Évaluation du risque toxique lié à la prévalence de trihalométhanes dans l'eau utilisée pour la dialyse

L'hémodialyse est une thérapeutique réservée aux sujets insuffisants rénaux en attente d'une greffe. Elle permet de recueillir dans un soluté aqueux les déchets que l'organisme ne peut plus évacuer par voie rénale. L'eau nécessaire à la préparation de ce dialysat représente un volume de 90 à 200 litres par séance et par sujet. Elle est obtenue en faisant subir à l'eau du réseau de distribution un traitement complémentaire. Celui-ci comporte en milieu spécialisé une chloration, un adoucissement par résines cationiques, une filtration sur colonne de charbon actif en grains et une osmose inverse.Les trihalométhanes sont probablement les sous-produits de chloration les plus répandus dans les eaux distribuées. Certains parmi eux sont cancérigènes chez l'animal et mutagènes in vitro. Chez l'homme, leurs effets à faibles doses et à long terme restent discutés. Compte tenu des importants volumes d'eau nécessaires à la pratique de l'hémodialyse, il nous a paru intéressant d'observer l'efflcacité du circuit de pré-traitement sur ces composés et d'évaluer les doses auxquelles sont exposés les patients qui bénéficient de cette thérapeutique.Des prélèvements ont été réalisés aux différentes étapes du pré-traitement, de façon hebdomadaire dans deux installations identiques, à la recherche de trihalométhanes. Ils permettent de constater que du chloroforme à une concentration moyenne de 10,5 llgA est encore présent en bout de chaîne. En tenant compte des volumes d'eau utilisés pour chaque séance, ceci signifie que les patients dialysés sont exposés, selon leur âge, à des doses pouvant atteindre jusqu'à dix fois la valeur préconisée dans l'eau potable par l'OMS. La moitié de ce chloroforme est susceptible de passer dans la circulation sanguine et d'exercer un effet toxique. Cette situation peut être corrigée par le choix d'une ressource en eau à charge organique faible, par un renouvellement fréquent du charbon actif et par l'utilisation de membranes en polyamides dans les modules d'osmose inverse. Ces résultats doivent amener à une réflexion plus générale sur la présenoe de sous-produits de la chloration et de micropolluants dans l'eau utilisée en dialyse. Ils doivent également inciter les cliniciens à rechercher, chez les dialysés les plus exposés, d'éventuels effets délétères liés à ces produits.Hemodialysis is an indispensable therapy for patients with chronic renal failure. Two or three times a week and over several years, their blood is dialyzed in an artificial kidney against a dialysis fluid called dialysate.Each time, 90 to 200 liters of this fluid will flow through the apparatus. Before being mixed with the dialysis concentrate, the water will be treated in order to eliminate harmful substances such as aluminum or endotoxins.Trihalomethanes (THM) are probably the most widespread chlorination byproducts of tap water. Most of them are known as carcinogens for animals and mutagens in vitro. Although their hepatotoxicity and nephrotoxicity are obvious after acute intoxication, their effects at low doses on human health have still not been clearly demonstrated.Considering the great amount of water required by hemodialysis patients, we found interested in determining wether the control of these substances by the hospital water treatment plant was efficient. We decided then to analyze weekly and during two months, the tap water of two hemodialysis departments for THM, before and after various forms of treatment. The treatment in both departments was the same and made up of four important stages: chlorination, softening, charcoal filtering and reverse osmosis.THM determinations were conducted using the headspace technique with a gas chromatograph equipped with a split injector and an [sup]63Ni electron capture detector.Our results show that chloroform and dichlorobromomethane were present in tap water. Their respective mean concentration in both department came to 56 µg/l and 5 µg/l. After chlorination and water softening, these figures had moderately but significantly increased. In the first department, thanks to new granular activated carbon, a large part of THM (especially dichlorobromomethane) had been removed. However after seven weeks, this treatment was no longer efficient and only 7% of the influent chloroform and 50% of the dichlorobromomethane could be removed. In the second department, the charcoal filter had already been working for more than one year at the beginning of our study. No decrease of the chloroform concentration had been observed and dichlorobromomethane had significantly increased. 80 to 90% of influent THM were removed after the double stage of reverse osmosis using polyamide membranes. With new granular activated carbon, the dialysis fluid only contains 1 µg/l of chloroform. But after seven weeks or more, it will reach an average of 10.5 g/l of chloroform and 1 µg/l of dichlorobromomethane. These figures are probably underestimated as our study was performed in winter and THM concentrations are less important during that season.These results mean that during a single session, 0.9 to 2.1 mg of chloroform will reach the artificial kidney. Depending on the weight of the patients, this exposure will be equivalent up to 10 times the value recommended by the World Health Organization (WHO) for drinking water.The last part of our study monitored the chloroform concentration in dialysate coming out the artificial kidney during an hemodialysis period. A significant decrease, reaching up to 45% of the influent amount, was observed. This result suggests that some of the chloroform must have crossed the dialysis membrane.According to all these results, we think that it would be of great interest to explore the metabolism of chloroform on hemodialysis patients and to search for eventual toxic effects. Practical advices to people in charge of water treatment plants in hemodialysis department would be to use raw water with low concentrations of humic materials, in order to restrict THM formation. The charcoal filter should be changed more often (probably after 6 or 7 weeks). Alternatively, ways could be found for rapid regeneration of charcoal for THM removal. Finally and according to previous studies, a polyamide membrane should be systematically used for reverse osmosis.Our study could eventually be completed by searching in the dialysis fluid any other chlorination by-products which are responsible to a large extent for tap water mutagenicity

An Adjustable Gas-Mixing Device to Increase Feasibility of In Vitro Culture of Plasmodium falciparum Parasites in the Field

A challenge to conducting high-impact and reproducible studies of the mechanisms of P. falciparum drug resistance, invasion, virulence, and immunity is the lack of robust and sustainable in vitro culture in the field. While the technology exists and is routinely utilized in developed countries, various factors–from cost, to supply, to quality–make it hard to implement in malaria endemic countries. Here, we design and rigorously evaluate an adjustable gas-mixing device for the in vitro culture of P. falciparum parasites in the field to circumvent this challenge. The device accurately replicates the gas concentrations needed to culture laboratory isolates, short-term adapted field isolates, cryopreserved previously non-adapted isolates, as well as to adapt ex vivo isolates to in vitro culture in the field. We also show an advantage over existing alternatives both in cost and in supply. Furthermore, the adjustable nature of the device makes it an ideal tool for many applications in which varied gas concentrations could be critical to culture success. This adjustable gas-mixing device will dramatically improve the feasibility of in vitro culture of Plasmodium falciparum parasites in malaria endemic countries given its numerous advantages

Multiplication rate variation in the human malaria parasite Plasmodium falciparum.

It is important to understand intrinsic variation in asexual blood stage multiplication rates of the most virulent human malaria parasite, Plasmodium falciparum. Here, multiplication rates of long-term laboratory adapted parasite clones and new clinical isolates were measured, using a newly standardised assay of growth from low starting density in replicate parallel cultures with erythrocytes from multiple different donors, across multiple cycles. Multiplication rates of long-term established clones were between 7.6 and 10.5 fold per 48 hours, with clone Dd2 having a higher rate than others (clones 3D7, HB3 and D10). Parasite clone-specific growth was then analysed in co-culture assays with all possible heterologous pairwise combinations. This showed that co-culture of different parasites did not affect their replication rates, indicating that there were no suppressive interactions operating between parasites. Multiplication rates of eleven new clinical isolates were measured after a few weeks of culture, and showed a spectrum of replication rates between 2.3 and 6.0 fold per 48 hours, the entire range being lower than for the long-term laboratory adapted clones. Multiplication rate estimates remained stable over time for several isolates tested repeatedly up to three months after culture initiation, indicating considerable persistence of this important trait variation

Widespread distribution of Plasmodium vivax malaria in Mauritania on the interface of the Maghreb and West Africa.

BACKGROUND: Plasmodium vivax is very rarely seen in West Africa, although specific detection methods are not widely applied in the region, and it is now considered to be absent from North Africa. However, this parasite species has recently been reported to account for most malaria cases in Nouakchott, the capital of Mauritania, which is a large country at the interface of sub-Saharan West Africa and the Maghreb region in northwest Africa. METHODS: To determine the distribution of malaria parasite species throughout Mauritania, malaria cases were sampled in 2012 and 2013 from health facilities in 12 different areas. These sampling sites were located in eight major administrative regions of the country, within different parts of the Sahara and Sahel zones. Blood spots from finger-prick samples of malaria cases were processed to identify parasite DNA by species-specific PCR. RESULTS: Out of 472 malaria cases examined, 163 (34.5 %) had P. vivax alone, 296 (62.7 %) Plasmodium falciparum alone, and 13 (2.8 %) had mixed P. falciparum and P. vivax infection. All cases were negative for Plasmodium malariae and Plasmodium ovale. The parasite species distribution showed a broad spectrum, P. vivax being detected at six of the different sites, in five of the country's major administrative regions (Tiris Zemmour, Tagant, Brakna, Assaba, and the capital Nouakchott). Most cases in Nouakchott were due to P. vivax, although proportions vary significantly among different health facilities in the city. In the northern town of Zouérat, all cases were due to P. vivax, whereas almost all cases in the south of the country were due to P. falciparum. All P. vivax cases tested were Duffy blood group positive. CONCLUSIONS: It is important that P. vivax is recognized to be a widespread cause of malaria in Mauritania, occurring in diverse regions. This should be noted by the World Health Organization, as it has significant implications for diagnosis, treatment and control of malaria in the northwestern part of Africa

Bimodality and alternative equilibria do not help explain long-term patterns in shallow lake chlorophyll-a

Since its inception, the theory of alternative equilibria in shallow lakes has evolved and been applied to an ever wider range of ecological and socioecological systems. The theory posits the existence of two alternative stable states or equilibria, which in shallow lakes are characterised by either clear water with abundant plants or turbid water where phytoplankton dominate. Here, we used data simulations and real-world data sets from Denmark and north-eastern USA (902 lakes in total) to examine the relationship between shallow lake phytoplankton biomass (chlorophyll-a) and nutrient concentrations across a range of timescales. The data simulations demonstrated that three diagnostic tests could reliably identify the presence or absence of alternative equilibria. The real-world data accorded with data simulations where alternative equilibria were absent. Crucially, it was only as the temporal scale of observation increased (>3 years) that a predictable linear relationship between nutrient concentration and chlorophyll-a was evident. Thus, when a longer term perspective is taken, the notion of alternative equilibria is not required to explain the response of chlorophyll-a to nutrient enrichment which questions the utility of the theory for explaining shallow lake response to, and recovery from, eutrophication.C.D.S. and T.A.D. would like to thank June and Derek Sayer for extraordinary support over many years. The authors of this work have been supported by a number of projects over the elephantine gestation period of this manuscript. These include support from the Poul Due Jensen Fonden, Danmarks Frie Forskningsfond Natur og Univers project GREENLAKES (No. 9040-00195B) and the UFM-funded project LTER_DK for Long Term Ecosystem Research in Denmark. In addition, support was provided by The European Union’s Horizon 2020 research and innovation programmes under grant agreement No 869296—The PONDERFUL Project”, TREICLAKE under grant agreement No 951963, and the AQUACOSM project and by the European Commission EU H2020- INFRAIA-project (No. 731065) and AQUACOSMplus (No. 871081). E.J. was also supported by the TÜBITAK outstanding researcher programme2232 (project 118C250) and AnaEE, Denmark. The work of D.G. was funded by the Fourth Period of Programme-oriented Funding, Helmholtz Association of German ResearchCentres, Research Field Earth and Environment.C.D.S. and T.A.D. would like to thank June and Derek Sayer for extraordinary support over many years. The authors of this work have been supported by a number of projects over the elephantine gestation period of this manuscript. These include support from the Poul Due Jensen Fonden, Danmarks Frie Forskningsfond Natur og Univers project GREENLAKES (No. 9040-00195B) and the UFM-funded project LTER_DK for Long Term Ecosystem Research in Denmark. In addition, support was provided by The European Union’s Horizon 2020 research and innovation programmes under grant agreement No 869296—The PONDERFUL Project”, TREICLAKE under grant agreement No 951963, and the AQUACOSM project and by the European Commission EU H2020- INFRAIA-project (No. 731065) and AQUACOSMplus (No. 871081). E.J. was also supported by the TÜBITAK outstanding researcher programme2232 (project 118C250) and AnaEE, Denmark. The work of D.G. was funded by the Fourth Period of Programme-oriented Funding, Helmholtz Association of German ResearchCentres, Research Field Earth and Environment

Inhibitory humoral responses to the Plasmodium falciparum vaccine candidate EBA-175 are independent of the erythrocyte invasion pathway

Plasmodium falciparum utilizes multiple ligand-receptor interactions for invasion. The invasion ligand EBA-175 is being developed as a major blood-stage vaccine candidate. EBA-175 mediates parasite invasion of host erythrocytes in a sialic acid-dependent manner through its binding to the erythrocyte receptor glycophorin A. In this study, we addressed the ability of naturally acquired human antibodies against the EBA-175 RII erythrocyte-binding domain to inhibit parasite invasion of ex vivo isolates, in relationship to the sialic acid dependence of these parasites. We have determined the presence of antibodies to the EBA-175 RII domain by enzyme-linked immunosorbent assay (ELISA) in individuals from areas of Senegal where malaria is endemic with high and low transmission. Using affinity-purified human antibodies to the EBA-175 RII domain from pooled patient plasma, we have measured the invasion pathway as well as the invasion inhibition of clinical isolates from Senegalese patients in ex vivo assays. Our results suggest that naturally acquired anti-EBA-175 RII antibodies significantly inhibit invasion of Senegalese parasites and that these responses can be significantly enhanced through limiting other ligand-receptor interactions. However, the extent of this functional inhibition by EBA-175 antibodies is not associated with the sialic acid dependence of the parasite strain, suggesting that erythrocyte invasion pathway usage by parasite strains is not driven by antibodies targeting the EBA-175/glycophorin A interaction. This work has implications for vaccine design based on the RII domain of EBA-175 in the context of alternative invasion pathways

Whole exome sequencing reveals pathogenic variants in MYO3A, MYO15A and COL9A3 and differential frequencies in ancestral alleles in hearing impairment genes among individuals from Cameroon

There is scarcity of known gene variants of hearing impairment (HI) in African populations. This knowledge deficit is ultimately affecting the development of genetic diagnoses. We used whole exome sequencing to investigate gene variants, pathways of interactive genes and the fractions of ancestral overderived alleles for 159 HI genes among 18 Cameroonian patients with non-syndromic HI (NSHI) and 129 ethnically matched controls. Pathogenic and likely pathogenic (PLP) variants were found in MYO3A, MYO15A and COL9A3, with a resolution rate of 50% (9/18 patients). The study identified significant genetic differentiation in novel population-specific gene variants at FOXD4L2, DHRS2L6, RPL3L and VTN between HI patients and controls. These gene variants are found in functional/co-expressed interactive networks with other known HI-associated genes and in the same pathways with VTN being a hub protein, that is, focal adhesion pathway and regulation of the actin cytoskeleton (P-values <0.05). The results suggest that these novel population-specific gene variants are possible modifiers of the HI phenotypes. We found a high proportion of ancestral allele versus derived at low HI patients-specific minor allele frequency in the range of 0.0–0.1. The results showed a relatively low pickup rate of PLP variants in known genes in this group of Cameroonian patients with NSHI. In addition, findings may signal an evolutionary enrichment of some variants of HI genes in patients, as the result of polygenic adaptation, and suggest the possibility of multigenic influence on the phenotype of congenital HI, which deserves further investigations

Identification of disease-causing genes using microarray data mining and gene ontology

Background: One of the best and most accurate methods for identifying disease-causing genes is monitoring gene expression values in different samples using microarray technology. One of the shortcomings of microarray data is that they provide a small quantity of samples with respect to the number of genes. This problem reduces the classification accuracy of the methods, so gene selection is essential to improve the predictive accuracy and to identify potential marker genes for a disease. Among numerous existing methods for gene selection, support vector machine-based recursive feature elimination (SVMRFE) has become one of the leading methods, but its performance can be reduced because of the small sample size, noisy data and the fact that the method does not remove redundant genes. Methods: We propose a novel framework for gene selection which uses the advantageous features of conventional methods and addresses their weaknesses. In fact, we have combined the Fisher method and SVMRFE to utilize the advantages of a filtering method as well as an embedded method. Furthermore, we have added a redundancy reduction stage to address the weakness of the Fisher method and SVMRFE. In addition to gene expression values, the proposed method uses Gene Ontology which is a reliable source of information on genes. The use of Gene Ontology can compensate, in part, for the limitations of microarrays, such as having a small number of samples and erroneous measurement results. Results: The proposed method has been applied to colon, Diffuse Large B-Cell Lymphoma (DLBCL) and prostate cancer datasets. The empirical results show that our method has improved classification performance in terms of accuracy, sensitivity and specificity. In addition, the study of the molecular function of selected genes strengthened the hypothesis that these genes are involved in the process of cancer growth. Conclusions: The proposed method addresses the weakness of conventional methods by adding a redundancy reduction stage and utilizing Gene Ontology information. It predicts marker genes for colon, DLBCL and prostate cancer with a high accuracy. The predictions made in this study can serve as a list of candidates for subsequent wet-lab verification and might help in the search for a cure for cancers

Temporal changes in Plasmodium falciparum reticulocyte binding protein homolog 2b (PfRh2b) in Senegal and The Gambia.

BACKGROUND: The Plasmodium falciparum reticulocyte binding protein homolog 2b (PfRh2b) is an important P. falciparum merozoite ligand that mediates invasion of erythrocytes by interacting with a chymotrypsin-sensitive "receptor Z". A large deletion polymorphism is found in the c-terminal ectodomain of this protein in many countries around the world, resulting in a truncated, but expressed protein. The varying frequencies by region suggest that there could be region specific immune selection at this locus. Therefore, this study was designed to determine temporal changes in the PfRh2b deletion polymorphism in infected individuals from Thiès (Senegal) and Western Gambia (The Gambia). It was also sought to determine the selective pressures acting at this locus and whether prevalence of the deletion in isolates genotyped by a 24-SNP molecular barcode is linked to background genotype or whether there might be independent selection acting at this locus. METHODS: Infected blood samples were sourced from archives of previous studies conducted between 2007 and 2013 at SLAP clinic in Thiès and from 1984 to 2013 in Western Gambia by MRC Unit at LSHTM, The Gambia. A total of 1380 samples were screened for the dimorphic alleles of the PfRh2b using semi-nested Polymerase Chain Reaction PCR. Samples from Thiès were previously barcoded. RESULTS: In Thiès, a consistent trend of decreasing prevalence of the PfRh2b deletion over time was observed: from 66.54% in 2007 and to 38.1% in 2013. In contrast, in Western Gambia, the frequency of the deletion fluctuated over time; it increased between 1984 and 2005 from (58.04%) to (69.33%) and decreased to 47.47% in 2007. Between 2007 and 2012, the prevalence of this deletion increased significantly from 47.47 to 83.02% and finally declined significantly to 57.94% in 2013. Association between the presence of this deletion and age was found in Thiès, however, not in Western Gambia. For the majority of isolates, the PfRh2b alleles could be tracked with specific 24-SNP barcoded genotype, indicating a lack of independent selection at this locus. CONCLUSION: PfRh2b deletion was found in the two countries with varying prevalence during the study period. However, these temporal and spatial variations could be an obstacle to the implementation of this protein as a potential vaccine candidate