5 research outputs found
Electrophoresis of PCR products from the co-amplification of GSTM1, GSTT1 and the internal control, CYP1A1 gene.
<p>Ethidium bromide stained 2% agarose gel. N = negative control, M = 100bp molecular weight marker. Lanes 2, 6, 7, 18 and 21 were positive for GSTT1 deletion genotype. Lanes 1, 11, 15, 17, 19, 20 and 22 had GSTM1 deletion genotype. Lanes 9 and 10 represented individuals with a deletion of both genotypes. Lanes 3, 4, 5, 8, 12, 13, 14 and 16 had both genes present.</p
Effect of GST polymorphism on GST enzyme activity.
<p>GST polymorphism was obtained by PCR amplification of the GSTM1 and GSTT1 genes, with CYP1A1 gene used as control. GST enzyme activity was assayed spectrophotometrically.</p
GST genotypes distribution in study population.
<p>GST genotypes distribution in study population.</p
Basic demographic data of study population stratified by ART use.
<p>Basic demographic data of study population stratified by ART use.</p
Characterisation of a rare, reassortant human G10P[14] rotavirus strain detected in Honduras
<div><p> BACKGROUND Although first detected in animals, the rare rotavirus strain G10P[14] has been sporadically detected in humans in Slovenia, Thailand, United Kingdom and Australia among other countries. Earlier studies suggest that the strains found in humans resulted from interspecies transmission and reassortment between human and bovine rotavirus strains. OBJECTIVES In this study, a G10P[14] rotavirus genotype detected in a human stool sample in Honduras during the 2010-2011 rotavirus season, from an unvaccinated 30-month old boy who reported at the hospital with severe diarrhea and vomiting, was characterised to determine the possible evolutionary origin of the rare strain. METHODS For the sample detected as G10P[14], 10% suspension was prepared and used for RNA extraction and sequence independent amplification. The amplicons were sequenced by next-generation sequencing using the Illumina MiSeq 150 paired end method. The sequence reads were analysed using CLC Genomics Workbench 6.0 and phylogenetic trees were constructed using PhyML version 3.0. FINDINGS The next generation sequencing and phylogenetic analyses of the 11-segmented genome of the G10P[14] strain allowed classification as G10-P[14]-I2-R2-C2-M2-A3-N2-T6-E2-H3. Six of the genes (VP1, VP2, VP3, VP6, NSP2 and NSP4) were DS-1-like. NSP1 and NSP5 were AU-1-like and NSP3 was T6, which suggests that multiple reassortment events occurred in the evolution of the strain. The phylogenetic analyses and genetic distance calculations showed that the VP7, VP4, VP6, VP1, VP3, NSP1, NSP3 and NSP4 genes clustered predominantly with bovine strains. NSP2 and VP2 genes were most closely related to simian and human strains, respectively, and NSP5 was most closely related to a rhesus strain. MAIN CONCLUSIONS The genetic characterisation of the G10P[14] strain from Honduras suggests that its genome resulted from multiple reassortment events which were possibly mediated through interspecies transmissions.</p></div