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The mechanisms of neurodegeneration in Alzheimer's disease (AD) remain under investigation. Alterations in the blood-brain barrier facilitate exchange of inflammatory mediators and immune cells between the brain and the periphery in AD. Here, we report analysis of phenotype and functions of polymorphonuclear neutrophils (PMN) in peripheral blood from patients with amnestic mild cognitive impairment (aMCI, n = 13), patients with mild AD (mAD, n = 15), and healthy elderly controls (n = 13). Results showed an increased expression of CD177 in mAD but not in healthy or aMCI patients. IL-8 stimulated increased expression of the CD11b integrin in PMN of healthy subjects in vitro but PMN of aMCI and mAD patients failed to respond. CD14 and CD16 expression was lower in PMN of mAD but not in aMCI individuals relative to controls. Only PMN of aMCI subjects expressed lower levels of CD88. Phagocytosis toward opsonized E. coli was differentially impaired in PMN of aMCI and mAD subjects whereas the capacity to ingest Dextran particles was absent only in mAD subjects. Killing activity was severely impaired in aMCI and mAD subjects whereas free radical production was only impaired in mAD patients. Inflammatory cytokine (TNFα, IL-6, IL-1β, IL-12p70) and chemokine (MIP-1α, MIP-1β, IL-8) production in response to LPS stimulation was very low in aMCI and nearly absent in mAD subjects. TLR2 expression was low only in aMCI. Our data showed a differentially altered capacity of PMN of aMCI and mAD subjects to respond to pathological aggression that may impact impaired responses associated with AD development.

Accumulating evidence suggests that the propagation of hyperphosphorylation of tau protein and the amyloid-β peptide can be mediated by extracellular vesicles (EVs) and be associated with the onset and the progression of Alzheimer's disease (AD). As EVs may transfer between the brain and the blood, we have thus hypothesized that the total plasma EVs (pEVs) may contain potential markers to predict the mild cognitive impairment (MCI) and AD progression. We have thus quantified AD-related proteins in isolated pEVs from controls, MCI and AD subjects. In pEVs, we observed early changes of total tau (tTau), amyloid precursor protein levels, and phospho-tau (pTau)-T181/tTau ratio from MCI subjects and late increases of Aβ42 and pTau-T181 levels from patients with moderate AD. Interestingly, abnormal amyloid precursor protein levels and pTau-T181/tTau ratio in pEVs demonstrated a high accuracy to define MCI and AD staging. Although larger samples sizes will be needed to generate well-powered investigations, these preliminary results highlighted the potential of AD-related proteins enriched in pEVs as a sensitive tool for differentiating patients with MCI to patients with AD and monitoring AD progression.

BACKGROUND AND RATIONALE FOR THE STUDY:The global plan to eradicate hepatitis C (HCV) led by the World Health Organisation (WHO) outlines the use of highly effective direct-acting antiviral drugs (DAAs) to achieve elimination by 2030. Identifying individuals with active disease and investigation of the breadth of diversity of the virus sub-Saharan Africa (SSA) is essential as genotypes in this region (where very few clinical trials have been carried out) are distinct from those found in other parts of the world. We undertook a population-based nested case-control study in Uganda and obtained additional samples from the Democratic Republic of Congo (DRC), to estimate the prevalence of HCV, assess strategies for disease detection using serological and molecular techniques, and characterise genetic diversity of the virus. Using next generation (NGS) and Sanger sequencing, we aimed to identify strains circulating in East and Central Africa. MAIN RESULTS:7751 Ugandan patients were initially screened for HCV and 20 PCR positive samples obtained for sequencing. Serological assays were found to vary significantly in specificity for HCV. HCV strains detected in Uganda included genotypes (g) 4k, 4p, 4q and 4s and a new unassigned genotype 7 HCV strain. Two additional unassigned g7 strains were identified in patients originating from DRC (one partial and one full ORF sequence). These g4 and 7 strains contain NS3 and NS5A polymorphisms associated with resistance to DAAs in other genotypes. Clinical studies are therefore indicated to investigate treatment response in infected patients. CONCLUSION:While HCV prevalence and genotypes have been well characterised in patients in well-resourced countries, clinical trials are urgently required in SSA where highly diverse g4 and 7 strains circulate. This article is protected by copyright. All rights reserved.

Alzheimerś disease (AD) is a progressive irreversible neurological brain disorder characterized by accumulation of amyloid-β, amyloid plaques, and neurofibrillary tangles. Inflammation and immune alterations have been linked to AD, suggesting that the peripheral immune system plays a role during the asymptomatic period of AD. NK cells participate in innate immune surveillance against intracellular pathogens and malignancy but their role in AD remains controversial. We have investigated changes in peripheral NK cell phenotypes and functions in amnestic mild cognitive impairment (aMCI, n = 10), patients with mild AD (mAD, n = 11), and healthy elderly controls (n = 10). Patients selected according to NINCDS-ADRDA criteria were classified using neuropsychological assessment tests. Phenotype analysis revealed differences in expression of CD16 (increased in mAD), NKG2A (decreased in aMCI), and TLR2 and TLR9 (both decreased in mAD). Functional assays revealed that NK cell killing activity and degranulation (CD107 expression) were unchanged in the three groups. In contrast, expression of the CD95 receptor was increased in aMCI and mAD. Granzyme B expression and cytokine production (TNFα, IFNγ) were increased in aMCI but not in mAD. CCL19- but not CCL21-dependent chemotaxis was decreased in aMCI and mAD, despite the fact that CCR7 expression was increased in aMCI. Our data suggest that the number of alterations observed in peripheral NK cells in aMCI represent an activation state compared to mAD patients and that may reflect an active immune response against a still to be defined aggression.

Background Clostridium difficile (CD) is the leading cause of health care–associated diarrhea and can result in asymptomatic carriage. Rates of asymptomatic CD colonization on hospital admission range from 1.4%-21%. The objective of this study was to evaluate host and bacterial factors associated with colonization on admission. Methods The Consortium de recherche quebecois sur le Clostridium difficile study provided data for analysis, including demographic information, known risk factors, and potential confounding factors, prospectively collected for 5,232 patients from 6 hospitals in Quebec and Ontario over 15 months from 2006-2007. Stool or rectal swabs were obtained for culture on admission. Pulsed-field gel electrophoresis was performed on the isolates. The presence of antibody against CD toxins A and B was measured. Results There were 212 (4.05%) patients colonized with CD on admission, and 5,020 patients were not colonized with CD. Multivariate logistic regression analysis showed that hospitalization within the last 12 months, use of corticosteroids, prior CD infection, and presence of antibody against toxin B were associated with colonization on admission. Of patients colonized on admission, 79.4% had non-NAP1, non-NAP2 strains. Conclusion There are identifiable risk factors among asymptomatic CD carriers that could serve in their detection and provide a basis for targeted screening.

Background Efficient viral load testing is needed for hepatitis C (HCV) surveillance and diagnosis. HCV viral load testing using dried blood spots (DBSs), made with a single drop of finger-prick whole blood on filter paper, is a promising alternative to traditional serum- or plasma-based approaches. Methods We adapted the Abbott Molecular m2000 instrument for high-throughput HCV viremia testing using DBSs with simple specimen processing and applied these methods to estimate the national burden of infection in the Democratic Republic of the Congo (DRC). We tested DBSs collected during the 2013-2014 DRC Demographic and Health Survey, including 1309 adults ≥40 years of age. HCV-positive samples underwent targeted sequencing, genotyping, and phylogenetic analyses. Results This high-throughput screening approach reliably identified HCV RNA extracted from DBSs prepared using whole blood, with a 95% limit of detection of 1196 (95% confidence interval [CI], 866-2280) IU/mL for individual 6-mm punches and 494 (95% CI, 372-1228) IU/mL for larger 12-mm punches. Fifteen infections were identified among samples from the DRC Demographic and Health Survey; the weighted country-wide prevalence of HCV viremia was 0.9% (95% CI, 0.3%-1.6%) among adults ≥40 years of age and 0.7% (95% CI, .6%-.8%) among human immunodeficiency virus-infected subjects. All successfully genotyped cases were due to genotype 4 infection. Conclusions DBS-based HCV testing represents a useful tool for the diagnosis and surveillance of HCV viremia and can easily be incorporated into specimen referral systems. Among adults ≥40 years of age in the DRC, 100000-200000 may have active infection and be eligible for treatment.

Clostridium difficile infection is the leading cause of health care-associated diarrhea, and the bacterium can also be carried asymptomatically. The objective of this study was to identify host and bacterial factors associated with health care-associated acquisition of C. difficile infection and colonization.We conducted a 15-month prospective study in six Canadian hospitals in Quebec and Ontario. Demographic information, known risk factors, potential confounding factors, and weekly stool samples or rectal swabs were collected. Pulsed-field gel electrophoresis (PFGE) was performed on C. difficile isolates to determine the genotype. Levels of serum antibodies against C. difficile toxins A and B were measured.A total of 4143 patients were included in the study; 117 (2.8%) and 123 (3.0%) had health care-associated C. difficile infection and colonization, respectively. Older age and use of antibiotics and proton-pump inhibitors were significantly associated with health care-associated C. difficile infection. Hospitalization in the previous 2 months; use of chemotherapy, proton-pump inhibitors, and H(2) blockers; and antibodies against toxin B were associated with health care-associated C. difficile colonization. Among patients with health care-associated C. difficile infection and those with colonization, 62.7% and 36.1%, respectively, had the North American PFGE type 1 (NAP1) strain.In this study, health care-associated C. difficile infection and colonization were differentially associated with defined host and pathogen variables. The NAP1 strain was predominant among patients with C. difficile infection, whereas asymptomatic patients were more likely to be colonized with other strains. (Funded by the Consortium de Recherche sur le Clostridium difficile.).

Heterosexual transmission of hepatitis C virus (HCV) is uncommon, with few studies undertaken in Central Africa. To determine the frequency of inter-spouse HCV transmission, cross-sectional studies of elderly individuals in Ebolowa, Cameroon and Nola, Central African Republic, in which, respectively, 24 and 83 long-term couples had been identified, were examined further. Blood samples were tested for antibody to HCV. Anti-HCV positive samples were genotyped by phylogenetic analysis of a fragment of the NS5B gene. In Nola, 4 out of 9 (44.4%) wives of anti-HCV positive husbands and 1 out of 74 (1.4%) wives of anti-HCV negative husbands were anti-HCV positive (P

The performances of commercial enzyme-linked immunosorbent assays (ELISAs) in detecting herpes simplex virus type 2 (HSV-2) antibodies have been inconsistent for African and human immunodeficiency virus (HIV)-positive populations. We compared the performances of the HerpeSelect and Kalon glycoprotein G2 ELISAs for patients with genital ulcer disease in Ghana and the Central African Republic. Sera from 434 women were tested with the HerpeSelect assay, and a subsample ( n = 199) was tested by the Kalon assay. Ulcer swabs and cervicovaginal lavage samples were tested for HSV-2 DNA by PCR. HSV-2-seronegative women with detectable genital HSV-2 DNA were retested for HSV-2 antibodies 14 and 28 days later by the two ELISAs. A total of 346 (80%) women were positive by HerpeSelect at baseline, and 225 (54%) had detectable genital (lesional or cervicovaginal) HSV-2 DNA. Sixty-six (19%) HerpeSelect-positive samples had low-positive index values (1.1 to 3.5), and 58% of these samples had detectable genital HSV-2 DNA. Global agreement between the two serological assays was 86%. Concordance was high (99%) for sera that were negative by HerpeSelect or had high index values (>3.5). Defining infection detected by HSV-2 DNA PCR and/or Kalon assay as true infection, 71% of sera with low-positive index values were associated with true HSV-2 infection. Twenty-five women were identified as having nonprimary first-episode genital HSV-2 infection. Rates of HSV-2 seroconversion at day 14 were 77% (10/13 patients) by HerpeSelect assay and 23% (3/13 patients) by Kalon assay, with four additional seroconversions detected by Kalon assay at day 28. HIV serostatus did not influence assay performance. Low index values obtained with the HerpeSelect assay may correspond to true HSV-2 infection, in particular to nonprimary first episodes of genital HSV-2 infection, and need to be interpreted in the context of clinical history.

We studied the association between herpes simplex virus type-2 (HSV-2) and HIV-1 viralload in plasma, semen, cervico-vaginal secretions and genital ulcers. Forty-seven (68%) men and 57 (80%) women were HSV-2 antibody positive, of whom 12 (26%, 95% confidence interval [CI] 20, 32) and five (8%, 95% CI 4, 12), respectively, had HSV-2 genital shedding detected by polymerase chain reaction. The mean HIV-1 seminal and cervico-vaginal viral loads did not differ significantly according to the presence of HSV-2 shedding. Eleven men and 15 women presented with genital ulcers; all ulcers were due to HSV-2. Ten men and nine women were followed up over six days: the mean (95% CI) HIV-1 log viral load copies/mL in the genital ulcers at baseline and final visits were 2.5 (2.3, 2.7) and 3.1 (2.0, 4.2) for men and 3.0 (2.6, 3.4) and 2.7 (2.3, 3.1) for women. These findings do not support the hypothesis that HSV-2 increases the HIV-1 viral load in genital secretions.

Second-generation surveillance for HIV includes measures of high-risk behaviors among the general adult population and sex workers (SW). Questionnaires are prone to social desirability biases because individuals minimize the frequency of behaviors not expected from them. The objective was to determine whether the prostate-specific antigen (PSA) could be used as a biological marker of unprotected intercourse. We measured the presence of PSA in vaginal secretions of women who were (n = 508) or were not (n = 658) SW presenting with vaginal discharge in health facilities of Ghana Togo Guinea and Mali. The cutoff for a positive assay was determined as = 0.4 µg/L based on a subsample of 95 non-SW claiming abstinence for 3 months. A positive PSA assay was correlated with infections with Neisseria gonorrhoeae Chlamydia trachomatis and Mycoplasma genitalium. Among non-SW a positive PSA was more common among those with HIV but less frequent in those better educated. Among SW and non-SW women from Ghana were less likely to have a positive PSA and had a lower prevalence of sexually transmitted infections than those from elsewhere. PSA can be used as a biological marker of unprotected intercourse allowing interventions to target efforts on those at highest risk. (authors)

The accuracy and usefulness of laboratory-developed real-time PCR procedures using a Light Cycler instrument (Roche Diagnostics) for detecting and quantifying human immunodeficiency virus type 1 (HIV-1) RNA and DNA as well as herpes simplex virus type 1 (HSV-1)/HSV-2 DNA in cervicovaginal secretions from women coinfected with HIV and HSV were evaluated. For HIV-1, the use of the NEC152 and NEC131 primer set and the NEC-LTR probe in the long terminal repeat gene allowed us to detect accurately the majority of HIV-1 subtypes of group M circulating in sub-Saharan Africa, including subtypes A, B, C, D, and G as well as circulating recombinant forms 02 and 11. The detection threshold of real-time PCR for HIV in cervicovaginal lavage samples was 5 copies per assay for both RNA and DNA; the intra- and interassay coefficients of variation of C T values were 1.30% and 0.69% (HIV-1 RNA) and 1.84% and 0.67% (HIV-1 DNA), respectively. Real-time PCR for HSV using primers and probe targeting the HSV DNA polymerase gene allowed both detection and quantification of HSV DNA and also differentiation between HSV-1 and HSV-2 genotypes. The detection threshold of real-time PCR for HSV was 5 copies per assay; the intra- and interassay coefficients of variation of C T values were 0.96% and 1.49%, respectively. Both manual and automated silica-based procedures were appropriate for combined extraction of HIV and HSV genomes from female genital secretions. Taken together, these findings indicate that real-time PCR may be used as a unique nucleic acid amplification procedure to detect and quantify HIV and HSV genomes in cervicovaginal secretions and thus to assess at reduced costs the genital shedding of both viruses in women included in intervention studies.

Objectives: (1) To assess the frequency of influenza and influenza-like illness (ILI) during pregnancy; (2) to explore the associated risk of adverse maternal and fetal outcomes. Methods: All women who delivered at the Sherbrooke University Hospital in the province of Quebec during the period of February 15 to April 30, 2002, were invited to complete a questionnaire. Medical records of participating women and of their infants were reviewed. Complement-fixing antibody titres against influenza A were measured in paired sera drawn during antenatal screening and at delivery. Results: The study participation rate was 96. 3%. Nine of 517 women stated that they had received the influenza vaccine during pregnancy, and only 3 of 47 women with a chronic pulmonary disease had been immunized. Overall, 464 ILI episodes were reported by 331 women, and a significant maternal complication occurred in 8 cases. ILI episodes resulted in medical consultations in 55.4% of cases, and in antibiotic prescriptions in 13.8% of cases. There was no indication of an increased frequency of adverse perinatal outcomes associated with ILI during pregnancy. Paired sera were available for 156 participants and seroconversion was observed in 6.6%. Conclusions: ILI during pregnancy was associated with health services utilization but complications were infrequent. More studies are needed to demonstrate the usefulness of routine vaccination of healthy pregnant women against influenza.

Objectives: (1) To assess risk factors for urethral infections with Chlamydia trachomatis , Neisseria gonorrhoeae , and Trichomonas vaginalis among male clients of female sex workers (FSWs) in Benin; (2) to study the validity of LED testing of male urine samples compared to a highly sensitive gold standard (PCR) for the diagnosis of urethral infections with the organisms cited above. Methods: Male clients of FSWs (n=404) were recruited on site at prostitution venues in Cotonou, Benin, between 28 May and 18 August 1998. A urine sample was obtained from each participant just before he visited the FSW, and tested immediately using a leucocyte esterase dipstick (LED) test. It was then tested for HIV using the Calypte EIA with western blot confirmation, and for C trachomatis, N gonorrhoeae , and T vaginalis by PCR. After leaving the FSW’s room, participants were interviewed about demographics, sexual behaviour, STI history and current symptoms and signs, and were examined for urethral discharge, genital ulcers, and inguinal lymphadenopathies. Results: STI prevalences were: C trachomatis , 2.7%; N gonorrhoeae , 5.4%; either chlamydia or gonorrhoea 7.7%; T vaginalis 2.7%; HIV, 8.4%. Lack of condom use with FSWs and a history of STI were independently associated with C trachomatis and/or N gonorrhoeae infection. Over 80% of these infections were in asymptomatic subjects. The overall sensitivity, specificity, positive and negative predictive values of the LED test for detection of either C trachomatis or N gonorrhoeae were 48.4%, 94.9%, 44.1%, and 95.7%, respectively. In symptomatic participants (n=22), all these parameters were 100% while they were 47.4%, 94.7%, 37.5%, and 96.4% in asymptomatic men (n=304). Conclusions: Since most STIs are asymptomatic in this population, case finding programmes for gonorrhoea and chlamydia could be useful. The performance characteristics of the LED test in this study suggest that it could be useful to detect asymptomatic infection by either C trachomatis or N gonorrhoeae in high risk men.

Background: Syphilis remains a major cause of fetal loss and neonatal mortality in sub-Saharan Africa. Guinea-Bissau is the epicenter of the HIV-2 epidemic, and little is known about the impact of HIV-2 on pregnancy. Goal: To understand better the impact of maternal syphilis and HIV-2 on the outcomes of pregnancy in Bissau, Guinea-Bissau. Study Design: Using a case-control design, maternal syphilis and HIV-2 seropositive results were examined in relation to adverse outcomes of pregnancy. From June 1997 to April 1998, women presenting to the Simao Mendes hospital of Bissau for a delivery or a spontaneous abortion were invited to participate in the study, and 1341 women were enrolled. The 743 control subjects were women who had delivered a term neonate with a birthweight greater than 2500 g that survived the first 28 days of life. The cases were classified into five groups of mothers according to the outcome of pregnancy: stillbirths (n = 185), spontaneous abortions (n = 89), premature deliveries (n = 256), small-for-gestation-age babies (n = 55), and neonatal deaths (n = 13). Results: Among the control subjects, the prevalences of serologic syphilis and HIV infection were 3.9% and 7.9%, respectively. Positive syphilis serology results together with a rapid plasma reagin titer of 1:16 or more were associated with delivery of a stillborn (adjusted odds ratio [AOR], 6.05) and premature delivery (AOR, 2.98). In Bissau, the population-attributable risk fraction of syphilis was 7.2% (95% CI, 2.2-11.9%) for stillbirths and only 2.4% (95% CI, 0-5.8%) for premature deliveries. Spontaneous abortions, delivery of a small-for-gestation-age baby, and neonatal deaths were not associated with positive syphilis serology results. None of these pregnancy outcomes was significantly associated with HIV-2 infection. Conclusions: The effect of syphilis on the outcome of pregnancy in West Africa is similar to that reported from other parts of Africa. However, because the prevalence of positive syphilis serology results is relatively low, the impact of screening for syphilis on the outcome of pregnancy is likely to be relatively modest. The findings from this study confirm the absence of association between HIV-2 and an adverse pregnancy outcome.

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

International audience; Pseudomonas aeruginosa and Staphylococcus aureus are the most prevalent pathogens in airway infections of cystic fibrosis (CF) patients. We studied how these pathogens coexist and interact with each other. Clinical isolates of both species were retrieved from adult CF patients. Culture supernatants from 63 P. aeruginosa isolates triggered a wide range of biofilm-stimulatory activities when added to the culture of a control S. aureus strain. The extent of biofilm formation by S. aureus was positively correlated to the levels of the 2-alkyl-4-(1H)-quinolones (AQs) Pseudomonas Quinolone Signal (PQS) and 2-heptyl-4-hydroxy quinoline N-oxide (HQNO) produced by the P. aeruginosa isolates. Supernatants from P. aeruginosa isogenic mutants deficient in PQS and HQNO production stimulated significantly less biofilm formation by S. aureus than that seen with the parental strain PA14. When studying co-isolated pairs of P. aeruginosa and S. aureus retrieved from patients showing both pathogens, P. aeruginosa supernatants stimulated less biofilm production by the S. aureus counterparts compared to that observed using the control S. aureus strain. Accordingly, some P. aeruginosa isolates produced low levels of exoproducts and also some of the clinical S. aureus isolates were not stimulated by their co-isolates or by PA14 despite adequate production of HQNO. This suggests that colonization of the CF lungs promotes some type of strain selection, or that co-existence requires specific adaptations by either or both pathogens. Results provide insights on bacterial interactions in CF.

Campylobacter infection is a leading cause of bacterial gastroenteritis worldwide, and most clinical cases appear as isolated, sporadic infections for which the source is rarely apparent. From July 2005 to December 2007 we conducted a prospective case-case study of sporadic, domestically-acquired Campylobacter enteritis in rural versus urban areas and a prevalence study of Campylobacter in animal and environmental sources in the Eastern Townships, Quebec. Isolates were typed using Multilocus Sequence Typing (MLST) to reinforce the case-case findings and to assign a source probability estimate for each human isolate. The risk of human campylobacteriosis was 1.89-fold higher in rural than urban areas. Unconditional multivariate logistic regression analysis identified two independent risk factors associated with human Campylobacter infections acquired in rural area: occupational exposure to animals (OR = 10.6, 95% CI: 1.2-91, p = 0.032), and household water coming from a private well (OR = 8.3, 95% CI: 3.4-20.4, p

OBJECTIVES--Methods to quantitate Chlamydia trachomatis have never been compared although it would be relevant to periodically evaluate the sensitivity of a detection system. We compared the sensitivity and reproducibility of culture, direct immunofluorescence and the polymerase chain reaction (PCR) to quantitate C trachomatis. METHODS--A competitive semiquantitative PCR procedure was developed. The number of inclusions in culture, particles by direct immunofluorescence and DNA copies by PCR were measured for 12 patient specimens. Variation was determined by measuring a sample 10 times for each method. RESULTS--Patient C trachomatis major outer membrane protein gene DNA was measured semiquantitatively by amplifying together with reference DNA. DNA molecules, particles and infectious units were quantitated in clinical samples with, on average, 595 DNA molecules and 87 immunofluorescent particles observed per inclusion-forming-unit. Similar coefficients of variation (47-52%) were observed for the 3 procedures. CONCLUSION--Competitive PCR and counting immunofluorescent particles provide reproducible and sensitive methods of quantitating C trachomatis.

Chlamydia trachomatis can be directly transmitted by sexual or perinatal contact and indirectly transmitted by flies or fomites. Whether distinct epidemiologic forces among human populations or biologic characteristics of the organism are responsible for the different routes of transmission is uncertain.To determine if ophthalmia neonatorum and trachoma are linked epidemiologically, 38 infants with ophthalmia and 277 children with trachoma were studied for evidence of C. trachomatis infection using culture, antigen and DNA detection tests. The study was performed in a trachoma endemic area of central Kenya.Of infants with ophthalmia neonatorum, 8% to 9% had microbiologic evidence of ocular C. trachomatis infection. Of the children with trachoma, 31% had evidence of chlamydial infection. Ninety-two percent of the 59 identified strains causing trachoma belonged to the classic trachoma serovars (A, B, Ba and C). Neither of the two chlamydial strains recovered from infants with ophthalmia was a trachoma serovar. Mothers rarely (3%) had cervical C. trachomatis infection.This study does not support a major role for perinatally transmitted C. trachomatis infection in trachoma epidemiology.Chlamydia trachomatis can be directly transmitted by sexual or perinatal contact and indirectly by flies or fomites. To determine if ophthalmia neonatorum and trachoma are linked epidemiologically, 38 infants with ophthalmia (mean age of 1.8 +or- 2.5 months) and 277 children with trachoma (3 months - 14 years old) were studied for evidence of C trachomatis infection using culture, antigen, and DNA detection tests. The study was performed in a trachoma endemic area in 2 rural health centers (Lare and Mutuwati) in the Meru District in central Kenya over a 2 1/2-year period between 1988-1991. 29 mothers of the 38 infants had tests for cervical C. trachomatis infection. 53 children with trachomatous inflammation (follicular) (TF) and 224 children with trachoma inflammation (intense) with or without TF were enrolled. Subjects were classified according to the World Health Organization trachoma grading scheme. 167 mothers of these 277 children were examined for evidence of cervical C trachomatis infection. Of infants with ophthalmia neonatorum, 8-9% had microbiologic evidence of ocular G trachomatis infection. Of the children with trachoma, 31% had evidence of chlamydial infection. 92% of the 59 identified strains causing trachoma belonged to the classic trachoma serovars (A, B, Ba and C). 3% of mothers had cervical C trachomatis infection. Among children with trachoma, 10% had positive ocular cultures for C. trachomatis, 24 had positive EIA tests, and 31% had positive PCR tests. Among infants with ophthalmia neonatorum, evidence of C. trachomatis infection was rare. 9% (2 of 22) had positive EIA tests, and 8% (2 of 26) had positive PCR tests. Among the mothers of children with trachoma, 3% had positive C. trachomatis cervical cultures and 2% had positive EIA tests. No mother of an infant with ophthalmia neonatorum had positive cervical tests for C. trachomatis. There was no evidence of a major role for perinatally transmitted C trachomatis infection in trachoma epidemiology.

Background Oral vancomycin (125 mg qid) is recommended as treatment of severe Clostridium difficile infection (CDI). Higher doses (250 or 500 mg qid) are sometimes recommended for patients with very severe CDI, without supporting clinical evidence. We wished to determine to what extent faecal levels of vancomycin vary according to diarrhoea severity and dosage, and whether it is rational to administer high-dose vancomycin to selected patients. Methods We recruited hospitalized adults suspected to have CDI for whom oral vancomycin (125, 250 or 500 mg qid) had been initiated. Faeces were collected up to 3 times/day and levels were measured with the AxSYM fluorescence polarization immunoassay. Results Fifteen patients (9 with confirmed CDI) were treated with oral vancomycin. Patients with ≥4 stools daily presented lower faecal vancomycin levels than those with a lower frequency. Higher doses of oral vancomycin (250 mg or 500 mg qid) led to consistently higher faecal levels (> 2000 mg/L), which were 3 orders of magnitude higher than the MIC90 of vancomycin against C. difficile. One patient receiving 125 mg qid had levels below 50 mg/L during the first day of treatment. Conclusions Faecal levels of vancomycin are proportional to the dosage administered and, even in patients with increased stool frequency, much higher than the MIC90. Patients given the standard 125 mg qid dosage might have low faecal levels during the first day of treatment. A loading dose of 250 mg or 500 mg qid during the first 24-48 hours followed by the standard dosage should be evaluated in larger studies, since it might be less disruptive to the colonic flora and save unnecessary costs.

BACKGROUND: Little is known about the impact of episodic treatment of herpes on human immunodeficiency virus type 1 (HIV-1). METHODS: Women from Ghana and the Central African Republic who had genital ulcers were enrolled in a randomized double-blind placebo-controlled trial of acyclovir plus antibacterials and were monitored for 28 days. Ulcer etiologies and detection of lesional HIV-1 RNA were determined by polymerase chain reaction (PCR). Cervicovaginal HIV-1 RNA and herpes simplex virus type 2 (HSV-2) DNA and plasma HIV-1 RNA were quantitated by real-time PCR. Primary analyses included 118 HIV-1-infected women with HSV-2 ulcers (54 of whom were given acyclovir and 64 of whom were given placebo). RESULTS: Acyclovir had little impact on (1) detection of cervicovaginal HIV-1 RNA (risk ratio [RR] 0.96; 95% confidence interval [CI] 0.8-1.2) at day 7 of treatment (2) the mean cervicovaginal HIV-1 RNA load (-0.06 log(10) copies/mL; 95% CI -0.4 to 0.3 log(10) copies/mL) at day 7 of treatment or (3) the plasma HIV-1 RNA load (+0.09 log(10) copies/mL; 95% CI -0.1 to 0.3 log(10) copies/mL) at day 14 of treatment. At day 7 women receiving acyclovir were less likely to have detectable lesional HIV-1 RNA (RR 0.70; 95% CI 0.4-1.2) or cervicovaginal HSV-2 DNA (RR 0.69; 95% CI 0.4-1.3) had a lower quantity of HSV-2 DNA (-0.99 log(10) copies/mL; 95% CI -1.8 to -0.2 log(10) copies/mL) and were more likely to have a healed ulcer (RR 1.26; 95% CI 0.9-1.9). CONCLUSION: Episodic therapy for herpes reduced the quantity of cervicovaginal HSV-2 DNA and slightly improved ulcer healing but it did not decrease genital and plasma HIV-1 RNA loads. TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT00158483 .

OBJECTIVE: To describe the epidemiology of HIV among core groups in Togo. METHODS: We enumerated sex workers (SWs) and conducted cross-sectional surveys of SWs and their clients in 2003 in Lome and in 2005 in the whole country. RESULTS: Sex work was concentrated in Lome which comprised 15% of the population but 52% of the 5397 SWs enumerated in Togo in 2005 and 68% of the estimated 101376 men who had bought sex in the year before the 2005 survey. HIV prevalence among SWs was highest in Lome (45.4% in 2005) and progressively decreased from south to north. A similar geographical pattern was seen for clients (8.3% were HIV infected in Lome in 2005) and had already been reported for pregnant women. In Lome the population attributable fraction of prevalent cases of HIV acquired during transactional sex was estimated at 32%; in the rest of the country this was only 2%. CONCLUSIONS: This is the first study quantifying sex work at a national level in Africa. Variations in HIV prevalence within Togo with a north-south gradient among SWs their clients and pregnant women may to a large extent reflect the concentration of the sex trade within Lome. Prostitution played only a modest a role in HIV dynamics outside Lome.

The combination of high sexually transmitted infection (STI) prevalence and low HIV prevalence has been described as the Indian Ocean paradox. To investigate current epidemiology of HIV and STI in the Comoros, we conducted cross-sectional surveys of a representative sample of the adult population, and convenience samples of female sex workers and male STI patients. Only one (0.025%) of 3990 community participants was HIV-infected, while 142 (3.6%) had treponemal antibodies. Treponemal antibodies were not associated with past genital ulcers, number of sexual partners or adverse outcomes of pregnancies; their prevalence did not increase with age and there was no concordance within couples. Thus, most individuals with treponemal antibodies were probably infected during childhood with a non-venereal treponematosis. Only 1/70 (1.4%) and 0/83 sex workers sampled in 2004 and 2005 were HIV-infected. The Comoros have been protected by their insular status, male circumcision and paucity of syphilis. HIV control should focus on sex workers.

OBJECTIVE:: To investigate correlates of herpes simplex virus type 2 (HSV-2) DNA and HIV-1 RNA among women with genital ulcer disease (GUD). DESIGN:: Baseline data from a randomized placebo-controlled trial of episodic herpes treatment in Ghana and the Central African Republic. METHODS:: GUD aetiology was determined by polymerase chain reaction (PCR) from a lesional swab. Real-time PCR was used to quantify HIV-1 RNA, and HSV-2 DNA in cervicovaginal lavages (CVL) and HIV-1 RNA in plasma. Genital infection was defined as the presence of virus in the lesion or CVL. RESULTS:: Of 441 women enrolled, 79.0% were HSV-2 seropositive, 46.6% were HIV-1 seropositive, and 50.0% had an HSV-2 ulcer. Among 180 HSV-2/HIV-1 co-infected women, cervicovaginal HIV-1 RNA was detected more frequently in women with HSV-2 ulcers (67.9%) or cervicovaginal HSV-2 DNA only (72.3%) compared with women without genital HSV-2 infection (42.4%) (P = 0.004). Women with genital HSV-2 infection had higher median cervicovaginal HIV-1-RNA loads (3.14 log10 copies/mL versus 2.10 log10 copies/mL; P = 0.003), higher plasma HIV-1-RNA loads (median 5.10 versus 4.65 log10 copies/mL; P = 0.07), and lower median CD4 cell counts) (198 versus 409 cells/mm, P = 0.03). Cervicovaginal HIV-1 RNA and HSV-2 DNA were significantly correlated after adjusting for plasma HIV-1 RNA and CD4 cell counts (P < 0.001) and a 10-fold increase in cervicovaginal HSV-2 DNA was associated with a 1.7-fold increase in plasma HIV-1 RNA (P = 0.003). CONCLUSION:: Genital HSV-2 infection is associated with increased cervicovaginal and plasma HIV-1 RNA among co-infected women with genital ulcers, independently of the level of immunodeficiency, highlighting the close interaction between these two viruses and the role of HSV-2 as a co-factor for the sexual transmission of HIV-1.

Clinical and subclinical genital herpes simplex virus type 2 (HSV-2) reactivations have been associated with increases in human immunodeficiency virus (HIV)-1 genital shedding. Whether HSV-2 shedding contributes to the selection of specific genital HIV-1 variants remains unknown. We evaluated the genetic diversity of genital and blood HIV-1 RNA and DNA in 14 HIV-1/HSV-2-co-infected women, including seven with HSV-2 genital reactivation, and seven without as controls. HIV-1 DNA and HIV-1 RNA env V1–V3 sequences in paired blood and genital samples were compared. The HSV-2 selection pressure on HIV was estimated according to the number of synonymous substitutions (dS), the number of non-synonymous substitutions (dN) and the dS/dN ratio within HIV quasi-species. HIV-1 RNA levels in cervicovaginal secretions were higher in women with HSV-2 replication than in controls (p0.02). Plasma HIV-1 RNA and genital HIV-1 RNA and DNA were genetically compartmentalized. No differences in dS, dN and the dS/dN ratio were observed between the study groups for either genital HIV-1 RNA or plasma HIV-1 RNA. In contrast, dS and dN in genital HIV-1 DNA were significantly higher in patients with HSV-2 genital reactivation (p

OBJECTIVE: To evaluate whether single-dose treatments are as effective as standard therapy in the syndromic management of vaginal discharge. METHODS: A randomized controlled effectiveness trial compared single-dose tinidazole plus fluconazole (TF) with treatment for 7 days with metronidazole plus 3 days of treatment with vaginal clotrimazole (MC) among 1570 women presenting with vaginal discharge at primary health care institutions in Ghana, Guinea, Mali and Togo. Participants were randomly allocated to one of the two treatments by research nurses or physicians using precoded envelopes. Effectiveness was assessed by symptomatic response on day 14. CLINICAL IDENTIFIER ClinicalTrials.gov NCT00313131. FINDINGS: The two treatment regimens had similar effectiveness: complete resolution was seen in 66% (TF) and 64% (MC) and partial resolution in 33% (TF) and 34% (MC) of participants (P = 0.26). Effectiveness was similar among subgroups with vulvovaginal candidiasis, Trichomonas vaginalis vaginitis or bacterial vaginosis. The two treatment regimens had a similar effectiveness among human immunodeficiency virus (HIV)-infected (TF: n = 76, 71% complete resolution, 28% partial; MC: n = 83, 72% complete resolution, 25% partial, P = 0.76) and HIV-uninfected women (TF: n = 517, 68% complete, 32% partial; MC: n = 466, 65% complete, 33% partial, P = 0.20). Cervical infections with Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium were uncommon among women not involved in sex work, were associated with bacterial vaginosis or T. vaginalis vaginitis, and did not alter response to treatment with agents active against vaginal infections. Four-fifths of women not relieved by a single dose of TF had a favourable response when MC was administered as second-line treatment. CONCLUSION: Single-dose TF is as effective as multiple-dose MC in the syndromic management of vaginal discharge, even among women with HIV-infection. Given its low price and easier adherence, TF should be considered as a first-line treatment for vaginal discharge syndrome.

Objectives: To identify the contribution of Mycoplasma genitalium to the aetiology of cervicitis in sub-Saharan Africa and its relative importance in the overall burden of sexually transmitted infections among female sex workers (FSW). Methods: The study population consisted of FSW recruited in Ghana and Benin during the initial visit of a randomised controlled trial. A questionnaire was administered, a pelvic examination carried out, and cervical samples obtained for detection of M genitalium, Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis. Clinical signs potentially indicating cervicitis were cervical discharge, pus on the cervical swab, bleeding after sampling, and inflammatory cervix. Results: Among 826 FSW, 26.3% were infected with M genitalium. N gonorrhoeae was strongly and independently associated with each of the four signs of cervicitis (adjusted odds ratios (AOR): 4.1 to 6.0). The AOR for C trachomatis were intermediate (1.3–4.1) and the AOR for M genitalium were lower (between 1.6 and 1.8) but statistically significant (p⩽0.05) for each sign. Conclusions:M genitalium is weakly associated with signs of cervicitis in west African FSW but is highly prevalent.

Objectives: To measure prevalence and risk factors for cervical infections among a large sample of women consulting for vaginal discharge in west Africa and to evaluate its syndromic management through a two visit algorithm. Methods: In 11 health centres in Benin, Burkina Faso, Ghana, Guinee, and Mali 726 women who presented with a vaginal discharge without abdominal pain and who denied being a sex worker (SW) were enrolled. Cervical samples were tested for the detection of Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) with polymerase chain reaction (PCR) assays. All participants were treated with single dose (2 g) metronidazole and clotrimazole cream for 3 days. They were randomised to be told either to come back on day 7 only if there was no improvement in the discharge (group A), or to come back on day 7 regardless of response to treatment (group B). Results: Overall, the prevalence of NG and CT was only 1.9% (14/726) and 3.2% (23/726) respectively. Risk factors previously recommended by the WHO were not associated with the presence of cervical infection, with the exception of the number of sex partners in the past 3 months. When taken together, these risk factors had a positive predictive value of only 6.4% to identify cervical infections. Prevalence of cervical infection was not higher in women who came back on day 7, regardless of the strategy used. Prevalence of NG/CT was lower in Ghana and Benin (5/280, 1.8%), where comprehensive interventions for SW have been ongoing for years, than in the three other countries (27/446, 6.1%, p = 0.01). Conclusions: NG and CT infections are uncommon in west African women who consult for vaginal discharge and who are not SW. Syndromic management of vaginal discharge should focus on the proper management of vaginitis. The control of gonococcal and chlamydial infection should be redesigned around interventions focusing on sex workers.

Objective: To evaluate the impact of Mycoplasma genitalium on the outcome of pregnancy. Methods: Cervical samples from women who had previously participated in a case-control study (designed to assess the impact of syphilis and HIV-2 on the outcome of pregnancy in Guinea-Bissau) were processed using a PCR assay to detect the presence of M genitalium. Controls were women who had delivered a term neonate with a birth weight over 2500 g. Cases were classified into four groups of mothers according to the outcome of pregnancy: stillbirths, spontaneous abortions, premature deliveries, and small for gestational age (SGA) babies. Results: Among the 1014 women included in this study, 6.2% were infected with M genitalium. M genitalium infection was not significantly associated with any of the adverse outcomes of pregnancy studied. Odds ratios (OR) for premature or SGA delivery in the presence of M genitalium infection were 1.37 (95% CI 0.69 to 2.60) and 0.44 (95% CI 0.01 to 2.75), respectively. For abortions and stillbirths, OR were respectively 0.61 (95% CI 0.07 to 2.51) and 1.07 (95% CI 0.42 to 2.42). Conclusion: M genitalium appears not to have a deleterious impact on the outcome of pregnancy.

Objectives: Male clients of female sex workers have rarely been specific targets for HIV/sexually transmitted diseases (STD) interventions in sub-Saharan Africa. We assessed the effectiveness of outreach methodology for contacting sexual partners of female sex workers for purposes of HIV/STD prevention in Cotonou, Benin. Design and methods: In collaboration with owners/managers, outreach personnel and female sex workers, 404 clients were recruited on-site at prostitution venues, and provided urine samples for leukocyte esterase dipstick (LED), STD and HIV testing before having sex with female sex workers. After having sex they underwent an interview and physical examination. No payment was made for study participation. Prostitution site personnel (n = 41) and boyfriends of female sex workers (n = 56) were also recruited. Results: In all 68% of the clients approached agreed to participate. On-site LED testing and free STD treatment were important factors in participation. HIV-1 prevalence was several-fold higher than in the general population in Cotonou, at 8.4, 12.2 and 16.1% in clients, personnel and boyfriends respectively, and was associated with increasing age and lack of condom use with female sex workers. Condom use rates by clients with female sex workers were non-negligible but sub-optimal, and low with regular partners. Approximately one-third of clients with regular partners also had other non-female sex worker sex partners. Boyfriends of female sex workers are of particular concern due to high numbers of partners, very low condom use rates and high HIV prevalence. Conclusions: Study findings indicate that male sex partners of female sex workers form a 'bridging population' for HIV/STD transmission both to female sex workers, as well as from female sex workers to the general population of women, particularly regular female partners.

Background The spectrum of bacteria associated with bacterial vaginosis (BV) has recently expanded through taxonomic changes and the use of molecular methods. These methods have yet to be used in large-scale epidemiological studies in Africa where BV is highly prevalent. Methods An analysis of samples obtained during a clinical trial of the management of vaginal discharge in four West African countries. Samples were available from 1555 participants; 843 (54%) had BV. Nucleic acids of 13 bacterial genera or species potentially associated with BV were detected through the polymerase chain reaction. Results The associations between various components of the vaginal flora were complex. Excluding Lactobacillus, the other 12 micro-organisms were all associated with each other at the p≤0.001 level. The prevalence of various bacterial genera or species varied according to age, sexual activity and HIV status. In multivariate analysis, the presence of Gardnerella vaginalis, Bifidobacterium, Megasphaera elsdenii, Dialister, Mycoplasma hominis, Leptotrichia, and Prevotella were independently associated with BV as was the absence of Lactobacillus and Peptoniphilus. However, Mobiluncus, Atopobium vaginae, Anaerococcus, and Eggerthella were not independently associated with BV. Unexpectedly, after treatment with a regimen that included either metronidazole or tinidazole, the proportion of patients with a complete resolution of symptoms by day 14 increased with the number of bacterial genera or species present at enrolment. Conclusions Numerous bacterial genera or species were strongly associated with each other in a pattern that suggested a symbiotic relationship. BV cases with a simpler flora were less likely to respond to treatment. Overall, the vaginal flora of West African women with BV was reminiscent of that of their counterparts in industrialized countries.

Chlamydia trachomatis serovar was determined by analysis of restriction fragment length polymorphism in the gene encoding the major outer membrane protein (MOMP) from 435 urogenital specimens. Of the specimens, 254 grew

OBJECTIVES--A method based on a nested polymerase chain reaction (PCR) was developed to detect and to type Chlamydia trachomatis from low titre samples by amplifying a large portion of the major outer membrane protein gene. The sensitivity of this procedure was evaluated in urogenital clinical samples in comparison with culture. SPECIMENS--A series of 787 urogenital specimens, including 37 (4.7%) positive by culture, together with 227 other samples that had been found to yield less than 25 chlamydial inclusions in culture were tested. METHODS--Samples were pelleted, resuspended in 1 mM NaOH, heated and amplified without further purification. After 40 cycles of PCR, 1 microliters of product was amplified by a further 30 cycles of PCR using a second set of primers nested within the initial pair. Positives were detected by agarose gel electrophoresis and confirmed by repeating the PCR analyses and determining the serovar of both amplified samples by restriction fragment length polymorphism. RESULTS--Nested PCR allowed detection of 96% and culture 77% of positives with only three samples repeatedly positive by PCR but considered false positives because a different serovar was identified in the two amplifications. Of culture-positive samples with less than 11 chlamydia inclusion-forming-units 97% could be detected by nested PCR and most still gave a positive signal when diluted hundred fold. CONCLUSIONS--Nested PCR provided the basis for a very sensitive C trachomatis detection and typing strategy. Repetition and typing positive samples facilitated detection of false-positive PCR specimens resulting from contamination of the PCR process or any reagent except the original sample.

Background Sub-Saharan Africa is the continent with the highest prevalence of Hepatitis C virus (HCV) infection. Genotype 2 HCV is thought to have originated from West Africa several hundred years ago. Mechanisms of transmission remain poorly understood. Methodology/Principal Findings To delineate mechanisms for HCV transmission in West Africa, we conducted a cross-sectional survey of individuals aged ≥50 years in Bissau, Guinea-Bissau. Dried blood spots were obtained for HCV serology and PCR amplification. Prevalence of HCV was 4.4% (47/1066) among women and 5.0% (27/544) among men. In multivariate analysis, the independent risk factors for HCV infection were age (baseline: 50–59 y; 60–69 y, adjusted odds ratio [AOR]: 1.67, 95% CI: 0.91–3.06; ≥70 y, AOR: 3.47, 95% CI: 1.89–6.39), belonging to the Papel, Mancanha, Balanta or Mandjako ethnic groups (AOR: 2.45, 95% CI:1.32–4.53), originating from the Biombo, Cacheu or Oio regions north of Bissau (AOR: 4.16, 95% CI: 1.18–14.73) and having bought or sold sexual services (AOR: 3.60, 95% CI: 1.88–6.89). Of 57 isolates that could be genotyped, 56 were genotype 2. Conclusions Our results suggest that transmission of HCV genotype 2 in West Africa occurs through sexual intercourse. In specific locations and subpopulations, medical interventions may have amplified transmission parenterally.

Infection with a simian retrovirus (STLV-I) closely related to human T-lymphotropic virus type I (HTLV-I) was investigated in non-human primates living in their native countries in Africa and Asia. Serum antibodies cross-reacting with HTLV-I antigens were detected in 85 of 567 non-human primates of 30 species. Seropositive animals were found among African green monkeys, olive baboons, Sykes' monkeys, mandrills and patas monkeys in several countries in Africa, and cynomolgus monkeys, Celebes macaques and siamangs in Indonesia. The frequency of seropositivity was much higher in adult than in young African green monkeys, cynomolgus monkeys and Celebes macaques. STLV-Is were isolated by establishing II lines of virus-producing lymphoid cells in the presence of interleukin-2 from 5 species of seropositive non-human primates, i.e. the African green monkey, Sykes' monkey, Celebes macaque, cynomolgus monkey and siamang. All these cell lines had T-cell markers and Tac antigen, and the cell lines from the African green monkey and Sykes' monkeys were Leu2a+ while those from other species were Leu3a+. These cell lines expressed viral antigens reacting with human sera from adult T-cell leukemia (ATL) patients and monoclonal antibodies (MAbs) against p19 and p24 of HTLV-I core proteins, and produced virus particles having RNA-dependent DNA polymerase activity. Cellular DNAs from these cell lines contained provirus sequences homologous to HTLV-I, shown by Southern blot hybridization. The restriction patterns of these provirus genomes were different from those of HTLV-I and were also dissimilar in the different species.

The auxotypes and serogroups of 250 consecutive isolates of Neisseria gonorrhoeae from Franceville, Gabon, including 33 penicillinase producing (PPNG) strains, were identified and the results tabulated for four periods. The PPNG and certain non-PPNG strains were isolated in clusters against a relatively stable pattern of non-PPNG biotypes. Most of the non-PPNG strains were non-requiring or proline requiring strains of serogroup WII or non-requiring strains that could not be grouped with commercially available antisera. No strain requiring arginine, hypoxanthine, and uracil was observed. The PPNG strains were all serogroup WI or ungroupable and non-requiring or proline or arginine dependent auxotypes. Only one non-PPNG strain required proline or arginine and was not serogroup WII.

Samples from 218 men with urethritis, 517 women with pelvic pain or pelvic pain and vaginal discharge, 218 women consulting for infertility, and 598 postpartal women were screened for Chlamydia trachomatis by culture and direct immunofluorescence. Chlamydiae were detected in 18% (39/218) of the men, 18% (45/252) of women with vaginal discharge, 14% (38/265) of those with pelvic pain, 10% (21/218) of infertile, and 10% (59/598) of postpartal women. A chlamydial prevalence of 18% (41) was observed in 229 postpartal women aged under 21, whereas only 5% (10) of 360 postpartal women over 21 had C trachomatis. In the other clinical groups, an age related decrease in prevalence was noted in women over 25. The direct immunofluorescence test correlated well with culture. The small difference in isolation between symptomatic and postpartal women indicates that women in this population do not seek medical attention for chlamydial infections and expose themselves to chlamydial salpingitis and infertility.Health workers took urethral samplers from 218 men with urethritis and 1850 women to screen for Chlamydia trachomatis using culture and immunofluorescence. All the men and women presented themselves at the Hospital Provincial de Franceville in semirural Gabon. 18% of men, 18% of women with vaginal discharge, 14% of women with pelvic pain, 10% of infertile women, and 10% of postpartum women tested positive for C. trachomatis infection. Postpartum women less than 21 years old had a significantly higher chlamydial prevalence than did older postpartum women (18 vs. 5%; p .05). Chlamydial prevalence among 21-25 year old postpartum women was significantly lower than among same age women presenting with other conditions (5% vs. 15% for infertile women, 21% for women with pelvic pain, and 2% for women with vaginal discharge; p .01). Except for postpartum women, older than 25 chlamydia prevalence decreased after age 25. The sensitivity of immunofluorescence ranged from 77% for infertile women to 93% for postpartum women. Its specificity ranged from 97% for women with discharge or pelvic pain to 100% for men. Thus, the direct immunofluorescence test corresponded well with culture. The isolation rates for asymptomatic women neared those of symptomatic women, indicating that most women with C. trachomatis infections have no or very mild symptoms and do no seek medical attention. This most likely jeopardizes their fertility.

Two isolates of simian retrovirus related to the human immunodeficiency virus (HIV) were obtained from apparently healthy mandrills, Papio (Mandrillus) sphinx, in western equatorial Africa. This virus, designated SIVMND (simian immunodeficiency virus from mandrills), appeared morphologically similar to HIV by electron microscopy, showed Mg2+-dependent reverse transcriptase activity, and induced cytopathic effect in human CD4-positive cells. Western blotting (immunoblotting) analyses revealed that the gag and pol products of SIVMND showed cross-reactivity with those of known HIVs and SIVs. Molecular clones covering full-length viral DNA were obtained from closed circular extrachromosomal DNA of SIVMND-infected cells. By clone-on-clone hybridization with known retroviruses of the HIV and SIV groups, SIVMND showed similar cross-hybridization with HIV-1, HIV-2, SIVAGM (African green monkey-derived SIV), and SIVMAC (rhesus macaque-derived SIV) in the gag and pol regions only at low stringency but not at high stringency, a result indicating that SIVMND is a new member of the HIV-SIV group. The existence of distinct SIVs in different monkey species suggest that recent interspecies transfer of HIV-SIV is unlikely in nature.

Placentas from 191 consecutive deliveries were cultured for Neisseria gonorrhoeae and other bacteria before being examined histologically. N gonorrhoeae was seen on direct microscopical examination and cultured from two placentas, one of which had chorioamnionitis and the other had a granulocytic invasion of the membranes. In both patients, rupture of the membranes had occurred two hours or less before delivery, and thus after infection. A third patient was also found to have light gonococcal infection of the placenta. Six infections with group B streptococci were identified, but only one correlated with chorioamnionitis. Chorioamnionitis was observed in 16 (8.4%) and granulocytic infiltration of the membranes in 18 (9.4%) of the deliveries.

Determinants of infertility were studied in 340 women in Eastern Gabon, an area situated in the "infertility belt" of Central Africa. Fallopian tube occlusion was diagnosed in 82.8% of cases, showing the importance of infection-related causes. Women with tubal occlusion did not differ significantly from women with normal tubes in obstetrical history or prevalence of Neisseria gonorrhoeae or Chlamydia trachomatis on endocervical culture. Antecedents of pelvic inflammatory disease or a pelvic mass were significantly more common in the group with tubal occlusion. This group also had a significantly higher prevalence of serum chlamydial antibodies at a titer of 1/64 or higher. Hormonal factors were found in 31.7% of women, a cervical factor in 29.0% and mechanical factors in 5.6%. No diagnosis could be made in 12.2% of cases. During the investigation, 4.4% of women became pregnant. The predominance of infectious related causes of infertility makes it imperative to focus resources on prevention programs of upper genital tract infections in women.Determinants of infertility were studied in 340 women in Eastern Gabon, an area situated in the "infertility belt" of Central Africa. Fallopian tube occlusion was diagnosed in 82.8% of cases, showing the importance of infection-related causes. Women with tubal occlusion did not differ significantly from women with normal tubes in obstetrical history or prevalence of Neisseria gonorrhea or Chlamydia trachomatis on endocervical culture. Antecedents of pelvic inflammatory disease or a pelvic mass were significantly more common in the group with tubal occlusion. This group also had a significantly higher prevalence of serum chlamydial antibodies at a titer of 1/64 or higher. Hormonal factors were found in 31.7% of women, a cervical factor in 29.0% and mechanical factors in 5.6%. No diagnosis could be made in 12.2% of cases. During the investigation, 4.4% of women became pregnant. The predominance of infectious related causes of infertility makes it imperative to focus resources on prevention programs of upper genital tract infections in women. The study sample is from a small semi-urban center of 25,000 inhabitants and consisted of all women consulting for infertility at the gynecology department of the Franceville hospital from January 1983 until December 1984.

The incidence and aetiology of ophthalmia neonatorum have been estimated over a 7-month period in Franceville, a semi-rural community in south-eastern Gabon. Chlamydia trachomatis was the most frequently observed pathogen, being isolated from 17 babies (2.7% of births), and Neisseria gonorrhoeae was recovered from 12 (1.6% of births). 5 of 17 cases of chlamydial conjunctivitis were in infants less than 5 d old as opposed to 9 in the typical 5 to 10-days-old group. As expected, most cases of gonococcal ophthalmia neonatorum occurred in the first 5 d of life with cases in older infants often not accompanied by a granulocytic response. Chlamydial conjunctivitis was usually unilateral whereas other cases were most frequently bilateral.

Objective Description of the epidemiology of HIV infection among sex workers (SW) in Accra, Ghana. Methods In all, 1013 SW working out of their homes ( seaters ) or finding customers in bars, hotels, brothels or on the street ( roamers ) were interviewed and tested for HIV. Results Overall, prevalence of HIV infection was nearly 50% (506 of 1013), varying from 26% (133 of 507) among the roamers to 74% (368 of 496) among the seaters. Profound differences were noted between these two categories of SW with regard to age, number of clients per day, price per instance of intercourse, condom use, and other characteristics. Respectively, 27% and 58% of roamers and seaters were infected with HIV within their first 6 months of sex work, despite a limited number of unprotected sex acts with seropositive clients. Independent risk factors for HIV infection varied between types of SW: age among the roamers; region of origin and duration of sex work among the seaters; number of clients per day, and presence of current or past genital ulcer and gonococcal cervicitis in both groups. Conclusion In Accra, considerable heterogeneity exists in the population of SWs. In both categories of SW, new recruits become rapidly infected with HIV after entering the trade. The 25-fold higher prevalence of HIV among SWs than in the general adult population suggests that in Accra, as in many cities of West Africa, a high fraction of new cases of HIV infection continue to be acquired from SWs. Intervention programs targeting SW should be an essential component of national AIDS control strategies. Special efforts should be made to identify and offer preventive services to new sex workers.

Of 35 women with acute salpingitis on laparoscopy, 86% had chlamydial antibodies at a titre of 1/16 or higher and 49% at a titre of 1/512 or higher. Geometric mean titres of chlamydial antibodies correlated significantly with a laparoscopic observation of chronic salpingitis (p less than 0.001), with a history of infertility (p less than 0.05), and with severe inflammation (p less than 0.10), but not with a history of salpingitis or a positive cervical culture for Chlamydia trachomatis. In the subgroup of infertile women, the geometric mean titre of antibodies to chlamydiae correlated significantly with the presence of chronic salpingitis (p less than 0.005). These data indicate that chlamydial infections play a major part in salpingitis and infertility in central Africa. In this area, which is known as the "infertility belt", programmes to control chlamydial infections should be implemented.