Identifying Recrudescent Plasmodium falciparum in Treated Malaria Patients by Real-time PCR and High Resolution Melt Analysis of Genetic Diversity.

Recurrent parasitaemia during follow up of clinical trials of antimalarial drug efficacy results from either recrudescence of parasites surviving treatment or from parasites newly emerging from the hepatic stage of infection. Nested PCR is used to distinguish these two possibilities and the technique is difficult to standardise. There is risk of both false positive and false negative results, leading to misclassification errors. The high-resolution melt (HRM) assay was developed with pairs of conserved primers targeting blocks of merozoite surface protein 1 and 2 (msp1 and msp2) genes, and polymorphisms were compared using sequence-confirmed Plasmodium falciparum DNA samples from laboratory isolates. In this study, the HRM dissociation profiles of msp1 and msp2 amplicons were determined and validated against parasite isolates from malaria patients. The msp1 and msp2 profiles of both laboratory and clinical isolates were reproducibly differentiated by HRM. These rapid assays are performed in a closed-tube system, and so avoid cross-contamination while increasing throughput, which are two major advantages. The HRM assays offer significant gains in simplicity, speed and interpretation of results, and reduced analysis cost, for studies that require discrimination of parasite clones. Assay performance in large-scale studies utilizing DNA samples derived from filter-paper bloodspots should now be evaluated

Etude ethnobotanique des plantes alimentaires utilisées en période de soudure dans les régions Sud du Mali

La période de soudure est celle pendant laquelle les stocks de céréales sont épuisés et les prochaines récoltes ne sont pas encore prêtes. Au Mali (Afrique de l’Ouest), elle varie d’août à septembre. Pendant cette période, les communautés locales utilisent les plantes sauvages comme aliment. L’objectif de la présente étude était de recenser les plantes sauvages alimentaires consommées dans les régions Sud du Mali (Kayes, Koulikoro, Sikasso et Ségou) pendant cette période. Des enquêtes ethnobotaniques utilisant les méthodes du focus group, d’enquête individuelle et d’interviews semi-structurées ont été menées. Au total, 454 personnes dont 338 hommes et 116 femmes soit respectivement 74,4% et 25,6% ont été interrogées. Les jeunes (15 - 30 ans), cultivateurs et Bambara étaient les plus nombreux. 87 plantes alimentaires appartenant à 44 familles ont été identifiées; les fruits de 56 plantes (62,2%), les feuilles de 43 plantes (47,8%) et les graines de 10 plantes (11,5%) sont consommées comme aliments pendant les périodes de soudure. Parkia biglobosa, Adansonia digitata et Vitellaria paradoxa sont les plantes les plus utilisées soit seules ou dans des plats comme additifs. Certaines de ces plantes servent également comme médicament et comme source de revenus pour les communautés locales.© 2016 International Formulae Group. All rights reserved.Mots clés: Plantes sauvages alimentaires, enquête ethnobotanique, période de soudure, Sud-MaliEnglish Title: Ethnobotanical study of food plants used in the period of soldering in the southern regions of MaliEnglish AbstractThe lean period is the one during which grain stocks are depleted and the next harvest is not yet ready. In Mali (West Africa), it varies from August to September. During this period, local communities use wild plants as food. The objective of this study was to identify wild food plants consumed in the southern regions of Mali (Kayes, Koulikoro, Segou and Sikasso) during this period. Ethnobotanical surveys using focus group methods, individual survey and semi-structured interviews were conducted. A Total of 454 respondents including 338 (74.4%) men and 116 (25.6%) women were interviewed. Yong farmers (15-30 years) from Bambara ethic group were the most numerous. 87 food plants belonging to 44 families have been identified; the fruit of 56 (62.2%) plants, the leaves of 43 (47.8%), and the seeds of 10 plants (11.5%) were consumed as food during lean periods. Parkia biglobosa, Adansonia digitata and Vitellaria paradoxa are the most used plants either alone or in dishes such as additives. Some of these plants are also used as a medicine and as a source of income for local communities.© 2016 International Formulae Group. All rights reserved.Keywords: wild food plants, ethnobotany survey, lean period, South Mal

MORINGA OLEIFERA LEAF POWDER FOR TYPE 2 DIABETES: A PILOT CLINICAL TRIAL

Background: Moringa oleifera Lam. (Moringaceae) leaves are commonly used for diabetes in Mali. This pilot clinical study aimed to evaluate its effect on post-prandial blood glucose in preparation for a larger trial. Methods: Diabetic patients and non-diabetic healthy volunteers (35 each) were asked to fast for 13 hours on three occasions. Blood glucose was measured before and after eating 100g of white bread (at 30, 60, 90, 120, 150 and 180 minutes). On their second and third study visits, they were given 1g and 2g respectively, of M. oleifera leaf powder, 30 minutes after eating the bread.  The mean paired reduction in blood glucose at each time interval and the incremental area under the curve were calculated. Results: Ingestion of Moringa powder had no effect on blood glucose in non-diabetic participants, but in diabetic patients, it lowered blood glucose at 90 minutes. There was a trend towards lower incremental area under the curve when diabetic patients took 2g of Moringa. No side-effects were reported by any participant. Conclusions: Moringa oleifera leaf powder reduced post-prandial glycaemia in diabetic patients. A larger study is needed to define the optimal dose and to assess whether this translates into longer-term benefits

Anesthesie au cours des ventriculocisternostomies au Mali : Une serie de 31 cas

Introduction La ventriculocisternostomie est une technique moderne du traitement des hydrocéphalies. Elle est peu fréquente en Afrique. Objectif:  Décrire la prise en charge anesthésique et l’évolution des patients opérés par ventriculocisternostomie. Patients et Méthode: Etude de cohorte prospective de 15 mois de janvier 2014 au 31 mars 2015. La saisie et l’analyse des données ont été effectuées par Microsoft word 2010 Epi info 3.5.3.fr. Résultats:  Les nourrissons prédominaient avec un sexe ratio de 1,81 en faveur du sexe masculin. Un antécédent de méningite ou d’infection respiratoire à répétition a été retrouvé chez 10 patients (32,2%). L’indication de la ventriculocisternostomie était une hydrocéphalie chez 30 patients (96, 8%). La classe ASA était II chez 19 patients (61,3%). L’intubation était prévue difficile chez 28 patients (90,3%).Tous les patients ont été opérés sous anesthésie générale. Une antibioprophylaxie a été faite chez tous les patients. La tachycardie isolée a été le seul évènement indésirable per opératoire observé chez 13 patients (41,9%). La durée de la chirurgie était de 62, 25 ± 20,9 minutes celle de l’anesthésie était de 93,5 ± 25,4 minutes. En postopératoire, une complication a été observée chez 7 patients (22,6%). Il s’agissait d’une méningite chez 3 patients (42,9%), d’une souffrance cérébrale, d’une obstruction de la stomie, d’un abcès cérébral et une paralysie du nerf III dans 14,3% chacune (1 patient). L’évolution était favorable chez 29 patients (93,5%). La durée médiane d’hospitalisation était de 3 jours. Conclusion:  Au Mali, la prise en charge anesthésique au cours de la ventriculocisternostomie s’adresse à une population pédiatrique avec un terrain précaire.   English title: Anesthesia during endoscopic third ventriculostomy in Mali: A series of 31 cases Introduction: Endoscopic Third Ventriculostomy (ETV) is a modern technique for the treatment of hydrocephalus. It is uncommon in Africa. Objective To describe ananesthesic management and the outcome of patients operated on by ETV. Patients and Methods Prospective cohort study over 15 months to January 2014 at 31 march 2015. The data entry and analysis were done by word office, Epi info 3.5.3.fr. Results: Infants predominated with a sex ratio of 1.81 in favor of men. A history of meningitis or recurrent respiratory infection was found in 10 patients (32.2%). The indication of ETV was hydrocephalus in 30 patients (96.8%). The ASA class was II in 19 patients (61.3%).  Intubation was expected to be difficult in 28 patients (90.3%). All patients were operated on under general anesthesia. Antibiotic  prophylaxis was done in all patients. Isolated tachycardia was the only peroperative adverse event observed in 13 patients (41.9%). The duration of the surgery was 62.25 ± 20.9 minutes that of the anesthesia was 93.5 ± 25.4 minutes. Postoperatively, a complication was observed in 7 patients (22.6%). It was meningitis in 3 patients (42.9%), brain pain, obstruction of the stoma, brain abscess and nerve III paralysis in 14.3% each (1 patient). The outcome was favorable in 29 patients (93.5%). The median hospital stay was 3 days. Conclusion:  In Mali, anesthetic management during ETV is aimed at a pediatric population with precarious terrain

Different Plasmodium falciparum clearance times in two Malian villages following artesunate monotherapy.

BACKGROUND: Artemisinin resistance described as increased parasite clearance time (PCT) is rare in Africa. More sensitive methods such as qPCR might better characterize the clearance phenotype in sub-Saharan Africa. METHODS: PCT is explored in Mali using light microscopy and qPCR after artesunate for uncomplicated malaria. In two villages, patients were followed for 28 days. Blood smears and spots were collected respectively for microscopy and qPCR. Parasitemia slope half-life was calculated after microscopy. Patient residual parasitemia were measured by qPCR. RESULTS: Uncorrected adequate clinical and parasitological responses (ACPR) observed in Faladje and Bougoula-Hameau were 78% and 92%, respectively (p=0.01). This reached 100% for both after molecular correction. Proportions of 24H microscopy positive patients in Faladje and Bougoula-Hameau were 97.2% and 72%, respectively (p<0.0001). Slope half-life was 2.8h in Faladje vs 2H in Bougoula-Hameau (p<0.001) and Proportions of 72H patients with residual parasitemia were 68.5% and 40% in Faladje and Bougoula-Hameau, respectively (p=0.003). The mean residual parasitemia was 2.9 in Faladje vs. 0.008 in Bougoula-Hameau (p=0.002). Although artesunate is efficacious in Mali, the longer parasite clearance time with submicroscopic parasitemia observed may represent early signs of developing P. falciparum resistance to artemisinins

Concordance of vaccination status and associated factors with incomplete vaccination: a household survey in the health district of Segou, Mali, 2019

Introduction:&nbsp;the region of Segou recorded 36.8% of children were incompletely vaccinated in 2018. In 2019, the district of Segou was one of the districts with the lowest vaccination coverage in the region, with 85.1% coverage for the three doses of the pentavalent vaccine and 85.4% for the measles vaccine. This study was initiated to better understand this low vaccination coverage, in the absence of specific studies on vaccination coverage in the district of Segou. Methods:&nbsp;a prospective cross-sectional study was conducted from May to August 2020 with 30 clusters. We performed Kappa coefficient, bivariate, and multiple logistic regression analysis. Results:&nbsp;findings showed that 18.46% (101/547) [15.44-21.93] of children were incompletely vaccinated. Mothers correctly reported the vaccination status of their children in 67.30% of cases (Kappa coefficient). Uneducated (OR[IC95%]=2.13[1.30-3.50]), living in rural area (OR[IC95%]=2.07[1.23-3.47]), lack of knowledge of Expanded Program on Immunization (EPI) target diseases (OR[IC95%]=2.37[1.52-3.68]), lack of knowledge of vaccination schedule (OR[IC95%]=3.33[1.90-5.81]) and lack of knowledge of the importance of vaccination (OR[IC95%]=3.6[2.35-6.32]) were associated with incomplete vaccination. In multivariate analysis, uneducated (ORa[IC95%&gt;]=1.68[1.004-2.810]) and lack of knowledge of the importance of vaccination were associated with incomplete vaccination (ORa[IC95%]=3.40[2.049-5.649]). Conclusion:&nbsp;findings showed a good concordance of the vaccination status. Living in a rural area, no education, lack of the knowledge of EPI target diseases, lack of the knowledge of vaccination schedule and lack of knowledge of the importance of vaccination were associated with incomplete vaccination

Quinine Treatment Selects the pfnhe-1 ms4760-1 Polymorphism in Malian Patients with Falciparum Malaria

Background. The mechanism of Plasmodium falciparum resistance to quinine is not known. In vitro quantitative trait loci mapping suggests involvement of a predicted P. falciparum sodium-hydrogen exchanger (pfnhe-1) on chromosome 13. Methods. We conducted prospective quinine efficacy studies in 2 villages, Kolle and Faladie, Mali. Cases of clinical malaria requiring intravenous therapy were treated with standard doses of quinine and followed for 28 days. Treatment outcomes were classified using modified World Health Organization protocols. Molecular markers of parasite polymorphisms were used to distinguish recrudescent parasites from new infections. The prevalence of pfnhe-1 ms4760-1 among parasites before versus after quinine treatment was determined by direct sequencing. Results. Overall, 163 patients were enrolled and successfully followed. Without molecular correction, the mean adequate clinical and parasitological response (ACPR) was 50.3% (n = 163). After polymerase chain reaction correction to account for new infections, the corrected ACPR was 100%. The prevalence of ms4760-1 increased significantly, from 26.2% (n = 107) before quinine treatment to 46.3% (n = 54) after therapy (P = .01). In a control sulfadoxine-pyrimethamine study, the prevalence of ms4760-1 was similar before and after treatment. Conclusions. This study supports a role for pfnhe-1 in decreased susceptibility of P. falciparum to quinine in the field.Howard Hughes Medical Institute [55005502]; Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health; European and Developing Countries Clinical Trials Partnership [EDCTP IP_07_31060_002]info:eu-repo/semantics/publishedVersio

Persistent Submicroscopic Plasmodium falciparum Parasitemia 72 Hours after Treatment with Artemether-Lumefantrine Predicts 42-Day Treatment Failure in Mali and Burkina Faso.

A recent randomized controlled trial, the WANECAM (West African Network for Clinical Trials of Antimalarial Drugs) trial, conducted at seven centers in West Africa, found that artemether-lumefantrine, artesunate-amodiaquine, pyronaridine-artesunate, and dihydroartemisinin-piperaquine all displayed good efficacy. However, artemether-lumefantrine was associated with a shorter interval between clinical episodes than the other regimens. In a further comparison of these therapies, we identified cases of persisting submicroscopic parasitemia by quantitative PCR (qPCR) at 72 h posttreatment among WANECAM participants from 5 sites in Mali and Burkina Faso, and we compared treatment outcomes for this group to those with complete parasite clearance by 72 h. Among 552 evaluable patients, 17.7% had qPCR-detectable parasitemia at 72 h during their first treatment episode. This proportion varied among sites, reflecting differences in malaria transmission intensity, but did not differ among pooled drug treatment groups. However, patients who received artemether-lumefantrine and were qPCR positive at 72 h were significantly more likely to have microscopically detectable recurrent Plasmodium falciparum parasitemia by day 42 than those receiving other regimens and experienced, on average, a shorter interval before the next clinical episode. Haplotypes of pfcrt and pfmdr1 were also evaluated in persisting parasites. These data identify a possible threat to the parasitological efficacy of artemether-lumefantrine in West Africa, over a decade since it was first introduced on a large scale