Delimiting allelic imbalance of TYMS by allele-specific analysis

Allelic imbalance of thymidylate synthase (TYMS) is attributed to polymorphisms in the 5′- and 3′-untranslated region (UTR). These polymorphisms have been related to the risk of suffering different cancers, for example leukemia, breast or gastric cancer, and response to different drugs, among which are methotrexate glutamates, stavudine, and specifically 5-fluorouracil (5-FU), as TYMS is its direct target. A vast literature has been published in relation to 5-FU, even suggesting the sole use of these polymorphisms to effectively manage 5-FU dosage. Estimates of the extent to which these polymorphisms influence in TYMS expression have in the past been based on functional analysis by luciferase assays and quantification of TYMS mRNA, but both these studies, as the association studies with cancer risk or with toxicity or response to 5-FU, are very contradictory. Regarding functional assays, the artificial genetic environment created in luciferase assay and the problems derived from quantitative polymerase chain reactions (qPCRs), for example the use of a reference gene, may have distorted the results. To avoid these sources of interference, we have analyzed the allelic imbalance of TYMS by allelic-specific analysis in peripheral blood mononuclear cells (PBMCs) from patients. Allelic imbalance in PBMCs, taken from 40 patients with suspected myeloproliferative haematological diseases, was determined by fluorescent fragment analysis (for the 3′-UTR polymorphism), Sanger sequencing and allelic-specific qPCR in multiplex (for the 5′-UTR polymorphisms). For neither the 3′- nor the 5′-UTR polymorphisms did the observed allelic imbalance exceed 1.5 fold. None of the TYMS polymorphisms is statistically associated with allelic imbalance. The results acquired allow us to deny the previously established assertion of an influence of 2 to 4 fold of the rs45445694 and rs2853542 polymorphisms in the expression of TYMS and narrow its allelic imbalance to 1.5 fold, in our population. These data circumscribe the influence of these polymorphisms in the clinical outcome of 5-FU and question their use for establishing 5-FU dosage, above all when additional genetic factors are not considered

After-school sports programmes and social inclusion processes in culturally diverse contexts: Results of an international multicase study

This research aimed to understand the role of after-school sports programs in social inclusion processes in culturally diverse contexts through a multicase study within two locations. The first location was in Spain where immigrant and Spanish students were enrolled, and the other was in Chile with Mapuche-Huilliche students, immigrant and Chilean students. The implemented programs at both sites were similar in their educational focus on socio-educational values, and teaching models (hybridization of teaching games for understanding and cooperative learning) that enhance social inclusion. Using individual and group interviews with teachers, sports coordinators, parents, and students, a qualitative approach was used to identify the factors that facilitate or hinder the social inclusion processes. In addition, the researchers used qualitative observations of the programs over six months using “notes logbook” to record their impressions during the observation process. Results indicated that the implemented sports programs successfully facilitated social inclusion processes, enabling the development of interpersonal skills and relationships between students from different cultural backgrounds. The previous training and experiences of teachers in culturally diverse contexts, and incorporation of traditional sporting games from all cultures, seems to be an important facilitator factor for the inclusion potential of the implemented programs