HIV enteropathy and 'Slim disease': Historical and current perspectives.

OBJECTIVES: Chronic diarrhoea and severe wasting associated with HIV infection were first described in East African patients as slim disease (SD) in 1985. The main histological features are flattening of the villi (villous atrophy) and crypt hyperplasia (elongated crypts), i.e., HIV enteropathy (HIVE). Selective loss of mucosal clusters of differentiation 4 (CD4)+ T helper (Th)17+ lymphocytes is the immunological hallmark of HIVE. This review explores (i) the historical background of HIVE and SD, (ii) the relationship between gut mucosal CD4+ Th17+ and intestinal-resident intra-epithelial gamma delta (IRIE) T lymphocytes in pathogenesis of HIVE, (iii) the role of cytokines in regulation of intestinal epithelial proliferation, and (iv) the role of antiretroviral therapy in HIVE. METHODS: Recent studies have highlighted the role of IRIE T lymphocytes, mostly CD8+, in regulating gut epithelial regeneration. CD4+Th17+ and IRIE T cells are necessary to maintain intestinal barrier integrity and mucosal antimicrobial immune defence. However, the immunological cross-talk between such lymphocyte sub-sets culminating in HIVE is uncertain. We undertook a narrative literature review under the headings 'HIVE', 'SD', and 'Highly active antiretroviral therapy (HAART). Relevant studies were located using the electronic search engines Google Scholar and PubMed from 1984 to 2022. RESULTS: Depletion of Th17+ cells in the lamina propria, attributed to low-level viraemia, is accompanied by concomitant increase in the density of gut mucosal IRIE T lymphocytes in AIDS. The latter express a broad range of cytokines (interferon-gamma, tumor necrosis factor-alpha, interleukin-17) and chemokines e.g., keratinocyte growth factor, post exposure to HIV-infected cells. Keratinocyte growth factor induces epithelial proliferation mainly in the crypts, leading to functional immaturity of enterocytes, reduced gut absorptive surface area and malabsorption in animal experiments. Of note, the absence of IRIE T cells is associated with a reduction in epithelial cell turnover. Patients with HIVE receiving early HAART show enhanced expression of mucosal repair genes and improvement of gut symptoms. CONCLUSION: Multiple lines of enquiry suggest HIVE is directly related to HIV infection and is a consequence of perturbations in mucosal CD4+Th17+ and IRIE T lymphocytes. The pathological result is enterocyte immaturity and dysfunction. SD whose main features are malabsorption, diarrhoea and weight loss, is a severe clinical expression of HIVE. A better understanding of immuno-pathogenesis of HIVE opens a window of opportunity for the potential use of immunotherapy in HIV disease and other T cell-mediated enteropathies.

Randomized controlled trial of home-based vs. hospital-based pulmonary rehabilitation in post COVID-19 patients.

BACKGROUND: Between 30% and 60% of people who have been infected with COVID-19 still had symptoms 3 months after the start of the disease. Prescribing a pulmonary rehabilitation program in rehabilitation facilities for post COVID-19 patients could help alleviate the symptoms. However, rehabilitation facilities known to provide good quality care to COVID-19 patients and all other patients, could become saturated by the rise in cases. Home-based rehabilitation is a potential solution that could be sustainable in the long term to avoid this saturation and/or a very long waiting list for patients. AIM: The aim of this study was to investigate whether home-based rehabilitation would have similar effects compared to inpatient rehabilitation on physical and respiratory variables in post COVID-19 patients. DESIGN: This is a randomized controlled trial. SETTING: Pulmonary rehabilitation facility. POPULATION: Seventeen post COVID-19 patients were randomized into two groups: inpatient pulmonary rehabilitation (IPR) or home-based pulmonary rehabilitation (HPR). METHODS: The comparison of the two rehabilitation methods relied on questionnaires, physical tests and the evaluation of several respiratory parameters. A 2-way Analysis of Variance (ANOVA) with repeated measures was performed to assess the effects of time (pre- vs. post-rehabilitation), group (IPR vs. HPR) and their interaction for all parameters. RESULTS: The main result of this study is that distance covered in the 6MWT (6MWD) shows significant improvements, between pre- and postrehabilitation program in both groups (+95 m in IPR group vs.+72 m in HPR group, P<0.001) with no significant interaction between time and group (P=0.420). CONCLUSIONS: These results suggest that home-based pulmonary rehabilitation would be as efficient as IPR to decrease physical sequelae in post COVID-19 patients. CLINICAL REHABILITATION IMPACT: It is possible to suggest both methods (home-based rehabilitation or inpatient pulmonary rehabilitation) according to the specificities of each patient and depending on hospital saturation. The choice of one or the other method should not be made to the detriment of the patient.

A novel bioactive glass/graphene oxide composite coating for a polyether ether ketone-based dental implant.

Polyether ether ketone (PEEK) is a biocompatible material that lacks antimicrobial activity and bioactivity; therefore, is not appropriate for use as a dental implant. To overcome these deficiencies, a novel composite coating of bioactive glass and graphene oxide was prepared. PEEK discs were polished, cleaned, and the surface treated with sulfuric acid for 15 min. The composite coating consisted of bioactive glass produced by the sol-gel route and doped with 0.75 wt% graphene oxide. X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy-energy dispersive spectroscopy analyses were employed to characterize the composite coating, and the coating adhesion strength quantified using a pull-off test. Cytotoxicity was assessed using osteoblast-like cells and gingival fibroblasts. The wettability of the coated and non-coated samples was determined by optical contact angle assessment, and bioactivity was assessed by immersion in simulated body fluid. The results revealed that the bioactive glass/graphene oxide composite coating, approximately 7 µm thick, was transparent, homogenous with few microcracks and microporosities, but adhered strongly and was not cytotoxic to either osteoblast-like cells or gingival fibroblasts. The wettability of the PEEK sample was increased to <20° after coating with the composite, and apatite formation was detectable after 14 days of immersion in simulated body fluid.

A method for multiplexed full-length single-molecule sequencing of the human mitochondrial genome.

Methods to reconstruct the mitochondrial DNA (mtDNA) sequence using short-read sequencing come with an inherent bias due to amplification and mapping. They can fail to determine the phase of variants, to capture multiple deletions and to cover the mitochondrial genome evenly. Here we describe a method to target, multiplex and sequence at high coverage full-length human mitochondrial genomes as native single-molecules, utilizing the RNA-guided DNA endonuclease Cas9. Combining Cas9 induced breaks, that define the mtDNA beginning and end of the sequencing reads, as barcodes, we achieve high demultiplexing specificity and delineation of the full-length of the mtDNA, regardless of the structural variant pattern. The long-read sequencing data is analysed with a pipeline where our custom-developed software, baldur, efficiently detects single nucleotide heteroplasmy to below 1%, physically determines phase and can accurately disentangle complex deletions. Our workflow is a tool for studying mtDNA variation and will accelerate mitochondrial research.

Prevalence of malaria among febrile patients and assessment of efficacy of artemether-lumefantrine and artesunate-amodiaquine for uncomplicated malaria in Dolisie, Republic of the Congo.

BACKGROUND: In the Republic of the Congo, malaria represents a major public health problem affecting all age groups. A regular surveillance of the current efficacy of first-line anti-malarial drugs is required in the face of possible emergence and spread of artemisinin-resistant Plasmodium falciparum strains in Africa. The purpose of this study was to determine the prevalence of malaria among febrile patients of all ages and assess the efficacy of artemether-lumefantrine (AL) and artesunate-amodiaquine (ASAQ) in Congolese children. METHODS: Febrile patients of all ages were initially screened for malaria by both rapid diagnostic test (RDT) and microscopy. Patients less than 12 years of age, with parasitaemia ≥ 1000 asexual parasites of P. falciparum/µL of blood, without any signs of severity, were enrolled in a therapeutic efficacy study and treated after obtaining their parents' (or legal guardian's) informed consent in two health centres in Dolisie. The patients were followed for 28 days in accordance with the 2009 World Health Organization standard protocol. If parasitaemia reappeared on or after day 7, the genetic profiles (genes expressing merozoite surface protein-1 [msp1], merozoite surface protein-2 [msp2], and glutamine-rich protein [glurp]) of pre-treatment and post-treatment isolates were compared by nested polymerase chain reaction (PCR) followed by capillary electrophoresis to make a distinction between recrudescence and re-infection. The clinical and parasitological outcome was analysed by the per-protocol method and Kaplan-Meier survival curves. RESULTS: A total of 994 febrile patients of all ages were screened by RDT and microscopy. Of 994 patients, 323 (32.5%) presented a positive RDT, and 266 (26.8%) were microscopy-positive. Based on microscopy as the reference diagnostic method, the sensitivity and the specificity of the RDT were 98.9 and 91.8%, respectively. The Cohen's kappa coefficient was 0.86. A total of 121 children aged less than 12 years (61 in AL treatment group and 60 in ASAQ treatment group) were included in therapeutic efficacy study. Before PCR correction, the proportions of adequate clinical and parasitological response were 96.6% for AL and 86.0% for ASAQ in the per-protocol population (P < 0.05). The PCR-corrected efficacy rates were 98.2% and 94.2% for AL and ASAQ, respectively (P > 0.05). Both treatments were well tolerated. CONCLUSIONS: AL and ASAQ remain highly effective for the first-line treatment of uncomplicated P. falciparum malaria in Dolisie. Despite high efficacy of first- and second-line treatment, there is a continuing need to scale up effective malaria preventive interventions and vector control strategies in the country. TRIAL REGISTRATION NUMBER: ACTRN12616001422415.

Longitudinal trajectories of mental health and loneliness for Australian adolescents with-or-without neurodevelopmental disorders: the impact of COVID-19 school lockdowns.

BACKGROUND: The impact of COVID-19 (SARS-CoV-2) pandemic school lockdowns on the mental health problems and feelings of loneliness of adolescents with neurodevelopmental disorders (NDDs) is hypothesized to be greater than that of their non-NDD peers. This two and a half year longitudinal study compared changes in the mental health and loneliness of Western Australian adolescents pre-COVID-19 (November 2018 and April 2019), immediately prior to COVID-19 school lockdowns (March 2020), and post schools reopening (July/August 2020). METHODS: An age-and-gender matched sample of 476 adolescents with-or-without NDDs completed online assessments for mental health and loneliness. RESULTS: Adolescents with NDDs reported elevated levels of adverse mental health across all four waves of data collection. These young people experienced little change in mental health problems and feelings of loneliness over time, and any increase during school lockdowns returned to, or fell below pre-COVID-19 levels once schools reopened. In comparison, adolescents without NDDs experienced significant increases from a low baseline in depression symptoms, externalizing symptoms, feelings of isolation, and having a positive attitude to being alone, and evidenced a significant decline in positive mental wellbeing. Quality of friendships were unaffected by COVID-19 school lockdowns for all adolescents regardless of NDD status. Of the adolescents with NDDs, those with Attention-Deficit/Hyperactivity Disorder reported a significant increase in positive mental wellbeing following school lockdowns. CONCLUSIONS: Adolescents with NDDs emerged relatively unscathed from COVID-19 school lockdowns and the short term impacts associated with these were not maintained over time. These findings should be considered in the context of this study's geographical location and the unpredictability of school lockdowns. Learning to live with school lockdowns into the future may be a critical element for further investigation in the context of interventions.

Neonatal Screening for Sickle Cell Disease in Congo.

INTRODUCTION: Sickle cell disease is an autosomal recessive inherited disorder due to the mutation of a gene coding for the globin beta chain. The aim of this study is to update the epidemiological data on hemoglobinoses, in particular sickle cell disease in newborns in Congo. MATERIALS AND METHODS: This was a descriptive cross-sectional study, conducted from October 1, 2019, to March 31, 2020, throughout the Congolese national territory. It involved all full-term newborns, without distinction of nationality, aged 5 days or less, and whose parents consented to participate in the study. The blood samples, taken at the heel and collected on Whatman blotting paper, were analyzed using the HPLC Variant NBS machine. RESULTS: In 2897 newborns (NN) screened, hemoglobin abnormalities were found in 603 NN (20.81%). The mean age of these newborns was 1 day (extremes 0 and 5 days). The male-to-female ratio was 1.03. Abnormal hemoglobins were mainly Hb S (n = 597 (97.71%)); Hb C (n = 5 (0.82%)); and variants (n = 7 (1.15%)). The national prevalence of major sickle cell (MSC) syndromes and sickle cell trait was 1.35% and 19.43%, respectively. The prevalence ranged from 1.77% to 2.56% for MSS in four departments and from 20.5% to 25.8% for the sickle cell trait in six other departments. CONCLUSION: Data on homozygous sickle cell disease remain consistent with previous studies. However, further studies should clarify the molecular anomalies of the variants observed in our samples.

Severe bilateral pleuropneumonia caused by Legionella sainthelensi: a case report.

BACKGROUND: Legionella spp. are ubiquitous freshwater bacteria responsible for rare but potentially severe cases of Legionnaires' disease (LD). Legionella sainthelensi is a non-pneumophila Legionella species that was first isolated in 1980 from water near Mt. St-Helens (USA). Although rare cases of LD caused by L. sainthelensi have been reported, very little data is available on this pathogen. CASE PRESENTATION: We describe the first documented case of severe bilateral pleuropneumonia caused by L. sainthelensi. The patient was a 35-year-old woman with Sharp's syndrome treated with long-term hydroxychloroquine and corticosteroids who was hospitalized for an infectious illness in a university hospital in Reunion Island (France). The patient's clinical presentation was complicated at first (bilateral pneumonia, multiloculated pleural effusion, then bronchopleural fistula) but her clinical condition eventually improved with the reintroduction of macrolides (spiramycin) in intensive care unit. Etiological diagnosis was confirmed by PCR syndromic assay and culture on bronchoalveolar lavage. CONCLUSIONS: To date, only 14 documented cases of L. sainthelensi infection have been described worldwide. This pathogen is difficult to identify because it is not or poorly detected by urinary antigen and molecular methods (like PCR syndromic assays that primarily target L. pneumophila and that have only recently been deployed in microbiology laboratories). Pneumonia caused by L. sainthelensi is likely underdiagnosed as a result. Clinicians should consider the possibility of non-pneumophila Legionella infection in patients with a compatible clinical presentation when microbiological diagnostic tools targeted L. pneumophila tested negative.

Molecular Insights Into Regulatory T-Cell Adaptation to Self, Environment, and Host Tissues: Plasticity or Loss of Function in Autoimmune Disease.

There has been much interest in the ability of regulatory T cells (Treg) to switch function in vivo, either as a result of genetic risk of disease or in response to environmental and metabolic cues. The relationship between levels of FOXP3 and functional fitness plays a significant part in this plasticity. There is an emerging role for Treg in tissue repair that may be less dependent on FOXP3, and the molecular mechanisms underpinning this are not fully understood. As a result of detailed, high-resolution functional genomics, the gene regulatory networks and key functional mediators of Treg phenotype downstream of FOXP3 have been mapped, enabling a mechanistic insight into Treg function. This transcription factor-driven programming of T-cell function to generate Treg requires the switching on and off of key genes that form part of the Treg gene regulatory network and raises the possibility that this is reversible. It is plausible that subtle shifts in expression levels of specific genes, including transcription factors and non-coding RNAs, change the regulation of the Treg gene network. The subtle skewing of gene expression initiates changes in function, with the potential to promote chronic disease and/or to license appropriate inflammatory responses. In the case of autoimmunity, there is an underlying genetic risk, and the interplay of genetic and environmental cues is complex and impacts gene regulation networks frequently involving promoters and enhancers, the regulatory elements that control gene expression levels and responsiveness. These promoter-enhancer interactions can operate over long distances and are highly cell type specific. In autoimmunity, the genetic risk can result in changes in these enhancer/promoter interactions, and this mainly impacts genes which are expressed in T cells and hence impacts Treg/conventional T-cell (Tconv) function. Genetic risk may cause the subtle alterations to the responsiveness of gene regulatory networks which are controlled by or control FOXP3 and its target genes, and the application of assays of the 3D organization of chromatin, enabling the connection of non-coding regulatory regions to the genes they control, is revealing the direct impact of environmental/metabolic/genetic risk on T-cell function and is providing mechanistic insight into susceptibility to inflammatory and autoimmune conditions.

Productive Syntheses of Privileged Scaffolds Inspired by the Recognition of a Diels-Alder Pattern Common to Three Classes of Natural Products.

Identification of a common Diels-Alder pattern in three classes of bioactive natural products led us to study the synthesis and cycloaddition of a new class of cyclic dienes readily available from ß,γ-unsaturated lactams. A practical and readily scalable route to the parent p-methoxybenzyl-protected 6- and 7-membered ß,γ-unsaturated lactams was developed. These were readily transformed into the corresponding O-silylated dienes, which were reacted with dimethyl and diethyl fumarate to yield stereoselectively highly functionalized bicyclic adducts. These exhibited unexpected and versatile transformations upon acid hydrolysis depending on the nature of the dienophile substituents and the acid catalyst. All reactions have been performed on multigram quantities. These transformations provide a convenient, economical, and easily scalable pathway for the rapid construction of functionally and stereochemically dense privileged scaffolds for the construction of libraries of natural products-inspired molecules of pharmacological relevance.

Physiological responses of wild zebra finches (Taeniopygia guttata) to heatwaves.

Desert birds inhabit hot, dry environments that are becoming hotter and drier as a consequence of climate change. Extreme weather such as heatwaves can cause mass-mortality events that may significantly impact populations and species. There are currently insufficient data concerning physiological plasticity to inform models of species' response to extreme events and develop mitigation strategies. Consequently, we examine here the physiological plasticity of a small desert bird in response to hot (mean maximum ambient temperature=42.7°C) and cooler (mean maximum ambient temperature=31.4°C) periods during a single Austral summer. We measured body mass, metabolic rate, evaporative water loss and body temperature, along with blood parameters (corticosterone, glucose and uric acid) of wild zebra finches (Taeniopygia guttata) to assess their physiological state and determine the mechanisms by which they respond to heatwaves. Hot days were not significant stressors; they did not result in modification of baseline blood parameters or an inability to maintain body mass, provided drinking water was available. During heatwaves, finches shifted their thermoneutral zone to higher temperatures. They reduced metabolic heat production, evaporative water loss and wet thermal conductance, and increased hyperthermia, especially when exposed to high ambient temperature. A consideration of the significant physiological plasticity that we have demonstrated to achieve more favourable heat and water balance is essential for effectively modelling and planning for the impacts of climate change on biodiversity.

Variation of prevalence of malaria, parasite density and the multiplicity of Plasmodium falciparum infection throughout the year at three different health centers in Brazzaville, Republic of Congo.

BACKGROUND: In the Republic of Congo, hot temperature and seasons distortions observed may impact the development of malaria parasites. We investigate the variation of malaria cases, parasite density and the multiplicity of Plasmodium falciparum infection throughout the year in Brazzaville. METHODS: From May 2015 to May 2016, suspected patients with uncomplicated malaria were enrolled at the Hôpital de Mfilou, CSI « Maman Mboualé¼, and the Laboratoire National de Santé Publique. For each patient, thick blood was examined and parasite density was calculated. After DNA isolation, MSP1 and MSP2 genes were genotyped. RESULTS: A total of 416, 259 and 131 patients with suspected malaria were enrolled at the CSI «Maman Mboualé¼, Hôpital de Mfilou and the Laboratoire National de Santé Publique respectively. Proportion of malaria cases and geometric mean parasite density were higher at the CSI «Maman Mboualé¼ compared to over sites (P-value <0.001). However the multiplicity of infection was higher at the Hôpital de Mfilou (P-value <0.001). At the Laboratoire National de Santé Publique, malaria cases and multiplicity of infection were not influenced by different seasons. However, variation of the mean parasite density was statistically significant (P-value <0.01). Higher proportions of malaria cases were found at the end of main rainy season either the beginning of the main dry season at the Hôpital de Mfilou and the CSI «Maman Mboualé¼; while, lowest proportions were observed in September and January and in September and March respectively. Higher mean parasite densities were found at the end of rainy seasons with persistence at the beginning of dry seasons. The lowest mean parasite densities were found during dry seasons, with persistence at the beginning of rainy seasons. Fluctuation of the multiplicity of infection throughout the year was observed without significance between seasons. CONCLUSION: The current study suggests that malaria transmission is still variable between the north and south parts of Brazzaville. Seasonal fluctuations of malaria cases and mean parasite densities were observed with some extension to different seasons. Thus, both meteorological and entomological studies are needed to update the season's periods as well as malaria transmission intensity in Brazzaville.

The Field Metabolic Rate, Water Turnover, and Feeding and Drinking Behavior of a Small Avian Desert Granivore During a Summer Heatwave.

Global environmental change is leading to an increase in the frequency, intensity, and duration of extreme weather events, so effective environmental management requires an understanding not only of the physiological response of organisms to increased mean temperatures, but also to extreme environmental conditions. To determine the physiological consequences of heatwaves on energy and water balance of arid-adapted zebra finches (Taeniopygia guttata), we measured field metabolic rate and water turnover rate of wild, free-living finches during a heatwave (consecutive days of maximum ambient temperature of 40-45°C) and during a cooler period (maximum ambient temperature of 28°C) during a summer drought. To understand how birds accommodated their energy and water requirements, we also monitored feeding and drinking behavior of zebra finches at the study site on hot and cold days over 2.5 months during the same summer. Zebra finches can accommodate heatwaves without major impacts on field energy or water turnover, even when the heatwave is superimposed on high summer temperatures and long-term drought, so long as drinking water is available. In fact, cooler periods may pose a greater energetic challenge than heatwaves during drought, when food availability is limited, due to the increased thermoregulatory cost of maintaining a high body temperature against a thermal gradient. Zebra finches avoided or limited activity during the most thermally challenging periods of the day. Their pre-emptive feeding and drinking in preparation for hours of relative inactivity at high ambient temperature, together with a high body water content and reduced midday activity and metabolic heat production, enabled zebra finches to maintain body mass during a heatwave. Predicting upcoming periods of unfavorably high ambient temperature, together with a high body water content, may be essential for survival by desert birds of extreme ambient temperature during heatwaves.

High prevalence of norovirus and rotavirus co-infection in children with acute gastroenteritis hospitalised in Brazzaville, Republic of Congo.

OBJECTIVE: To evaluate the clinical severity of diarrhoea associated to viral co-infection in children with acute gastroenteritis. METHODS: About 461 children under five years hospitalised with acute diarrhoea (266 males and 187 females) were enrolled in the study. Using stool samples, rotavirus and adenovirus infections were investigated by ELISA, and norovirus infections by nested duplex RT-PCR. We assessed social, demographic, clinical and behavioural conditions that might influence the occurrence of rotavirus, adenovirus and norovirus infections. RESULTS: Mono-viral infection was detected in 49% and mixed viral infection in 12% of patients. The prevalence of mixed infection was neither dependent on age nor sex. Three samples were infected with all three viruses. A significant association was found between fever (axillary temperature> 37.5 °C) and rotavirus-norovirus dual infection (aOR (CI 95%) = 2.1 (1.14-3.84), P = 0.016; aOR (CI 95%) = 0.37 (0.19-0.73), P = 0.004). Mixed infection was the most common during the dry season from June to October (71.4% versus 54.7%, P = 0.023). CONCLUSION: Co-infection with both rotavirus and norovirus is common in under-five hospitalised children but does not contribute to the severity of the disease.

OBJECTIF: Evaluer la sévérité clinique de la diarrhée associée à la coinfection virale chez les enfants atteints de gastroentérite aiguë. MÉTHODES: 461 enfants de moins de cinq ans hospitalisés pour une diarrhée aiguë (266 garçons et 187 filles) ont été inclus dans l'étude. Sur des échantillons de selles, les infections à rotavirus et à adénovirus ont été investiguées par ELISA et les infections à norovirus par RT-PCR duplex imbriqué. Nous avons évalué les conditions sociales, démographiques, cliniques et comportementales susceptibles d'influencer la survenue d'infections à rotavirus, adénovirus et norovirus. RÉSULTATS: Une infection mono virale a été détectée chez 49% des patients et une infection virale mixte chez 12% des patients. La prévalence des infections mixtes ne dépendait ni de l'âge ni du sexe. Trois échantillons étaient infectés par tous les trois virus. Une association significative a été observée entre la fièvre (température axillaire > 37,5 °C) et la double infection rotavirus-norovirus (aOR (IC95%) = 2,1 (1,14-3,84), P = 0,016; aOR (IC95%) = 0,37 (0,19-0,73), P = 0,004). Les infections mixtes étaient les plus courantes pendant la saison sèche de juin à octobre (71,4% contre 54,7%, P = 0,023). CONCLUSION: La coinfection à la fois par le rotavirus et par le norovirus est fréquente chez les enfants de moins de cinq ans hospitalisés, mais ne contribue pas à la sévérité de la maladie.

Distribution of the cytochrome P450 CYP2C8*2 allele in Brazzaville, Republic of Congo.

BACKGROUND: Cytochrome P450 (CYP) enzymes are essential in the metabolism of most drugs used today. Single nucleotide polymorphism(s) occurring in CYP genes can adversely affect drug pharmacokinetics, efficacy, and safety. Individuals carrying the CYP2C8*2 c.805A > T (CYP2C8*2; rs11572103) allele have impaired amodiaquine metabolism, increased risk of amodiaquine-related adverse events, and may promote the selection of drug-resistant parasite strains. This study investigated the distribution of the CYP2C8*2 allele in Brazzaville, Republic of Congo, where artesunate + amodiaquine is used as the second-line treatment for uncomplicated Plasmodium falciparum malaria. METHODS: A total of 285 febrile children visiting the Marien Ngouabi paediatric hospital were genotyped for CYP2C8*2 using PCR-restriction fragment length polymorphism (PCR-RFLP). The allele frequencies and genotype distribution were determined. RESULTS: The CYP2C8*2 allele was successfully genotyped in 75% (213/285) of the study participants. The CYP2C8*2A allele had a frequency of 63%, whereas the CYP2C8*2T allele had a frequency of 37%. Genotypes CYP2C8*2AA (rapid metabolizer), CYP2C8*2AT (intermediate metabolizer), and CYP2C8*2TT (poor metabolizer) were observed in 44%, 38%, and 18% of the investigated participants, respectively. CONCLUSIONS: This study gives the first description of CYP2C8*2 allele distribution in the Republic of Congo and highlights the potential risk of amodiaquine-related adverse events. Information from this study will be beneficial during pharmacovigilance investigations.

Sub-microscopic Plasmodium falciparum infections in matched peripheral, placental and umbilical cord blood samples from asymptomatic Congolese women at delivery.

In malaria-endemic areas, most pregnant women are susceptible to asymptomatic Plasmodium falciparum infections. We present here the results of a cross-sectional study conducted in Madibou, a southern district of Brazzaville in the Republic of Congo, between March 2014 and April 2015. The main aim was to characterize P. falciparum infections. Blood samples corresponding to peripheral, placental and cord from 370 asymptomatic malaria women at delivery were diagnosed for plasmodium infection by thick blood smears (microscopic infection). Sub-microscopic infection was detected by PCR, using the MSP-2 gene as marker. Microscopic infections were detected in peripheral, placental and cord blood samples with a prevalence of respectively 7.3% (27/370), 2.7% (10/370) and 0%. The negative samples were submitted to sub-microscopic detection, with respective prevalence of 25.4% (87/343), 16.7% (60/360) and 9.4% (35/370) (P < 0.001). We further investigated the genetic diversity of the parasite by characterizing MSP2 allelic families 3D7 (24 distinct alleles) and FC27 (20 distinct alleles). The total number of alleles for these two families were 31, 25 and 19 in peripheral, placental and cord samples respectively. The 3D7 MSP-2 was the predominant allelic family. The multiplicity of infections (MOI) in peripheral (mean 1.4 ± 0.01; range 1-4), placental (mean 1.2 ± 0.01; range 1-3) and cord samples (1.4 ± 0.01; range 1-3) were similar (P = 0.9) and are unaffected by age, gravidity or sulfadoxine-pyrimethamine. These results shown a high prevalence of sub-microscopic infection and a high genetic diversity of Plasmodium falciparum strains in Congo. Age, gravidity and doses of preventive treatment based on sulfadoxine-pyrimethamine do not interfere with the multiplicity of infections.

Cytochrome P450 CYP2B6*6 distribution among Congolese individuals with HIV, Tuberculosis and Malaria infection.

BACKGROUND: The cytochrome P450 CYP2B6*6 (CYP2B6 c.516G>T; rs3745274) is one of the genetic factors that alters the drug metabolism in antimalarial, antiretroviral and TB first-line drugs. In Central African populations, the distribution of the CYP2B6*6 variant is poorly documented. This study investigated the distribution of CYP2B6 c.516G>T variant among Congolese individuals. METHODS: A total of 418 patients with HIV-1 mono-infection, HIV-1 and Tuberculosis coinfection and symptomatic P. falciparum malaria were genotyped for the CYP2B6 c.516G>T SNP using Restriction Fragment Length Polymorphism (RFLP). The allele frequencies and genotype distributions were determined. RESULTS: The CYP2B6 c.516G>T was successfully analysed in 69% (288/418) of the study participants. Among the investigated individuals, the distribution of the major allele CYP2B6*G was 45% and the minor CYP2B6*T allele was 55%. Significant differences in genotype distribution were also observed among the studied individuals. The CYP2B6*GG (rapid metabolizer) genotype was observed in 17% (49/288) followed by CYP2B6*GT (intermediate metabolizer) 55% (159/288) and CYP2B6*TT (poor metabolizers) 28% (80/288). CONCLUSION: This study contributes to increasing understanding on population pharmacogenetics and may help policy makers regulate treatment guidelines in the Congolese population with a high burden of HIV, Malaria and TB.

An update on glucose-6-phosphate dehydrogenase deficiency in children from Brazzaville, Republic of Congo.

BACKGROUND: Malaria transmission-blocking anti-malarial drugs, such as primaquine, offers an effective strategy for reducing the incidence of falciparum malaria. However, this drug induces haemolytic anaemia among glucose-6-phosphate dehydrogenase (G6PD) deficient individuals. The distribution of G6PD deficiency in Brazzaville, Republic of Congo and the association of G6PD deficiency with haemoglobin levels and blood cell counts were investigated. METHODS: A total of 212 febrile children were recruited for this study. Plasmodium falciparum diagnosis was conducted by microscopy and nested PCR. Sanger sequencing was used to assess G6PD deficiency by detecting 202G>A (rs1050828) and 376A>G (rs1050829) single nucleotide polymorphisms. RESULTS: Two hundred and twelve children were successfully genotyped for G6PD variants. Overall, 13% (27/212) of the children were G6PD deficient and 25% (25/100) females were heterozygous (11 BA- and 14 A+A-). The remaining 160 children had a normal G6PD genotype. The mean red blood and mean platelet counts were significantly lower in hemizygous male (G6PD A-) participants than in normal male (G6PD A+ or B) participants (p < 0.05). CONCLUSION: This study gives an update on G6PD deficiency among Congolese children. Understanding the distribution of G6PD deficiency in other geographical regions is recommended before primaquine is adopted in the malaria control programme.

No polymorphisms in K13-propeller gene associated with artemisinin resistance in Plasmodium falciparum isolated from Brazzaville, Republic of Congo.

BACKGROUND: In the Republic of Congo, artemisinin-based combinations have been recommended for the treatment of uncomplicated malaria since 2006. However, the emergence of resistant parasites again these combinations in Southeast Asia is a threat for the control of this disease, especially in sub-Saharan Africa where the weight of the disease is important. Indeed, polymorphisms in Plasmodium falciparum K13-propeller gene have been involved in variations of drug sensitivity of Plasmodium falciparum to artemisinin-based combinations. The aim of the current study is to determine the prevalence of mutations of this gene in isolates collected in three health centers in Brazzaville. METHODS: From May 2015 to May 2016, a total of 131, 259 and 416 samples from patients with suspected malaria were collected at the Laboratoire National de Santé Publique, Hôpital de Mfilou, and the CSI «Maman Mboualé¼ respectively. After DNA isolation, genotyping and sequencing of Plasmodium falciparum K13-propeller were performed in positive Plasmodium falciparum isolates identified after msp-2 gene genotyping. RESULTS: All 806 samples collected were msp-2 genotyped and Plasmodium falciparum infections were confirmed in 287 samples with 43, 85, 159 samples from Laboratoire National de Santé Publique, Hôpital de Mfilou, and the CSI «Maman Mboualé¼ respectively. Of these 287 msp-2 positives samples, K13-propeller nested PCR products were successfully obtained from 145 (50.52%) isolates and sequences were generated from 127(87.58%) nested products. None of mutations that were associated with ACTs resistance in Southeast Asia were detected on the samples from three different study sites from Brazzaville. However, one mutation type was observed at position 578, where alanine was substituted by serine (A578S) in two isolates (1.57%, 2/127), those from the Hôpital de Mfilou. No mutation was found in isolates from the two other sites. CONCLUSION: The current study shows a very limited polymorphism in the K13-propeller gene in isolates from the Republic of Congo and K13 polymorphisms associate with ACT resistance are not present in this country. However, permanent and large surveillance of resistant parasite population using K13-propeller gene is recommended.