endocervix was performed for C . trachomatis and N . gonorrhea . RESULTS: On univariate analysis, young age (less than 25 years), unmarried marital status, black race, and the presence of leukorrhea were all associated with increased rates of testing positive for C . trachomatis and N . gonorrhea (P <.05) . We stratified patients into three groups: 1) no evidence of bacterial vaginosis or leukorrhea; 2) evidence of either bacterial vaginosis or leukorrhea, but not both; and 3) evidence of both bacterial vaginosis and leukorrhea . Using logistic regression analysis to control for age, marital status, and race, we found an odds ratio of 3.8 (95% confidence interval 1.3, 11.6) for a positive microbiologic test for either C . trachomatis or N . gonorrhea in women with the combination of bacterial vaginosis and leukorrhea . CONCLUSION: In this group of high-risk women seen in an urgent care facility, the presence of both bacterial vaginosis and leukorrhea was associated with an increased risk of cervical infection . Future prospective studies should evaluate whether empiric treatment of high-risk women with these findings is justified and cost-effective.

Arch Pediatr Adolesc Med, 2002 Jun, 156(6), 588 - 91
What sexually transmitted disease screening method does the adolescent prefer? Adolescents' attitudes toward first-void urine, self-collected vaginal swab, and pelvic examination; Serlin M et al.; OBJECTIVE: To assess sexually active adolescents' attitudes toward 3 screening collection techniques for detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis using first-void urine (FVU), self-collected vaginal swab specimens, and pelvic examination with clinician-collected endocervical swab specimens . DESIGN: Participants completed a preexamination health survey, provided FVU and self-collected vaginal swab samples, and had a pelvic examination with endocervical swab specimen collection . In a confidential postexamination interview, patients ranked the 3 screening techniques according to preference and responded to qualitative positive and negative descriptors to evaluate each technique . SETTING: San Francisco area health maintenance organization and university clinics . PARTICIPANTS: A convenience sample of 155 ethnically diverse females aged 12 to 21 years, who were sexually active and were to have a pelvic examination . MAIN OUTCOME MEASURES: Adolescents' preferences for and evaluations of 3 sexually transmitted disease screening techniques . RESULTS: Participants preferred the FVU test for sexually transmitted disease screening over the pelvic examination and the self-administered vaginal swab test (P<.001) . These results were consistent when controlling for potentially mitigating experiences, including previous pelvic examination, tampon or condom use, and prior pregnancy . In evaluating what they liked and disliked about each of the 3 screening methods, participants described the FVU most positively, the pelvic examination most negatively, and the vaginal swab technique slightly less positively than the FVU . CONCLUSION: Most sexually active adolescents attending clinics for pelvic examination prefer to be screened for sexually transmitted diseases first by the FVU, second by the self-collected vaginal swab test, and last by the pelvic examination.

J Clin Microbiol, 2002 Jun, 40(6), 2037 - 40
Antimicrobial susceptibility of Neisseria gonorrhoeae isolated in Bangladesh (1997 to 1999): rapid shift to fluoroquinolone resistance; Rahman M et al.; Periodic monitoring of antimicrobial susceptibility of Neisseria gonorrhoeae is essential for early detection of emergence of drug resistance . A total of 343 gonococcal strains isolated from high-risk and general populations in Bangladesh from 1997 to 1999 were studied . The MICs of penicillin, tetracycline, ciprofloxacin, ceftriaxone, and spectinomycin for the isolates were determined by the agar dilution method . Of the isolates from 1997, 9% were resistant (MIC >or= 1.0 microg/ml) to ciprofloxacin, while 41 and 49% of the isolates from 1998 and 1999, respectively, were resistant to ciprofloxacin . Of the N . gonorrhoeae isolates from 1998 and 1999, 1.2 and 3.6%, respectively, both were penicillinase producing and displayed plasmid-mediated tetracycline resistance.

Sex Transm Dis, 2002 Jun, 29(6), 331 - 4
Phthirus pubis as a predictor for chlamydia infections in adolescents; Pierzchalski JL et al.; BACKGROUND: Providers not skilled in the evaluation of sexually transmitted infections (STIs) may treat pubic lice infestation without considering other organisms . This study compared the rate of chlamydia and gonorrhea infections in adolescents with and without pubic lice . GOAL: The goals of the study were to compare the rate of chlamydia and gonorrhea infections between adolescents infested or not infested with pubic lice and to evaluate lice infestation as a predictor for concurrent chlamydia or gonorrhea infection . STUDY DESIGN: A retrospective chart review of sexually active adolescents at a juvenile detention center in the Midwest between July 1998 and June 2000 was conducted . The index group was 62 adolescents with pubic lice screened for concurrent STIs . The control group included 201 randomly selected adolescents without pubic lice who underwent STI screening . RESULTS: In the entire study population (263 subjects), there were 60 cases of chlamydia (23% of all subjects) and 29 cases of gonorrhea (11%) . Neisseria gonorrhoeae infection was present in 18% of index subjects and 9% of controls . Chlamydia trachomatis infection was noted in 39% of index subjects and 18% of controls . Pubic lice infestation predicted C trachomatis infection (odds ratio = 3.31) . CONCLUSIONS: Pubic lice infestation is predictive of a concurrent C trachomatis infection in this population . Adolescents infested with pubic lice should be screened for other STIs, including chlamydia and gonorrhea.

Sex Transm Dis, 2002 Jun, 29(6), 324 - 30
Syndromic versus laboratory-based diagnosis of cervical infections among female sex workers in Benin: implications of nonattendance for return visits; Mukenge-Tshibaka L et al.; BACKGROUND: The syndromic diagnostic approach is the most realistic and cost-effective strategy for controlling sexually transmitted infections (STIs) in the developing world . Its potential advantages should be evaluated . GOAL: The goal of the current study was to examine whether the syndromic approach might diagnose more cases of cervicitis due to Neisseria gonorrhoeae or Chlamydia trachomatis than laboratory tests . STUDY DESIGN: The participants were 481 female sex workers in Benin, screened for STIs and treated on the basis of the clinical findings . They were asked to return to the clinic within 10 days for laboratory test results and appropriate treatment when necessary . RESULTS: The prevalence of cervical infections was 24.5% . In comparison to the gold standard, the sensitivity of the syndromic diagnosis approach for the detection of N gonorrhoeae/C trachomatis infections was 48.3%; that of the locally performed laboratory tests was 74.6% . However, the sensitivity of the laboratory tests dropped to 28.8% when it was taken into consideration that 57.6% of the infected women did not return to the clinic within 10 days . CONCLUSIONS: The syndromic diagnosis approach should continue to be used for female sex workers in Benin because returning for treatment is problematic . Presumptive treatment at their initial visit could be a complement to this approach, given the high prevalence of cervicitis in this population.

Sex Transm Dis, 2002 Jun, 29(6), 319 - 23
A recent increase in the incidence of male urethritis in France; Massari V et al.; BACKGROUND: At-risk sexual behaviors appear to have increased recently in France, possibly because of the reassuring efficacy of highly active antiretroviral treatments . The objective of this study was to assess the pattern of change . GOAL: The goal was to analyze trends in the incidence of male urethritis, a marker of at-risk sexual behavior, between 1989 and 2000 . STUDY DESIGN: The general practitioners of the Sentinelles Network reported, through an online computer system, all cases of male urethritis they diagnosed, together with the patients' age, presence of discharge, sexual preference, history of sexually transmitted disease, prescriptions, and results of microbiologic findings . RESULTS: After falling markedly between 1989 and 1995, the incidence of urethritis increased slightly in 1996 and then stabilized . The percentage of homosexual/bisexual men was higher among the cases than in the general population . Homosexual/bisexual men were younger than heterosexual men with urethritis; they were more likely to have a discharge and a history of sexually transmitted disease and had more sex partners . The main causative organisms were chlamydiae (18%) and Neisseria gonorrhoeae (13%) . CONCLUSION: The increasing incidence of urethritis in France calls for a readjustment of preventive strategies.

Clin Infect Dis, 2002 Jun 15, 34(12), E74 - 6 Epub 2002 May 24.
Wound infection with Neisseria weaveri and a novel subspecies of pasteurella multocida in a child who sustained a tiger bite; Capitini CM et al.; A 7-year-old girl developed a wound infection as a result of a tiger bite she sustained . DNA sequence analysis revealed that the causative organisms were Neisseria weaveri and what is, to our knowledge, a previously undescribed subspecies of Pasteurella multocida, for which we propose the designation "Pasteurella multocida subspecies tigris subspecies nov."

J Microbiol Methods, 2002 Aug, 50(3), 249 - 62
A novel approach to study variable heavy chain gene usage in response to the capsular polysaccharide of Neisseria meningitidis serogroup C; Srivastava N et al.; The molecular mechanisms involved in the relatively poor immune response in the elderly are not clearly understood . Qualitative aspects of the immune response could be a possible explanation for the differential response to T-independent antigens in young adults and elderly . This study is directed towards elucidating the differential usage of variable heavy chain by young adult and elderly derived sequences in response to the capsular polysaccharide of Neisseria meningitidis serogroup C . We currently report findings of a preliminary study designed to test the feasibility of a novel approach to isolate antigen-specific B cells . Paramagnetic beads coated with an anti-idiotypic antibody, which mimics the capsular polysaccharide of N . meningitidis serogroup C, were used to select B cells . Analysis of the gene usage data indicates some unexpected differences in the use of variable chain heavy chain in the case of young adult versus elderly sequences . The elderly derived sequences use a more diverse array of V(H) gene families in contrast to the young adult sequences, where the V(H) gene family usage is restricted . Nearly half the young adult sequences utilize V(H)3-15 germline sequence while only 25% of the elderly sequences use this germline sequence . There were interesting differences in the types of JH chain and the composition and length of CDR3 utilized by the two groups . Together, these significant differences may contribute towards the poor immune response to T-independent antigens in the elderly . These data validate the techniques used for these studies and suggest that it is pertinent to use this approach towards future investigations to elucidate gene usage in response to an antigen.

Biochim Biophys Acta, 2002 Jun 7, 1576(1-2), 39 - 44
The abundant class of nemis repeats provides RNA substrates for ribonuclease III in Neisseriae; De Gregorio E et al.; About 2% of the Neisseria meningitidis genome is made up by nemis, short DNA sequences which feature long terminal inverted repeats (TIRs) . Most nemis are interspersed with single-copy DNA and are found at close distance from cellular genes . In this work, we demonstrate than RNAs spanning nemis of different length and sequence compositions are specifically cleaved at hairpins formed by nemis termini by total cellular lysates derived from both Escherichia coli and Neisseria lactamica strains . The use of cellular extracts from E . coli strains impaired in the activity of known ribonucleases let to establish that cleavage at nemis TIRs is specifically mediated by the endoribonuclease RNase III . Data set the base for the identification of all of the neisserial genes that are regulated by RNase III because of their physical association with nemis DNA.

Mol Cell Probes, 2002 Apr, 16(2), 153 - 8
Integration host factor interactions with Neisseria gene sequences: correlation between predicted binding sites and in vitro binding of Neisseria -derived IHF protein; Hill SA et al.; Putative integration host factor (IHF) binding sites are frequently being identified in Neisseria gene sequences on the basis of similarity to a degenerate Escherichia coli -derived consensus binding sequence . In this report, three different Neisseria genetic systems that contain predicted IHF binding sites were assessed for IHF binding through gel retardation analysis . The results show a positive correlation between the identification of a predicted Neisseria IHF binding site and in vitro binding of Neisseria -derived IHF protein .

Antimicrob Agents Chemother, 2002 Jun, 46(6), 1671 - 3
High prevalence of high-level ciprofloxacin resistance in Neisseria gonorrhoeae in Tel Aviv, Israel: correlation with response to therapy; Dan M et al.; A survey of the drug susceptibilities of gonococcal isolates from 100 consecutive patients attending clinics in the Tel Aviv area of Israel during the period from February to September 2000 has shown a 61% rate of ciprofloxacin resistance (MICs, > or =1 microg/ml); 51 isolates were highly resistant (MICs, > or =4 microg/ml) . Ciprofloxacin-resistant strains were more prevalent among isolates with chromosomally mediated resistance to penicillin and tetracycline . Therapeutic failures with ciprofloxacin correlated with MICs of > or =1 mg/ml (P < 0.01) . The high rate of ciprofloxacin resistance coinciding with a sharp rise in the incidence of gonorrhea precludes the use of fluoroquinolone drugs for the empirical treatment of gonococcal infections acquired in Israel.

MMWR Morb Mortal Wkly Rep, 2002 May 10, 51(18), 389 - 92
Nonoxynol-9 spermicide contraception use--United States, 1999; Mass spectrometric strategy for the characterization of lipooligosaccharides from Neisseria gonorrhoeae 302 using FTICR; Department of Chemistry, University of California, Berkeley 94720, USAThe lipooligosaccharides (LOS) of Neisseria gonorrhoeae 302 were profiled using Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) . Using techniques developed in this laboratory, the topology and some of the linkages of the LOS were determined . Mass spectrometric analysis in the negative ion mode yielded a glycoform of the composition: Hex3 Hep2 Hxn1 PEA1 KDO2 DPLA . The composition was confirmed through exact mass measurements, which showed only a 2 ppm error between the exact mass and theoretical mass . Although the core structure has been postulated previously, the positioning of the three hexose moieties were in question for this particular strain of N . gonorrhoeae . Topology assignment was performed through collision-induced dissociation analysis of the O-deacylated glycoform in the negative ion mode followed by submission to the saccharide topology analysis tool (STAT) computer program, which confirmed the topology assignment . It was found that the three hexoses were added to the Hep{I} of the conserved core of N . gonorrhoeae in a linear fashion, while Hep{II} remains unbranched . Linkage position analysis was performed through application of a mild acid hydrolysis technique followed by collision-induced dissociation of the sodiated precursor ions, yielding a 1 --> 4 linkage between the terminating and penultimate hexoses.

Infect Immun, 2002 Jun, 70(6), 2752 - 7
A lytic transglycosylase of Neisseria gonorrhoeae is involved in peptidoglycan-derived cytotoxin production; Cloud KA et al.; Neisseria gonorrhoeae releases soluble fragments of peptidoglycan during growth . These molecules are implicated in the pathogenesis of various forms of gonococcal infection . A major peptidoglycan fragment released by gonococci is identical to the tracheal cytotoxin of Bordetella pertussis and has been shown to kill ciliated fallopian tube cells in organ culture . Previous studies indicated that a unique lytic peptidoglycan transglycosylase (AtlA) was responsible for some, but not all, of the peptidoglycan-derived cytotoxin (PGCT) production in certain gonococcal strains . To examine the role of other putative lytic transglycosylases in PGCT production, we made a deletion mutation in a gonococcal gene exhibiting similarity with genes encoding lytic transglycosylases from other bacterial species . The gonococcal mutant was viable and grew normally, but it was less autolytic than the wild-type strain in stationary-phase culture and under nongrowth conditions . The gonococcal mutant was reduced in peptidoglycan turnover, and the profile of the released products showed a reduction in monomeric peptidoglycan . Proportionally more multimeric fragments were released . These results suggest that this gonococcal gene (ltgA) encodes a lytic peptidoglycan transglycosylase and that it is responsible for a significant proportion of the PGCT released by N . gonorrhoeae.

Biosci Biotechnol Biochem, 2002 Mar, 66(3), 508 - 15
Purification and characterization of an endo-1,4-beta-glucanase from Neisseria sicca SB that hydrolyzes beta-1,4 linkages in cellulose acetate; Moriyoshi K et al.; An enzyme catalyzing hydrolysis of beta-1,4 bonds in cellulose acetate was purified 18.3-fold to electrophoretic homogeneity from a culture supernatant of Neisseria sicca SB, which can assimilate cellulose acetate as the sole carbon and energy source . The molecular mass of the enzyme was 41 kDa and the isoelectric point was 4.8 . The pH and temperature optima of the enzyme were 6.0-7.0 and 60 degrees C . The enzyme catalyzed hydrolysis of water-soluble cellulose acetate (degree of substitution, 0.88) and carboxymethyl cellulose . The Km and Vmax for water-soluble cellulose acetate and carboxymethyl cellulose were 0.242% and 2.24 micromol/min/mg, and 2.28% and 12.8 micromol/min/mg, respectively . It is estimated that the enzyme is a kind of endo-1,4-beta-glucanase (EC 3.2.1.4) from the substrate specificity and hydrolysis products of cellooligosaccharides . The enzyme and cellulose acetate esterase from Neisseria sicca SB degraded water-insoluble cellulose acetate by synergistic action.

East Afr Med J, 2001 May, 78(5), 259 - 61
Antimicrobial sensitivity of Neisseria gonorrhoea in Gondar, Ethiopia; Tadesse A et al.; OBJECTIVES: To determine the antimicrobial susceptibility pattern of Neisseria gonorrheae and to obtain epidemiological information on resistance of Neisseria gonorrheae in Gondar town . DESIGN: A cross sectional study . SETTING: Gondar Health Centre . SUBJECTS: One hundred and seventy eight male patients presented with urethral discharge and who volunteered to participate in the study were included . RESULTS: A total of 142 strains of N . gonorrhoeae were isolated from 168 cultures received . The sensitivity of gonococcal isolates ranges from 98.6% to chloramphenicol to 7.7% to cotrimoxazole . Multiple drug resistance was reported in 87.5% of isolates and only four isolates were sensitive to all antibiotics . One strain of N . gonorrheae was resistant to as many as eight antibiotics (tetracycline, penicillin, ampicillin, kanamicin, methicilin, carbenicillin, cotrimoxazole and ceftriaxone) . More than eighty five per cent of the isolated strains were penicillinase-producing Neisseria gonorrhoeae (PPNG) . CONCLUSION: Gonococcal resistance is a significant public health problem in Gondar region and the drugs recommended for treatment of gonococcal cases by the national sexually transmitted diseases (STDs) control programme need to be revised.

Emerg Infect Dis, 2002 May, 8(5), 462 - 6
Clonal groupings in serogroup X Neisseria meningitidis; Gagneux S et al.; The genetic diversity of 134 serogroup X Neisseria meningitis isolates from Africa, Europe, and North America was analyzed by multilocus sequence typing and pulsed-field gel electrophoresis . Although most European and American isolates were highly diverse, one clonal grouping was identified in sporadic disease and carrier strains isolated over the last 2 decades in the United Kingdom, the Netherlands, Germany, and the United States . In contrast to the diversity in the European and American isolates, most carrier and disease isolates recovered during the last 30 years in countries in the African meningitis belt belonged to a second clonal grouping . During the last decade, these bacteria have caused meningitis outbreaks in Niger and Ghana . These results support the development of a comprehensive conjugate vaccine that would include serogroup X polysaccharide.

J Infect Dis, 2002 May 15, 185(10), 1525 - 9 Epub 2002 Apr 30.
Gonococcal cervicitis is associated with reduced systemic CD8+ T cell responses in human immunodeficiency virus type 1-infected and exposed, uninfected sex workers; Kaul R et al.; Neisseria gonorrhoeae cervicitis and human immunodeficiency virus (HIV) type 1 frequently coinfect core transmitter populations, such as female sex workers . Gonococcal cervicitis is associated with increased viral shedding and plasma viremia in HIV-1-infected women and increased HIV-1 susceptibility in uninfected women . We studied the influence of gonococcal cervicitis on CD8(+) interferon (IFN)-gamma responses to HIV-1 and cytomegalovirus (CMV) epitopes in HIV-1-infected and in highly-exposed, persistently seronegative (HEPS) female sex workers . In HIV-1-infected women, gonococcal cervicitis was associated with reduced IFN-gamma responses in bulk CD8(+) lymphocyte populations, and intracellular cytokine staining, combined with class I major histocompatibility complex (MHC)-peptide tetramer studies, demonstrated reduced IFN-gamma production by HIV-1 epitope-specific CD8(+) lymphocytes . In HEPS sex workers, cervicitis was associated with the transient loss of systemic HIV-1-specific CD8(+) responses and with reduced function of CMV-specific CD8(+) lymphocytes . Impaired function of virus-specific CD8(+) lymphocytes may partly explain the deleterious effects of gonococcal cervicitis on HIV-1 immune control and susceptibility.

Przegl Epidemiol, 2001, 55 Suppl 3, 63 - 7
{Usefulness of plasma C-reactive protein (CRP) estimation in patients with bacterial sepsis}; Kepa L et al.; Between 1997-2000 there were 30 patients with bacterial sepsis treated at Department of Infectious Diseases of Silesian Medical University School in Bytom . Neisseria meningitidis (23.33%), Streptococcus pneumoniae (20%) and Staphylococcus aureus (3.33%) were most frequent etiological factors of the disease . The etiology of sepsis was not revealed in 53.34% of cases . Clinical state of patients was assessed using SSS and SAPS II score . All subjects were divided into two groups: group I--patients with septic syndrome symptoms (11 cases), group II--patients without septic syndrome symptoms (19 cases) . The aim of the study was usefulness of plasma C-reactive protein estimation in the course of bacterial sepsis monitoring . Plasma CRP concentration was assessed by immunoturbidometric method (using RANDOX, UK kits) . Blood samples were taken on th 1., 4 . and 10 . day of therapy . Plasma CRP levels were statistically significantly higher in patients of the group I in samples of the first and the second assay . CRP serum concentration correlated with severity of a patient's clinical state . Type of sepsis's etiological factor did not exert influence on plasma CRP level . In fatal cases high plasma CRP concentration was recorded during the whole time of hospitalization, whereas in cases of recovery fast decrease of this protein level was noted . The obtained result indicate, that estimation of plasma CRP concentration in patients with bacterial sepsis may be helpful in evaluation of clinical state severity, monitoring of the disease course as well as therapy efficacy.

Protein Eng, 2002 Apr, 15(4), 297 - 304
Model of Vibrio cholerae toxin coregulated pilin capable of filament formation; Chattopadhyaya R et al.; A complete three-dimensional model (RCSB001169; PDB code 1qqz ) for the Vibrio cholerae toxin coregulated pilus protein (TcpA), including residues 1-197, is presented . We have used the crystal structure of the Neisseria gonorrhoeae pilin (PilE), available biochemical data about TcpA, variations in the primary sequences of TcpA among various Vibrio cholerae strains and secondary structure prediction, hydrophilicity, surface probability and antigenicity plots for TcpA to build our model . In our TcpA model, the first 137 residues possess a structure similar to the PilE, but the remainder is different . Though the ladle shape is still preserved, TcpA possesses a larger ladle head or globular domain compared to PilE . Using this model, it has been possible to identify two kinds of conserved residues: (i) those forming the core of the TcpA monomer and (ii) those involved in the monomer-monomer interactions leading to fibre formation . Residues on the fibre exterior, important in the mediation of bacterium (pilus)-bacterium (pilus) and bacterium (pilus)-host interactions, show more variability in comparison to those of (i) and (ii).

Proc Natl Acad Sci U S A, 2002 Apr 30, 99(9), 6103 - 7
Mutator clones of Neisseria meningitidis in epidemic serogroup A disease; Richardson AR et al.; Serogroup A Neisseria meningitidis has repeatedly caused widespread epidemics of meningitis and septicemia throughout the 20th century . Recently, in a limited collection of strains, epidemic serogroup A isolates were found to have elevated mutation rates that was caused by defects in mismatch repair pathways . To ascertain the role of these mutators in the epidemic spread of this serogroup, the prevalence of hypermutability in a collection of 95 serogroup A N . meningitidis invasive isolates was determined . Overall mutability in Neisseriae can be described by measuring both missense mutation rates as well as phase variation frequencies of "contingency loci." Fifty-seven percent of serogroup A isolates possessed elevated mutability, which could be divided into two classes: intermediate and high level . Eleven of 20 high-level mutators, with phase variation rates >100-fold higher than wild-type isolates, were defective in mismatch repair . Ten of the 34 intermediate mutators possessing >10-fold increases in phase variation rates could be partially complemented by a wild-type mutL allele . A high prevalence of mutators in epidemic isolates indicates that hypermutability may play a major role in the transmission of this pathogen . The added diversity derived from increased phase variation rates may allow fixation of mutator alleles more frequently during epidemic spread.

J Endotoxin Res, 2002, 8(1), 47 - 58
Phase variable changes in genes lgtA and lgtC within the lgtABCDE operon of Neisseria gonorrhoeae can modulate gonococcal susceptibility to normal human serum; Shafer WM et al.; The alpha-chain of the core oligosaccharide of the lipo-oligosaccharide (LOS) produced by Neisseria gonorrhoeae can undergo reversible and rapid changes in structure due to phase-variable production of certain enzymes employed in the biosynthesis of the lacto-N-neotetraose structure . Five of these enzymes are encoded by the lgtABCDE operon, and polynucleotide tracts within three of these genes (lgtA, lgtC and lgtD) can be substrates for slipped-strand mispairing events that lead to nucleotide insertions or deletion events which result in variable production of their respective gene products . We now report that phase-variable synthesis of the lgtA and lgtC gene products in strain FA19 results in the production of elongated LOS alpha-chains and that the presence of these LOS species can result in gonococci being sensitive to the bacteriolytic action of serum-antibody and complement . Hence, phase variation within the lgtABCDE operon can significantly impact the ability of gonococci to subvert this important host defense system.

EMBO J, 2002 May 1, 21(9), 2122 - 31
Export of autotransported proteins proceeds through an oligomeric ring shaped by C-terminal domains; Veiga E et al.; An investigation was made into the oligomerization, the ability to form pores and the secretion-related properties of the 45 kDa C-terminal domain of the IgA protease (C-IgAP) from Neisseria gonorrhoeae . This protease is the best studied example of the autotransporters (ATs), a large family of exoproteins from Gram-negative bacteria that includes numerous virulence factors from human pathogens . These proteins contain an N-terminal passenger domain that em bodies the secreted polypeptide, while the C-domain inserts into the outer membrane (OM) and trans locates the linked N-module into the extracellular medium . Here we report that purified C-IgAP forms an oligomeric complex of approximately 500 kDa with a ring-like structure containing a central cavity of approximately 2 nm diameter that is the conduit for the export of the N-domains . These data overcome the previous model for ATs, which postulated the passage of the N-module through the hydrophilic channel of the beta-barrel of each monomeric C-domain . Our results advocate a secretion mechanism not unlike other bacterial export systems, such as the secretins or fimbrial ushers, which rely on multimeric complexes assembled in the OM.

EMBO J, 2002 May 1, 21(9), 2076 - 86
Crystal structure of murine sCEACAM1a{1,4}: a coronavirus receptor in the CEA family; Tan K et al.; CEACAM1 is a member of the carcinoembryonic antigen (CEA) family . Isoforms of murine CEACAM1 serve as receptors for mouse hepatitis virus (MHV), a murine coronavirus . Here we report the crystal structure of soluble murine sCEACAM1a{1,4}, which is composed of two Ig-like domains and has MHV neutralizing activity . Its N-terminal domain has a uniquely folded CC' loop that encompasses key virus-binding residues . This is the first atomic structure of any member of the CEA family, and provides a prototypic architecture for functional exploration of CEA family members . We discuss the structural basis of virus receptor activities of murine CEACAM1 proteins, binding of Neisseria to human CEACAM1, and other homophilic and heterophilic interactions of CEA family members.

Curr Opin Infect Dis, 2001 Feb, 14(1), 45 - 51
New tests for bacterial sexually transmitted diseases; Morse SA; Recent advances in diagnostic tests for sexually transmitted diseases include the development of a synthetic Venereal Disease Research Laboratory reagent that will improve the sensitivity and stability of nontreponemal serologic tests for syphilis . A second generation user friendly and high throughput nucleic acid amplification test for Chlamydia trachomatis and Neisseria gonorrhoeae has also been developed.

Trends Microbiol, 2002 May, 10(5), 227 - 32
How do extracellular pathogens cross the blood-brain barrier?
Nassif X, Bourdoulous S, Eugene E, Couraud PO.
Bacterial invasion of the meninges can occur as a consequence of bloodstream invasion by some bacterial pathogens . Bacteria enter the central nervous system following a direct interaction with the luminal side of the cerebral endothelium, which constitutes the blood-brain barrier . To breach the barriers protecting the brain, extracellular pathogens must cross a monolayer of tight junction-expressing endothelial or epithelial cells . The limited number of pathogens capable of crossing these tight barriers and invading the meninges suggests that they display very specific attributes . For Neisseria meningitidis, type IV pili have been identified as being essential for meningeal invasion and it is believed other, as-yet-unidentified factors are also involved.

Int J STD AIDS, 2002 May, 13(5), 323 - 5
Observations on the microbiology of urethritis in black South African men; Taylor-Robinson D et al.; The occurrence of Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium was determined by molecular techniques in urine specimens from 182 black South African men who had symptoms and/or overt signs of urethritis . Eighty-six (47.3%) of these men were infected with N . gonorrhoeae . There were 185 men without overt evidence of urethritis, 16 (8.6%) of whom were also infected with N . gonorrhoeae . Of the 96 men who had non-gonococcal urethritis, 14 (14.6%) were infected with C . trachomatis, 16 (16.7%) with M . genitalium and only one with both microorganisms . In comparison, 15 (8.9%) of 169 men without overt urethritis and without N . gonorrhoeae were infected with C . trachomatis and 15 (8.9%) with M . genitalium, proportions that were about half the size of those in the group with overt urethritis.

Mol Microbiol, 2002 Mar, 43(6), 1379 - 86
Enhanced bacterial virulence through exploitation of host glycosaminoglycans; Menozzi FD et al.; Present in the extracellular matrix and membranes of virtually all animal cells, proteoglycans (PGs) are among the first host macromolecules encountered by infectious agents . Because of their wide distribution and direct accessibility, it is not surprising that pathogenic bacteria have evolved mechanisms to exploit PGs for their own purposes, including mediating attachment to target cells . This is achieved through the expression of adhesins that recognize glycosaminoglycans (GAGs) linked to the core protein of PGs . Some pathogens, such as Bordetella pertussis and Chlamydia trachomatis, may express more than one GAG-binding adhesin . Bacterial interactions with PGs may also facilitate cell invasion or systemic dissemination, as observed for Neisseria gonorrhoeae and Mycobacterium tuberculosis respectively . More-over, pathogenic bacteria can use PGs to enhance their virulence via a shedding of PGs that leads to there lease of effectors that weaken the host defences.The exploitation of PGs by pathogenic bacteria is thus a multifaceted mechanistic process directly related to the potential virulence of a number of microorganisms.

Nippon Hinyokika Gakkai Zasshi, 2002 Mar, 93(3), 450 - 6
{The clinical evaluation of 414 cases of male urethritis}; Hayakawa T et al.; PURPOSE: We reviewed the clinical data of male urethritis diagnosed as sexually transmitted disease with the aim of revealing its clinical features . PATIENTS AND METHODS: In a total number of 414 males diagnosed as having acute urethritis transmitted by sexual activities between January and December in 2000, clinical data were collected and analyzed . In addition, questionnaire concerning the awareness of pharyngeal infection was obtained . RESULTS: Their ages ranged from 16 to 60 years with a mean of 31 years . The source of infection was a commercial sexual worker (CSW) in 288 (69.6%) out of 414 cases . In 241 cases in whom an infection route was determined, 199 cases (82.6%) were considered to have an infection through oral sex . Microbiological examinations isolated Neisseria gonorrhea alone in 206 (49.8%), Chlamydia tracomatis alone in 47 (11.3%) and both in 46 (11.1%) cases . In our study, 57.5% and 20.8% of Neisseria gonorrhea strains were resistant to penicillin and new quinolones, respectively . Based on questionnaire, 174 (57.8%) out of 301 cases were not aware of possible infection from the pharynx through oral sex . CONCLUSION: The remarkable prevalence of gonococcal urethritis might be in part due to the lacking of awareness of infection through oral sex and sexual activities without a condom . Instruction on the prevention as well as the actual situation of urethritis is needed for general population with a broad age spectrum . The appropriate use of antimicrobial agents with sufficient susceptibility and less inductivity of drug-resistance is also of clinical importance.

Am J Obstet Gynecol, 2002 Apr, 186(4), 690 - 5
Endometritis: the clinical-pathologic syndrome; Eckert LO et al.; OBJECTIVE: The purpose of this study was to evaluate histologically proved endometritis as a clinical syndrome that is distinct from laparoscopically confirmed salpingitis . STUDY DESIGN: This was a cross-sectional study of 152 women in an urban hospital with a suspected pelvic inflammatory disease . All women provided a standardized medical history and underwent physical examination, endometrial biopsy, and laparoscopy . We defined endometritis by the presence of plasma cells in endometrial stroma and neutrophils in the endometrial epithelium . RESULTS: Of 152 women who were enrolled, 43 women had neither endometritis nor salpingitis; 26 women had endometritis alone without salpingitis, and 83 women had salpingitis . Those women with endometritis alone more often had douched recently, had a current intrauterine device, and were in menstrual cycle day 1 to 7, compared with women with no endometritis or salpingitis (P =.007,.04,.005, respectively) or women with acute salpingitis (P =.03,.01,.02, respectively) . Infection with Neisseria gonorrhoeae and/or Chlamydia trachomatis was found more frequently in women with endometritis alone than in women with no endometritis or salpingitis (P <.001) and less frequently than in women with salpingitis (P =.05) . Lower quadrant, adnexal, cervical motion, rebound tenderness, peritonitis, tenderness score, fever, and laboratory abnormalities that indicated inflammation and detection of gonorrheal or chlamydial infection were significantly less common in women with endometritis alone than in women with salpingitis but were somewhat more common in women with endometritis alone than among women with no salpingitis or endometritis . CONCLUSION: Among women with suspected pelvic inflammatory disease, the histopathologic manifestations of endometritis were associated with clinical manifestations, infection, and specific risk factors that were intermediate in frequency between women with salpingitis and women with neither endometritis nor salpingitis.

Akush Ginekol (Sofiia), 1999, 38(1), 60 - 3
{Problems of diagnosis and treatment of cervicitis}; Borisov I et al.; Infectious cervicitis is a common disease in women of reproductive age . The symptoms are not characteristic and they are often neglected by the patient and the physician . Cervicitis is a very important clinical entity, because the infection can ascend to the genital tract . The most common causes for the infectious cervicitis are Chlamydia trachomatic and Neisseria gonorrhoeae . A variety of methods for the precise diagnosis must be employed--gynecologic examination, colposcopy, pathomorphology and cytology and bacteriological examination as well . Therapeutic modalities must include coverage for both Chlamydia trachomatis and Neisseria gonorrhoeae . Bacteriological test of cure is mandatory.

Curr Opin Infect Dis, 2000 Feb, 13(1), 47 - 51
Sexually transmitted infections in teenagers; James NJ et al.; Prevention of sexually transmitted infection in teenagers is a high priority . The introduction of non-invasive methods for diagnostic testing of Chlamydia trachomatis and Neisseria gonorrhoeae has enabled screening for these sexually transmitted micro-organisms in a range of community-based settings among populations at high risk, such as teenagers, who do not readily access existing sexual health services.

Cent Afr J Med, 2001 Mar, 47(3), 57 - 64
Determinants of reproductive tract infections among asymptomatic women in Harare, Zimbabwe; Mbizvo EM et al.; OBJECTIVES: To determine the prevalence of and risk factors for reproductive tract infections (RTI) among asymptomatic women attending primary health care clinics in Harare, Zimbabwe . DESIGN: A cross sectional study . MAIN OUTCOME MEASURES: Prevalent RTI, HIV and socio-demographic data . SETTING: Two primary health care clinics in Harare . SUBJECTS: 393 consecutive, consenting women aged 15 to 49 years, presenting at the clinics for antenatal care, family planning, or bringing their children to attend preventive care clinics . RESULTS: More than half of the women had at least one ongoing RTI . Voluntary reporting of symptoms was almost non-existent . Prevalence of HIV was 29.3% and that of classical STIs was 15.4% Trichomoniasis vaginalis, 3.9% syphilis, 3.9% Chlamydia trachomatis and 1.8% Neisseria gonorrhoea . Bacteria vaginosis was diagnosed in 30.3% of the women and candidiasis in 25.4% . Presence of any RTI was significantly associated with lower level of education, sexual debut < 20 years, a non-monogamous partner, and use of a condom during the last sexual encounter . Independent factors associated with cervical infections were young age (< 20 years), being unmarried, current dysuria, warts, clinical signs of purulent or yellowish discharge, lower abdominal pain and cervical friability . Vaginitis was associated with sexual debut or marriage < 20 years, a partner who was a frequent traveller, history of STI, and presence of abnormal vaginal discharge . CONCLUSION: In view of the high prevalence of RTI and HIV and the low reporting of symptoms by infected women, it is recommended that strategies that improve women's awareness of urogenital symptoms be adopted, along with pro-active inquiry of genital symptoms by the health care workers, and treatment of RTIs, in order to reduce the unnecessary burden of RTIs.

Ethiop Med J, 1999 Jul, 37(3), 181 - 7
Increasing incidence of resistance to antimicrobials in Sidamo; Belihu A et al.; This study compares the prevailing sensitivity of bacterial isolates to common antimicrobials during the two-year 1985-87, 1990-92 and 1996-97 periods at Yirga Alem Hospital in southern Ethiopia . All specimens were from patients attending the hospital . We studied 1371 specimens, 337,671 and 363 specimens from the periods 1985-87, 1990-92 and 1996-97, respectively . The study confirms earlier observations of widespread resistance to commonly used antibiotics . Significant increases in the rate of resistance during this thirteen-year period were observed for Neisseria gonorrhoea against erythromycin, trimethoprim-sulphamethoxazole and tetracyclines, for Escherichia coli against ampicillin and trimethoprim-sulphamethoxazole and for Proteus species against ampicillin . We observed a decrease in the prevalence of resistance for Klebsiella and Proteus species against chloramphenicol.

J Infect Chemother, 2002 Mar, 8(1), 81 - 6
A remarkable reduction in the susceptibility of Neisseria gonorrhoeae isolates to cephems and the selection of antibiotic regimens for the single-dose treatment of gonococcal infection in Japan; Tanaka M et al.; We compared the susceptibilities of 100 Neisseria gonorrhoeae isolates obtained from January to June 2000 to a variety of antimicrobial agents, including various cephems, with the susceptibilities of 55 isolates obtained from May to December 1995 . In 11 of the 15 cephems tested, the MIC50 (minimum inhibitory concentration for 50% of isolates) ratios, calculated by dividing the MIC50 for isolates from 2000 by that for isolates from 1995, showed 4-fold to 16-fold differences . The MIC90 ratios of cefodizime and cefditoren showed only a 1-fold and 2-fold difference, respectively . However, the MIC90 ratios of the remaining 13 cephems showed 4-fold to 64-fold differences . We selected the most favorable antibiotic regimens for the single-dose treatment of gonorrhea in Japan according to the antigonococcal activity and pharmacokinetics, based on the criterion (therapeutic time) proposed by Moran and Levine i.e., the regimen should produce a concentration in the serum or plasma at least four times that of the MIC90 value for at least 10 h . Of the 20 single-dose regimens evaluated, only ceftriaxone 1 g intravenously and cefodizime 1 g intravenously were found to have a therapeutic time of more than 10 h for the isolates from 2000 . Our results indicated that the susceptibilities of the isolates from 2000 to all the cephems tested, except for cefditoren and cefodizime, decreased remarkably in comparison with the susceptibilities of the isolates from 1995, while parenteral ceftriaxone 1 g and cefodizime 1 g appeared to be the most favorable single-dose regimens for the treatment of gonococcal infections in Japan.

Biochemistry, 2002 Apr 23, 41(16), 5075 - 85
Mechanistic studies of a retaining alpha-galactosyltransferase from Neisseria meningitidis; Ly HD et al.; Lipopolysaccharyl alpha-galactosyltransferase from Neisseria meningitidis catalyzes the transfer of a galactosyl moiety from the activated donor UDP-Gal to glycoconjugates to yield an elongated saccharide product with net retention of anomeric configuration relative to the donor substrate . Through kinetic analyses in which the concentrations of both substrates are independently varied and through inhibition studies with dead-end analogues of both substrates and with the oligosaccharide product, we have demonstrated that this enzyme follows an ordered bi-bi kinetic mechanism . Various aspects of the chemical mechanism including the possible formation of a covalent glycosyl-enzyme intermediate were also probed using an assortment of strategies . While the results of these investigations were unable to clearly delineate the chemical mechanism of this enzyme, they provide important insights into the catalytic machinery surrounding the events involved in catalysis.

EMBO J, 2002 Apr 15, 21(8), 1916 - 29
VDAC and the bacterial porin PorB of Neisseria gonorrhoeae share mitochondrial import pathways; Muller A et al.; The human pathogen Neisseria gonorrhoeae induces host cell apoptosis during infection by delivering the outer membrane protein PorB to the host cell's mitochondria . PorB is a pore-forming beta-barrel protein sharing several features with the mitochondrial voltage-dependent anion channel (VDAC), which is involved in the regulation of apoptosis . Here we show that PorB of pathogenic Neisseria species produced by host cells is efficiently targeted to mitochondria . Imported PorB resides in the mitochondrial outer membrane and forms multimers with similar sizes as in the outer bacterial membrane . The mitochondria completely lose their membrane potential, a characteristic previously observed in cells infected with gonococci or treated with purified PorB . Closely related bacterial porins of non-pathogenic Neisseria mucosa or Escherichia coli remain in the cytosol . Import of PorB into mitochondria in vivo is independent of a linear signal sequence . Insertion of PorB into the mitochondrial outer membrane in vitro depends on the activity of Tom5, Tom20 and Tom40, but is independent of Tom70 . Our data show that human VDAC and bacterial PorB are imported into mitochondria by a similar mechanism.

Eur J Clin Microbiol Infect Dis, 2002 Feb, 21(2), 97 - 101
Relationship of Ureaplasma urealyticum biovar 2 to nongonococcal urethritis; Povlsen K et al.; The aim of this study was to determine the frequency of Ureaplasma urealyticum biovars 1 and 2 among 340 men with or without nongonococcal urethritis (NGU) attending a venereal disease clinic in Sweden . NGU was defined by the presence of at least four polymorphonuclear leukocytes per microscopic field (x1,000 magnification) on a smear in which Neisseria gonorrhoeae could not be detected . Ureaplasma urealyticum was detected by polymerase chain reaction, and biovar determination was performed directly on the amplicons by liquid hybridization with biovar-specific probes . Patients with NGU were younger, had had more sexual partners, and exhibited symptoms of urethritis more often than patients without NGU . Ureaplasma urealyticum was detected with the same frequency among patients with and among those without NGU . Among patients with NGU, Ureaplasma urealyticum-positive men were more frequently symptomatic than Ureaplasma urealyticum-negative men . Among patients without NGU, Ureaplasma urealyticum-positive men had had more sexual partners than Ureaplasma urealyticum-negative men . Ureaplasma urealyticum biovar 2 was detected more often among patients with NGU than among those without (P=0.012) . Logistic regression analysis was performed using detection of biovar 2 as the response variable and the following four variables as explanatory variables: presence or absence of NGU, symptoms of urethritis, number of partners, and age < or = 24 years . The only association found was that between Ureaplasma urealyticum biovar 2 and age < or = 24 years . More studies should be conducted to determine the possible pathogenic impact of Ureaplasma urealyticum biovar 2.

Nat Struct Biol, 2002 May, 9(5), 348 - 52
The mirrored methionine sulfoxide reductases of Neisseria gonorrhoeae pilB; Lowther WT et al.; Methionine sulfoxide reductases (Msr) protect against oxidative damage that can contribute to cell death . The tandem Msr domains (MsrA and MsrB) of the pilB protein from Neisseria gonorrhoeae each reduce different epimeric forms of methionine sulfoxide . The overall fold of the MsrB domain revealed by the 1.85 A crystal structure shows no resemblance to the previously determined MsrA structures from other organisms . Despite the lack of homology, the active sites show approximate mirror symmetry . In each case, conserved amino acid motifs mediate the stereo-specific recognition and reduction of the substrate . Unlike the MsrA domain, the MsrB domain activates the cysteine or selenocysteine nucleophile through a unique Cys-Arg-Asp/Glu catalytic triad . The collapse of the reaction intermediate most likely results in the formation of a sulfenic or selenenic acid moiety . Regeneration of the active site occurs through a series of thiol-disulfide exchange steps involving another active site Cys residue and thioredoxin . These observations have broad implications for modular catalysis, antibiotic drug design and continuing longevity studies in mammals.

J Immunol, 2002 Apr 15, 168(8), 4078 - 86
Regulation of the mannan-binding lectin pathway of complement on Neisseria gonorrhoeae by C1-inhibitor and alpha 2-macroglobulin; Gulati S et al.; We examined complement activation by Neisseria gonorrhoeae via the mannan-binding lectin (MBL) pathway in normal human serum . Maximal binding of MBL complexed with MBL-associated serine proteases (MASPs) to N . gonorrhoeae was achieved at a concentration of 0.3 microg/ml . Preopsonization with MBL-MASP at concentrations as low as 0.03 microg/ml resulted in approximately 60% killing of otherwise fully serum-resistant gonococci . However, MBL-depleted serum (MBLdS) reconstituted with MBL-MASP before incubation with organisms (postopsonization) failed to kill at a 100-fold higher concentration . Preopsonized organisms showed a 1.5-fold increase in C4, a 2.5-fold increase in C3b, and an approximately 25-fold increase in factor Bb binding; enhanced C3b and factor Bb binding was classical pathway dependent . Preopsonization of bacteria with a mixture of pure C1-inhibitor and/or alpha(2)-macroglobulin added together with MBL-MASP, all at physiologic concentrations before adding MBLdS, totally reversed killing in 10% reconstituted serum . Reconstitution of MBLdS with supraphysiologic (24 microg/ml) concentrations of MBL-MASP partially overcame the effects of inhibitors (57% killing in 10% reconstituted serum) . We also examined the effect of sialylation of gonococcal lipooligosaccharide (LOS) on MBL function . Partial sialylation of LOS did not decrease MBL or C4 binding but did decrease C3b binding by 50% and resulted in 80% survival in 10% serum (lacking bacteria-specific Abs) even when sialylated organisms were preopsonized with MBL . Full sialylation of LOS abolished MBL, C4, and C3b binding, resulting in 100% survival . Our studies indicate that MBL does not participate in complement activation on N . gonorrhoeae in the presence of "complete" serum that contains C1-inhibitor and alpha(2)-macroglobulin.

Microbiology, 2002 Apr, 148(Pt 4), 909 - 22
LuxS: its role in central metabolism and the in vitro synthesis of 4-hydroxy-5-methyl-3(2H)-furanone; Winzer K et al.; Many bacteria produce extracellular molecules which function in cell-to-cell communication . One of these molecules, autoinducer 2 (AI-2), was first described as an extracellular signal produced by Vibrio harveyi to control luciferase expression . Subsequently, a number of bacteria have been shown to possess AI-2 activity in their culture supernatants, and bear the luxS gene product, which is required for AI-2 synthesis . In Porphyromonas gingivalis, luxS and pfs, encoding a 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTA/SAH'ase), form an operon, suggesting that S-adenosylhomocysteine (SAH) or 5'-methylthioadenosine (MTA) serves as a substrate for AI-2 production . Cell-free extracts of Escherichia coli MG1655, but not DH5alpha (which carries a luxS frame-shift mutation) were capable of generating AI-2 activity upon addition of SAH, but not MTA . S-Ribosyl-homocysteine (RH) derived from SAH also served as a substrate in E . coli MG1655 extracts . RH-supplemented cell-free extracts of Pseudomonas aeruginosa, a bacterium that lacks luxS, only generated AI-2 activity following the introduction of a plasmid containing the Por . gingivalis pfs-luxS operon . In addition, defined in vitro systems consisting of the purified LuxS proteins from Por . gingivalis, E . coli, Neisseria meningitidis or Staphylococcus aureus converted RH to homocysteine and a compound that exhibits AI-2 activity.4-Hydroxy-5-methyl-3(2H)-furanone was identified by mass spectrometry analysis as a major product formed in this in vitro reaction . In E . coli MG1655, expression of T3SH {the bacteriophage T3 S-adenosylmethionine (SAM) hydrolase} significantly reduced AI-2 activity in culture supernatants, suggesting that AI-2 production is limited by the amount of SAH produced in SAM-dependent transmethylase reactions . The authors suggest that the LuxS protein has an important metabolic function in the recycling of SAH . They also show that Ps . aeruginosa is capable of removing AI-2 activity, implying that this molecule may act as a nutrient . In many bacteria AI-2 may in fact represent not a signal molecule but a metabolite which is released early and metabolized in the later stages of growth.

Curr Infect Dis Rep, 2002 Apr, 4(2), 144 - 147
Update on Quinolone Resistance in Neisseria gonorrhoeae; Zenilman JM; Quinolones are widely used for treating gonococcal infections, typically in single-dose, oral regimens . However, in the 1990s, quinolone-resistant Neisseria gonorrhoeae emerged, potentially compromising the utility of this drug class . In the past year, these strains have widely disseminated, accounting for over half of isolates in parts of Southeast Asia . The molecular mechanism of resistance has been localized to multiple mutations in genes coding for the bacterial DNA gyrase and topoisomerase enzymes . These mutations accumulate until the minimum inhibitory concentration is 4.0 g/mL or more, which in clinical studies appears to be the threshold for clinical treatment failure . Quinolone-resistant N . gonorrhoeae is independent from other plasmid- and chromosomally-mediated resistance determinants; nearly all isolates to date have been sensitive to cephalosporins and spectinomycin . Nevertheless, designing public health strategies to contain quinolone-resistant N . gonorrhoeae will be difficult.

Proteomics, 2001 Nov, 1(11), 1378 - 89
Combined automated PCR cloning, in vitro transcription/translation and two-dimensional electrophoresis for bacterial proteome analysis; Norais N et al.; The most popular approach for proteomics analysis is based on the combination of two-dimensional gel electrophoresis and mass spectrometry (MS) . Although very effective, the method suffers from a number of limitations, the most serious one being the necessity to have expensive and sophisticated instrumentation requiring handling by skilled personnel . Here we propose an alternative approach which may offer some advantages over the current methods, at least for some specific applications . The method is based on two-dimensional gel separation of radiolabeled synthetic proteins derived from transcription/translation reactions of linear polymerase chain reaction amplified genes . The gel is autoradiographed and this is superimposed on the sample gel whose protein spots have to be identified . Matching between autoradiographs and sample gel spots allows immediate protein identification . The method has been validated identifying six proteins from a membrane protein preparation of Neisseria meningitidis MC58 strain . All proteins were correctly identified as judged by confirmation analysis with MS . The approach is particularly useful when a specific subset of proteins needs to be identified in a complex protein mixture.

Przegl Epidemiol, 2001, 55(4), 443 - 9
{Recurrent purulent bacterial meningoencephalitis}; Janeczko J et al.; During the period of 25 years there were 55 patients treated in our Institute because of recurrent purulent bacterial meningoencephalitis(rpbme) . This group consisted of 42 males (76%) and 13 (24%) females, the prevalent number (53%) of patients being under 21 years of age . The diagnosis of rpbme was based on the commonly accepted criteria and confirmed by the laboratory results of CSF examination . The cause of the recurrences was established considering the skull X-ray examination, CT and MRI . The evaluation of the clinical status was based on the Glasgow Coma Score (GCS) . During the first hospitalisation, severe or critic clinical status was noted in 42 patients (76%) and moderate in 13 (24%) . The subsequent recurrences were mostly moderate, rarely severe or mild . The number of recurrences varied from 1 to 9 . During the first hospitalisation, the etiologic factor was detected in 39 patients (71%), i.e . Streptococcus pneumoniae in 28 (51%), Neisseria meningitidis in 8 (14%), Pseudomonas aeruginosa and Staphylococcus aureus in 2 and 1 patients respectively . In 37 patients (67%) rpbme developed following cranial trauma, in 18 cases (33%) with single or comminuted fractures of the anterior cranial fossa (in 4 cases accompanied by CSF nasal exsudate) . In 4 it followed neurosurgical intervention, in 3 it accompanied recurrent purulent highmorities, in 1 case--after removal of the nasal polyps and subsequent CSF nasal exsudate, and in 1 patient with recurrent mastoiditis . In 6 cases (11%) the cause of the recurrences remained unelucidated . The clinical signs and symptoms, diagnostic difficulties and the causative treatment of rpbme are discussed . In the authors' opinion, surgical treatment of the communication between the CSF and the external environment prevents the recurrences and is the only successful way of treatment . Special attention is drawn to the great diagnostic value of CT and MRI . The use of other modern techniques, e.g . positron emission tomography (PET) is recommended, because it is useful not only in the functional evaluation of the cerebral tissue after the injury, but also in assessing the dynamics of pathologic changes.

Sex Transm Dis, 2002 Apr, 29(4), 242 - 5
Surveillance of antibiotic resistance of Neisseria gonorrhoeae isolates in China, 1993-1998; Ye S et al.; BACKGROUND: The prevalence of sexually transmitted diseases (STDs) has been increasing in China since the 1980s . Because gonorrhea is the most frequently reported STD there, information on the antimicrobial susceptibility of Neisseria gonorrhoeae will aid in its control . GOAL: To investigate the antimicrobial susceptibility of N gonorrhoeae isolates in China and to provide data for formulation of treatment guidelines and control policies . METHODS: The agar dilution technique was used to determine antimicrobial susceptibility, and acidimetric method was used to test for penicillinase-producing N gonorrhoeae . RESULTS: A total of 3186 gonococcal isolates were tested during the 6-year study period . The rate of resistance to penicillin was 66.70%; 8.14% of isolates were penicillinase-producing N gonorrhoeae . The percentage of tetracycline-resistant isolates was 92.03%, and that of highly tetracycline-resistant isolates was 4.65% . The rate of resistance to ciprofloxacin was also relatively high (34.25%) . The rates of resistance to spectinomycin and ceftriaxone were 0.44% and 0.57% . CONCLUSIONS: The gonococcal isolates in China are relatively highly resistant to penicillin, tetracycline, and ciprofloxacin, but most of them are still susceptible to spectinomycin and ceftriaxone . Standardized treatment of gonorrhea is needed to prevent further spread of resistant gonococcal strains.

Sex Transm Dis, 2002 Apr, 29(4), 216 - 21
Screening for Neisseria gonorrhoeae and Chlamydia trachomatis at entertainment venues among men who have sex with men; Debattista J et al.; BACKGROUND: To assess the prevalence and increase awareness of Neisseria gonorrhoeae and Chlamydia trachomatis infections among men who have sex with men, a screening program was conducted at three inner-city homosexual entertainment venues and one community function . STUDY DESIGN: Each venue was accessed twice over a 3-month period between March and June 2000 . First-catch urine specimens were collected for analysis by polymerase chain reaction (PCR), and throat swabs were collected for culture and PCR . RESULTS: A total of 202 men were tested during the 7 screening nights, including 16 who were tested more than once . From the 186 men tested for the first time, 184 urine specimens were collected, of which 8 (4.3%; 95% CI, 1.9-7.8%) were PCR-positive for C trachomatis and 1 (0.5%; 95% CI, 0-2.1%) was PCR-positive for N gonorrhoeae . Of the 161 throat swabs collected, none were positive for C trachomatis or N gonorrhoeae . CONCLUSIONS: C trachomatis is a potentially significant pathogen in this population of urban homosexual men . Screening programs such as these are valuable as health-promotion exercises.

J Antimicrob Chemother, 2002 Apr, 49(4), 661 - 6
Antibiotic guidelines and antibiotic use in adult bacterial meningitis in The Netherlands; van de Beek D et al.; In The Netherlands, national guidelines for the treatment of adult patients with bacterial meningitis were introduced in October 1997 . In 1998 we began a prospective, nationwide study to evaluate the compliance with these consensus-based guidelines . In addition, we evaluated whether the recommended initial treatment provides adequate microbiological coverage . From October 1998 to January 2000, 365 adults with bacterial meningitis were identified using information from The Netherlands Reference Laboratory for Bacterial Meningitis; 263 patients were classified into four categories depending on patient's age and underlying health status . In the first category, patients 16-60 years without risk factors, Neisseria meningitidis was the most common pathogen (53%); 62 of 127 patients (49%) received treatment in compliance with the guidelines . In the second and third categories, patients >60 years without risk factors and those with risk factors independently of age, Streptococcus pneumoniae caused 61% and 58% of cases, respectively . Compliance in these categories was about 17% . Overall, 33% of patients received treatment in compliance with the guidelines . The microbiological coverage of patients treated in compliance and not in compliance with the guidelines was 98% and 93%, respectively . In conclusion, 1 year after national consensus-based guidelines for the initial treatment of adult patient with bacterial meningitis were introduced in The Netherlands, only one-third of Dutch physicians were adhering to the guidelines . The microbiological coverage for the patients who were treated in compliance with the guidelines was almost complete (98%).

FEBS Lett, 2002 Feb 27, 513(2-3), 299 - 304
Purification and characterisation of the BIOH protein from the biotin biosynthetic pathway; Tomczyk NH et al.; Conversion of pimeloyl-coenzyme A (CoA) to biotin in Escherichia coli requires at least four enzymes encoded by genes in the bio operon . One gene, bioH, which is not present in the bioABFCD operon, is required for the synthesis of pimeloyl-CoA but its exact role in formation of this intermediate is unknown . To investigate this further, we have overexpressed and purified the bioH gene products from both E . coli (BIOH EC) and Neisseria meningitis (BIOH NM) in E . coli . When purified BIOH was incubated with excess CoA and analysed by electrospray mass spectrometry a species of mass corresponding to a BIOH:CoA complex was observed . Mutation of a conserved serine residue to alanine (BIOH EC S82A) did not prevent CoA binding . This is the first report of the purification of BIOH and the observation of a small molecule bound to the protein provides clues to its role in pimeloyl-CoA synthesis.

Expert Rev Mol Diagn, 2001 Nov, 1(4), 445 - 55
Molecular diagnostic testing for infectious diseases using TMA technology; Hill CS; Molecular diagnostic tests based on nucleic acid amplification technologies (NAATs) have become widely established in clinical microbiology laboratories in recent years . The acceptance of these tests has been driven by the development of more accurate and less labor-intensive commercial assay kits by diagnostic manufacturers . Infectious disease diagnostic assays using transcription-mediated amplification (TMA) NAAT have become increasingly popular in many clinical microbiology laboratories . Recent technology developments have improved the performance and simplified the use of the TMA assays . These new technologies have been applied to the development of multiplex TMA tests to improve the testing accuracy for organisms, such as Chlamydia trachomatis and Neisseria gonorrhoeae in clinical microbiology laboratories . TMA tests for HIV-1 and HCV have also led to improvements in blood bank testing which can improve the safety of the public blood supply.

J Cell Biol, 2002 Mar 18, 156(6), 951 - 7 Epub 2002 Mar 18.
CD46 is phosphorylated at tyrosine 354 upon infection of epithelial cells by Neisseria gonorrhoeae; Lee SW et al.; The Neisseria type IV pilus promotes bacterial adhesion to host cells . The pilus binds CD46, a complement-regulatory glycoprotein present on nucleated human cells (Kallstrom et al., 1997) . CD46 mutants with truncated cytoplasmic tails fail to support bacterial adhesion (Kallstrom et al., 2001), suggesting that this region of the molecule also plays an important role in infection . Here, we report that infection of human epithelial cells by piliated Neisseria gonorrhoeae (GC) leads to rapid tyrosine phosphorylation of CD46 . Studies with wild-type and mutant tail fusion constructs demonstrate that Src kinase phosphorylates tyrosine 354 in the Cyt2 isoform of the CD46 cytoplasmic tail . Consistent with these findings, infection studies show that PP2, a specific Src family kinase inhibitor, but not PP3, an inactive variant of this drug, reduces the ability of epithelial cells to support bacterial adhesion . Several lines of evidence point to the role of c-Yes, a member of the Src family of nonreceptor tyrosine kinases, in CD46 phosphorylation . GC infection causes c-Yes to aggregate in the host cell cortex beneath adherent bacteria, increases binding of c-Yes to CD46, and stimulates c-Yes kinase activity . Finally, c-Yes immunoprecipitated from epithelial cells is able to phosphorylate the wild-type Cyt2 tail but not the mutant derivative in which tyrosine 354 has been substituted with alanine . We conclude that GC infection leads to rapid tyrosine phosphorylation of the CD46 Cyt2 tail and that the Src kinase c-Yes is involved in this reaction . Together, the findings reported here and elsewhere strongly suggest that pilus binding to CD46 is not a simple static process . Rather, they support a model in which pilus interaction with CD46 promotes signaling cascades important for Neisseria infectivity.

Emerg Infect Dis, 2002 Feb, 8(2), 188 - 94
Broad-range bacterial detection and the analysis of unexplained death and critical illness; Nikkari S et al.; Broad-range rDNA polymerase chain reaction (PCR) provides an alternative, cultivation-independent approach for identifying pathogens . In 1995, the Centers for Disease Control and Prevention initiated population-based surveillance for unexplained life-threatening infections (Unexplained Death and Critical Illness Project {UNEX}) . To address the causes of UNEX cases, we examined 59 specimens from 46 cases by using broad-range bacterial 16S rDNA PCR and phylogenetic analysis of amplified sequences . Specimens from eight cases yielded sequences from Neisseria meningitidis (cerebrospinal fluid from two patients with meningitis), Streptococcus pneumoniae (cerebrospinal fluid from one patient with meningitis2 and pleural fluid from two patients with pneumonia), or Stenotrophomonas maltophilia (bone marrow aspirate from one patient with pneumonia) . Streptococcus pneumoniae rDNA sequence microheterogeneity was found in one pleural fluid specimen, suggesting the presence of multiple strains . In conclusion, known bacterial pathogens cause some critical illnesses and deaths that fail to be explained with traditional diagnostic methods.

Infect Immun, 2002 Apr, 70(4), 2108 - 20
Specific entry of Helicobacter pylori into cultured gastric epithelial cells via a zipper-like mechanism; Kwok T et al.; Although Helicobacter pylori has generally been considered an extracellular pathogen, a number of in vitro infection experiments and biopsy examinations have shown that it is capable of occasionally entering mammalian host cells . Here, we characterized this entry process by using AGS cells as a host cell model . In gentamicin protection-invasion assays, the number of H . pylori colonies recovered was lower than that for Salmonella enterica serovar Typhimurium X22, Escherichia coli expressing InvA, and Yersinia enterocolitica YO:9 grown at 25 degrees C but higher than that for Neisseria gonorrhoeae VP1 and Y . enterocolitica YO:9 grown at 37 degrees C . At the ultrastructural level, the entry process was observed to occur via a zipper-like mechanism . Internalized H . pylori was bound in tight LAMP-1-containing vacuoles in close association with condensed filamentous actin and tyrosine phosphorylation signals . Wortmannin, a potent inhibitor of phosphatidylinositol 3-kinase, and calphostin C, an inhibitor of protein kinase C, both inhibited the entry of H . pylori in a sensitive and dose-dependent manner; however, the level of entry was enhanced by sodium vanadate, an inhibitor of tyrosine phosphatases and ATPases . Furthermore, the cytokine tumor necrosis factor alpha antagonized the entry of H . pylori into AGS cells . Collectively, these results demonstrate that the entry of H . pylori into AGS cells occurs via a zipper-like mechanism which involves various host signal transduction events.

Infect Immun, 2002 Apr, 70(4), 1715 - 23
Carcinoembryonic antigen family receptor recognition by gonococcal Opa proteins requires distinct combinations of hypervariable Opa protein domains; Bos MP et al.; Neisserial Opa proteins function as a family of adhesins that bind heparan sulfate proteoglycan (HSPG) or carcinoembryonic antigen family (CEACAM) receptors on human host cells . In order to define the CEACAM binding domain on Opa proteins, we tested the binding properties of a series of gonococcal (strain MS11) recombinants producing mutant and chimeric Opa proteins with alterations in one or more of the four surface-exposed loops . Mutagenesis demonstrated that the semivariable domain, present in the first loop, was completely dispensable for CEACAM binding . In contrast, the two hypervariable (HV) regions present in the second and third loops were essential for binding; deletion of either domain resulted in loss of receptor recognition . Deletion of the fourth loop resulted in a severe decrease in Opa expression at the cell surface and could therefore not be tested for CEACAM binding . Chimeric Opa variants, containing combinations of HV regions derived from different CEACAM binding Opa proteins, lost most of their receptor binding activity . Some chimeric variants gained HSPG binding activity . Together, our results indicate that full recognition of CEACAM receptors by Opa proteins requires a highly coordinate interplay between both HV regions . Furthermore, shuffling of HV regions may result in novel HSPG receptor binding activity.

J Pediatr Adolesc Gynecol, 2002 Feb, 15(1), 37 - 41
Use of ligase chain reaction for laboratory identification of Chlamydia trachomatis and Neisseria gonorrhoeae in adolescent women; Braverman PK et al.; STUDY OBJECTIVE: To compare the ligase chain reaction (LCR) with culture for the detection of Neisseria gonorrhoeae (GC) and with culture and direct fluorescent antibody (DFA) for identification of Chlamydia trachomatis (CT) in cervical specimens from adolescent women . DESIGN: A prospective study of test performance . SETTING: Two urban, hospital-based adolescent clinics . PARTICIPANTS: Adolescent women aged 12-22 yr undergoing pelvic examination for routine sexually transmitted disease (STD) screening or symptoms suggestive of an STD . MAIN OUTCOME MEASURES: LCR results were considered to be true positives if confirmed by culture and/or DFA (CT only) . Discrepant LCR results were confirmed by testing an alternative locus . RESULTS: With 538 subjects, LCR for CT had a sensitivity of 98.4% (61/62) and specificity of 96.4% (459/476) prior to resolution and a sensitivity of 98.6% (70/71) and specificity of 99.6% (459/461) after resolution . With 1225 subjects, LCR for GC had a sensitivity of 90.0% (54/60) and specificity of 99.4% (1158/1165) prior to resolution and a sensitivity of 90.6% (58/64) and specificity of 100% (1158/1158) after resolution . CT culture alone identified 80% of the true positives and DFA alone only identified 72% . GC culture alone identified 94% of the true positives . CONCLUSIONS: LCR is an extremely sensitive and specific rapid test, utilizing a single swab and convenient room-temperature storage and transport of specimens . LCR testing of cervical specimens for CT in adolescent women is a better test for detecting CT infection than culture or DFA . LCR testing for cervical GC infection may provide an advantage over culture in circumstances in which optimal transport conditions and viability of the organism cannot be assured.

Int J STD AIDS, 2002 Apr, 13(4), 238 - 45
Risk behaviours and prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae genital infections among Montreal street youth; Haley N et al.; We estimated the prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae genital tract infections among 302 Montreal street youth (223 boys) and identified associated risk factors . Study participants, 14-25 years old (average 20.9 years), meeting specific criteria for homelessness, were recruited in street youth agencies . Participation included a structured interview and provision of a urine specimen . Among sexually active youth, (n = 300) 30.0% had more than five heterosexual partners and 13.0% had at least one homosexual partner (last year), 10.7% had received money in exchange for sex (last six months) and 47.0% reported sexual relations resulting in pregnancy (lifetime) . Among all youths, 82.1% had used at least one type of illicit drug, and 30.1% injected drugs at least once (last six months) . The prevalence of C . trachomatis infection was 6.6% (95% CI 4.1-10.0%) . Prevalence did not vary significantly by sex, age or any other variable, except history of pregnancy (10.4% among youth with history of pregnancy vs 3.6% among others, P = 0.02) . No cases of N . gonorrhoeae infection were found.

J Cell Sci, 2002 Mar 15, 115(Pt 6), 1231 - 41
Microvilli-like structures are associated with the internalization of virulent capsulated Neisseria meningitidis into vascular endothelial cells; Eugene E et al.; Bacterial pathogens are internalized into non-phagocytic cells either by a zipper mechanism involving a direct contact between a bacterial ligand and a cellular receptor or a trigger mechanism secondary to the formation of membrane ruffles . Here we show that internalization of capsulated Neisseria meningitidis within endothelial cells following type IV pilus-mediated adhesion is associated with the formation of cellular protrusions at the site of bacterial attachment . These protrusions, like microvilli, are highly enriched in ezrin and moesin, two members of the ERM (ezrin/radixin/moesin) family, whereas vinculin and paxillin are absent . ERM-binding transmembrane proteins, such as CD44, and cortical actin polymerization colocalized within these membrane protrusions . Expression of dominant-negative ezrin largely prevented cortical actin polymerization, thus confirming the role of this molecule in bacteria-induced cytoskeletal modifications . Moreover, using selective inhibitors and dominant-negative mutants of the Rho family GTPases, we show that bacteria-induced actin polymerization required the activation of both Rho and Cdc42 but not of Rac1 . Whereas GTPase inhibition dramatically reduced actin polymerization at the site of bacterial attachment, ezrin recruitment was not affected, indicating that bacterial adhesion promotes ezrin recruitment independently of the activity of the Rho-GTPases . Furthermore, GTPase inhibition largely reduced N . meningitidis entry into endothelial cells without affecting adhesion . We thus propose that following pilus-mediated adhesion, capsulated N . meningitidis recruit ERM-binding transmembrane proteins, as well as ezrin and moesin, and that both Rho and Cdc42 are critical for the subsequent cytoskeletal modifications responsible for the formation of microvilli-like cellular protrusions and bacterial internalization.

Res Microbiol, 2002 Jan-Feb, 153(1), 1 - 6
Genomic analyses of bacterial respiratory and cytochrome c assembly systems: Bordetella as a model for the system II cytochrome c biogenesis pathway; Kranz RG et al.; An analysis of thirty-three genomes of selected bacteria for the presence of specific respiratory pathways and cytochrome c biogenesis systems has led to observations on respiration and biogenesis . A table summarizing these results is presented . The data suggested that Bordetella pertussis would be an excellent genetic model to study the System II cytochrome c biogenesis pathway . These observations are discussed and the results of genetic studies on System II biogenesis in B . pertussis are presented as a case for the power of comparative genomics . System II is present in organisms as diverse as Helicobacter, Neisseria, Porphyromonas, mycobacteria, cyanobacteria, and plants (chloroplasts), indicating this pathway's prominence and that horizontal transfer of system II (and/or System I) must have occurred on multiple occasions.

J Clin Microbiol, 2002 Mar, 40(3), 758 - 61
Evaluation of dry and wet transported intravaginal swabs in detection of Chlamydia trachomatis and Neisseria gonorrhoeae infections in female soldiers by PCR; Gaydos CA et al.; Screening women for sexually transmitted diseases (STD) in nonclinic settings is highly desirable because many infections are asymptomatic . This is especially true for military women, for whom logistical, social, and other job-related obstacles present barriers to accessing medical care . We assessed the accuracy of intravaginal swabs transported by mail in a wet versus a dry state for PCR (Amplicor CT/NG test) detection of chlamydia and gonorrhea infections in a cross-sectional study of 793 active-duty military women attending an STD clinic . PCR tests of vaginal swabs (wet and dry) were compared to local clinical methods used on cervical swabs . Standard wet vaginal swab PCR testing detected more chlamydia (11.6%) than cervical enzyme immunoassay (9.3%) . For detection of chlamydia using wet swabs, the sensitivity and specificity compared with adjudicated true positives were 94.6% (87 of 92) and 99.3% (696 of 701), respectively . Comparing dry swabs to true-positives for chlamydia, the sensitivity was 91.3% (84 of 92) and the specificity was 99.3% (696 of 701) . Standard wet vaginal swab PCR detected more gonorrhea (3.3%) than routine cervical culture (2.1%) . The sensitivity and specificity of PCR testing of wet swabs compared to true-positives (infected patients) were 96.3% (26 of 27) and 98.2% (752 of 766) for gonorrhea, respectively . For gonorrhea, the sensitivity and specificity of dry swabs compared to true-positives (infected patients) were 88.9% (24 of 27) and 98.3% (753 of 766), respectively . PCR testing of wet and dry transported intravaginal swabs to detect chlamydia and gonorrhea infections was an accurate diagnostic method for military women.

Int Rev Immunol, 2001, 20(2), 251 - 61
Development and characterization of anti-idiotype based peptide and DNA vaccines which mimic the capsular polysaccharide of Neisseria meningitidis serogroup C; Westerink MA et al.; Anti-idiotypic antibodies and peptides which mimic antigens may offer an alternate strategy for converting a thymus-independent (TI) antigen to a thymus dependent (TD) antigen . We have developed an anti-idiotype based peptide mimic of the capsular polysaccharide of N . meningitidis serogroup C (MCPS) which induces a T-dependent protective immune response in mice . Subsequent studies have demonstrated that immunization of severe combined immunodeficient mice reconstituted with human B cells respond to immunization with the MCPS peptide mimic with bactericidal anti-polysaccharide directed antibody response . We hypothesized that administration of a DNA vaccine resulting in endogenous expression of this carbohydrate peptide mimic would induce anti-MCPS antibodies.

Int Rev Immunol, 2001, 20(2), 229 - 50
Strategies for mimicking Neisserial saccharide epitopes as vaccines; Gulati S et al.; Monoclonal antibody (mAb) 2C7 recognizes a conserved and widely expressed oligosaccharide (OS) epitope on Neisseria gonorrhoeae . This OS epitope evokes a significant bactericidal and opsonic immune response after natural infection and vaccination . The OS epitope structure represents an excellent target for a potential protective gonococcal vaccine . Because carbohydrate antigens are T-cell independent, inducing weak antibody responses, OS molecules are not useful immunogens . We developed and examined two different strategies to mimic the 2C7 OS epitope: (i) an anti-idiotope (mAb CA1); and (ii) a peptide (PEP-1) . These surrogate immunogens elicited antibody responses in mice (CA1 and PEP-1) and rabbits (CA1) that were bactericidal in vitro against gonococci . Both CA1 and PEP-1 are true immunologic mimics of OS and may form a basis for the development of vaccine candidates for human immunization against N . gonorrhoeae.

Int Rev Immunol, 2001, 20(2), 201 - 20
Molecular mimetics of Neisseria meningitidis serogroup B polysaccharide; Moe GR et al.; Strains of Neisseria meningitidis serogroup B (NmB) are an important cause of meningitis and sepsis . Efforts to develop a NmB vaccine have been hampered by poor immunogenicity of the polysaccharide capsule, which cross-reacts with host polysialic acid, and the danger of eliciting autoantibodies . To investigate the potential of molecular mimetics to circumvent these problems, we prepared murine monoclonal antibodies (mAbs) against the N-propionyl derivative (N-Pr) of NmB polysaccharide . Several mAbs were found that reacted with capsular polysaccharide epitopes, which were distinct from host polysialic acid . These mAbs also passively conferred protection against experimental bacteremia . We used these mAbs to screen novel independently folding peptide phage display libraries, and pools of combinatorial small molecules, each consisting of approximately 30 to approximately 700 small molecules of diverse composition . To date, several mimetic candidates have been identified . One is a peptide selected from a library of independently folding alphabeta peptides, and others are peptoid dimers or trimers selected from the small molecule pools . The peptoids contain an indan-type of ring system, and some of them also contain a large hydrophobic group such as oleyl amine or dehydroabietyl amine, and a positively charged group at the amino-terminus . Both the alphabeta peptide from the phage library, and the peptoids from the small molecule pools, inhibit binding of the mAbs to N-Pr NmB polysaccharide . Future studies will focus on the structure/activity relationship of these mimetics, and the development of immunogens that may be capable of eliciting anticapsular antibody without autoantibody activity.

Scand J Infect Dis, 2002, 34(1), 11 - 3
Neisseria meningitidis with decreased susceptibility to penicillin in Istanbul, Turkey; Punar M et al.; This study was conducted to estimate the rate of decreased susceptibility to penicillin (MIC > 0.06-1 microg/ml) in Neisseria meningitidis isolates in Istanbul, Turkey . A total of 30 isolates collected during a 1-y period from patients with meningitis and from nasopharyngeal carriers were tested for penicillin and cefotaxime susceptibility using the E-test . Two out of 12 (17%) clinical isolates and 11/18 (61%) nasopharyngeal isolates showed decreased susceptibility to penicillin with MICs in the range 0.094-1.0 microg/ml, giving an overall resistance of 43% (n = 13) . These data show that continued surveillance of trends in antimicrobial susceptibility of N . meningitidis is important for detecting the emergence of N . meningitidis strains with MICs > 1 microg/ml which may pose serious therapeutic problems.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Sep-Oct, (5), 101 - 8
{Stress-inducible bacterial proteins and virulence}; Basnak'ian IA et al.; Different species of pathogenic bacteria, including Salmonella, Neisseria, Listeria and Francisella have been used to demonstrate relationship between the synthesis of stressor induced proteins by cells and the phenotypic manifestation of their virulence . The impact of such external factors as high temperature, low pH, osmolarity, substrate limitation, the content of active forms of oxygen, etc . is accompanied by the synthesis of different stressor induced proteins playing a complex role . Under unfavorable environmental conditions the synthesis of these proteins ensures the survival of the infective agents . Under conditions of a macroorganism synthesis of some stressor induced proteins promotes the survival of infective agents and their resistance to the action of humoral and cell-mediated protective factors of the host . As is known, the expression of virulence genes is not constitutive . The expression of these genes greatly depends on environmental conditions and its induction is determined by extra- or intracellular location of the infective agent . Several systems of the regulation of bacterial pathogenicity factors have been described that are relatively not numerous, conservative and respond to external signals . The relevance of a number of stressor induced proteins of bacteria to virulence associated factors is discussed.

Scand J Infect Dis, 2001, 33(12), 909 - 13
D- and L-lactate in rabbit and human bacterial meningitis; Wellmer A et al.; Increased total CSF lactate is an important indicator differentiating bacterial from aseptic meningitis . Bacteria can produce D- and L-lactate; mammalian cells produce only L-lactate . We measured D- and L-lactate production of Streptococcus pneumoniae, Staphylococcus aureus, Neisseria meningitidis and Escherichia coli in vitro, of S . pneumoniae and E . coli in rabbit experimental meningitis and of various common pathogens in CSF from patients with bacterial meningitis . Despite marked in vitro production of D-lactate by S . aureus (maximum: 4.59 mmol/l; i.e . 34.9% of total lactate), N . meningitidis (4.62 mmol/l; i.e . 98.1%) and E . coli (3.14 mmol/l; i.e . 97.2%), minimal amounts were measured in human S . aureus (0.38 mmol/l; i.e . 1.3% of total lactate) or N . meningitidis (0.28 mmol/l; i.e . 3.9%) and experimental E . coli meningitis (0.75 mmol/l; i.e . 4.4%) . In only 9 of 54 human CSF samples did D-lactate exceed 0.15 mmol/l . S . pneumoniae did not produce significant amounts of D-lactate in vitro (maximum: 0.55 mmol/l; i.e . 2.7% of total lactate), in experimental meningitis (0.18 mmol/l; i.e . 3%) or in human cases of meningitis (0.28 mmol/l; i.e . 1.9%) . In conclusion, increased total CSF lactate in meningitis consists mainly of L-lactate and originates predominantly from host cells . CSF D-lactate is of limited diagnostic value.

J Antimicrob Chemother, 2002 Mar, 49(3), 545 - 7
What about antibiotic resistance in Neisseria lactamica?
Arreaza L, Salcedo C, Alcala B, Vazquez JA.
The in vitro activity of penicillin, ampicillin, cefotaxime, ceftriaxone, rifampicin and ciprofloxacin against 286 Neisseria lactamica isolates was determined by agar dilution and the category of susceptibility was analysed in accordance with the criteria used for Neisseria meningitidis . All isolates were considered to have intermediate susceptibility to penicillin . A total of 1.7% of the isolates were resistant to ampicillin but all were susceptible to cefotaxime and ceftriaxone . Rifampicin MICs ranged between 0.12 and 2 mg/L . Six isolates (2.1%) showed decreased susceptibility to ciprofloxacin.

Clin Microbiol Infect, 1998 Jan, 4(11), 627 - 633
Incidence, epidemiology and evolution of reduced susceptibility to ciprofloxacin in Neisseria gonorrhoeae in Korea; Lee K et al.; OBJECTIVE: To verify the decrease of susceptibility to ciprofloxacin in Neisseria gonorrhoeae, determine the size of the recently reported new beta-lactamase plasmid and explain the high prevalence of penicillinase-producing Neisseria gonorrhoeae (PPNG) . METHODS: Gonococci were isolated from prostitutes in Korea . Antimicrobial susceptibility was tested by NCCLS disk diffusion and agar dilution methods . Plasmid was isolated by an alkaline lysis method . Patterns of Nhel-digested genomic DNA were compared after pulsed-field gel electrophoresis (PFGE) . RESULTS: The minimum inhibitory concentration of ciprofloxacin for 50% of the isolates rose from 0.015 mg/L in 1993 to 0.12 mg/L in 1996 . The proportion of PPNG remained at 70% or over during the 5-year period . The size of a novel beta-lactamase plasmid, first reported in 1994, was determined to be approximately 3.2 MDa, and 48% of the PPNG isolates contained it . Twelve of 50 isolates had the same PFGE pattern and nine others another pattern . CONCLUSION: The rapid decrease of fluoroquinolone-susceptible gonococci suggests that in the near future the drug may become less useful for gonorrhea treatment . The new 3.2-MDa plasmid may have been introduced as a result of the recent increase in overseas travel . The PFGE pattern suggests that high prevalence of PPNG may be due to dissemination of a few resistant clones among the high-risk groups.

Mol Genet Genomics, 2002 Feb, 266(6), 962 - 72 Epub 2001 Dec 11.
Recombinational error and deletion formation in Neisseria gonorrhoeae: a role for RecJ in the production of pilE (L) deletions; Hill SA et al.; Genetic linkage within Neisseria gonorrhoeae populations is in equilibrium, yet the physical linkage map indicates a relatively stable chromosome structure, despite an apparently vast potential for mispairing between repeated sequences (e.g . between the multiple pil or opa alleles, or through mispairing of any of the numerous small repeated sequences that are liberally scattered throughout the chromosome) . Therefore, the stability of the physical linkage map suggests that aberrant recombination between repeated sequences is a rare event . This study was undertaken to explore some of the parameters that may govern deletion events between short direct oligonucleotide repeats, using a chromosomal locus that appears to be especially prone to deletions (the pilin expression locus; pilE) . In this report, we demonstrate that deletion formation at pilE occurs primarily through recombinational error following a pilE/pilS interaction; illegitimate (i.e . RecA-independent) events can occur, but they are infrequent . In contrast, when genetically engineered opa deletion substrates were constructed and placed in the chromosome, deletions at the opa loci were infrequent even under rec(+) conditions . A model is presented in which the gonococcal RecA and RecJ proteins promote pilE deletions through a recombination event that is templated or stabilised by a pilE/pilS interaction.

Int J STD AIDS, 2002 Mar, 13(3), 178 - 80
A retrospective study of the clinical effectiveness of the treatment of genital co-infection with N . gonorrhoeae and C . trachomatis in Coventry; Das S et al.; Concomitant infection of Neisseria gonorrhoeae and Chlamydia trachomatis is frequent and it is common practice to prescribe ancillary treatment for chlamydial infection when infection with N . gonorrhoeae is suspected or confirmed . In Coventry cases are treated as they are diagnosed . Our objective was to determine the clinical effectiveness of treating gonorrhoea and chlamydial infection separately in cases of co-infection . Case notes of co-infection with N . gonorrhoeae and C . trachomatis diagnosed in Coventry GU clinic from March 1989 to February 2000 were reviewed retrospectively . There were 1250 episodes of gonorrhoea, 4127 of chlamydial infections, and both infections were found in 332 cases . The two infections were treated in 322 cases and in 235 cases were treated separately . Ten cases did not come back for treatment of chlamydial infection, which is less than one case per year and 0.2% of total chlamydial infection in 11 years . On the other hand, 918 (73%) of total number of gonorrhoea patients did not have to take unnecessary treatment for chlamydial infection . In some clinical settings co-infection with N . gonorrhoeae and C . trachomatis could be treated separately with significant success and in the long run this might prevent development of antibiotic resistance of C . trachomatis infection.

Int J STD AIDS, 2002 Mar, 13(3), 174 - 7
The sac-4 gene of Neisseria gonorrhoeae correlates with gonococcal subtype not co-existing chlamydial infection; Gretton S et al.; We examined the hypothesis that the sac-4 gene of Neisseria gonorrhoeae varies with gonococcal subtype and that this could account for an earlier report that sac-4 increased the likelihood of co-infection with Chlamydia trachomatis . A polymerase chain reaction (PCR) assay was used to determine the prevalence of sac-4 in 435 gonococcal isolates . The prevalence of sac-4 was analysed in relation to chlamydial co-infection, serovar, auxotype, gender and sexual orientation . Although the prevalence of sac-4 was higher in isolates from patients with chlamydial co-infection (55%) than in those without chlamydial co-infection (42%) the difference was not significant (P<0.05) . Statistically significant differences in association with sac-4 were, however, shown between various serovars and auxotypes . Dual classification based on auxotype/serovar (A/S) classes showed highly significant differences in sac-4 prevalence between groups: 95% in NR/1B18 and 8% in P/1B2 (P<0.001) . Sac-4 was also significantly less common (P<0.05) in isolates from homosexual men (35%) than from heterosexual men (49%) or women (49.5%) . Sac-4 appears to have an epidemiological association with gonococcal auxotype and serovar rather than a direct association with chlamydial co-infection.

J Immunol, 2002 Mar 1, 168(5), 2424 - 32
Response to Neisseria gonorrhoeae by cervicovaginal epithelial cells occurs in the absence of toll-like receptor 4-mediated signaling; Fichorova RN et al.; Toll-like receptors (TLRs) have recently been identified as fundamental components of the innate immune response to bacterial pathogens . We investigated the role of TLR signaling in immune defense of the mucosal epithelial cells of the lower female genital tract . This site provides first line defense against microbial pathogens while remaining tolerant to a complex biosystem of resident microbiota . Epithelial cells derived from normal human vagina, ectocervix, and endocervix expressed mRNA for TLR1, -2, -3, -5, and -6 . However, they failed to express TLR4 as well as MD2, two essential components of the receptor complex for LPS in phagocytes and endothelial cells . Consistent with this, endocervical epithelial cells were unresponsive to protein-free preparations of lipooligosaccharide from Neisseria gonorrhoeae and LPS from Escherichia coli . However, they were capable of responding to whole Gram-negative bacteria and bacterial lysates, as demonstrated by NF-kappaB activation and proinflammatory cytokine production . The presence of soluble CD14, a high-affinity receptor for LPS and other bacterial ligands, enhanced the sensitivity of genital tract epithelial cells to both low and high concentrations of bacteria, suggesting that soluble CD14 can act as a coreceptor for non-TLR4 ligands . These data demonstrate that the response to N . gonorrhoeae and other Gram-negative bacteria at the mucosal surface of the female genital tract occurs in the absence of endotoxin recognition and TLR4-mediated signaling.

Diagn Microbiol Infect Dis, 2002 Feb, 42(2), 85 - 9
A real-time PCR assay for the detection of Neisseria gonorrhoeae by LightCycler; Whiley DM et al.; The detection of Neisseria gonorrhoeae by the polymerase chain reaction (PCR) is now recognized as a sensitive and specific method of diagnosing infection by the organism . In this Study 152 urine specimens were examined for N . gonorrhoeae by a real-time PCR method using the LightCycler platform and results were compared to an "in-house" PCR assay using an ELISA-based detection method . N . gonorrhoeae DNA was detected in 29 (19%) specimens by LightCycler PCR (LC-PCR) and in 31 (20%) specimens by the "in house" PCR method . The LightCycler assay proved to be specific and 94% sensitive when compared to the "in house" PCR method . These features combined with the rapid turn-around time for results makes the LC-PCR particularly suitable for the detection of N . gonorrhoeae in a routine clinical laboratory.

Curr Infect Dis Rep, 2002 Feb, 4(1), 7 - 16
Treatment of Bacterial Meningitis; Tunkel AR et al.; The therapeutic approach to acute bacterial meningitis has changed in recent years as a result of changes in in vitro susceptibility of many meningeal pathogens to previously standard antimicrobial therapy . Given the emergence of strains of Streptococcus pneumoniae that are resistant to penicillin and the cephalosporins, the combination of vancomycin plus a third-generation cephalosporin is recommended as empiric therapy for suspected or proven pneumococcal meningitis, pending results of in vitro susceptibility testing . Strains of Neisseria meningitidis with reduced susceptibility to penicillin have also been described, although most patients with these resistant strains have recovered with standard penicillin therapy . Although the third-generation cephalosporins have greatly improved outcome in patients with meningitis caused by aerobic gram-negative bacilli, many organisms in this group are now resistant to these drugs; the carbapenems and fluoroquinolones may be effective alternative agents and have been successfully used in small case series . Further surveillance of the in vitro antimicrobial susceptibility patterns of meningeal pathogens is critical for future recommendations in the treatment of bacterial meningitis.

Nat Immunol, 2002 Mar, 3(3), 229 - 36 Epub 2002 Feb 19.
Neisserial binding to CEACAM1 arrests the activation and proliferation of CD4+ T lymphocytes; Boulton IC et al.; Infection with Neisseria gonorrhoeae can trigger an intense inflammatory response, yet there is little specific immune response or development of immune memory . In addition, gonorrhea typically correlates with a transient reduction in T lymphocyte counts in blood, and these populations recover when gonococcal infection is resolved . Such observations suggest that the gonococci have a suppressive effect on the host immune response . We report here that N . gonorrhoeae Opa proteins were able to bind CEACAM1 expressed by primary CD4+ T lymphocytes and suppress their activation and proliferation . CEACAM1 bound by gonococcal Opa52 associated with the tyrosine phosphatases SHP-1 and SHP-2, which implicates the receptor's ITIM (immunoreceptor tyrosine-based inhibitory motif) in this effect.

Antimicrob Agents Chemother, 2002 Mar, 46(3), 769 - 77
Mutations in ponA, the gene encoding penicillin-binding protein 1, and a novel locus, penC, are required for high-level chromosomally mediated penicillin resistance in Neisseria gonorrhoeae; Ropp PA et al.; Chromosomally mediated penicillin resistance in Neisseria gonorrhoeae occurs in part through alterations in penicillin-binding proteins (PBPs) and a decrease in outer membrane permeability . However, the genetic and molecular mechanisms of transformation of a penicillin-susceptible strain of N . gonorrhoeae to high-level penicillin resistance have not been clearly elucidated . Previous studies suggested that alterations in PBP 1 were involved in high-level penicillin resistance . In this study, we identified a single amino acid mutation in PBP 1 located 40 amino acids N terminal to the active-site serine residue that was present in all chromosomally mediated resistant N . gonorrhoeae (CMRNG) strains for which MICs of penicillin were > or = 1 microg/ml . PBP 1 harboring this point mutation (PBP 1*) had a three- to fourfold lower rate of acylation (k2/K') than wild-type PBP 1 with a variety of beta-lactam antibiotics . Consistent with its involvement in high-level penicillin resistance, replacement of the altered ponA gene (ponA1) in several CMRNG strains with the wild-type ponA gene resulted in a twofold decrease in the MICs of penicillin . Surprisingly, transformation of an intermediate-level penicillin-resistant strain (PR100; FA19 penA4 mtr penB5) with the ponA1 gene did not increase the MIC of penicillin for this strain . However, we identified an additional resistance locus, termed penC, which was required along with ponA1 to increase penicillin resistance of PR100 to a high level (MIC = 4 microg/ml) . The penC locus by itself, when present in PR100, increases the MICs of penicillin and tetracycline twofold each . These data indicate that an additional locus, penC, is required along with ponA1 to achieve high-level penicillin resistance.

Ann Clin Lab Sci, 2002 Winter, 32(1), 61 - 4
Comparison of M4 and M4RT media for transporting cervical swab samples for PCR detection of Chlamydia trachomatis and Neisseria gonorrhoeae; Aslanzadeh J et al.; In a prospective study, M4RT medium was compared to the traditional M4 medium to transport cervical swab specimens for Neisseria gonorrhoeae/Chlamydia trachomatis (NG/CT) PCR testing using the Roche COBAS Amplicor . Two cervical swab samples were collected from 270 consecutive patients screened for NG/CT in a satellite facility . The swabs were placed individually in M4RT and M4 medium and were immediately refrigerated, transported to the laboratory on wet ice, and stored at 2 to 8 degrees C until the PCR testing was performed within 7 da of collection . Seven of the cervical swab samples transported in M4 or M4RT were PCR positive for CT . Two additional samples transported in M4RT and a third swab transported in M4 were CT PCR positive . These samples were PCR negative in the alternative medium . Similarly, 12 of the cervical swabs transported in M4 or M4RT were NG PCR positive . Three additional swabs transported in M4 media were NG PCR positive . Initially, 2 of these samples when transported in M4RT were NG PCR equivocal and were considered NG PCR positive on repeat testing . Similarly, 2 additional swab samples transported in M4 RT media were NG PCR positive . These samples, when transported in M4 media, were NG PCR equivocal or negative . However, on repeat testing the equivocal sample was considered NG PCR positive . We conclude M4 and M4RT transport media are equally reliable for transporting cervical swab samples for NG/CT PCR testing . M4RT medium is more convenient to use, as it did not require refrigeration until it was inoculated with the clinical sample.

EMBO J, 2002 Feb 15, 21(4), 560 - 71
Distinct mechanisms of internalization of Neisseria gonorrhoeae by members of the CEACAM receptor family involving Rac1- and Cdc42-dependent and -independent pathways; Billker O et al.; Opa adhesins of pathogenic Neisseria species target four members of the human carcinoembryonic antigen-related cellular adhesion molecule (CEACAM) family . CEACAM receptors mediate opsonization-independent phagocytosis of Neisseria gonorrhoeae by human granulocytes and each receptor individually can mediate gonococcal invasion of epithelial cells . We show here that gonococcal internalization occurs by distinct mechanisms depending on the CEACAM receptor expressed . For the invasion of epithelial cell lines via CEACAM1 and CEACAM6, a pathogen-directed reorganization of the actin cytoskeleton is not required . In marked contrast, ligation of CEACAM3 triggers a dramatic but localized reorganization of the host cell surface leading to highly efficient engulfment of bacteria in a process regulated by the small GTPases Rac1 and Cdc42, but not Rho . Two tyrosine residues of a cytoplasmic immune receptor tyrosine-based activating motif of CEACAM3 are essential for the induction of phagocytic actin structures and subsequent gonococcal internalization . The granulocyte-specific CEACAM3 receptor has properties of a single chain phagocytic receptor and may thus contribute to innate immunity by the elimination of Neisseria and other CEACAM-binding pathogens that colonize human mucosal surfaces.

Int J STD AIDS, 2002 Feb, 13(2), 124 - 30
Molecular typing of Neisseria gonorrhoeae from hospital and community isolates by restriction fragment length polymorphism; Bindu MP et al.; Strains of Neisseria gonorrhoeae are generally characterized by auxotyping, serotyping, plasmid profile, antibiotic sensitivity and deoxyribonucleic acid (DNA) amplification fingerprinting . The aim of this study was to analyse the generation of restriction fragment length polymorphism (RFLP) patterns by BgIII digestion of total genomic DNA of N . gonorrhoeae isolated from the community (n =30) and the hospital (n =15) and to establish an association with serogrouping and antibiogram . The RFLP patterns produced by BgIII restriction digestion showed 7 different patterns among 30 community isolates and 9 different patterns among 15 hospital isolates . 66.7% of isolates belonged to serogroup WI . Penicillin resistance was observed in 46.7% of community isolates and 66.7% hospital isolates . However, penicillinase producing N . gonorrhoeae (PPNG) were lower in the community (6.6%) than in the hospital isolates (53.3%) . PPNG strains were more often seen in serogroup WI . This is the first Indian report on RFLP genotype pattern in N . gonorrhoeae . We noted differences in RFLP genotypes of the community (RFLP types 1, 2, 3, 4, 5 and 7) and hospital strains (RFLP types 6 and 8), while no differences in the serogroup were observed . Ciprofloxacin resistance was 20.0% and 26.6% in the community and hospital isolates, respectively . Ceftriaxone emerges as the current drug of choice for an effective policy of antibiotic treatment of gonorrhoea through syndromic management in developing countries.

Int J STD AIDS, 2002 Feb, 13(2), 109 - 14
One year of Neisseria gonorrhoeae isolates in Sweden: the prevalence study of antibiotic susceptibility shows relation to the geographic area of exposure; Berglund T et al.; The aim of this study was to compare epidemiological data with antibiotic susceptibility patterns, so as to characterize the risk of infection with a highly resistant Neisseria gonorrhoeae strain . N . gonorrhoeae strains isolated in Sweden from February 1998 through January 1999 were tested for antibiotic susceptibility . Epidemiological data were received from each clinician reporting a case of gonorrhoea and these data were linked to the N . gonorrhoeae strains . A total of 348 N . gonorrhoeae isolates, representing 89% of all Swedish cases diagnosed during the 12-month period, were tested for antibiotic susceptibility . Of all isolates, 24% were beta-lactamase-producing, and 18% had decreased susceptibility to ciprofloxacin (MIC>0.064 mg/l) . All isolates were fully susceptible to ceftriaxone and spectinomycin . More than 99% of the isolates were fully susceptible to azithromycin . The antibiotic susceptibility varied with the places where patients were exposed to infection . When exposed in Asia, 63% of the isolates showed reduced susceptibility to ciprofloxacin, compared with 0-8.5% of the isolates from patients exposed in other places (RR=8.5, P<0.001) . Ciprofloxacin cannot be recommended as the first choice of treatment if the place of exposure was in Asia.

Oncol Rep, 2002 Mar-Apr, 9(2), 267 - 76
Cancer vaccines: an update with special focus on ganglioside antigens; Bitton RJ et al.; Vaccine development is one of the most promising and exciting fields in cancer research; numerous approaches are being studied to developed effective cancer vaccines . The aim of this form of therapy is to teach the patient's immune system to recognize the antigens expressed in tumor cells, but not in normal tissue, to be able to destroy these abnormal cells leaving the normal cells intact . In other words, is an attempt to teach the immune system to recognize antigens that escaped the immunologic surveillance and are by it, therefore able to survive and, in time, disseminate . However each research group developing a cancer vaccine, uses a different technology, targeting different antigens, combining different carriers and adjuvants, and using different immunization schedules . Most of the vaccines are still experimental and not approved by the US or European Regulatory Agencies . In this work, we will offer an update in the knowledge in cancer immunology and all the anticancer vaccine approaches, with special emphasis in ganglioside based vaccines . It has been demonstrated that quantitative and qualitative changes occur in ganglioside expression during the oncogenic transformation . Malignant transformation appears to activate enzymes associated with ganglioside glycosylation, resulting in altered patterns of ganglioside expression in tumors . Direct evidence of the importance of gangliosides as potential targets for active immunotherapy has been suggested by the observation that human monoclonal antibodies against these glycolipids induce shrinkage of human cutaneous melanoma metastasis . Thus, the cellular over-expression and shedding of gangliosides into the interstitial space may play a central role in cell growth regulation, immune tolerance and tumor-angiogenesis, therefore representing a new target for anticancer therapy . Since 1993 researchers at the University of Buenos Aires and the University of Quilmes (Argentina), have taken part in a project carried out by the <Centro de Inmunologia Molecular> (CIM) from La Havana, Cuba, to developed new strategies for specific active immunotherapy . The project included two ganglioside based vaccines and one anti-idiotypic vaccine . We focused on two antigens: first GM3, an ubiquitous antigen which is over-expressed in several epithelial tumor types; and a second one, N-Glycolyl-GM3 a more <tumor specific> molecule, not being expressed in normal tissues and recently found in several neoplastic cells, in particular breast, melanoma and neuroectodermal cancer cells . We developed two vaccines, one with each antigen, both using proteins derived from the outer membrane proteins (OMP) of Neisseria Meningitidis B, as carriers . We developed also the 1E10 vaccine; an anti-idiotype vaccine designed to mimic the N-Glycolyl-GM3 gangliosides . This monoclonal antibody is an Ab2-type-antibody which recognizes the Ab1 antibody called P3, the latter is a monoclonal antibody that specifically recognizes gangliosides as antigens . Since 1998 we initiated a clinical development program for these three compounds . Results of the phase I clinical trials proved that the three vaccines were safe and able to elicit specific antibody responses . In addition we were able to demonstrate the activation of the cellular arm of the immune response in patients treated with the GM3 vaccine . Although phase I trials are not designed to evaluate antitumor efficacy, it was encouraging to observe tumor shrinkage in some patients treated both with the GM3 and N-Glycolyl-GM3 vaccines . We have already begun a phase II program in several neoplastic diseases, with all three vaccines.

AIDS, 2002 Feb 15, 16(3), 463 - 70
Decline in the prevalence of HIV and sexually transmitted diseases among female sex workers in Cotonou, Benin, 1993-1999; Alary M et al.; BACKGROUND: Within an ongoing HIV/STD prevention project aimed at female sex workers (FSW) in Cotonou, Benin, we evaluated time trends in HIV and STD prevalences from 1993 to 1999 . DESIGN: Three serial cross-sectional surveys were conducted in 1993 (n = 374), 1995-1996 (n = 365), and 1998-1999 (n = 591) . A questionnaire was administered to the FSW and they were screened for HIV, syphilis, Neisseria gonorrhoeae, and Chlamydia trachomatis . RESULTS: The mean percentage of condom use with clients in the week preceding the interview increased from 62.2% in 1993 to 80.7% in 1998-1999 (P = 0.0001) . The prevalence of all infections decreased significantly (all P < 0.02; chi-square for trend) over time: HIV from 53.3% in 1993 to 40.6% in 1998-1999; syphilis from 8.9 to 1.5%; gonorrhoea from 43.2 to 20.5%; and chlamydia from 9.4 to 5.1% . However, the mean age of FSW decreased from 31.0 to 28.4 years between 1993 and 1998-1999 . Moreover, the country of origin of these women changed dramatically over time: the proportion of Ghanaian women decreased from 66.3% in 1993 to 21.6% in 1998-1999 when the predominant group became Nigerian (38.0%) . When controlling for age and country of origin, HIV prevalence was stable over time (P = 0.71), whereas the downward trend remained significant for syphilis and gonorrhoea (both P < 0.001), and was present but not significant for chlamydia (P = 0.13) . CONCLUSION: These data suggest that the time trends in HIV and STD prevalences are partly due to the changing sex work milieu, but that the intervention also had an impact . Prevention programmes aimed at FSW should be highly prioritized.

Expert Opin Pharmacother, 2002 Feb, 3(2), 147 - 57
Current concepts in the management of gonorrhoea; Tapsall J; The incidence of gonorrhoea is again rising in developed countries and a high disease rate has been maintained in less developed regions for a number of years . The need not only for treatment of the individual but also for control of gonorrhoea at a community level has increased significantly following recognition of its role in the amplification of human immunodeficiency virus (HIV) transmission . A sustainable decrease in the incidence of gonorrhoea and other sexually transmitted diseases (STDs) requires an integrated approach combining improved prevention, better diagnosis and optimal treatment . Effective antibiotic treatment is an essential element of this approach . However, antibiotic treatment of gonorrhoea has been severely hampered by the development of antibiotic resistance in Neisseria gonorrhoeae, to the extent that many therapies are no longer effective . Those treatments that retain acceptable efficacy are often unaffordable where they are most needed . Penicillins and tetracyclines should no longer be used in gonococcal disease, there are limitations on the effectiveness of newer macrolides and spectinomycin and in many parts of the world quinolones have been withdrawn from schedules for the treatment of gonorrhoea . Of all the current agents used to treat all forms of gonococcal disease, only the third generation cephalosporins (most notably ceftriaxone) have retained their efficacy; however, decreased susceptibility to these antibiotics has also appeared . Continuing reliance on antibiotic treatment for controlling gonorrhoea in the absence of other necessary approaches will see a further deterioration in the situation . In these circumstances the possibility that gonorrhoea will be untreatable becomes more real.

Tidsskr Nor Laegeforen, 2001 Nov 20, 121(28), 3306 - 9
{Treatment of acute bacterial meningitis}; Skrede S et al.; BACKGROUND: Acute bacterial meningitis requires immediate antimicrobial therapy . MATERIALS AND METHODS: Guidelines to antimicrobial treatment of children and adults with acute bacterial meningitis are presented . RESULTS: The most common agents causing acute bacterial meningitis are Streptococcus agalactiae in children less than one month of age, and Streptococcus pneumoniae and Neisseria meningitidis in individuals more than one month of age . If the causative bacterial agent is not known, children below one month of age should be given ampicillin and gentamicin, whereas older children and adults should be given benzylpenicillin in combination with either cefotaxime or ceftriaxone . We suggest treatment with specific antibiotic regimens in cases of known aetiology.

J Clin Microbiol, 2002 Feb, 40(2), 641 - 4
Comparison of Digene hybrid capture 2 and conventional culture for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in cervical specimens; Darwin LH et al.; Digene's Hybrid Capture 2 (HC2) CT/GC, CT-ID, and GC-ID DNA tests were evaluated by comparison to traditional culture methods for detecting Chlamydia trachomatis and Neisseria gonorrhoeae infections in 669 cervical specimens from high-risk female populations attending two sexually transmitted disease clinics . For detection of either or both infections, the HC2 CT/GC test algorithm had 93.8% sensitivity and 95.9% specificity compared to those of culture . After resolution of discrepant results by direct fluorescent-antibody (DFA) staining or PCR assay, the relative sensitivity and specificity of the HC2 CT/GC test algorithm increased to 94.8 and 99.8%, while the values for culture were 83.6% (McNemar's P value, 0.0062) and 100%, respectively . For detection of the individual pathogens, the relative sensitivities for the HC2 CT-ID and GC-ID tests were 97.2 and 92.2% and the specificities were greater than 99% compared to culture adjucated by DFA staining and PCR . Test performance varied at the two clinics: the HC2 CT/GC algorithm, CT-ID, and GC-ID tests had significantly higher sensitivities (McNemar's P value, <0.05) than that of culture for the population at one clinic as well as for the combined populations . At the other clinic, the HC2 tests performed as well as culture.

J Immunol, 2002 Feb 15, 168(4), 1533 - 7
Cutting edge: Immune stimulation by neisserial porins is toll-like receptor 2 and MyD88 dependent; Massari P et al.; The immunopotentiating activity of neisserial porins, the major outer membrane protein of the pathogenic Neisseria, is mediated by its ability to stimulate B cells and up-regulate the surface expression of B7-2 . This ability is dependent on MyD88 and Toll-like receptor (TLR)2 expression, as demonstrated by a lack of a response by B cells from MyD88 or TLR2 knockout mice to the porins . Using previously described TLR2-dependent reporter constructs, these results were confirmed and were shown to be due to induction of NF-kappaB nuclear translocation . This is the first demonstration of known vaccine adjuvant to stimulate immune cells via TLR2.

J Biol Chem, 2002 Apr 5, 277(14), 12016 - 22 Epub 2002 Jan 25.
Characterization of the methionine sulfoxide reductase activities of PILB, a probable virulence factor from Neisseria meningitidis; Olry A et al.; PILB has been described as being involved in the virulence of bacteria of Neisseria genus . The PILB protein is composed of three subdomains . In the present study, the central subdomain (PILB-MsrA), the C terminus subdomain (PILB-MsrB), and the fused subdomain (PILB-MsrA/MsrB) of N . meningitidis were produced as folded entities . The central subdomain shows a methionine sulfoxide reductase A (MsrA) activity, whereas PILB-MsrB displays a methionine sulfoxide reductase B (MsrB) activity . The catalytic mechanism of PILB-MsrB can be divided into two steps: 1) an attack of the Cys-494 on the sulfur atom of the sulfoxide substrate, leading to formation of a sulfenic acid intermediate and release of 1 mol of methionine/mol of enzyme and 2) a regeneration of Cys-494 via formation of an intradisulfide bond with Cys-439 followed by reduction with thioredoxin . The study also shows that 1) MsrA and MsrB display opposite stereoselectivities toward the sulfoxide function; 2) the active sites of both Msrs, particularly MsrB, are rather adapted for binding protein-bound MetSO more efficiently than free MetSO; 3) the carbon Calpha is not a determining factor for efficient binding to both Msrs; and 4) the presence of the sulfoxide function is a prerequisite for binding to Msrs . The fact that the two Msrs exhibit opposite stereoselectivities argues for a structure of the active site of MsrBs different from that of MsrAs . This is further supported by the absence of sequence homology between the two Msrs in particular around the cysteine that is involved in formation of the sulfenic acid derivative . The fact that the catalytic mechanism takes place through formation of a sulfenic acid intermediate for both Msrs supports the idea that sulfenic acid chemistry is a general feature in the reduction of sulfoxides by thiols.

J Infect Chemother, 2001 Dec, 7(4), 218 - 23
Association between proline-requiring auxotype and fluoroquinolone resistance in Neisseria gonorrhoeae isolated in Japan; Tsunoe H et al.; We examined the association between auxotype and fluoroquinolone resistance in Neisseria gonorrhoeae isolated in Fukuoka, Japan, and investigated whether the prevalence of fluoroquinolone-resistant N . gonorrhoeae isolates was caused by the dissemination of the same clone in the community . We examined the antimicrobial susceptibility of 294 N . gonorrhoeae, isolates obtained during three different periods in Fukuoka, Japan, and 89 isolates of N . gonorrhoeae, classified by the presence of amino-acid substitutions in the quinolone resistance-determining regions (QRDRs) of GyrA and ParC proteins, to various agents, and we examined the auxotypes of the isolates . In 22 isolates with amino-acid substitutions within QRDRs in GyrA and ParC, pulsed-field gel electrophoresis analysis was performed . The proportion of fluoroquinolone-resistant isolates (ciprofloxacin, minimum inhibitory concentration {MIC} > or = 1 microg/ml) in 1998 (23.9%) was significantly higher than that in 1992-1993 (0%) . The proportion of proline-requiring isolates increased significantly, from 4.4% in 1992-1993 to 54.5% in 1998 . The ciprofloxacin MIC90 for the proline-requiring isolates were 32- and 128-fold, respectively, higher than those for the prototrophic isolates and the arginine-requiring isolates . The proportion of isolates with amino-acid substitutions within the QRDRs in GyrA and ParC in the proline-requiring group (55.5%) was significantly higher than that in the prototrophic group (0%) . Of the 22 isolates with amino-acid substitutions within the QRDRs in GyrA and ParC, 16 showed the same pulsed-field gel electrophoresis pattern . These results suggest that a close association exists between the increase in the proline-requiring isolates and the increase in the fluoroquinolone-resistant isolates in the gonococci isolated in Fukuoka, and that the prevalence of fluoroquinolone-resistant N . gonorrhoeae isolates with GyrA and ParC substitutions may be mainly caused by the dissemination of a single clone in the community.

J Infect Chemother, 2001 Sep, 7(3), 175 - 9
Comparison of Neisseria gonorrhoeae isolates from the genital tract and pharynx of two gonorrhea patients; Saika T et al.; Neisseria gonorrhoeae was isolated simultaneously from urethral and pharyngeal specimens of two gonorrhea patients . The pair of isolates from one of the two patients were identical in auxotype, pulsed-field gel electrophoresis pattern, and antimicrobial susceptibility, which indicated that both sites in that patient were infected with the same strain of N . gonorrhoeae . However, the two isolates from the other patient had different properties, indicating that the two sites in the second patient were infected with different strains . The gonococcal infections in these patients failed to respond to initial treatment with sparfloxacin or sulbactam/ampicillin, because the causative strains of N . gonorrhoeae were resistant to the respective antibiotics . Variable patterns and routes of gonococcal infection have recently been discovered in individual patients, suggesting that specimens for bacterial isolation should be taken not just from one site but from various sites that might be infected . This method may contribute to the successful treatment and epidemiological investigation of gonococcal infections.

J Bacteriol, 2002 Feb, 184(4), 919 - 27
Roles of the recJ and recN genes in homologous recombination and DNA repair pathways of Neisseria gonorrhoeae; Skaar EP et al.; The paradigm of homologous recombination comes from Escherichia coli, where the genes involved have been segregated into pathways . In the human pathogen Neisseria gonorrhoeae (the gonococcus), the pathways of homologous recombination are being delineated . To investigate the roles of the gonococcal recN and recJ genes in the recombination-based processes of the gonococcus, these genes were inactivated in the N . gonorrhoeae strain FA1090 . We report that both recN and recJ loss-of-function mutants show decreased DNA repair ability . In addition, the recJ mutant was decreased in pilus-dependent colony morphology variation frequency but not DNA transformation efficiency, while the recN mutant was decreased in DNA transformation efficiency but not pilus-dependent variation frequency . We were able to complement all of these deficiencies by supplying an ectopic functional copy of either recJ or recN at an irrelevant locus . These results describe the role of recJ and recN in the recombination-dependent processes of the gonococcus and further define the pathways of homologous recombination in this organism.

Commun Dis Intell, 2001 Nov, 25(4), 274 - 6
Surveillance of antibiotic resistance in Neisseria gonorrhoeae in the WHO Western Pacific Region, 2000; Identification of bacterial genes required for in-vivo survival; University Department of Paediatrics, John Radcliffe Hospital, Oxford, UK . christoph.tang@paediatrics.oxford.ac.uk

Genetic approaches used for in-vivo studies of bacterial pathogenesis are providing insights into how bacteria disrupt host defences and exploit host molecules for their own advantage . Signature tagged mutagenesis (STM) provides a means of identifying the genes involved in the process of infection, particularly those genes that are important for bacterial proliferation in-vivo . In this review, the application of STM to the understanding of bacterial pathogenesis and findings from work on three human pathogens, Salmonella typhimurium, Mycobacterium tuberculosis and Neisseria meningitidis, are discussed . The next challenge is to understand how these and other genes influence the infective process at the molecular and cellular levels and to design novel interventions to block the progression of disease.

Int J STD AIDS, 2002 Jan, 13(1), 39 - 45
Determinants of gonorrhoea infection among STD clinic attenders in Trinidad--I: sociodemographic factors, knowledge, risk perception and history of STD; Castor D et al.; We determined the prevalence of Neisseria gonorrhoeae and describe sociodemographic factors, knowledge and sexually transmitted disease (STD) history associated with gonococcal infection among 991 STD clinic attenders in Trinidad, West Indies (WI) . The prevalence of gonorrhoea was 25% . Predictors for gonorrhoea were: male gender (odds ratios {OR} 3.5), age 15-19 (OR 2.5) and 20-29 years (OR 2.1), unawareness of the term STD (OR 1.5), self-perceived risk for STDs (OR 1.7) and no history of syphilis (OR 2.0) . When the data were analysed by gender, significant associations were identified between gonococcal infection and age <30 years, single status, and self-perceived risk for STDs for males . Among females, significant associations existed between gonorrhoea and age <30 years, and unawareness of the term STD . This study is novel in describing sociodemographic characteristics of STD clinic attenders, and risk markers associated with gonococcal infection in Trinidad.

Clin Infect Dis, 2002 Feb 15, 34(4), 519 - 22 Epub 2002 Jan 07.
Trichomonas vaginalis is associated with pelvic inflammatory disease in women infected with human immunodeficiency virus; Moodley P et al.; We assessed the association between the causative agents of vaginal discharge and pelvic inflammatory disease (PID) among women attending a rural sexually transmitted disease clinic in South Africa; the role played by coinfection with human immunodeficiency virus type 1 (HIV-1) was studied . Vaginal and cervical specimens were obtained to detect Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, and bacterial vaginosis . HIV-1 infection was established by use of serum antibody tests . A total of 696 women with vaginal discharge were recruited, 119 of whom had clinical PID . Patients with trichomoniasis had a significantly higher risk of PID than did women without trichomoniasis (P=.03) . PID was not associated with any of the other pathogens . When the patients were stratified according to HIV-1 status, the risk of PID in HIV-1-infected patients with T . vaginalis increased significantly (P=.002); no association was found in patients without HIV-1 . T . vaginalis infection of the lower genital tract is associated with a clinical diagnosis of PID in HIV-1-infected women.

Infect Immun, 2002 Feb, 70(2), 909 - 20
Intracellular survival of Neisseria gonorrhoeae in male urethral epithelial cells: importance of a hexaacyl lipid A; Post DM et al.; Neisseria gonorrhoeae is a strict human pathogen that invades and colonizes the urogenital tracts of males and females . Lipooligosaccharide (LOS) has been shown to play a role in gonococcal pathogenesis . The acyl transferase MsbB is involved in the biosynthesis of the lipid A portion of the LOS . In order to determine the role of an intact lipid A structure on the pathogenesis of N . gonorrhoeae, the msbB gene was cloned and sequenced, a deletion and insertion mutation was introduced into N . gonorrhoeae, and the mutant strain was designated 1291A11K3 . Mass spectrometric analyses of 1291A11K3 LOS determined that this mutation resulted in a pentaacyl rather than a hexaacyl lipid A structure . These analyses also demonstrated an increase in the phosphorylation of lipid A and an increase in length of the oligosaccharide of a minor species of the msbB LOS . The interactions of this mutant with male urethral epithelial cells (uec) were examined . Transmission and scanning electron microscopy studies indicated that the msbB mutants formed close associations with and were internalized by the uec at levels similar to those of the parent strain . Gentamicin survival assays performed with 1291A11K3 and 1291 bacteria demonstrated that there was no difference in the abilities of the two strains to adhere to uec; however, significantly fewer 1291A11K3 bacteria than parent strain bacteria were recovered from gentamicin-treated uec . These studies suggest that the lipid A modification in the N . gonorrhoeae msbB mutant may render it more susceptible to innate intracellular killing mechanisms when internalized by uec.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 561 - 5
Inducible, but not constitutive, resistance of gonococci to hydrophobic agents due to the MtrC-MtrD-MtrE efflux pump requires TonB-ExbB-ExbD proteins; Rouquette-Loughlin C et al.; The MtrC-MtrD-MtrE efflux pump possessed by Neisseria gonorrhoeae is very similar to the MexA-MexB-OprM efflux pump of Pseudomonas aeruginosa . Because the antimicrobial resistance property afforded by the MexA-MexB-OprM efflux pump also requires the TonB protein, we asked whether a similar requirement exists for the gonococcal efflux pump . Unlike earlier studies with P . aeruginosa, we found that constitutive levels of gonococcal resistance to hydrophobic antimicrobial agents (i.e., Triton X-100 {TX-100}) did not require the TonB, ExbB, or ExbD protein . However, inducible levels of TX-100 resistance in gonococci had an absolute requirement for the TonB-ExbB-ExbD system, suggesting that such resistance in gonococci has an energy requirement above and beyond that required for constitutive pump activity.

Biochim Biophys Acta, 2001 Dec 19, 1568(3), 205 - 15
Exploring computational lead optimisation with affinity constants obtained by surface plasmon resonance for the interaction of PorA epitope peptides with antibody against Neisseria meningitidis; Fischer MJ et al.; LUDI is a program used for de novo structure-based design of ligands and can predict binding of ligands quantitatively using a scoring function . Here we evaluate LUDI in a lead optimisation study with ligands for the antibody MN12H2, that has been raised against outer membrane protein PorA epitope P1.16 of Neisseria meningitidis . The ligands were synthetic peptides that are derived from the smallest binding epitope (182)DTNNN(186) . LUDI's fragment building rules are used for the proposal of new peptide-ligands for MN12H2 and were focused on replacements of Asp(186) in the epitope . Accordingly, a series of peptides was synthesised with isosteric mutations . The interaction of the peptides with MN12H2 was analysed with a surface plasmon resonance competition assay yielding equilibrium binding constants in solution (K(S)) . The binding affinity seems to be largely determined by entropy, and the side chain of Asn(186) is sensitive for charge, inversion, hydrophobicity and size . Head-to-tail cyclisation of the peptide in a nine-amino-acid ring gives little reduction in affinity . It is concluded that the scoring function of LUDI does not help in optimisation of the peptide lead for MN12H2 binding . Other more elaborate molecular mechanics calculations show similar results . This implies that our current knowledge of molecular recognition is insufficient for explaining a case of peptide-protein binding, where the design process requires subtle changes in structure (from lead finding to lead optimisation).

Mikrobiol Z, 2001 Sep-Oct, 63(5), 21 - 6
{ATPase activity of Neisseria gonorrhoeae and Staphylococcus aureus strains}; Skliar TV et al.; It has been established that membrane vesicles of gonococci possess ATPase activity within 0.05-0.14 mmol of ATP during 60 min per 1 mg of protein and for staphylococcus--within 0.07-0.19 mmol of ATP for 60 min per 1 mg of protein . The identity in all the kinetic parameters is observed for all the studied gonococcus strains: Na+ and Ca2+ ions inhibit ATPase activity within 12-19%; Mg2+ ions increase the ATPase activity 2-2.5 times . Dicyclohexylcarboimide, a specific ATPase inhibitor, suppresses ATPase activity by 50-70% . ATPase activity in the strains of bacteria containing antibiotic-resistant plasmids is established to increase 1.2-2.8 times for gonococcus and 1.2-2.7 for staphylococcus as compared to sensitive ones.

Microbiology, 2002 Jan, 148(Pt 1), 229 - 36
A broad-host-range vector of incompatibility group Q can work as a plasmid vector in Neisseria meningitidis: a new genetical tool; Takahashi H et al.; Plasmid pHT128, a derivative of the broad-host-range IncQ vector pGSS33, was successfully introduced into Neisseria meningitidis . Under optimal conditions, pHT128 was transferred from Escherichia coli to N . meningitidis by triparental conjugation at a frequency of 10(-5)-10(-6) . The copy number of pHT128 in N . meningitidis was almost the same as in E . coli, in which the copy number of IncQ plasmids per chromosome is estimated to be 10 . pHT128 was maintained as an episome in N . meningitidis in the presence of chloramphenicol, a marker of the plasmid . It was also shown that an opc or pilE1 gene cloned on pHT128 could be expressed in N . meningitidis under control of the tac promoter and could complement a mutation of opc or pilE1, respectively . In addition, the conjugational introduction of pHT128 into N . meningitidis was demonstrated to be independent of natural transformation competence . All the results indicate that pHT128 is a useful vector for N . meningitidis as a new genetical tool.

Int J STD AIDS, 2001 Dec, 12(12), 829 - 30
Gonococcal ophthalmia treated with ciprofloxacin; Price LM et al.; Neisseria gonorrhoeae infection in the eye and its treatment with ciprofloxacin is presented.

Sex Transm Dis, 2002 Jan, 29(1), 50 - 8
The Bali STD/AIDS Study: evaluation of an intervention for sex workers; Ford K et al.; BACKGROUND: Prostitution has been an important factor in the spread of HIV infection in Asia . Interventions need to be developed to reduce the risk of transmission of sexually transmitted infections in this area . GOALS: To educate female sex workers about sexually transmitted infections and assess the impact of the educational intervention . STUDY DESIGN: Brothel areas in Denpasar, Bali, Indonesia, were divided into areas of high and low program (interventional) effort . The intervention included educational sessions for sex workers, treatment of sex workers for sexually transmitted disease (STD), condom distribution, and printed information for clients of the sex workers . A high-effort area was one in which a more intensive educational intervention occurred . A clinic was available for STD treatment in both areas . Behavioral surveys and STD testing were used to evaluate the programs . Six hundred female sex workers participated in behavioral surveys and STD examinations every 6 months for four rounds of data collection . Each round, about half of the women were new to the study . A total of 1586 women participated in at least one evaluation round . Changes were evaluated in AIDS knowledge, STD knowledge, and condom use, as well as in the prevalence of Neisseria gonorrhoeae, Chlamydia trachomatis, syphilis, and Trichomonas vaginalis . Testing for HIV was conducted anonymously . RESULTS: Improvements were noted in the knowledge of sex workers about AIDS and STDs and in the reduction of some bacterial STDs . Women who remained in the study area for more than one round had increased knowledge of HIV infection/STDs and condom use and had reduced levels of syphilis, gonorrhea, and trichomonas infection (P < 0.01) . The additional education received by women in the high-effort program area was associated with a reduction in the prevalence of syphilis . Prevalence of HIV remained low throughout the study . The high level of turnover of female sex workers contributed to the maintenance of significant levels of STDs in this population . CONCLUSIONS: Developers of HIV/STD prevention programs for sex workers need to consider the mobility of the sex worker population . Interventions combining behavioral and medical approaches can contribute to prevention of these diseases.

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