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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.

Using 42 isolates contributed by laboratories in Canada, The Netherlands, the United Kingdom, and the United States, we compared the results of analyses done with seven Clostridium difficile typing techniques: multilocus variable-number tandem-repeat analysis (MLVA), amplified fragment length polymorphism (AFLP), surface layer protein A gene sequence typing ( slpA ST), PCR-ribotyping, restriction endonuclease analysis (REA), multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). We assessed the discriminating ability and typeability of each technique as well as the agreement among techniques in grouping isolates by allele profile A (AP-A) through AP-F, which are defined by toxinotype, the presence of the binary toxin gene, and deletion in the tcdC gene. We found that all isolates were typeable by all techniques and that discrimination index scores for the techniques tested ranged from 0.964 to 0.631 in the following order: MLVA, REA, PFGE, slpA ST, PCR-ribotyping, MLST, and AFLP. All the techniques were able to distinguish the current epidemic strain of C. difficile (BI/027/NAP1) from other strains. All of the techniques showed multiple types for AP-A (toxinotype 0, binary toxin negative, and no tcdC gene deletion). REA, slpA ST, MLST, and PCR-ribotyping all included AP-B (toxinotype III, binary toxin positive, and an 18-bp deletion in tcdC ) in a single group that excluded other APs. PFGE, AFLP, and MLVA grouped two, one, and two different non-AP-B isolates, respectively, with their AP-B isolates. All techniques appear to be capable of detecting outbreak strains, but only REA and MLVA showed sufficient discrimination to distinguish strains from different outbreaks.

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)

To reliably immobilize different biomoieties on surfaces of III-V semiconductors is one of the most critical issues in the development of biodetector devices based on the optical/electronic properties of these materials. Herein we demonstrate the successful immobilization of avidin, a robust and well-studied protein, on a (001) GaAs surface. The immobilization was investigated via specific binding to biotin, which was connected to the GaAs surface through commercially available long- or short-chain amino group terminated alkanethiols (HS(CH2)11NH2 or HS(CH2)2NH2), or through a biotinylated thiol synthesized in our laboratory. The immobilization performance was evaluated by photoluminescence and fluorescence microscopy measurements. We found that the biotinylated thiol mixed with a diluent thiol provides the highest avidin immobilization efficiency.

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.

Summary Background Toxins A and B are the primary virulence factors of Clostridium difficile . Since 2002, an epidemic of C difficile -associated disease with increased morbidity and mortality has been present in Quebec province, Canada. We characterised the dominant strain of this epidemic to determine whether it produces higher amounts of toxins A and B than those produced by non-epidemic strains. Methods We obtained isolates from 124 patients from Centre Hospitalier Universitaire de Sherbrooke in Quebec. Additional isolates from the USA, Canada, and the UK were included to increase the genetic diversity of the toxinotypes tested. Isolate characterisation included toxinotyping, pulsed-field gel electrophoresis (PFGE), PCR ribotyping, detection of a binary toxin gene, and detection of deletions in a putative negative regulator for toxins A and B ( tcdC ). By use of an enzyme-linked immunoassay, we measured the in-vitro production of toxins A and B by epidemic strain and non-dominant strain isolates. Findings The epidemic strain was characterised as toxinotype III, North American PFGE type 1, and PCR-ribotype 027 (NAP1/027). This strain carried the binary toxin gene cdtB and an 18-bp deletion in tcdC . We isolated this strain from 72 patients with C difficile -associated disease (58 [67%] of 86 with health-care-associated disease; 14 [37%] of 38 with community-acquired disease). Peak median (IQR) toxin A and toxin B concentrations produced in vitro by NAP1/027 were 16 and 23 times higher, respectively, than those measured in isolates representing 12 different PFGE types, known as toxinotype 0 (toxin A, median 848 μg/L [IQR 504–1022] vs 54 μg/L [23–203]; toxin B, 180 μg/L [137–210] vs 8 μg/L [5–25]; p Interpretation The severity of C difficile -associated disease caused by NAP1/027 could result from hyperproduction of toxins A and B. Dissemination of this strain in North America and Europe could lead to important changes in the epidemiology of C difficile -associated disease.

OBJECTIVE: To delineate the population attributable fraction (PAF) of transactional sex in prevalent cases of HIV infection in the male adult population of Accra, Ghana. DESIGN AND METHODS: Cross-sectional study of clients who visited a sex worker (SW), of boyfriends of SW and of male personnel in prostitution venues. A questionnaire was administered and urine obtained for detection of anti-HIV antibodies. The PAF of prevalent HIV acquired from SW was calculated using a combination of data from this survey of clients, from on-going SW surveys, the national HIV surveillance system and the national census. RESULTS: HIV prevalence was 4.9% (8/162) among clients of mobile SW, 15.8% (53/335) among clients of home-based SW, 17.5% (10/57) among personnel and 32.1% (9/28) among boyfriends. A condom was used in 90% of intercourses, according to clients. Non-use of a condom was clustered in selected locations and independently associated with older age of client, frequency of intercourse with SW in the last year and current urethritis. Among the male population of Accra aged 15-59 years, 84% of prevalent cases of HIV were attributable to transactional sex. A sensitivity analysis showed that under various assumptions PAF varied between 47% and 100%. CONCLUSIONS: In Accra, approximately four-fifths of prevalent cases of HIV in adult males were acquired from SW. Comprehensive interventions providing education, condoms and treatment for sexually transmitted diseases for SW and their clients should be approached as other public health priorities and provided in all cities, large and small, of West Africa.

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.

The diagnostic performances of the clinical case definition of influenza virus infection based on the combination of fever and cough and of two rapid influenza diagnostic tests, the Directigen Flu A+B test (Directigen; BD Diagnostic Systems, Sparks, Md.) and the QuickVue influenza test (QuickVue; Quidel, San Diego, Calif.), were compared to those of viral culture and an in-house reverse transcription (RT)-PCR during the 2000-2001 flu season. Two hundred consecutive nasopharyngeal aspirates were analyzed from 192 patients, including 122 adults and 70 children. Viral culture identified influenza virus A in 16 samples and influenza virus B in 55 samples, whereas RT-PCR identified influenza virus A in 21 samples and influenza virus B in 64 samples. When RT-PCR was used as the reference standard, the likelihood ratios for a positive test were 40.0 for Directigen, 8.6 for QuickVue, and 1.4 for the combination of fever and cough, whereas the likelihood ratios for a negative test were 0.22, 0.16, and 0.48, respectively. Our study suggests that (i) the poor specificity (35 to 58%) and the poor positive predictive value (41 to 60%) of the clinical case definition of influenza preclude its use for prediction of influenza virus infections during epidemics, especially when infection control decision making in the hospital setting is considered; (ii) Directigen has a higher diagnostic yield than QuickVue but is associated with a larger number of invalid results; (iii) the sensitivities of the rapid diagnostic tests are significantly lower with samples from adults than with samples from children, with the rates of false-negative results reaching up to 29%; and (iv) RT-PCR detects more cases of influenza than viral culture, and this greater accuracy makes it a more useful reference standard.

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.

Alzheimer disease (AD) is a progressive irreversible neurological brain disorder. 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 and neutrophils (PMN) 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 and PMN 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) for NK and CD14 (increased in MCI). 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. Similarly, PMN phagocytosis and free radical production were differentially modulated in MCI and mAD patients. 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.

An outbreak of Ebola virus disease (EVD) began in Guinea in December 2013, and by 21 November 2014, 15,531 cases had been identified with 5,429 associated deaths primarily in Guinea, Liberia, and Sierra Leone [1, 2]. Case fatality rates during the current outbreak are around 70 % among patients with known outcome [1]. As treatment options are limited and largely supportive, preventative measures are paramount in the control of this disease. Rapid and reliable diagnosis of EVD is critically important for patient management, infection prevention, and control measures, and optimization of use of healthcare resources. This article briefly reviews new developments in the diagnosis of EVD.

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

Can populations self organize a crisis response? This is a field report on the first two efforts in a continuing series of exercises termed “Exercise24 or x24.” The first Exercise 24 focused on Southern California, while the second (24 Europe) focused on the Balkan area of Eastern Europe. These exercises attempted to demonstrate that self-organizing groups can form and respond to a crisis using low-cost social media and other emerging web technologies. Over 10,000 people participated in X24 while X24 Europe had over 49,000 participants. X24 involved people from 79 nations while X24 Europe officially included participants from at least 92 countries. Exercise24 was organized by a team of workers centered at the SDSU Viz Center including significant support from the US Navy as well as other military and Federal organizations. Dr. George Bressler, Adjunct Faculty member at the Viz Center led both efforts. Major efforts from senior professionals EUCOM and NORTHCOM contributed significantly to the preparation for and success of both X24 and especially X24 Europe. This paper presents lessons learned and other experiences gained through the coordination and performance of Exercise24.

A reverse transcriptase and polymerase chain reaction (RT-PCR)-based assay for G-typing of bovine rotaviruses (BRV) was compared with a monoclonal antibody-based immunoassay (MAbs-ELISA) in the characterization of BRV field strains obtained from calves in different regions of Quebec between 1992 and 1994. The strains were analysed for two G types (G6 and G10) which are the most predominate BRV field strains. Fecal samples positive for BRV by polyacrylamide gel electrophoresis (n = 74) were typed by both methods revealing 77% correlation. RT-PCR detected 10 more G6 and 2 more G10 serotypes than MAbs-ELISA. Nine of the 12 discrepant samples could be cultivated and were confirmed as G6 (8) or G10 (1) by both methods. RT-PCR was able to efficiently detect artificial mixes of G6 and G10 and detected two mixed field infections. Four additional infections considered as mixed by MAbs-ELISA and as only G6 by RT-PCR were possibly MAbs-ELISA cross-reactions. RT-PCR provided a very sensitive method for typing BRV field isolates.

The events of September 11, 2001, the Indian Ocean tsunami in 2004, and Hurricane Katrina in 2005 awakened American policymakers to the importance of the need for emergency management. This paper explains how a cloud computing environment can support social networks and logistical coordination on a global scale during crises. Basic cloud computing functionality is covered to show how social networks can connect seamlessly to work together with profound interoperability. Lastly, the benefits of a cloud computing solution is presented as the most cost-effective, efficient, and secure method of communication during a disaster response, with the unique capability of being able to support a global community through its massive scalability.

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.

The authors have investigated the effect of chemotaxis on immobilization of bacteria on the surface of biofunctionalized GaAs (001) samples. Escherichia coli K12 bacteria were employed to provide a proof-of-concept of chemotaxis-enhanced bacterial immobilization, and then, these results were confirmed using Legionella pneumophila. The recognition layer was based on a self-assembled monolayer of thiol functionalized with specific antibodies directed toward E. coli or L. pneumophila, together with the enzyme beta-galactosidase (β-gal). The authors hypothesized that this enzyme together with its substrate lactose would produce a gradient of glucose which would attract bacteria toward the biochip surface. The chemotaxis effect was monitored by comparing the number of bacteria bound to the biochip surface with and without attractant. The authors have observed that β-gal plus lactose enhanced the immobilization of bacteria on our biochips with a higher effect at low bacterial concentrations. At 100 and 10 bacteria/ml, respectively, for E. coli and L. pneumophila, the authors observed up to 11 and 8 times more bacteria bound to biochip surfaces assisted with the chemotaxis effect in comparison to biochips without chemotaxis. At 10(4) bacteria/ml, the immobilization enhancement rate did not exceed two times.

Staphylococcus aureus small-colony variants (SCVs) are persistent pathogenic bacteria characterized by slow growth and, for many of these strains, an increased ability to form biofilms and to persist within host cells. The virulence-associated gene expression profile of SCVs clearly differs from that of prototypical strains and is often influenced by SigB rather than by the agr system. One objective of this work was to confirm the role of SigB in the control of the expression of virulence factors involved in biofilm formation and intracellular persistence of SCVs. This study shows that extracellular proteins are involved in the formation of biofilm by three SCV strains, which, additionally, have a low biofilm-dispersing activity. It was determined that SigB activity modulates biofilm formation by strain SCV CF07-S and is dominant over that of the agr system without being solely responsible for the repression of proteolytic activity. On the other hand, the expression of fnbA and the control of nuclease activity contributed to the SigB-dependent formation of biofilm of this SCV strain. SigB was also required for the replication of CF07-S within epithelial cells and may be involved in the colonization of lungs by SCVs in a mouse infection model. This study methodically investigated SigB activity and associated mechanisms in the various aspects of SCV pathogenesis. Results confirm that SigB activity importantly influences the production of virulence factors, biofilm formation and intracellular persistence for some clinical SCV strains.

After the 2007 Southern California wildfire events, event-assessment of the efficacy of spreadsheets and paper forms raised the question of whether alternative tools could have achieved greater efficiencies in the logistical support of command centers, the sites from which the local utility’s electric restoration personnel were deployed. In this paper, the authors examine what approach would have enabled personnel working on the logistics of the command center effort to have easier-to-use, faster-to-access, command center data stored in, and provided via, a catastrophe resilient platform other than the traditional company computer network. Additionally, the capability to store basic command center requirements from previous emergency responses, thereby saving time during the next emergency, was examined.

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.

Emergency management is a diverse field. Effective disaster management involves knowledge of various subjects as well as work experience in all aspects related to mitigation, planning, response, and recovery efforts. One field not being fully exploited by disaster management is the use of geospatial tools in the form of Geographic Information Systems (GIS), cartography, and geovisualization. One reason for this is that many emergency managers are not fully aware of the assistance GIS can lend to effectively manage disaster situations. All functions of emergency management have a strong geographic component. Where is the earthquake epicenter? Where is the damage? Where does the dam inundation run and who/what is in that path? Where is the area of road closures? The questions asking “where” are endless in effective emergency management and range from the mitigation stage through to the recovery stage. For example, a tsunami may inundate only a certain portion of the region, therefore, it is important to have mitigation and planning efforts concentrated in those regions. It is also important to know what businesses, housing, and populations are in the affected areas. The integration of geospatial tools for risk assessment, mitigation, planning, response, and recovery efforts is emerging as an effective and potentially invaluable resource for answering such questions in regards to emergency management.

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.

International audience; BACKGROUND: The simultaneous emergence of human immunodeficiency virus (HIV)-1 group M and HIV-2 into human populations, circa 1921-1940, is attributed to urbanization and changes in sexual behavior. We hypothesized that the initial dissemination of HIV-1, before sexual transmission predominated, was facilitated by the administration, via reusable syringes and needles, of parenteral drugs against tropical diseases. As proxies for highly lethal HIV-1, we investigated risk factors for hepatitis C virus (HCV) and human T cell lymphotropic virus 1 (HTLV-1) infections, blood-borne viruses compatible with prolonged survival, in an area known in 1936-1950 as the most virulent focus of African trypanosomiasis. METHODS: Cross-sectional survey of individuals 55 years and older in Mbimou land and Nola, Central African Republic. Dried blood spots were used for HCV and HTLV-1 serologic testing and nucleic acid detection. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were measured by logistic regression. RESULTS: The only risk factor for HCV genotype 4 infection was treatment of trypanosomiasis before 1951 (OR, 3.13; 95% CI, 1.38-7.09). HTLV-1 infection was associated with having received 2 injections of pentamidine for trypanosomiasis chemoprophylaxis (adjusted OR, 2.03; 95% CI, 1.01-4.06) and with transfusions (adjusted OR, 2.82; 95% CI, 1.04-7.67). From historical data, we predicted that 59% of Mbimous 65 years and older would report treatment for trypanosomiasis before 1951; only 11% did so. CONCLUSIONS: Treatment of trypanosomiasis before 1951 may have caused iatrogenic HCV transmission. Population-wide half-yearly intramuscular pentamidine for trypanosomiasis chemoprophylaxis in 1947-1953 may have caused iatrogenic HTLV-1 transmission. These and other interventions against tropical diseases could have iatrogenically transmitted SIV(cpz), jump-starting the HIV-1 epidemic. The excess mortality among patients with trypanosomiasis treated before 1951 supports this hypothesis.

In the quest for the development of an all-optical biosensor for rapid detection and typing of viral pathogens, we investigate bio- sensing architectures that take advantage of strong photolumines- cence emission from III-V quantum semiconductors QS. One of the key elements in the development of such a biosensor is the ability to attach various analytes to GaAs—a material of choice for capping III-V QS of our interest. We report on the study of biofunctionalization of GaAs 001 with polyethylene-glycol PEG thiols and the success- ful immobilization of influenza A virus. A diluted solution of biotiny- lated PEG thiols in OH-terminated PEG thiols is used to form a net- work of sites for the attachment of neutravidin. Biotinylated polyclonal influenza A antibodies are applied to investigate the pro- cess of the immobilization of inactivated influenza A virus. The suc- cessful immobilization is demonstrated using atomic force micros- copy and fluorescence microscopy measurements. © 2009 Society of Photo

The molecular epidemiology of hepatitis C virus (HCV) in the Central African Republic (CAR) is poorly documented. Thus, we conducted phylogenetic analyses of NS5B gene sequences from 58 HCV-infected inhabitants of a remote area of south-west CAR, which indicated that 48 (82.8 %) were infected with genotype 4 (HCV-4), five (8.6 %) with genotype 2 and five (8.6 %) with genotype 1. HCV-4 strains were highly heterogeneous, containing previously described subtypes 4k (48 %), 4c (27 %), 4r (4 %), 4f (4 %) and unclassified subtypes (17 %). To estimate the epidemic history of these HCV-4 strains, an evolutionary analysis using the coalescent approach was used. The estimated date of the most recent common ancestor of the CAR HCV-4 strains was 1539 (95 % confidence intervals, 1317–1697). They exhibited a rapid, exponential spread from 1935 to 1965, simultaneously with what was recently reported in neighbouring Cameroon and Gabon. The hypothesis of a massive iatrogenic transmission during interventions for the control of endemic tropical diseases is discussed.

This study aimed to assess the importance of quantitatively detecting Campylobacter spp. in environmental surface water. The prevalence and the quantity of Campylobacter spp., thermotolerant coliforms, and Escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the Eastern Townships, Québec, Canada, were determined. Overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for Campylobacter spp., thermotolerant coliforms, and E. coli , respectively. There were weak correlations between the weekly distributions of Campylobacter spp. and thermotolerant coliforms (Spearman's ρ coefficient = 0.27; P = 0.008) and between the quantitative levels of the two classes of organisms (Kendall tau-b correlation coefficient = 0.233; P < 0.0001). Well water samples from the Eastern Townships were also tested. Five (10%) of 53 samples from private surface wells were positive for Campylobacter jejuni , of which only 2 were positive for thermotolerant coliforms. These findings suggest that microbial monitoring of raw water by using only fecal indicator organisms is not sufficient for assessing the occurrence or the load of thermophilic Campylobacter spp. Insights into the role of environmental water as sources for sporadic Campylobacter infection will require genus-specific monitoring techniques.

Background Staphylococcus aureus and Pseudomonas aeruginosa are often found together in the airways of cystic fibrosis (CF) patients. It was previously shown that the P. aeruginosa exoproduct 4-hydroxy-2-heptylquinoline-N- oxide (HQNO) suppresses the growth of S. aureus and provokes the emergence of small-colony variants (SCVs). The presence of S. aureus SCVs as well as biofilms have both been associated with chronic infections in CF. Results We demonstrated that HQNO stimulates S. aureus to form a biofilm in association with the formation of SCVs. The emergence of SCVs and biofilm production under HQNO exposure was shown to be dependent on the activity of the stress- and colonization-related alternative sigma factor B (SigB). Analysis of gene expression revealed that exposure of a prototypical S. aureus strain to HQNO activates SigB, which was leading to an increase in the expression of the fibronectin-binding protein A and the biofilm-associated sarA genes. Conversely, the quorum sensing accessory gene regulator (agr) system and the α-hemolysin gene were repressed by HQNO. Experiments using culture supernatants from P. aeruginosa PAO1 and a double chamber co-culture model confirmed that P. aeruginosa stimulates biofilm formation and activates SigB in a S. aureus strain isolated from a CF patient. Furthermore, the supernatant from P. aeruginosa mutants unable to produce HQNO induced the production of biofilms by S. aureus to a lesser extent than the wild-type strain only in a S. aureus SigB-functional background. Conclusions These results suggest that S. aureus responds to HQNO from P. aeruginosa by forming SCVs and biofilms through SigB activation, a phenomenon that may contribute to the establishment of chronic infections in CF patients.

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 .

Understanding the origin and nature of hepatitis C virus (HCV) genetic diversity is critical for improving treatment and vaccine design, and such diversity is the sole source of information about the virus' epidemic history prior to its identification 20 years ago. In this paper, we study the molecular epidemiology of HCV genotype 2 in its region of endemic origin, west and central Africa. Our analysis includes 56 new and highly diverse HCV isolates sampled from infected individuals in Guinea-Bissau. By combining phylogenetic, geographical and epidemiological information, we find a previously unappreciated geographical structure in the diversity of HCV genotype 2, pointing to a history of eastwards spatial spread from the west African coast to Cameroon that took place over several centuries. Molecular clock analysis dates the common ancestor of HCV in Guinea-Bissau to 1470 (1414-1582). The phylogenetic position of isolates from Madagascar and Martinique suggests a role for the historical slave trade in the global dissemination of HCV and of the epidemic subtypes 2a and 2c. Coalescent-based estimates of epidemic growth indicate a rapid 20th-century spread of HCV genotype 2 in Cameroon that is absent in Guinea-Bissau. We discuss this contrast in the context of possible parenteral HCV exposure during public-health campaigns undertaken during the colonial era.

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.

Molecular strain typing is essential for deciphering the epidemiology of Campylobacter jejuni infections. We applied two different methods, multilocus sequence typing (MLST) and analysis of the flaA short variable repeat (SVR), to 289 isolates (163 human, 56 chicken, 34 raw milk, and 36 environmental water isolates) collected in the province of Québec, Canada, over 3 years; in addition, the analysis included the pulsed-field gel electrophoresis (PFGE) typing results for a subset of 131 isolates studied previously. MLST defined 96 sequence types (STs) and 20 clonal complexes (CCs), including 49 STs (73 isolates, 25%) and 39 alleles not previously documented in an international database. The frequency of new STs was significantly higher among water isolates than among isolates from other sources (18/36 [50%] and 55/253 [22%], respectively; P < 0.001). Nine of the 10 most prevalent CCs included isolates from humans and at least one other source; five CCs comprised exclusively or mostly human and chicken isolates. However, water and milk were the predominant nonhuman sources among the remaining CCs, suggesting that sporadic C. jejuni infections in humans may frequently arise from sources other than chickens. All three typing systems were discriminatory (discriminatory index > 0.9). Among 131 isolates analyzed by PFGE, each of the 20 types represented by two or more isolates corresponded to a single CC. In contrast, among the 14 most prevalent types detected by analysis of the flaA SVR (5 to 27 isolates each), 8 (57%) included isolates that represented multiple different CCs. The basis for these discordant results was uncertain. Antimicrobial resistance was randomly distributed among the CCs and appeared to be more closely related to the source of an isolate than its genotype. Although MLST is labor-intensive and expensive, it remains the single best method for the genotyping of C. jejuni isolates and deciphering the epidemiologic relationships among isolates.

In 1957, Chlamydia trachomatis (non-genital serovars A, B, Ba and C) was recognized as the aetiological agent of trachoma.1 C trachomatis genital serovars D to K cause ophthalmia neonatorum and conjunctivitis in older children and adults. Trachoma remains endemic in many countries of sub-Saharan Africa, the Middle East and other areas of the developing world.2 Nucleic acid amplification testing (NAAT) represented a major advance in the diagnosis of C trachomatis infection among patients with trachoma, cell culture being rarely available in endemic countries. Even using this most sensitive technology, however, one fifth of patients with severe trachoma and up to one half of those with milder infections do not have …

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.

This study compares the occurrence of antimicrobial resistance to erythromycin, ciprofloxacin, and tetracycline among 384 Campylobacter jejuni isolates from humans (245), fresh whole retail chickens (56), raw milk (33), and environmental water (41) collected between 2000 and 2003 in Quebec, Canada. Resistance to ciprofloxacin was significantly more frequent in human isolates acquired abroad than in those acquired locally (50 versus 5.9%; P < 0.001); ciprofloxacin resistance was almost absent in water, chicken, and raw milk isolates. In contrast, resistance to erythromycin was significantly more common in chicken than in locally acquired human isolates (16 versus 3.0%, respectively; P < 0.001); no erythromycin resistance was found among water, raw milk, and human isolates acquired abroad. Resistance to tetracycline was significantly more common in chicken and human isolates acquired locally (58.9 and 45.8%, respectively) than in raw milk and water isolates (9.1 and 7.3%, respectively, P < 0.001). Tetracycline resistance was also observed in 44.4% of human isolates acquired abroad. No human isolate was resistant to both ciprofloxacin and erythromycin, but one chicken isolate was resistant to all three antimicrobial agents. Our results suggest that from 2000 to 2003 in Quebec, antimicrobial resistance remained stable among locally acquired C. jejuni human clinical isolates and might even have decreased. However, the high erythromycin resistance rate observed among chicken isolates is concerning because of the risk of transmission of such isolates to humans. Additional studies are needed to monitor trends in antimicrobial resistance among food, environment, and human C. jejuni isolates as well as antibiotic use in animals.

Objective Type 2 diabetes mellitus (T2DM) is one of the most prevalent chronic inflammatory diseases of the elderly. Its development is related to the alteration of the immune system with aging characterized by immunosenescence and inflamm-aging. In turn, T2DM also alters the immune response. As a consequence, older people with T2DM are more susceptible to influenza and to its complications as compared with healthy controls. Vaccination against influenza has shown poor efficacy in the older population and even less efficacy in patients with diabetes. We studied here the antibody response to vaccination in healthy and diabetic elderly participants. Research design and methods In 2 groups of elderly participants (healthy N=119 and T2DM N=102), we measured the immunogenicity of influenza vaccine by hemagglutination inhibition assays. We assessed several blood and functional parameters as potential predictors of the vaccine efficacy. Results We found no difference between antibody responses in diabetic elderly compared with healthy elderly. Among the biological and functional determinants, the cytomegalovirus (CMV) serostatus played a more prominent role in determining the magnitude of response. We concluded that in addition to age and diabetic status, immunological history such as CMV status should be taken into account. None of the other biological or functional parameters studied could be reliably linked to the vaccine antibody response in older adults who are not frail including those with well-controlled diabetes. Conclusions Our data strongly suggest that influenza vaccine should be administered to elderly patients with T2DM; however, the immune determinants of the antibody response to influenza vaccination should be further investigated.

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

Alzheimer's disease (AD) is the most common form of dementia. Amyloïd plaques accumulation and microbial infections have been proposed as an important components of AD aetiology. Defects in amyloïd-β clearance by monocyte/macrophage/microglia, imply that the immune system may participate in development and progression of AD. Phenotype and functional analysis of polymorphonuclear neutrophils (PMN) in peripheral blood were performed by flow cytometry. Our cohort of patients comprised amnestic Mild Cognitive Impairment (aMCI) (n = 10), mild stage AD (mAD) (n = 10) and healthy elderly controls (n = 10). aMCI patients had memory impairment but no functional disorders. However, these patients have a 50% chance of developing AD within 3 years following the diagnostic. PMN number was unchanged between the three groups. In contrast, the CD177+ population was significantly increased in mAD patients (p < 0.05). Phagocytosis of opsonized bacteria was less in aMCI and AD patients than controls. This observation was associated in aMCI with a decrease in CD33 (siglec 3) and complement receptor for C5A expression. Decreased expression of CD14 (a pattern recognition receptor), CD15 (carbohydrate adhesion molecule) and CD16 (FcγRIII) expression was found in mAD patients. Our results describe for the first time alterations of neutrophils in aMCI patients. PMN analysis may provide a useful tool in early identification of AD patients.

We have investigated the deposition of biotinylated nano-beads on the surface of GaAs. The deposition procedure involved either direct coating of (001) GaAs with nano-beads, or binding the nano-beads with avidin immobilized on the surface of (001) GaAs through the interface of biotin and the NH 2 terminal group of 11-amino-1-undecanethiol (HS(CH 2 ) 11 NH 2 ). The efficiency of binding was tested by washing the samples in a solution of a commercial detergent and by subjecting them to a deionized water ultrasonic bath. The results indicate that nano-beads deposited directly on the surface of (001) GaAs withstand the detergent washing test but they are easily removed by ultrasonic washing. In contrast, the nano-beads attached to (001) GaAs through the avidin-biotin-thiol interface survive the ultrasonic washing tests.

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.