Simplifying the detection of MUTYH mutations by high resolution melting analysis

<p>Abstract</p> <p>Background</p> <p><it>MUTYH</it>-associated polyposis (MAP) is a disorder caused by bi-allelic germline <it>MUTYH </it>mutation, characterized by multiple colorectal adenomas. In order to identify mutations in <it>MUTYH </it>gene we applied High Resolution Melting (HRM) genotyping. HRM analysis is extensively employed as a scanning method for the detection of heterozygous mutations. Therefore, we applied HRM to show effectiveness in detecting homozygous mutations for these clinically important and frequent patients.</p> <p>Methods</p> <p>In this study, we analyzed phenotype and genotype data from 82 patients, with multiple (>= 10) synchronous (19/82) or metachronous (63/82) adenomas and negative <it>APC </it>study (except one case). Analysis was performed by HRM-PCR and direct sequencing, in order to identify mutations in <it>MUTYH </it>exons 7, 12 and 13, where the most prevalent mutations are located. In monoallelic mutation carriers, we evaluated entire <it>MUTYH </it>gene in search of another possible alteration. HRM-PCR was performed with strict conditions in several rounds: the first one to discriminate the heteroduplex patterns and homoduplex patterns and the next ones, in order to refine and confirm parameters. The genotypes obtained were correlated to phenotypic features (number of adenomas (synchronous or metachronous), colorectal cancer (CRC) and family history).</p> <p>Results</p> <p><it>MUTYH </it>germline mutations were found in 15.8% (13/82) of patients. The hot spots, Y179C (exon 7) and G396D (exon 13), were readily identified and other mutations were also detected. Each mutation had a reproducible melting profile by HRM, both heterozygous mutations and homozygous mutations. In our study of 82 patients, biallelic mutation is associated with being a carrier of ≥10 synchronous polyps (p = 0.05) and there is no association between biallelic mutation and CRC (p = 0.39) nor family history (p = 0.63). G338H non-pathogenic polymorphism (exon 12) was found in 23.1% (19/82) of patients. In all cases there was concordance between HRM (first and subsequent rounds) and sequencing data.</p> <p>Conclusions</p> <p>Here, we describe a screening method, HRM, for the detection of both heterozygous and homozygous mutations in the gene encoding <it>MUTYH </it>in selected samples of patients with phenotype of MAP. We refine the capabilities of HRM-PCR and apply it to a gene not yet analyzed by this tool. As clinical decisions will increasingly rely on molecular medicine, the power of identifying germline mutations must be continuously evaluated and improved.</p

The making of energy evidence: How exclusions of Social Sciences and Humanities are reproduced (and what researchers can do about it)

Overland and Sovacool (2020) and Baum and Bartkowski (2020), in this journal, have provided important insights on the neglect of Social Sciences and Humanities (SSH) in energy and sustainability research. In this response, we develop this conversation further, arguing that the commissioning and funding of energy research can be understood as part of a process of making evidence, which is co-constructive with policy-making. This helps us analyse how exclusions of SSH within the energy research-policy landscape are reproduced, and, crucially, to identify opportunities for change. We draw on concepts from Science and Technology Studies regarding the co-construction of knowledges and policies; epistemic communities; and the active role of expectations and imaginaries around energy evidence, and we apply these to empirical material from workshops with EU policyworkers, and analysis of key documents relating to European energy research and policy. We explore ways that SSH are excluded through expectations around: i) the research enterprise and purpose of evidence; ii) the contributions of different SSH communities, iii) how different epistemic communities should work together; and iv) validity and rigour. Finally, we offer some reflections for research professionals and research funding organisations who wish to integrate SSH more meaningfully into energy research and policy, including suggestions around actors; documents; and the processes of reviewing and monitoring that are involved in the making of energy evidence

Exposure to the Environmental Endocrine Disruptor TCDD and Human Reproductive Dysfunction: Translating Lessons from Murine Models

Humans and other animals are exposed to a wide array of man-made toxicants, many of which act as endocrine disruptors that exhibit differential effects across the lifespan. In humans, while the impact of adult exposure is known for some compounds, the potential consequences of developmental exposure to endocrine disrupting chemicals (EDCs) is more difficult to ascertain. Animal studies have revealed that exposure to EDCs prior to puberty can lead to adult reproductive disease and dysfunction. Specifically, in adult female mice with an early life exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), we demonstrated a transgenerational occurrence of several reproductive diseases that have been linked to endometriosis in women. Herein, we review the evidence for TCDD-associated development of adult reproductive disease as well as known epigenetic alterations associated with TCDD and/or endometriosis. We will also introduce new "Organ-on-Chip" models which, combined with our established murine model, are expected to further enhance our ability to examine alterations in gene-environment interactions that lead to heritable disease

Deliverable 3.1: Matrix for integration of learning cases and framework of analysis

The SHAPE-ID project was scheduled to organise six learning case workshops across Europe between December 2019 and May 2020 to enable stakeholders to explore best practices in interdisciplinary and transdisciplinary research (IDR/TDR) with an emphasis on research involving the Arts, Humanities and Social Sciences (AHSS). The first three of these workshops – held in Dublin in December 2019, Edinburgh in January 2020 and Turin in February 2020 – took place as planned. The remaining three – intended to take place in Bilbao in March, in Warsaw in April and in Zurich in May 2020 – were postponed due to the COVID-19 outbreak. Due to the ongoing and uncertain situation with COVID-19, the SHAPE-ID consortium considered the organisation of in-person meetings no longer feasible and decided to reorganise the workshops in a remote setting. This choice, besides ensuring the protection of all participants’ health while reducing further delays to the project, has presented the opportunity to experiment with interdisciplinary/ transdisciplinary learning activities in a virtual environment. Indeed, despite the adversity, there is an opportunity to be seized in organising online events. Exploring the potential of online techniques for working and developing interdisciplinary and transdisciplinary collaborations is a necessary step considering that personal mobility and gatherings will not resume at previous levels of frequency and intensity soon. In particular, the role of the Arts and Humanities in this scenario is essential because the technology-driven process of moving meetings online will challenge most of the tacit and emotional aspects of human interaction, such as informal and private communication, the emotional effects of collaboration, and all those non-visible elements of social exchange that need to be considered and influence the outcome of collaborative research. We therefore had the opportunity to learn how to design, prepare and conduct online workshops – taking advantage of the necessary delay to research suitable methodologies and redesign already well-planned in-person workshops – and evaluate their efficacy compared to the traditional face-to-face workshops. This document reports on the workshops, organising the findings into a coherent framework in order to feed into a second report, D3.3 – Recommendations and measures to maximise IDR impact on society. The current report is structured as follows: Section 1: Integration of challenge-oriented learning journeys Section 2: Overview of the six workshops’ outcomes (Dublin, Edinburgh, Turin, Zurich, Warsaw, Bilbao) Section 3: Detailed reports for each of the six workshops Section 4: Conclusions on IDR learning cases tackling societal challenges and missions In addition, Appendix 1 includes the full set of six workshops evaluation reports and Appendix 2 the full list of participants at all learning workshops