Development of reverse-transcription PCR techniques to analyse the density and sex ratio of gametocytes in genetically diverse Plasmodium chabaudi infections

We have developed cross-genotype and genotype-specific quantitative reverse-transcription PCR (qRT-PCR) assays to detect and quantify the number of parasites, transmission stages (gametocytes) and male gametocytes in blood stage Plasmodium chabaudi infections. Our cross-genotype assays are reliable, repeatable and generate counts that correlate strongly (R(2)s > 90%) with counts expected from blood smears. Our genotype-specific assays can distinguish and quantify different stages of genetically distinct parasite clones (genotypes) in mixed infections and are as sensitive as our cross-genotype assays. Using these assays we show that gametocyte density and gametocyte sex ratios vary during infections for two genetically distinct parasite lines (genotypes) and present the first data to reveal how sex ratio is affected when each genotype experiences competition in mixed-genotype infections. Successful infection of mosquito vectors depends on both gametocyte density and their sex ratio and we discuss the implications of competition in genetically diverse infections for transmission success

Predictability of IL-28B-polymorphism on protease-inhibitor-based triple-therapy in chronic HCV-genotype-1 patients: A meta-analysis

AIM: To investigate the predictability of interleukin-28B single nucleotide polymorphism rs12979860 with respect to sustained virological response (SVR) in chronically hepatitis C virus (HCV) genotype-1 patients treated with a protease-inhibitor and pegylated interferon-$\alpha$ (Peg-INF-$\alpha$) based triple-therapy. METHODS: We searched PubMed, the Cochrane Library and Web of Knowledge for studies regarding the interleukin 28B (IL-28B)-genotype and protease-inhibitor based triple-therapy. Ten studies with 2707 patients were included into this meta-analysis. We used regression methods in order to investigate determinants of SVR. RESULTS: IL-28B-CC-genotype patients achieved higher SVR rates (odds 5.34, CI: 3.81-7.49) than IL-28B-non-CC-genotype patients (1.88, CI: 1.43-2.48) receiving triple-therapy. The line of therapy (treatment-na\"ive or -experienced for Peg-INF-$\alpha$) did not affect the predictive value of IL-28B (p=0.1). IL-28B-CC-genotype patients treated with protease inhibitor-based triple-therapy consisting of Boceprevir, Simeprevir, Telaprevir or Vaniprevir showed odds of 1.86, 9.77, 4.51 and 0.89, respectively. The odds for CC genotype patients treated with Faldaprevir cannot be quantified, as only a single study with a 100% SVR rate was available. CONCLUSION: IL-28B-SNP predicts the outcome for chronic HCV genotype-1 patients receiving protease inhibitor-based triple-therapy. The predictive value varies between the different protease inhibitors.Comment: 21 pages, 3 figures, 1 tabl

PON1 status does not influence cholinesterase activity in Egyptian agricultural workers exposed to chlorpyrifos.

Animal studies have shown that paraoxonase 1 (PON1) genotype can influence susceptibility to the organophosphorus pesticide chlorpyrifos (CPF). However, Monte Carlo analysis suggests that PON1 genotype may not affect CPF-related toxicity at low exposure conditions in humans. The current study sought to determine the influence of PON1 genotype on the activity of blood cholinesterase as well as the effect of CPF exposure on serum PON1 in workers occupationally exposed to CPF. Saliva, blood and urine were collected from agricultural workers (n=120) from Egypt's Menoufia Governorate to determine PON1 genotype, blood cholinesterase activity, serum PON1 activity towards chlorpyrifos-oxon (CPOase) and paraoxon (POase), and urinary levels of the CPF metabolite 3,5,6-trichloro-2-pyridinol (TCPy). The PON1 55 (P≤0.05) but not the PON1 192 genotype had a significant effect on CPOase activity. However, both the PON1 55 (P≤0.05) and PON1 192 (P≤0.001) genotypes had a significant effect on POase activity. Workers had significantly inhibited AChE and BuChE after CPF application; however, neither CPOase activity nor POase activity was associated with ChE depression when adjusted for CPF exposure (as determined by urinary TCPy levels) and stratified by PON1 genotype. CPOase and POase activity were also generally unaffected by CPF exposure although there were alterations in activity within specific genotype groups. Together, these results suggest that workers retained the capacity to detoxify chlorpyrifos-oxon under the exposure conditions experienced by this study population regardless of PON1 genotype and activity and that effects of CPF exposure on PON1 activity are minimal

Grain Characteristics, Chemical Composition, and Functional Properties of Rye (Secale cereale L.) As Influenced by Genotype and Harvest Year

Grain characteristic, chemical composition, and functional properties of rye were measured in 19 different cultivars grown in one location in up to 3 years. The cultivars included 8 adapted hybrids, 7 adapted population cultivars, and 4 nonadapted population cultivars. The results showed a significant influence of both harvest year and genotype on grain characteristics, chemical composition, and functional properties of the grain. Multivariate data analysis confirmed that the variations in the data were explained by yearly and genotype differences. Calculations of variance components showed that the variations in plant height, harvest yield, and protein content were mainly due to genotype differences and to a lesser extent to differences among harvest years. The kernel weight, hardness index, and content of dietary fiber components, however, were more strongly influenced by the harvest year than by the genotype. Differences in starch properties measured by falling number (FN), amylograph peak viscosity, and temperature at peak viscosity were more strongly influenced by harvest year. The water absorption was strongly influenced by genotype effects, compared to yearly differences. FN and amylograph peak temperature were positively correlated (r = 0.94). No correlation was found between the water absorption and the relative proportion of water-extractable arabinoxylan (AX) compared to the total AX content. However, the degree of ferulic acid cross-linking showed a negative correlation (r = -0.70) with the water absorption

Growth and sensory characteristics of alternative genotype broilers reared in organic orchards

The effects of age, sex and genotype on growth and sensory characteristics in organically produced broilers, when kept considerably longer before slaughtering, were examined. A total of 450 birds consisting of three genotypes, Light Sussex, New Hampshire, and the commercial strain I 657 were inserted at Fejoe Research Orchard. Half the birds were slaughtered at 91 days of age and half at 120 days of age. I 657 was significantly heavier compared to the slower growing breeds and Light Sussex was significantly heavier than New Hampshire. Males were significantly heavier than females across genotype, but weight ratios differed with genotype. No overall effect of genotype was found on the flavour or smell of the breast meat, but the commercial strain went towards a tougher and less tender consistency at 120 days of age, whereas the opposite was the case for the slower growing pure breeds. The positive flavour of salt was significantly improved at 120 days of age across genotype and age, with females having saltier flavour than the males. The positive flavour of sweet corn was improved in the meat from the males, but the positive smell of sweet corn was significantly improved in both males and females. No negative consequences of age were found