Molecular Markers of In Vivo Plasmodium vivax Resistance to Amodiaquine Plus Sulfadoxine-Pyrimethamine: Mutations in pvdhfr and pvmdr1

Background. Molecular markers for sulfadoxine-pyrimethamine (SP) resistance in Plasmodium vivax have been reported. However, data on the molecular correlates involved in the development of resistance to 4-aminoquinolines and their association with the in vivo treatment response are scarce. Methods. We assessed pvdhfr (F57L/I, S58R, T61M, S117T/N, and I173F/L) and pvmdr1 (Y976F and F1076L) mutations in 94 patients who received amodiaquine (AQ) plus SP in Papua New Guinea (PNG). We then investigated the association between parasite genotype and treatment response. Results. The treatment failure (TF) rate reached 13%. Polymorphisms in pvdhfr F57L, S58R, T61M, and S117T/N and in pvmdr1 Y976F were detected in 60%, 67%, 20%, 40%, and 39% of the samples, respectively. The single mutant pvdhfr 57 showed the strongest association with TF (odds ratio [OR], 9.04; P=.01). The combined presence of the quadruple mutant pvdhfr 57L+58R+61M+117T and pvmdr1 mutation 976F was the best predictor of TF (OR, 8.56; P=.01). The difference in TF rates between sites was reflected in the genetic drug-resistance profile of the respective parasites. Conclusions. The present study identified a new molecular marker in pvmdr1 that is associated with the in vivo response to AQ+SP. We suggest suitable marker sets with which to monitor P. vivax resistance against AQ+SP in countries where these drugs are use

A systematic review and meta-analysis of evidence for correlation between molecular markers of parasite resistance and treatment outcome in falciparum malaria

<p>Abstract</p> <p>Background</p> <p>An assessment of the correlation between anti-malarial treatment outcome and molecular markers would improve the early detection and monitoring of drug resistance by <it>Plasmodium falciparum</it>. The purpose of this systematic review was to determine the risk of treatment failure associated with specific polymorphisms in the parasite genome or gene copy number.</p> <p>Methods</p> <p>Clinical studies of non-severe malaria reporting on target genetic markers (SNPs for <it>pfmdr1</it>, <it>pfcrt</it>, <it>dhfr</it>, <it>dhps</it>, gene copy number for <it>pfmdr1</it>) providing complete information on inclusion criteria, outcome, follow up and genotyping, were included. Three investigators independently extracted data from articles. Results were stratified by gene, codon, drug and duration of follow-up. For each study and aggregate data the random effect odds ratio (OR) with 95%CIs was estimated and presented as Forest plots. An OR with a lower 95^thconfidence interval > 1 was considered consistent with a failure being associated to a given gene mutation.</p> <p>Results</p> <p>92 studies were eligible among the selection from computerized search, with information on <it>pfcrt </it>(25/159 studies), <it>pfmdr1 </it>(29/236 studies), <it>dhfr </it>(18/373 studies), <it>dhps </it>(20/195 studies). The risk of therapeutic failure after chloroquine was increased by the presence of <it>pfcrt </it>K76T (Day 28, OR = 7.2 [95%CI: 4.5–11.5]), <it>pfmdr1 </it>N86Y was associated with both chloroquine (Day 28, OR = 1.8 [95%CI: 1.3–2.4]) and amodiaquine failures (OR = 5.4 [95%CI: 2.6–11.3, p < 0.001]). For sulphadoxine-pyrimethamine the <it>dhfr </it>single (S108N) (Day 28, OR = 3.5 [95%CI: 1.9–6.3]) and triple mutants (S108N, N51I, C59R) (Day 28, OR = 3.1 [95%CI: 2.0–4.9]) and <it>dhfr</it>-<it>dhps </it>quintuple mutants (Day 28, OR = 5.2 [95%CI: 3.2–8.8]) also increased the risk of treatment failure. Increased <it>pfmdr1 </it>copy number was correlated with treatment failure following mefloquine (OR = 8.6 [95%CI: 3.3–22.9]).</p> <p>Conclusion</p> <p>When applying the selection procedure for comparative analysis, few studies fulfilled all inclusion criteria compared to the large number of papers identified, but heterogeneity was limited. Genetic molecular markers were related to an increased risk of therapeutic failure. Guidelines are discussed and a checklist for further studies is proposed.</p

Détection des polymorphismes nucléotidiques pour la surveillance des résistances de Plasmodium falciparum

LYON1-BU Santé (693882101) / SudocSudocFranceF

Déterminisme génomique et post-génomique de la résistance de Plasmodium falciparum aux antipaludéens

L'impact médical du paludisme est la conséquence de la résistance de P.falciparum à la plupart des antipaludéens. L'étude de la résistance repose sur les tests in vivo et les tests in vitro. Leurs contraintes respectives limitent leur capacité à produire des résultats rapides. L'objectif de ce travail est de développer une méthode permettant de distinguer les espèces plasmodiales, de mesurer la sensibilité du parasite et de détecter les mutations. La PCR en temps réel nous a permis de déterminer la sensibilité à la chloroquine de souches de culture, la charge parasitaire de patients traités par antipaludéens et d'identifier les 4 espèces plasmodiales. L'utilisation de sondes d'hybridation a permis d'établir le polymorphisme de 217 échantillons collectés sur anticoagulant ou sur papier buvard pour 3 loci du gène pfcrt et 5 loci du gène pfmdr1. L'utilisation de cette méthode doit permettre d'établir une base comportant les données essentielles à la définition rapide d'une politique thérapeutique de première et deuxième intention.LYON1-BU Santé (693882101) / SudocPARIS-BIUP (751062107) / SudocSudocFranceF

Characterisation of polyglutamylases in trypanosomatids

International audienceMicrotubules are subject to post-translational modifications, which are thought to have crucial roles in the function of complex microtubule-based organelles. Among these, polyglutamylation was relatively recently discovered, and was related to centrosome stability, axonemal maintenance and mobility, and neurite outgrowth. In trypanosomatids, parasitic protozoa where microtubules constitute the essential component of the cytoskeleton, the function of polyglutamylated microtubules is unknown. Here, in order to better understand the role of this conserved but highly divergent post-translational modification, we characterised glutamylation and putative polyglutamylases in these parasites. We showed that microtubules are intensely glutamylated in all stages of the cell cycle, including interphase. Moreover, a cell cycle-dependent gradient of glutamylation was observed along the cell anteroposterior axis, which might be related to active growth of the microtubule 'corset' during the cell cycle. We also identified two putative polyglutamylase proteins (among seven analysed here) which appeared to be clearly and directly involved in microtubule polyglutamylation in in vitro activity assays. Paradoxically, in view of the importance of tubulins and of their extensive glutamylation in these organisms, RNA interference-based knockdown of all these proteins had no effect on cell growth, suggesting either functional redundancy or, more likely, subtle roles such as function modulation or interaction with protein partners

Advantages and limits of real-time PCR assay and PCR-restriction fragment length polymorphism for the identification of cutaneous Leishmania species in Tunisia.

International audienceCutaneous leishmaniasis (CL), a public health problem in Tunisia, is associated to three species: Leishmania (L.) infantum, L. major and L. killicki. Accurate and sensitive procedures for the diagnostic of Leishmania infection and for species identification are required to enable adequate treatment and appropriate control measures. Several PCR-methods are applied for the diagnosis and the identification of Leishmania parasites such as PCR-restriction fragment length polymorphism (PCR-RFLP), DNA sequencing, hybridization probes and real-time PCR (RT-PCR). In this study, PCR-RFLP and RT-PCR were performed on skin scrapings from 27 patients with confirmed CL by microscopic examination, in order to compare their usefulness and efficiency for identification of Leishmania species in routine diagnostic laboratories. Identification of Leishmania species was successfully achieved in 96.3% and 81.5% respectively. Agreement between using internal transcribed spacer 1 (ITS1)-PCR-RFLP and kDNA-RT-PCR assays was 70% (19/27). Characterization problems using RT-PCR were mainly due to the difficulties in analyzing the melting temperatures. ITS1-PCR-RFLP and kDNA-RT-PCR presented an interesting alternative to conventional methods for the identification of Leishmania parasites from clinical samples. Both PCR assays can be used in a routine diagnostic, however, further prospective studies including largest sampling, are required to determine their performances in a routine use

Marine protected areas in fisheries management: Synthesis on the state of the art

83 pages☞The international context shows that many MPAs overall objectives, identified in international Conventions, werenot achieved in 2012 and consequently were repeated: “By 2020, at least … 10 per cent of coastal and marine areas,especially areas of particular importance for biodiversity and ecosystem services, are conserved through effectivelyand equitably managed, ecologically representative and well-connected systems of protected areas and other effectivearea-based conservation measures, and integrated into the wider landscapes and seascapes” (CBD Strategic Plan2011-2020, Goal C, Target 11, CBD CoP10, Decision X2, Nagoya 2010).The present document aims to present the results of ananalysis of the world literature, draw lessons from learnedexperiences, and share relevant recommendationswith regard to situations experienced in the SRFC zone onthe best ways and means to use MPAs as fisheries managementtools.Through the future programs of the SRFC and States ofthe region, the challenges will be the best utilization ofthe recommendations contained therein in order to, onthe one hand, improve the relations between the governancesof coastal management, fisheries, and MPAs and,on the other hand, to develop solutions enabling a betterintegration of the concerns of fisheries in the managementof MPAs and of the MPAs in the management of fisheries..The work, conducted by a group of experts from the Universityof Brest (UMR Amure) – Agrocampus from Rennes(fisheries) – the IUCN-CEM Fisheries Expert Group coordinatedby BRL Engineering and EBCD and supported bymany international contributions, has highlighted a seriesof illustrative examples and case-studies on MPAs inthree main documents:■ A “technical report“ in french representing the State ofthe Art made up with 4 volumes dealing with “Governance“,“socio-economic and bio-economic modelling“,“bio-ecological and bioecological modelling“, and “Elementof Reflection for the SRFC and its partners“ in supportto the regional Workshop;■ The present “Synthesis Report“ ;■ A ten-page note summarizing the outcomes of thestudy (www.spcsrp.org).These works as well as the supporting document andthoughts they produced enabled the holding of a regionalworkshop in December 2011 in Dakar during which theoutcomes of the review were presented. Exchanges withseveral representatives of fisheries and environmentalinstitutions and other stakeholders throughout the subregion(fishermen, funders, scientists, etc.,) led to confirmationof the relevance of the supporting documents ofthe Workshop and specified further the recommendationspresented in this document.☞Three main questions are asked in this study:■ Are MPAs tools preferable to conventional fishery managementtools when it comes to promoting :(1) theparticular protection of certain areas, habitats or, species;(ii) the, allocation of resources; and (iii) the participationof communities in the decision-making process?■ What lessons have been learned on the effects of MPAson fishing and on the tools associated to the measurementof these effects?■ Are there any lessons to be drawn from the internationalexperience on governance of MPAs in relation to fishingthat could lead to improvements in management

Les aires marines protégées dans la gestion des pêches.: Synthèse de l' état de l'art.

83 pagesLe contexte international indique que de nombreux objectifs globaux des AMP n’ont pas pu être atteint en 2012comme prévu dans les conventions internationales et sont donc reconduits : « En 2020, au moins… 10% des zones côtièreset marines revêtant une importance particulière… seront conservées par des systèmes d’AMP, gérés eficacement,écologiquement représentatifs et correctement connectés, ainsi que par d’autres mesures de conservationspatiales efficaces, et intégrés dans des paysages… marins plus vastes (CdP 10, 2010, Nagoya, Décision X/2, Plan Stratégique2011-2020).Le présent document vise à disposer d’une analyse de lalittérature mondiale afin de tirer les enseignements etpartager des recommandations pertinentes aux situationsrencontrées dans la région CSRP sur les meilleursvoies et moyens d’utiliser des AMP gérées en tant qu’outilsd’aménagement des pêches.Les enjeux à travers les programmes futurs de la CSRP etdes Etats de la région seront de valoriser ces travaux afind’améliorer les relations entre les gouvernances de la gestioncôtière, des pêches, et des AMP mais aussi de développerdes solutions permettant de mieux intégrer lespréoccupations de la pêche dans la gestion des AMP etdes AMP dans la gestion des pêches.Le travail fourni par un groupe d’expert, composé de l’universitéde Brest (UMR Amure)-Agrocampus (halieutique)-UICN-EBCD, coordonné par BRL ingénierie et appuyé parde nombreuses contributions internationales, a mis en valeurune série d’exemples illustratifs et de cas d’études ausein de 3 documents principaux :■ Un “rapport technique“ représentant le corps de l’étatde l’art constitué en 4 volumes autour d’un volet “Gouvernance“,d’un volet “socioéconomie et modélisationbioéconomique“, d’un volet “bioécologie et modélisationbiologique“ et d’un volet “Elément de réflexion pour laCSRP et ses partenaires» en appui à l’Atelier régional“ ;■ Le présent “rapport de synthèse“ ;■ Une note de 10 pages qui résume les conclusions del’étude (www.spcsrp.org).Ces travaux ainsi que le document d’appui et de réflexionsur les recommandations ont permis la réalisation d’unAtelier régional en décembre 2011 à Dakar durant lequelont été présentés les résultats. Les échanges avec de nombreuxreprésentants d’institutions des pêches et de l’environnementet d’acteurs de toute la sous-région(pêcheurs, financeurs, scientifiques,…) ont permis de validerla pertinence des documents d’appui à l’Atelier et depréciserles recommandations qui sont présentées dans ce document.☞Trois grandes questions sont abordées dans cet ouvrage :■ Est-ce que les AMP sont des instruments préférablesaux instruments conventionnels de gestion despêches lorsqu’il s’agit de promouvoir la protectionparticulière de certaines zones, habitats, espèces,l’allocation des ressources, la participation descommunautés à la décision ?■ Quels sont les enseignements sur les effets, les outilsassociés à la mesure des effets des AMP sur la pêche ?■ Existe-t-il des enseignements à tirer de l’expérienceinternationale en matière de gouvernance des AMPen lien avec la pêche et permettant des améliorationsdans la gestion

Good Efficacy of Artemether-Lumefantrine for Uncomplicated Falciparum Malaria in Eastern Sumba, East Nusatenggara, Indonesia

Aim: to evaluate the safety and efficacy of a fixed combination of artemether-lumefantrine for likely use against failures of the artesunate-amodiaquine first line therapy. Methods: the study was an open label single arm uncontrolled trial. we evaluated the safety and efficacy of standard artemether-lumefantrine therapy in 59 subjects with uncomplicated malaria caused by Plasmodium falciparum on the island of Sumba in eastern Indonesia. No treatment failures occurred up to day 35. One subject had recurrent parasitemia on day 42 that showed a genotype consistent with recrudescence. The efficacy of this therapy was thus estimated to be 98.3% (95% confidence interval=95%-100%). Descriptive analysis was done using the SPSS 12 computer software. Results: two hundred and thirteen P. falciparum patients met the inclusion criteria for in vivo efficacy study, 79 were given artemether-lumefantrine and 134 were treated under another protocol with artesunate-amodiaquine or sulfadoxine-pyrimethamine. Among 79 eligible subjects, 59 successfully completed the 42-day test. As expected, the mean PCT was longer than the mean FCT, i.e. 1.34+-0.67 (95% CI 1.21–1.47) and 1.05+-0.05 (95% CI 0.95–1.15) days, respectively. On day 3 of treatment, both fever and asexual stage of P. falciparum disappeared in all subjects. Observation until Day 35 showed that all of the 59 subjects treated with artemether-lumefantrine were cured. Conclusion: the findings of this uncontrolled study suggest good safety and efficacy of artemether-lumefantrine for treatment of uncomplicated falciparum malaria on Sumba Island in the Lesser Sundas archipelago of eastern Indonesia