Schistosome infection is a major public health concern affecting millions of people living in tropical regions of Africa, Asia, and South America. Schistosomes cause mild clinical symptoms in most subjects, whereas a small proportion of individuals presents severe clinical disease (as periportal fibrosis (PPF)) that may lead to death. Severe PPF results from an abnormal deposition of extracellular matrix proteins in the periportal spaces due to a chronic inflammation triggered by eggs and schistosome Ags. Extracellular matrix protein production is regulated by a number of cytokines, including IFN-γ. We have now screened putative polymorphic sites within this gene in a population living in an endemic area for Schistosoma mansoni. Two polymorphisms located in the third intron of the IFN-γ gene are associated with PPF. The IFN-γ +2109 A/G polymorphism is associated with a higher risk for developing PPF, whereas the IFN-γ +3810 G/A polymorphism is associated with less PPF. The polymorphisms result in changes in nuclear protein interactions with the intronic regions of the gene, suggesting that they may modify IFN-γ mRNA expression. These results are consistent with the results of previous studies. Indeed, PPF is controlled by a major locus located on chromosome 6q22-q23, closely linked to the gene encoding the α-chain of the IFN-γ receptor, and low IFN-γ producers have been shown to have an increased risk of severe PPF. Together, these observations support the view that IFN-γ expression and subsequent signal transduction play a critical role in the control of PPF in human hepatic schistosome infection (S. mansoni).

Highly sensitive and field deployable molecular diagnostic tools are critically needed for detecting submicroscopic, yet transmissible levels of malaria parasites prevalent in malaria endemic countries worldwide. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated in comparison with thick blood smear microscopy, an antigen-based rapid diagnostic test (RDT), and an in-house RT-PCR targeting the same RT-LAMP transcript. The optimized assay detected Plasmodium falciparum infections in as little as 0.25ng of total parasite RNA, and exhibited a detection limit of 0.08 parasites/ μL when tested directly on infected whole blood lysates, or ~0.0008 parasites/ μL when using RNA extracts. Assay positivity was observed as early as eight minutes from initiation of the RT-LAMP and in most cases the reaction was complete before twenty minutes. Clinical evaluation of the assay on 132 suspected malaria cases resulted in a positivity rate of 90% for RT-LAMP using extracted RNA, and 85% when using whole blood lysates. The positivity rates were 70% for P. falciparum-specific RDT, 83% for RT-PCR, and 74% for thick blood smear microscopy (Mean parasite density = 36,986 parasites/ μL). Concordance rates between the developed RT-LAMP and comparator tests were greater than 75%, the lowest being with light microscopy (78%, McNemar’s test: P = 0.0002), and the highest was with RT-PCR (87%, McNemar’s test: P = 0.0523). Compared to reference RT-PCR, assay sensitivity was 90% for RT-LAMP on whole blood, and 96% for RT-LAMP using corresponding RNA extracts. Electricity-free heaters were further developed and evaluated in comparison with a battery-operated isothermal amplification machine for use with the developed test in resource-limited settings. Taken together, the data highlight the benefits of targeting high abundant RNA transcripts in molecular diagnosis, as well as the potential usefulness of the developed RT-LAMP-assay in malaria diagnosis in low to high parasite density settings.

Malaria vectors control is essentially based on the use of insecticides against adult mosquitoes. However because of the development of resistance to insecticides, there is now a renewed interest in the management of larval sources. The aim of the present study was to map and characterize the breeding sites of Anopheles coluzzii in the Wouri river estuary in Cameroon. Larval surveys were carried out between December 2013 and August 2014 in rural areas on the island of Manoka and urban area in Youpwe at Douala. Culicidae breeding sites identified were georeferenced and mapped. Their larval productivity was evaluated by the method of "dipping" and their physicochemical parameters measured by spectrophotometry and oximetry. Culicidae collected larvae were reared in the insectarium to the adult stage. Adult mosquitoes were subjected to morphological identification and those belonging to the Anopheles gambiae complex have subsequently been subjected to molecular identification by the PCR-RFLP technique. A total of 240 breeding sites were geo-referenced in the two sites, including 10 types. Abandoned containers and pools were the most frequent breeding sites respectively in Manoka and in Youpwe. After morphological and molecular identification, eleven mosquito species have been identified. Anopheles coluzzii and Culex quinquefasciatus were the most frequent species respectively in Manoka and in Youpwe. Mosquito density was higher in managed gutters and canoes respectively in Manoka and in Youpwe. Culex and Aedes genus were more frequent in the hollow palm and water wells respectively in Manoka and Youpwe. The productivity of breeding sites varied according to the physicochemical parameters. Species richness varied according to the type of breeding site. Anopheles coluzzii was observed for the first time in Cameroon in water storage containers, tires, discarded containers and canoes. This study highlighted diversity in the type of breeding site of An. coluzzii in the Wouri estuary, suggesting the adaptation of this species in its environment. These results could be used to develop an antilarval control strategy in Manoka and in Youpwe.

Hepatic periportal fibrosis (PPF), associated with portal hypertension, is a major pathological consequence of infections with Schistosoma mansoni and Schistosoma japonicum. Indeed, affected subjects may die from portal hypertension. Previous studies have indicated that tumor necrosis factor alpha (TNF-␣) may aggravate fibrosis. We therefore investigated whether PPF was associated with certain polymorphisms of the TNF-␣ gene. Four polymorphisms (TNF-␣ ؊376 G/A, ؊308 G/A, ؊238 G/A, and ؉488 G/A) were investigated in two Sudanese populations living in an area in which S. mansoni is endemic. These polymorphisms were analyzed for 105 Sudanese subjects with various grades of PPF, from mild to advanced; all subjects were from two neighboring villages (Taweela and Umzukra). They were then analyzed for 70 subjects with advanced liver disease and for 345 matched controls from the Gezira region. We found no evidence of associations between these four polymorphisms and PPF in both of these studies. Thus, these four polymorphisms, two of which (TNF-␣ ؊376 and ؊308) were found to increase TNF-␣ gene transcription, are unlikely to have a major effect on PPF progression in these populations. However, this result does not exclude the possibility that these polymorphisms have a minor effect on PPF development.

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