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Transcript Expression Analysis of Putative Trypanosoma brucei GPI-Anchored Surface Proteins during Development in the Tsetse and Mammalian Hosts

Human African Trypanosomiasis is a devastating disease caused by the parasite Trypanosoma brucei. Trypanosomes live extracellularly in both the tsetse fly and the mammal. Trypanosome surface proteins can directly interact with the host environment, allowing parasites to effectively establish and maintain infections. Glycosylphosphatidylinositol (GPI) anchoring is a common posttranslational modification associated with eukaryotic surface proteins. In T. brucei, three GPI-anchored major surface proteins have been identified: variant surface glycoproteins (VSGs), procyclic acidic repetitive protein (PARP or procyclins), and brucei alanine rich proteins (BARP). The objective of this study was to select genes encoding predicted GPI-anchored proteins with unknown function(s) from the T. brucei genome and characterize the expression profile of a subset during cyclical development in the tsetse and mammalian hosts. An initial in silico screen of putative T. brucei proteins by Big PI algorithm identified 163 predicted GPI-anchored proteins, 106 of which had no known functions. Application of a second GPI-anchor prediction algorithm (FragAnchor), signal peptide and trans-membrane domain prediction software resulted in the identification of 25 putative hypothetical proteins. Eighty-one gene products with hypothetical functions were analyzed for stage-regulated expression using semi-quantitative RT-PCR. The expression of most of these genes were found to be upregulated in trypanosomes infecting tsetse salivary gland and proventriculus tissues, and 38% were specifically expressed only by parasites infecting salivary gland tissues. Transcripts for all of the genes specifically expressed in salivary glands were also detected in mammalian infective metacyclic trypomastigotes, suggesting a possible role for these putative proteins in invasion and/or establishment processes in the mammalian host. These results represent the first large-scale report of the differential expression of unknown genes encoding predicted T. brucei surface proteins during the complete developmental cycle. This knowledge may form the foundation for the development of future novel transmission blocking strategies against metacyclic parasites

Spatial Assessment of Heavy Metals in Surface Soil from Klang District (Malaysia): An Example from a Tropical Environment

Heavy metals (Al, Cd, Co, Cr, Cu, Fe, Pb, Zn) in surface soil of Klang district were determined and multivariate analysis was used to understand their potential sources. The total and bioavailability of concentrations were used in identifying the potential risks to the ecology and human health. The means for the total heavy metal concentrations were found to be in the order of Fe > Al > Zn > Pb > Cu > Cr > Co > Cd, while the means for the bioavailability concentrations were found to be in the order of Al > Fe > Zn > Cu > Co > Cd > Pb > Cr. Principal Component Analysis showed Principal Component 1 as being of natural origin whereas Principal Components 2, 3, and 4 were associated with mixed anthropogenic sources, such as traffic and industrial emissions, organic matter, and granulometric fractions. Potential ecological risk assessment indicated an overall low ecological risk. Spatial assessment of non-carcinogenic risks showed that the Hazard Index values were more than one in Johan Setia, due to biomass burning of peat swamps exploited for agricultural development. While for spatial assessment of carcinogenic risks, the Lifetime Cancer Risk values were in the limit (1 × 10−5), indicating low cancer inducing risks. Nevertheless, with intense development pressure in the Klang district could overlap pollution inputs in the future