Association between Zika virus and fetopathy: a prospective cohort study in French Guiana.

OBJECTIVE: To establish the incidence of fetal central nervous system (CNS) anomalies (including microcephaly), signs of congenital infection and fetal loss in pregnant women infected with Zika virus (ZIKV) and non-infected pregnant women in western French Guiana. METHODS: This prospective cohort study was conducted between 1 January and 15 July 2016. We evaluated and compared clinical and fetal ultrasound examinations of 301 pregnant women with biological confirmation of ZIKV infection and 399 pregnant women who were negative for ZIKV infection. RESULTS: Overall, the total number of fetuses with CNS involvement was higher in the infected than in the control group (9.0% vs 4.3%; relative risk, 2.11 (95% CI, 1.18-4.13)). Anomalies of the corpus callosum and presence of cerebral hyperechogenicities were significantly more common in the infected group. There was an increased risk of microcephaly in the infected compared with the control group (1.7% vs 0.3%; relative risk, 6.63 (95% CI, 0.78-57.83)), although this was not statistically significant. When the mother was infected during the first or second trimester, there was a greater risk of severe CNS involvement, more signs of infection and intrauterine fetal death than with infection in the third trimester. The rate of vertical transmission in the exposed group was 10.9%. CONCLUSION: ZIKV infection during pregnancy is associated with a significant risk of fetal CNS involvement and intrauterine fetal death, particularly when infection occurs during the first or second trimesters. Microcephaly was not present in every case of congenital ZIKV syndrome that we observed. Until more is known about this disease, it is paramount to evaluate suspected cases by detailed neurosonography on a monthly basis, paying particular attention to the corpus callosum and the presence of hyperechogenic foci. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Highly efficient and sensitive membrane-based concentration process allows quantification, surveillance, and sequencing of viruses in large volumes of wastewater.

Wastewater-based epidemiology is experiencing exponential development. Despite undeniable advantages compared to patient-centered approaches (cost, anonymity, survey of large populations without bias, detection of asymptomatic infected peoples…), major technical limitations persist. Among them is the low sensitivity of the current methods used for quantifying and sequencing viral genomes from wastewater. In situations of low viral circulation, during initial stages of viral emergences, or in areas experiencing heavy rains, the extremely low concentrations of viruses in wastewater may fall below the limit of detection of the current methods. The availability during crisis and the cost of the commercial kits, as well as the requirement of expensive materials such as high-speed centrifuge, can also present major blocks to the development of wastewater-based epidemiological survey, specifically in low-income countries. Thereby, highly sensitive, low cost and standardized methods are still needed, to increase the predictability of the viral emergences, to survey low-circulating viruses and to make the results from different labs comparable. Here, we outline and characterize new protocols for concentrating and quantifying SARS-CoV-2 from large volumes (500 mL-1 L) of untreated wastewater. In addition, we report that the methods are applicable for monitoring and sequencing. Our nucleic acid extraction technique (the routine C: 5 mL method) does not require sophisticated equipment such as automatons and is not reliant on commercial kits, making it readily available to a broader range of laboratories for routine epidemiological survey. Furthermore, we demonstrate the efficiency, the repeatability, and the high sensitivity of a new membrane-based concentration method (MBC: 500 mL method) for enveloped (SARS-CoV-2) and non-enveloped (F-specific RNA phages of genogroup II / FRNAPH GGII) viruses. We show that the MBC method allows the quantification and the monitoring of viruses in wastewater with a significantly improved sensitivity compared to the routine C method. In contexts of low viral circulation, we report quantifications of SARS-CoV-2 in wastewater at concentrations as low as 40 genome copies per liter. In highly diluted samples collected in wastewater treatment plants of French Guiana, we confirmed the accuracy of the MBC method compared to the estimations done with the routine C method. Finally, we demonstrate that both the routine C method processing 5 mL and the MBC method processing 500 mL of untreated wastewater are both compatible with SARS-CoV-2 sequencing. We show that the quality of the sequence is correlated with the concentration of the extracted viral genome. Of note, the quality of the sequences obtained with some MBC processed wastewater was improved by dilutions or enzyme substitutions suggesting the presence of specific enzyme inhibitors in some wastewater. To the best of our knowledge, our MBC method is one of the first efficient, sensitive, and repeatable method characterized for SARS-CoV-2 quantification and sequencing from large volumes of wastewater.

S-OtrH3N2 viruses: use of sequence data for description of the molecular characteristics of the viruses and their relatedness to previously circulating H3N2 human viruses.

Emergence of influenza viruses from the animal reservoir is a permanent challenge. The rapid description and immediate sharing of information on these viruses is invaluable for influenza surveillance networks and for pandemic preparedness. With the help of data generated from the World Health Organization Collaborating Centre for Reference and Research on Influenza at the United States Centers for Disease Control and Prevention, we provide here information on the swine­origin triple reassortant influenza A(H3N2) viruses detected in human cases in the north-east of the United States.

Highly heterogeneous temperature sensitivity of 2009 pandemic influenza A(H1N1) viral isolates, northern France.

We assayed the temperature sensitivity of 2009 pandemic influenza A(H1N1) viral isolates (n=23) and seasonal influenza A(H1N1) viruses (n=18) isolated in northern France in 2007/08 and 2008/09. All isolates replicated with a similar efficiency at 34 °C and 37 °C, and with a lower efficiency at 40 °C. The pandemic viral isolates showed a stronger heterogeneity in their ability to grow at the highest temperature, as compared with the seasonal isolates. No statistically significant difference in temperature sensitivity was observed between the pandemic viral isolates from severe and mild cases of influenza. Our data point to the impact of temperature sensitivity on the genetic evolution and diversification of the pandemic influenza A(H1N1) virus since its introduction into the human population in April 2009, and call for close surveillance of this phenotypic marker related to host and tissue tropism.

Metal enriched quasi-ultrafine particles from stainless steel gas metal arc welding induced genetic and epigenetic alterations in BEAS-2B cells.

Recent evidence has supported welding fume (WF)-derived ultrafine particles (UFP) could be the driving force of their adverse health effects. However, UFP have not yet been extensively studied and are currently not included in present air quality standards/guidelines. Here, attention was focused on the underlying genetic and epigenetic mechanisms by which the quasi-UFP (Q-UFP, i.e., ≤ 0.25 µm) of the WF emitted by gas metal arc welding-stainless steel (GMAW-SS) exert their toxicity in human bronchial epithelial BEAS-2B cells. The Q-UFP under study showed a monomodal size distribution in number centered on 104.4 ± 52.3 nm and a zeta potential of -13.8 ± 0.3 mV. They were enriched in Fe > Cr > Mn > Si, and displayed a relatively high intrinsic oxidative potential. Dose-dependent activation of nuclear factor erythroid 2-related factor 2 and nuclear factor-kappa B signaling pathway, glutathione alteration, and DNA, protein and lipid oxidative damage were reported in BEAS-2B cells acutely (1.5 and 9 µg/cm2, 24 h) or repeatedly (0.25 and 1.5 µg/cm2, 3 × 24 h) exposed to Q-UFP (p < 0.05). Alterations of the Histone H3 acetylation were reported for any exposure (p < 0.05). Differentially regulated miRNA and mRNA indicated the activation of some critical cell signaling pathways related to oxidative stress, inflammation, and cell cycle deregulation towards apoptosis. Taken together, these results highlighted the urgent need to better evaluate the respective toxicity of the different metals and to include the Q-UFP fraction of WF in current air quality standards/guidelines relevant to the occupational settings.

Hydroxychloroquine lung pharmacokinetics in critically ill patients with COVID-19.

Different dosage regimens of hydroxychloroquine (HCQ) have been used to manage COVID-19 (coronavirus disease 2019) patients, with no information on lung exposure in this population. The aim of our study was to evaluate HCQ concentrations in the lung epithelial lining fluid (ELF) in patients infected with SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), the virus that causes COVID-19. This was a retrospective, observational, multicentre, pharmacokinetic study of HCQ in critically ill COVID-19 patients. No additional interventions or additional samples compared with standard care of these patients were conducted in our teaching hospital. We included all intubated COVID-19 patients treated with crushed HCQ tablets, regardless of the dosage administered by nasogastric tube. Blood and bronchoalveolar lavage samples (n = 28) were collected from 22 COVID-19 patients and total HCQ concentrations in ELF were estimated. Median (interquartile range) HCQ plasma concentrations were 0.09 (0.06-0.14) mg/L and 0.07 (0.05-0.08) mg/L for 400 mg × 1/day and 200 mg × 3/day, respectively. Median HCQ ELF concentrations were 3.74 (1.10-7.26) mg/L and 1.81 (1.20-7.25) for 400 mg × 1/day and 200 mg × 3/day, respectively. The median ratio of ELF/plasma concentrations was 40.0 (7.3-162.7) and 21.2 (18.4-109.5) for 400 mg × 1/day and 200 mg × 3/day, respectively. ELF exposure is likely to be underestimated from HCQ concentrations in plasma. In clinical practice, low plasma concentrations should not induce an increase in drug dosage because lung exposure may already be high.

Automatic sample production by depositing solutions on filters for the organization of proficiency tests.

This article describes a device intended to produce replicas on filters by liquid deposition of anion or metal solutions. Schematically, the filters are housed in cassettes labelled automatically by means of a code. An automatic arm takes each cassette, reads the code, and deposits the amount of element required. Weighing before and after deposition allows the amount deposited to be accurately checked and determined. This automated system allows the production of replicas with high deposition regularity, replica dispersion for the most part being <1%. The samples produced can be used during proficiency tests where the assigned value is determined either by the participants or by the organizer.

Census and analysis of persistent false-negative results in serological diagnosis of human immunodeficiency virus type 1 group O infections.

Human immunodeficiency viruses (HIV) have a high level of genetic diversity. The outlier variants of HIV type 1 (HIV-1) group O are distantly related to HIV-1 group M. Their divergence has an impact on serological diagnosis, with a risk of false-negative results. In this study, we report 20 failure cases, involving patients with primary or chronic infection, in France and Cameroon between 2001 and 2008. Our results indicate that some assays detected group O infection much less efficiently than others. Two major reasons for these false-negative results were identified: the presence or absence of a group O-specific antigen (and the designed sequence) for the detection of antibodies and the greater envelope variability of group O than of group M strains. This study highlights the complexity of screening for these divergent variants and the need to evaluate test performance with a large panel of strains, due to the extensive diversity of group O variants.

TYMSTR, a putative chemokine receptor selectively expressed in activated T cells, exhibits HIV-1 coreceptor function.

BACKGROUND: Chemokines bind to specific receptors and mediate leukocyte migration to sites of inflammation. Recently, some chemokine receptors, notably CXCR4 and CCR5, have been shown to be essential fusion factors on target cells for infection by human immunodeficiency virus (HIV); the chemokines bound by these receptors have also been shown to act as potent inhibitors of HIV infection. Here, we describe the isolation of a novel, putative chemokine receptor. RESULTS: We have isolated the cDNA for a putative human chemokine receptor, which we have termed TYMSTR (T-lymphocyte-expressed seven-transmembrane domain receptor). The TYMSTR gene is localized to human chromosome 3 and encodes a protein that has a high level of identity with chemokine receptors. TYMSTR mRNA was selectively expressed in interleukin-2-stimulated T lymphocytes but not in freshly isolated lymphocytes and leukocytes or related cell lines. The natural ligand for TYMSTR was not identified among 32 human chemokines and other potential ligands. Cells co-expressing TYMSTR and human CD4 fused with cells expressing envelope glycoproteins of macrophage (M)-tropic HIV-1 as well as T-cell line (T)-tropic HIV-1 isolates. Addition of infectious, T-tropic HIV-1 particles to TYMSTR/CD4-expressing cells resulted in viral entry and proviral DNA formation. CONCLUSIONS: Our findings demonstrate that TYMSTR, in combination with CD4, mediates HIV-1 fusion and entry. The high-level expression of TYMSTR in CD4(+) T lymphocytes and the selectivity of this receptor for T-tropic and M-tropic HIV-1 strains indicates that TYMSTR might function as HIV coreceptor at both early and late stages of infection.

IL-10-producing T cells suppress immune responses in anergic tuberculosis patients.

The lethality of Mycobacterium tuberculosis remains the highest among infectious organisms and is linked to inadequate immune response of the host. Containment and cure of tuberculosis requires an effective cell-mediated immune response, and the absence, during active tuberculosis infection, of delayed-type hypersensitivity (DTH) responses to mycobacterial antigens, defined as anergy, is associated with poor clinical outcome. To investigate the biochemical events associated with this anergy, we screened 206 patients with pulmonary tuberculosis and identified anergic patients by their lack of dermal reactivity to tuberculin purified protein derivative (PPD). In vitro stimulation of T cells with PPD induced production of IL-10, IFN-gamma, and proliferation in PPD(+) patients, whereas cells from anergic patients produced IL-10 but not IFN-gamma and failed to proliferate in response to this treatment. Moreover, in anergic patients IL-10-producing T cells were constitutively present, and T-cell receptor-mediated (TCR-mediated) stimulation resulted in defective phosphorylation of TCRzeta and defective activation of ZAP-70 and MAPK. These results show that T-cell anergy can be induced by antigen in vivo in the intact human host and provide new insights into mechanisms by which M. tuberculosis escapes immune surveillance.

First report of Litomosa spp. (Nematoda: Filarioidea) from Malagasy bats; review of the genus and relationships between species.

The presence of the filarial genus Litomosa in Malagasy bats is demonstrated by the finding of L. goodmani n. sp. from Miniopterus gleni and Litomosa sp. (male unknown) from M. manavi, both in the Special Reserve of Ankarana. These materials are compared to the 22 Litomosa species, including two Indian species originally placed in the genus Litomosoides, L. fotedari (Gupta and Trivedi, 1989) n. comb. and L. tewarii (Gupta and Trivedi, 1989) n. comb., and the new taxon L. seurati n. sp. (= L. beaucournui Bain, 1966 pro parte), type-host Rhinolophus ferrum-equinum, Algeria, distinguished by the narrow area rugosa and the female caudal extremity with two conspicuous points, instead of several small ones. The Malagasy material belongs to a group of species close to the type, L. filaria, which have a male area rugosa composed of cuticular bosses and microfilariae folded within the sheath, and which are parasitic in Vespertilionidae, Hipposideridae and Rhinolophidae from Africa and Europe. The two Malagasy species resemble L. seurati n. sp., L. beshkovi Jancev, 1971, L. chiropterum Ortlepp, 1932, L. adami Petit, 1980 and L. ottavianii Lagrange et Bettini, 1948, with the enlarged third segment of the buccal capsule. L. goodmani n. sp. is distinct with its small size and female caudal extremity with a single point, which is suppressed in old mature worms; the females of Litomosa sp. have two conical points. Relationships among Litomosa species appear to be dependent upon both the chiropteran host groups and the geographical region.

[Zika virus outbreak in Latin America: what are the challenges for French Guiana in April 2016?]. / Épidémie de virus Zika en Amérique latine : quels enjeux pour la Guyane française en avril 2016 ?

Started in 2015 in Brazil, an outbreak linked to a little known arbovirus, Zika virus spread throughout Latin America. This virus, considered until recently as responsible of only mild symptoms, made mention of previously unsuspected complications, with severe neurological manifestations in adults and malformations of the central nervous system, including microcephaly, in newborns of mother infected during the pregnancy. While the continent is more accustomed to the succession of arbovirus epidemics, suspected complications and the many unknowns keys of the latter arriving raise many public health issues. French Guiana, a French territory located in the north-east of the continent, combines both European level of resources and climate and issues specific to the Amazon region and Latin America. We discuss here the issues for 2016 Zika virus epidemic in our region, many of them are generalizable to neighboring countries.

[Effect of 2 beta-blockers on arterial hypertension during pregnancy. Results of a prospective study on 56 pregnant hypertensive women treated with atenolol and labetalol]. / Action de deux bêta-bloquants sur l'hypertension artérielle pendant la grossesse. Résultats d'une étude prospective sur 56 femmes enceintes hypertendues traitées par Aténolol et Labétalol.

This study reports the results that were obtained in 56 cases of arterial hypertension in pregnancy solely by beta-blocking with Atenolol or Labetalol. Any pregnant woman whose arterial blood pressure rises to or exceeds 140/90 mm mercury in two successive examinations at intervals of 8 days with rest is considered to be hypertensive. As soon as treatment is started mothers' supervision is assured regularly by clinical and biological examinations and the dose of drug is adapted to each case. Fetal monitoring is ensured by ultrasound, cardiac rhythm tracings and hormone estimations. As far as the newborn is concerned, blood sugar and electrocardiogram measurements are taken to add to the normal examination at birth. Finally plasma levels of beta-blockers are estimated at birth in the mother and in the cord blood. The analysis of these results shows: for the mother: a fairly constant antihypertensive effect which is about the same for either drug in pregnancy. Further complementary injection therapy was needed, however, in 8 cases in labour. There were alterations in the method of delivery and in particular the Caesarean section rate rose to 12.5% and induction had to be carried out more frequently, triggered off by the slightest sign of fetal distress. As far as the child was concerned: 2 died in utero, the Apgar score was comparable to a control series, there was no bradycardia or broncho-spasm or teratogenic effect, mean weight at birth was significantly higher with Labetalol (3280 g +/- 555) than with Atenolol (2750 g +/- 630), the blood sugar levels at birth were in six cases lower than 1.4 mmol (0.25 g/l) but these were easily pu right by transfusion. The plasma levels of beta-blockers showed that there was a linear relationship between the maternal and fetal concentrations which confirmed that the two molecules pass through the placenta. This study confirms therefore that it is worth while using beta-blockers in cases of hypertension in pregnancy so long as careful observation is carried out, and it seems that the alpha constituent of Labetalol has advantages over the other.

[Seroprevalence of viral hepatitis B in the city of Mahajanga, Madagascar in 1999]. / Séroprévalence de l'hépatite virale B dans la ville de Mahajanga à Madagascar en 1999.

A seroepidemiological study was carried out to assess the prevalence of hepatitis B virus (HBV) in the city of Mahajanga, Madagascar in July 1999. A total of 654 serum samples were collected from randomly selected subjects over the age of 2 years. The ELISA technique was used to detect serum markers of HBV infection. Prevalence rates were 14.2 p. 100 for HBs, 1.4 p. 100 for HBe antigens, and 49.5 p. 100 for HBV infection as defined by the presence of at least one serum markers. HBs antigens were detected in 20.8 p. 100 of children between the ages of 2 and 4 years and 15.3 p. 100 of women of childbearing age. In the latter age group, 5.6 p. 100 demonstrated HBe antigens, which are considered as a replication marker. The findings of this study are in agreement with those of a previous study in Madagascar and indicate that an expanded program of immunization against hepatitis B virus is warranted.

[Genetic variability of Poliovirus: typing of strains isolated in Madagascar by restriction fragment length polymorphism (RFLP) assay]. / Variabilité génétique des Poliovirus: étude du polymorphisme de la longueur des fragments de restriction enzymatique de souches isolées à Madagascar.

The differentiation of the vaccine or wild origin of Poliovirus at the laboratory is an important step towards the process of the poliomyelitis eradication. We report herein the results obtained from Poliovirus types 3 and 2, isolated in Madagascar in 1997 and 2002 from healthy children and cases of acute flaccid paralysis, respectively. The technique used is based on the amplification of genome (RT-PCR), followed by Restriction Fragment Length Polymorphism assay (RFLP), performed in 3 different regions of the genome. In the capsid region (VP3-VP1 and VP1-2A), RFLP analysis allowed us to differentiate without ambiguity the wild or vaccine origin of the Poliovirus type 3, and to identify Vaccine-Derived Poliovirus (VDPV) type 2. In the noncapsid region, including the RNA polymerase and 3' non coding region (3Dpol-3' NTR), the VDPV were found to be recombinant with other Enteroviruses. These results confirm that RFLP assay is a reliable tool for intratypic differentiation and to study the genetic drift and recombination of Poliovirus.

[New method of molecular typing in human Enteroviruses: characterization of Madagascar "untypable" strains]. / Nouvelle méthode de typage moléculaire des Entérovirus humains: caractérisation des souches malgaches "non sérotypables".

Enteroviruses, members of the family Picornaviridae, are responsible for a wide variety of diseases and represent a major public health hazard. Typing of non polio enterovirus (NPEV) infection is traditionally based on a serum neutralization assay. However, this method is time-consuming, labor-intensive, expensive, and may fail to identify antigenic variation. A new molecular typing involving partial sequencing of the genome has been recently developed. In this study, 46 NPEV strains were analyzed, including 37 antigenicaly "untypeable" viruses. Partial sequencing of the C-end of the viral capsid protein VP1 and pairwise identity with the prototype strains allow us to assign a serotype for all "untypeable" viruses. The results show a large number and wide variety of Coxsackieviruses A which belong to the HEV-C species and also Echoviruses and Coxsackieviruses B of the HEV-B species. This method may be useful to identify all NPEV serotypes in Madagascar and to assess the possible impact of circulating NPEV populations, as we enter the final stage of poliomyelitis eradication.

[Prospective randomized study comparing the efficacy and tolerance of clindamycin-gentamycin versus metronidazole-gentamycin in acute utero-adnexal infections in hospitalized patients]. / Essai prospectif randomisé comparant l'efficacité et la tolérance de la clindamycine-gentamycine versus métronidazole-gentamycin dans les infections utéro-annexielles aiguës chez des patientes hospitalisées.

The efficacy and safety of two antibiotic combination (clindamycin + gentamycin [C + G] versus metronidazole + gentamycin [M + G]) have been compared in 45 in-patients suffering from pelvic inflammatory disease in a clinical prospective randomized trial. The rates of clinical and bacteriological recovery reached respectively 85.7% and 71.4% for C + G group compared to 83.3% and 78.6% for M + G group (no significant differences). Side effects (vomiting, gastralgia and vaginal mycosis) developed in four occasions in each group. The most frequently isolated organisms were chlamydiae, E. coli and Neisseria gonorrheae (around 50% of overall isolated organisms). Due to the lack of significant differences between the two antibiotic combinations, the final choice will depend on potential risks generated by these products.