Rapid genotyping of the human renin (REN) gene by the LightCycler® instrument: Identification of unexpected nucleotide substitutions within the selected hybridization probe area

Preeclampsia is a serious disorder affecting nearly 3% of all in the Western world. It is associated with hypertension and proteinuria, and several lines of evidence suggest that the renin-angiotensin system (RAS) may be involved in the development of hypertension at different stages of a preeclamptic pregnancy. In this study, we developed rapid genotyping assays on the LightCycler® instrument to allow the detection of genetic variants in the renin gene (REN) that may predispose to preeclampsia. The method is based on real-time PCR and allele-specific hybridization probes, followed by fluorescent melting curve analysis to expose a change in melting temperature (Tm). Ninety-two mother-father-child triads (n=276) from preeclamptic pregnancies were genotyped for three haplotype-tagging single nucleotide polymorphisms (htSNPs) in REN. All three htSNPs (rs5705, rs1464816 and rs3795575) were successfully genotyped. Furthermore, two unexpected nucleotide substitutions (rs11571084 and rs61757041) were identified within the selected hybridization probe area of rs1464816 and rs3795575 due to aberrant melting peaks. In conclusion, genotyping on the LightCycler® instrument proved to be rapid and highly reproducible. The ability to uncover additional nucleotide substitutions is particularly important in that it allows the identification of potentially etiological variants that might otherwise be overlooked by other genotyping methods.publishedVersio

Maternal angiotensinogen (AGT) haplotypes, fetal renin (REN) haplotypes and risk of preeclampsia; estimation of gene-gene interaction from family-triad data

Background Preeclampsia is a debilitating disorder affecting approximately 3% of pregnant women in the Western world. Although inconclusive, current evidence suggests that the renin-angiotensin system may be involved in hypertension. Therefore, our objective was to determine whether the genes for placental renin (REN) and maternal angiotensinogen (AGT) interact to influence the risk of preeclampsia. Methods Three haplotype-tagging SNPs (htSNPs) covering REN (rs5705, rs1464818, and rs3795575) and another three covering AGT (rs2148582, rs2478545 and rs943580) were genotyped in 99 mother-father-child triads of preeclampsia pregnancies. We estimated relative risks (RR) conferred by maternal AGT and fetal REN haplotypes using HAPLIN, a statistical software designed to detect multi-marker transmission distortion among triads. To assess a combined effect of maternal AGT and fetal REN haplotypes, the preeclamptic triads were first stratified by presence/absence of maternal AGT haplotype C-T-A and tested for an effect of fetal REN across these strata. Results We found evidence that mothers carrying the most frequent AGT haplotype, C-T-A, had a reduced risk of preeclampsia (RR of 0.4, 95% CI = 0.2-0.8 for heterozygotes and 0.6, 95% CI = 0.2-1.5 for homozygotes). Mothers homozygous for AGT haplotypes t-c-g and C-c-g appeared to have a higher risk, but only the former was statistically significant. We found only weak evidence of an overall effect of fetal REN haplotypes and no support for our hypothesis that an effect of REN depended on whether the mother carried the C-T-A haplotype of AGT (p = 0.33). Conclusion Our findings indicate that the mother's AGT haplotypes affect her risk for developing preeclampsia. However, this risk is not influenced by fetal REN haplotypes.publishedVersio

Rapid genotyping of the human renin (REN) gene by the LightCycler® instrument: Identification of unexpected nucleotide substitutions within the selected hybridization probe area

Preeclampsia is a serious disorder affecting nearly 3% of all in the Western world. It is associated with hypertension and proteinuria, and several lines of evidence suggest that the renin-angiotensin system (RAS) may be involved in the development of hypertension at different stages of a preeclamptic pregnancy. In this study, we developed rapid genotyping assays on the LightCycler® instrument to allow the detection of genetic variants in the renin gene (REN) that may predispose to preeclampsia. The method is based on real-time PCR and allele-specific hybridization probes, followed by fluorescent melting curve analysis to expose a change in melting temperature (Tm). Ninety-two mother-father-child triads (n=276) from preeclamptic pregnancies were genotyped for three haplotype-tagging single nucleotide polymorphisms (htSNPs) in REN. All three htSNPs (rs5705, rs1464816 and rs3795575) were successfully genotyped. Furthermore, two unexpected nucleotide substitutions (rs11571084 and rs61757041) were identified within the selected hybridization probe area of rs1464816 and rs3795575 due to aberrant melting peaks. In conclusion, genotyping on the LightCycler® instrument proved to be rapid and highly reproducible. The ability to uncover additional nucleotide substitutions is particularly important in that it allows the identification of potentially etiological variants that might otherwise be overlooked by other genotyping methods

Chronic Myelogenous Leukemia with the e6a2 <i>BCR-ABL</i> and Lacking Imatinib Response: Presentation of Two Cases

The &lt;i&gt;BCR-ABL&lt;/i&gt; fusion gene represents the hallmark of chronic myelogenous leukemia (CML) and is derived from a translocation between chromosome 9 and 22. The majority of CML patients have a breakpoint in the major &lt;i&gt;BCR&lt;/i&gt; region of the &lt;i&gt;BCR&lt;/i&gt; gene giving rise to e13a2 or e14a2 &lt;i&gt;BCR-ABL&lt;/i&gt; transcripts. Occasionally, other &lt;i&gt;BCR&lt;/i&gt; breakpoints occur. The current report describes two e6a2 CML patients with imatinib treatment failure and unusual disease progression. One patient was Philadelphia chromosome positive and one was Philadelphia chromosome negative with an atypical &lt;i&gt;BCR-ABL&lt;/i&gt; rearrangement, ins (22;9).</jats:p

The relation between travel behaviour, ICT usage and social networks. The design of a web based survey

The relatively recent developments in Information and Telecommunication Technologies (ICT) arose the interest of both researchers and policymakers about how these technologies could shape travel. Several hypotheses have been put forward relating the use of ICT and travel behaviour. These include substitution of travel; complementarity or induction; modification or neutrality. This topic has been the subject of relevant research efforts, with a strong focus on the role of telecommuting as a way to reduce travel and its negative environmental impacts. The relation between ICT usage and other motives of travel have not been so thoroughly researched. One of these is social travel. More recent technological developments, in particular the development of the Web 2.0 and social media, as well as the dissemination of smartphones and related web applications increase the potential usefulness of ICT to influence social travel behaviour. Particularly relevant in this context is the study of the role of social networks and their characteristics in influencing mobility patterns. The work presented here is the design of a web-based survey aimed at studying the relations between ICT and social media usage, social networks and social travel. The survey was designed to be applied to university students, because they are stronger users of social media and ICT devices. Specifically, it aims to investigate a series of potential research questions related with the effect of ICT on social travel, its interaction with social networks, and, how the perception of its usefulness influences its usage. The survey structure and its relations with the previous literature in these subjects are presented. Finally, the results from a pilot already implemented in 3 cities - Lisbon, Granada and Zagreb are presented and its role on the validation of the survey structure is discussedThis work was developed under the auspices of the COST Action TU 1305 – Social Networks and Travel Behavio