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Studies of the effects of electromagnetic waves on Saccharomyces cerevisiae emphasize the need to develop instrumented experimental systems ensuring a characterization of the exposition level to enable unambiguous assessment of their potential effects on living organisms. A bioreactor constituted with two separate compartments has been designed. The main element (75% of total volume) supporting all measurement and control systems (temperature, pH, agitation, and aeration) is placed outside the exposure room whereas the secondary element is exposed to irradiation. Measurements of the medium dielectric properties allow the determination of the electromagnetic field at any point inside the irradiated part of the reactor and are consistent with numerical simulations. In these conditions, the growth rate of Saccharomyces cerevisiae and the ethanol yield in aerobic conditions are not significantly modified when submitted to an electromagnetic field of 900 and 2400 MHz with an average exposition of 6.11 V.m −1 and 3.44 V.m −1 respectively. [ABSTRACT FROM AUTHOR]

High performance assays are essential for the serological diagnosis of recent and past Zika virus (ZIKV) infections but few are presently available. We used two commercially available NS1 antigen-based enzyme-linked immunoassays to study the kinetics of anti-ZIKV IgM and IgG in 15 ZIKV-infected patients for up to 180 days after clinical onset. The Diapro assay detected anti-ZIKV IgM reactivity more frequently (100%) and for longer (median 53 days) than did the Euroimmun assay (60%; 13 days, P < 0.005). Both assays detected anti-ZIKV IgG reactivity 11 days after clinical onset in all subjects. ZIKV IgG reactivity decreased in 3 subjects, suggesting long-term false-negative results with the Euroimmun assay. Existing anti-Dengue antibodies seem to modify the detection of ZIKV IgG but the specificity of the immunoassays was not assessed. These enzyme-linked immunoassays were user-friendly and provided results rapidly in our hands but they need further assessment before being widely used for diagnosis or public health surveys. [ABSTRACT FROM AUTHOR]

Persistent infection with high-risk human papillomavirus (HPV-HR) is a recognized cause of cervical cancer. The aimof this studywas to compare analytical and clinical performances of the Cobas®HPV and Anyplex™ II HPV28 assays for HPV detection and genotyping. A total of 94 cervical samples were tested. For HPV-HR, the results agreed very well (94.68%),with 100% agreement when detecting CIN2+. The Anyplex™ II HPV28 assay detected more genotypes than the Cobas® HPV Test, but their clinical performances were similar. [ABSTRACT FROM AUTHOR]

Abstract: The ability to detect and quantify HIV and HCV genomes is important for checking spermatozoid preparation protocols also known as “sperm washing”. But no commercial assay is available. A method was developed for detecting HIV and HCV in semen fractions using the COBAS Ampliprep™ and COBAS Taqman™ instruments. It will detect fewer than 200 copies of HIV RNA per ml of semen plasma and fewer than 200 copies/3×106 semen cells. The sensitivity for HCV is similar at more than 200IU/ml and below 200IU/3×106 semen cells. No inhibitor of PCR amplification was detected. This automated protocol permits a convenient, standardized testing for HIV and HCV in semen. The performance is the same as that of the previous generation of automated assays but the cost and operating time are both reduced. [Copyright &y& Elsevier]

To enable serodiscordant couples to reproduce while minimizing the risk of infecting the female partner and to demonstrate the feasibility of including these couples in an insemination program.Prospective follow-up study.Ambulatory IUIs.Fifty-six HIV-1–serodiscordant couples with a male infected partner.Sperm cell separation by the gradient density method followed by the swim-up method, virological detection, and IUI after ovarian stimulation.Pregnancy rate, HIV-1 test, and p24 antigenemia in women.Two hundred thirteen IUIs were performed, and 37 pregnancies occurred, resulting in 33 births and 2 ongoing pregnancies. Fifty percent of couples have a child. No women were infected.Our study demonstrates the efficiency of an IUI program with prepared and virologically tested spermatozoa in serodiscordant couples with an HIV-1–infected male partner, allowing the couples to have children without transmission of the virus to the female partner. [Copyright &y& Elsevier]

Phylogenetic analysis of human immunodeficiency virus type 1 (HIV-1) pol gene is a useful method for subtyping European strains of HIV-1. The suitability of this method for genetically diverse African strains was evaluated by comparing HIV-1 subtyping of Cameroon strains using a long fragment of the pol gene sequence to the findings obtained using env gene sequences. When the pol gene could not be amplified, the reverse transcriptase (RT) or the protease (PR) genes were used. Phylogenetic analysis of the env C2/V3 gene sequences of 60 HIV-1 isolates showed 52 to be subtype A, 2 subtype G, plus one each of subtypes C, F2 and H, with 3 subtypes not determined. A long fragment of the pol gene was amplified successfully and sequenced in 23% of cases. The RT region was amplified for 42% of the samples that could not be typed by analysing the long fragment, and the PR gene was amplified for 40% of them. Thus, 63% of samples were typable. Env and pol gene subtypings were in agreement in 86% of cases. It is concluded that the phylogenetic analysis of pol gene sequences is not a practical method for HIV-1 subtyping in areas of high subtype diversity, despite the good agreement between the env and pol gene subtypings. However, it can be a useful method for HIV-1 subtyping, provided that the gene is amplifiable. [Copyright &y& Elsevier]

• Severe and protracted monkeypox virus infection can occur in a patient with AIDS. • In this setting, tecovirimat might improve symptoms and decrease viral replication. • Randomized controlled trials in humans are underway. • Patients who are diagnosed with monkeypox should be routinely tested for HIV. A patient aged 28 years who is immunocompromised and living with HIV/AIDS became infected with the monkeypox virus (MPXV). His clinical condition deteriorated for 37 days, with fever, skin lesions, and diarrhea before going to the infectious diseases department, where his severe, protracted infection was treated with tecovirimat for 14 days. His condition rapidly improved, and the skin lesions decreased, as did the MPXV loads, with no adverse events. This case indicates that tecovirimat might be effective for treating patients who are immunocompromised and are infected with MPXV. [ABSTRACT FROM AUTHOR]

Abstract Zika (ZIKV), Dengue (DENV) and Chikungunya viruses (CHIKV) co-circulate in the same geographical areas during the same seasonal period through the same biting arthropods. Therefore a rapid sensitive and specific molecular assay for these viruses would be a considerable help in the disease management and the epidemiological survey. We developed a one-step multiplex real-time PCR for the simultaneous detection of these viruses. Intra and inter-reproducibilities varied from 0.41% to 3.29% and from 1.13% to 4.93% for each virus respectively. The specificity was 100%. Whole blood, plasma and urines were used for comparison with commercially available monoplex assays (RealStar® kits, Altona Diagnostics GmbH, Hamburg, Germany). The concordance was 96%, 92.9% and 95.7% for ZIKV, DENV and CHIKV respectively. No cross reaction and no PCR inhibition were observed for any of the clinical samples. This test can thus be used as a rapid molecular assay for ZIKV, DENV1–4 and CHIKV infections. Highlights • The multiplex assay detects three viral targets: ZIKV, DENV and CHIKV. • The multiplex assay includes an internal control (GAPDH). • The multiplex assay is suitable for use with whole blood specimens. [ABSTRACT FROM AUTHOR]

Background: Evidence of human sexual transmission during Zika virus emergence is a matter of concern, particularly in procreation, but to date, kinetics of seminal shedding and the effects of infection on human reproductive function have not been described. To investigate the effects of Zika virus infection on semen and clearance of Zika virus from semen and body fluids, we aimed to study a cohort of Zika virus-infected men.Methods: This prospective observational study recruited men presenting with acute Zika virus infection at Pointe-à-Pitre University Hospital in Guadeloupe, French Caribbean, where a Zika virus outbreak occurred between April and November, 2016. Blood, urine, and semen were collected at days 7, 11, 20, 30, 60, 90, and 120 after symptom onset, and semen characteristics, such as total sperm count, sperm motility, vitality, and morphology, and reproductive hormone concentrations, such as testosterone, inhibin, follicle-stimulating hormone, and luteinising hormone, were assessed. At days 7, 11, and 20, semen was processed to isolate motile spermatozoa. Zika virus RNA was detected by RT-PCR using whole blood, serum, urine, seminal plasma, semen cells, and motile spermatozoa fractions. Zika virus was isolated from different sperm fractions on Vero E6 cultures.Findings: 15 male volunteers (mean age 35 years [SD 5; range 25-44) with acute Zika virus infection and positive Zika virus RNA detection in blood or urine were enrolled. Total sperm count was decreased from median 119 × 106 spermatozoa (IQR 22-234) at day 7 to 45·2 × 106 (16·5-89·6) at day 30 and 70 × 106 (28·5-81·4) at day 60, respectively, after Zika virus infection. Inhibin values increased from 93·5 pg/mL (IQR 55-162) at day 7 to 150 pg/mL (78-209) at day 120 when total sperm count recovered. In motile spermatozoa obtained after density gradient separation, Zika virus RNA was found in three of 14 patients at day 7, four of 15 at day 11, and four of 15 at day 20, and replication-competent virus was found in the tested patient. Seminal shedding kinetics seemed heterogeneous among patients. Whole blood was the fluid most frequently positive for Zika virus RNA (62 of 92 samples) and three patients remained positive at day 120.Interpretation: Semen alterations early after acute Zika virus infection might affect fertility and could be explained by virus effects on the testis and epididymis. Frequency of shedding and high viral load in semen, together with the presence of replicative virus in a motile spermatozoa fraction, can lead to Zika virus transmission during sexual contact and assisted reproduction procedures. Whole blood seems to be the best specimen for Zika virus RNA detection, diagnosis, and follow-up.Funding: Agence de la Biomédecine/Agence Régionale de Santé de la Guadeloupe/Inserm-REACTing. [ABSTRACT FROM AUTHOR]

Objective: To study sperm parameters in patients infected with human immunodeficiency virus (HIV)-1 and to analyze mitochondrial DNA (mtDNA) in sperm according to the HIV treatment. Design: Observational study. Setting: University-affiliated teaching hospital. Patient(s): Thirty-two patients infected with HIV-1 and 31 noninfected healthy men provided semen samples. Intervention(s): None. Main Outcome Measure(s): After DNA extraction, mtDNA level was assessed using real-time polymerase chain reaction (PCR) (LightCycler) in whole semen and in selected spermatozoa from 90% density centrifugation gradients. For each sample, mtDNA and β-globin gene sequences were amplified and PCR products were quantified. The mtDNA-to-β-globin ratio expressed the number of mtDNA copies per cell. Result(s): Compared with the control group, several sperm parameters were altered in patients with HIV. The number of mtDNA copies per cell in whole semen was increased in HIV-infected patients (6.3 ± 6.3 vs. 3.5 ± 3.2). However, there was no statistically significant difference in mtDNA copy number in the spermatozoa obtained after density gradient centrifugation. The number of nucleoside analogue reverse transcriptase inhibitors (NRTI) taken by patients during treatment significantly influenced the mtDNA level in sperm (1 NRTI 7.6 ± 8.1, 2 NRTIs 7.0 ± 5.1, 3 NRTIs 3.2 ± 2.1). Conclusion(s): Using a specific method to measure sperm mtDNA, we demonstrated a decrease of mtDNA copies in spermatozoa after use of NRTIs with known mitochondrial toxicity. [ABSTRACT FROM AUTHOR]

Abstract: Objective: Assisted reproduction techniques can minimize the risk of HIV female contamination when the male partner is HIV-infected. The aim of this study was to investigate the efficiency of sperm washing and intrauterine insemination (IUI) in these couples. Study design: Retrospective comparative study. Eighty-four HIV-1 serodicordant couples underwent 294 IUI. The control group was composed of 90 couples (320 IUI cycles) with donor sperm. Spermatozoa from HIV-1 infected male partner were prepared and tested for HIV-1 according to sperm washing method. Spermatozoa from HIV-1 and donor male were frozen before IUI. IUI were performed after ovarian stimulation. Main outcomes measures were pregnancy rate per cycle and baby take-home rate per couples. Results: Although the pregnancy rate and baby take-home rate were higher in IUI with sperm washing than in IUI using donor sperm (18.0 versus 14.7 and 52.4 versus 41.1, respectively), the differences were not statistically significant. In serodiscordant couples, blood estradiol levels under ovarian stimulation and total motile sperm inseminated were a determining factor in achieving pregnancy. No female HIV-1 contamination occurred. Conclusion: This study demonstrates that sperm washing and IUI are highly effective in enabling serodiscordant couples with an HIV-1 infected male partner to have a child. [Copyright &y& Elsevier]

Abstract: Hepatitis C virus (HCV) infection in Cameroon is characterized by widespread seropositivity and great virus genetic diversity (3 genotypes and over 10 subtypes). A total of 244 HCV NS5B sequences of 382–405bp long (95 type 1, 58 type 2, and 91 type 4) were phylogenetically analyzed to estimate the history of the HCV epidemic in Cameroon. The newly developed Bayesian coalescent approach was used to infer the history of each HCV type. The estimated dates of the most recent common ancestors (MRCA) for genotypes 1 (1500; 95% confidence interval (95% CI): 1300–1650) and 4 (1500; 95% CI: 1350–1700) were in the same range, while the date for genotype 2 MRCA (1600; 95% CI: 1400–1750) was slightly more recent. The mean genetic distance between HCV genotype 1 sequences was greater than that of HCV type 4 sequences, itself greater than that of HCV type 2 sequences. The initial infected populations of all three genotypes did not grow until recently, when they grew exponentially. The growth rate has now begun to slow, with a less steep exponential growth curve. The period of exponential growth of all the three genotypes was between 1920 and 1960. These results (i) confirm that HCV genotypes 1 and 4 have produced long-term endemics, (ii) suggest that genotype 2 was introduced into Cameroon more recently, and (iii) indicate that the exponential spread of the three genotypes between 1920 and 1960 coincided with the mass campaign against trypanosomiasis and mass vaccinations in Cameroon. [Copyright &y& Elsevier]

Abstract: Conventional tests for antibody to Hepatitis C virus (HCV) and HCV RNA require considerable time before results are available, remain very expensive and are not adapted to many sub-Saharan African countries where HCV is endemic. The aim of this study was to evaluate the accuracy of an algorithm consisting of two HCV rapid tests to diagnose and predict HCV viremia in patients in Cameroon. Three hundred and twenty nine plasma samples were screened by two HCV rapid tests (ImmunoComb® II HCV, PBS Orgenics and Hexagon® HCV, Human). Previous evaluation of these samples for HCV antibodies (anti-HCV) by conventional third generation ELISA, considered as a reference test, indicated that 168 were anti-HCV negative and 161 positive. Among the 161 anti-HCV positive plasma, 114 (71%) were HCV RNA-positive by RT-PCR assay. The ImmunoComb® II HCV test provided the more sensitive detection of anti-HCV (sensitivity: 99.4% with a 95% CI=96–100%). Surprisingly, the second HCV rapid test, Hexagon® HCV, showed a high capacity to identify non-viremic subjects amongst anti-HCV positive cases (93.6% [95% CI: 82–99%]). These results suggest an algorithm using ImmunoComb® II HCV as a first test to screen anti-HCV positive subjects, and Hexagon® HCV as a second test to discriminate between viremic and non-viremic HCV seropositive subjects. [Copyright &y& Elsevier]

: ObjectiveTo study seminal excretion of human immunodeficiency virus type 1 (HIV-1) during 4 years of follow-up in an HIV-1–infected patient, the relationship between high viral excretion and inflammatory status of semen, and the efficiency of sperm processing methods in obtaining spermatozoa with undetectable RNA and proviral DNA levels.: DesignCase report.: SettingUniversity hospital and research group on human fertility.: Patient(s)One HIV-1–infected patient.: Intervention(s)Paired blood and semen samples were obtained during 4 years of follow-up.: Main outcome measure(s)CD4 cell count; blood and seminal plasma viral load; and HIV-1 RNA and proviral DNA in different cell fractions obtained during sperm processing, as measured by the density gradient method and the swim-up method; sperm parameters; and polymorphonuclear granulocyte count.: Result(s)Shedding of HIV-1 in semen was intermittent. The highest seminal viral loads were associated with a markedly increased polymorphonuclear granulocyte count, which reflects inflammation of the genital tract. Spermatozoa with undetectable levels of HIV-1 RNA or DNA were obtained regardless of the viral load in semen.: Conclusion(s)In an HIV-1–infected man with intermittent seminal viral excretion, sperm processing was effective in obtaining spermatozoa without detectable HIV-1 genomes. [Copyright &y& Elsevier]

Objective: To report the effects of ribavirin plus pegylated interferon on semen parameters and sperm DNA integrity in a man given treatment for hepatitis C.Design: Case report.Setting: University-affiliated teaching hospital.Intervention(s): None.Patient(s): A 37-year-old man given treatment with ribavirin and pegylated interferon for hepatitis C.Main Outcome Measure(s): Semen parameters (sperm count, motility, round cells), sperm protamination state measured by sperm chromatin structure assay, and sperm DNA fragmentation measured by terminal uridine nick-end labeling assay.Result(s): The percentage of progressive spermatozoa and the number of motile sperm per ejaculate decreased during treatment. The round cell/spermatozoa ratio, which reflects spermatogenic abnormality, increased from 2.6% +/- 1.4% to 23.6% +/- 13.0% during treatment and returned to baseline value 4 months later. Moreover, the sperm DNA fragmentation index (as measured by sperm chromatin structure assay) increased very markedly during treatment (from 14.5% before to 69.2% at 7 months of treatment) and remained elevated 8 months later.Conclusion(s): This study reports for the first time not only quantitative but also qualitative alterations of spermatogenesis with DNA packaging abnormalities and emphasizes the need for prospective clinical studies. While the results of other studies are awaited, the alterations that persisted 8 months after treatment argue for a longer contraception period after treatment discontinuation in men. [ABSTRACT FROM AUTHOR]