A repeat-primed PCR assay for pentanucleotide repeat alleles in spinocerebellar ataxia type 37

Spinocerebellar ataxia 37 (SCA37) is caused by an (ATTTC)n insertion in a polymorphic ATTTT repeat in the non-coding region of DAB1. The non-pathogenic alleles have a configuration [(ATTTT)7-400], whereas pathogenic alleles have a complex structure of [(ATTTT)60-79(ATTTC)31-75(ATTTT)58-90]. Molecular diagnosis of SCA37 is laborious because about 7% of the pentanucleotide repeat alleles in DAB1 are larger than 30 units and, thus, fail to amplify with standard PCR conditions, resulting in apparently homoallelism or in complete lack of PCR amplification in several cases. The molecular test currently available requires long-range PCR and sequencing analysis for the detection and characterization of these large alleles. We developed a simple assay capable of rapidly detecting the presence or absence of large pentanucleotide repeat sizes. This assay is based on repeat-primed PCR followed by high-throughput capillary electrophoresis. Combining the standard PCR with RP-PCR allows completion of the diagnosis in more than 80% of individuals, minimizing the number of samples that require long-range PCR followed by Sanger sequencing analysis. This assay meets many of the requirements for pre-screening of large cohorts of affected individuals.This work was funded by Fundo Europeu de Desenvolvimento Regional-FEDER funds through the COMPETE 2020—Operational Programme for Competitiveness and Inter- nationalisation (POCI), Portugal 2020, and by funding from FCT— Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tec- nologia e Inovação, Portugal, in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145- FEDER-007274); by Grant PTDC/SAU-GMG/098305/2008, from FCT, to I.S. J.R.L. was supported by scholarships from Grant PTDC/ GMG-SAU/098305/2008, FCT, PEst- C/SAU/LA0002/2013 and EMBO (ASTF494-2015). C.L.O. was supported by a scholarship from PEst-C/SAU/LA0002/2013. This work was also funded by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (FEDER), Portugal, that supports the Norte-01-0145-FEDER-000008—Porto Neurosciences and Neurologic Disease Research Initiative at I3S

Stress-Related Growth Scale-Short Form: A Portuguese validation for cancer patients

Abstract background/objective: Cancer can be extremely disruptive, triggering high levels of distress, and at the same time transformative, promoting perceptions of positive life changes and growth. This study aims to analyze the psychometric proprieties of the Stress-Related Growth Scale Short-Form (SRGS-SF) in cancer patients. Method: 209 Cancer patients heterogeneous in disease stage and diagnosis completed: clinical and sociodemographic information, Distress Thermometer, Positive and Negative Affect Schedule, Visual-analogue Scale of Perceived Positive Life Changes, and Stress-Related Growth Scale-Short Form. Results: The analysis of internal structure pointed to an one-dimensional scale, with high reliability (.92) measured through the McDonald`s omega coefficient. Validity was also evidenced through significant correlations with other variables. Conclusions: The Portuguese version of the SRGS-SF seems to present the necessary psychometric proprieties to be considered a valid and reliable short tool, to assess perceptions of growth following cancer and contribute to targeted and integrative psycho-oncological interventions.info:eu-repo/semantics/publishedVersio

Como sabemos o que sabemos por que acreditamos nisso : an?lise de um modelo de ensino sobre ci?ncia a partir de pr?ticas cient?ficas e epist?micas escolares.

Programa de P?s-Gradua??o em Educa??o. Departamento de Educa??o, Instituto de Ci?ncias Humanas e Sociais, Universidade Federal de Ouro Preto.Nesta pesquisa consideramos que aprender ci?ncia significa compreender os produtos cient?ficos, incluindo os fundamentos das alega??es cient?ficas e os processos envolvidos na produ??o, comunica??o, avalia??o e legitima??o do conhecimento. Para isso a literatura da ?rea de Educa??o em Ci?ncias direciona para um ensino que promova a socializa??o dos estudantes em pr?ticas an?logas aquelas das comunidades cient?ficas. Nessa perspectiva, os estudantes precisam ter um papel ativo em comunidades de pr?tica, de modo que compreendam suas reponsabilidades como produtores de conhecimento e tenham autoridade epist?mica para tal. Indo nessa dire??o, o objetivo da pesquisa centrase em analisar como as pr?ticas cient?ficas escolares no contexto do desenvolvimento de uma sequ?ncia did?tica (SD) possibilitam a emerg?ncia de pr?ticas epist?micas escolares. A pesquisa ? do tipo qualitativa e utiliza a videograva??o de sala de aula e anota??es de campo da pesquisadora no desenvolvimento da SD como instrumentos para constru??o dos dados. A SD foi aplicada em uma turma com 18 estudantes matriculados na disciplina de Est?gio Supervisionado no Ensino de Ci?ncias Biol?gicas I da Universidade Federal de Ouro Preto. Na an?lise dos dados a turma foi tratada como o caso e este foi organizado em epis?dios A partir dos dados conclu?mos que (i) a esfera social, incluindo as normas sociais para produ??o social do conhecimento, implicam no desenvolvimento da argumenta??o, das pr?ticas cient?ficas escolares, das pr?ticas epist?micas escolares, e nos conhecimentos conceitual, processual e epist?mico; (ii) a perspectiva ?hands-on? no contexto escolar interfere na ocorr?ncia das pr?ticas epist?micas; (iii) a vis?o epistemol?gica dos estudantes influ?ncia no desenvolvimento das pr?ticas epist?micas; (iv) os movimentos realizados pelo professor contribuem para a ocorr?ncia das pr?ticas epist?micas escolares. Como implica??es para o ensino ressaltamos a utiliza??o das pr?ticas cient?ficas interconectadas e com foco na avalia??o do conhecimento. Como implica??es para a pesquisa destacamos a necessidade de aprofundar em estudos com foco no estabelecimento das normas sociais na sala de aula, j? que elas influenciaram diretamente no desenvolvimento das pr?ticas cient?ficas e epist?micas escolares e na argumenta??o. As reflex?es advindas deste trabalho evidenciaram que a distin??o das pr?ticas cient?ficas e epist?micas para o contexto de ensino ? importante porque possibilita a compreens?o de como as pr?ticas epist?micas surgem no ensino, considerando que para isto as pr?ticas cient?ficas n?o se limitem ao fazer pr?tico e sejam experienciadas de forma metacognitiva e reflexiva.In this research, we consider that learning science means understanding scientific products, including the fundamentals of scientific claims, and the processes involved in the production, communication, evaluation and legitimation of knowledge. For that, the literature in the area of Science Education directs to a teaching that promotes the socialization of students in practices similar to those of the scientific communities. In this perspective, students need to take an active role in communities of practice, so that they understand their responsibilities as producers of knowledge and have epistemic authority to do so. Going in this direction, the aim of the research focuses on analyzing how school scientific practices in the context of the development of a teaching sequence (TS) enable the emergence of school epistemic practices. The research is qualitative and uses the videography of the classroom and field notes of the researcher in the development of TS as instruments for the construction of the data. The TS was applied in a class with 18 students in the subject of Supervised Internship in the Teaching of Biological Sciences I of the Federal University of Ouro Preto. The class was treated as the case in the analysis of the data. The case was organized in episodes. From the data we concluded that (i) the social sphere, including the social norms for the social production of knowledge, implies the development of argumentation, school scientific practices, school epistemic practices, and conceptual, procedural and epistemic knowledge; (ii) the ?hands-on? perspective in the school context interferes with the occurrence of school epistemic practices; (iii) the epistemological view of students influences the development of school epistemic practices; (iv) the movements made by the teacher contribute to the occurrence of school epistemic practices. As implications for teaching, we emphasize the use of interconnected scientific practices and with a focus on knowledge evaluation. As implications for the research, we emphasize the need to develop more studies focusing on the establishment of social norms in the classroom, since they directly influenced the development of scientific and epistemic school practices and argumentation. The reflections from this work showed that the distinction between scientific and epistemic practices for the teaching context is important, as it allows to understand how epistemic practices in teaching arise, considering that, for this purpose, scientific practices are not limited to ?doing science ?. The scientific practices practiced must be experienced in a metacognitive and reflective way

Data-driven inference for the spatial scan statistic

<p>Abstract</p> <p>Background</p> <p>Kulldorff's spatial scan statistic for aggregated area maps searches for clusters of cases without specifying their size (number of areas) or geographic location in advance. Their statistical significance is tested while adjusting for the multiple testing inherent in such a procedure. However, as is shown in this work, this adjustment is not done in an even manner for all possible cluster sizes.</p> <p>Results</p> <p>A modification is proposed to the usual inference test of the spatial scan statistic, incorporating additional information about the size of the most likely cluster found. A new interpretation of the results of the spatial scan statistic is done, posing a modified inference question: what is the probability that the null hypothesis is rejected for the original observed cases map with a most likely cluster of size k, taking into account only those most likely clusters of size k found under null hypothesis for comparison? This question is especially important when the p-value computed by the usual inference process is near the alpha significance level, regarding the correctness of the decision based in this inference.</p> <p>Conclusions</p> <p>A practical procedure is provided to make more accurate inferences about the most likely cluster found by the spatial scan statistic.</p

Mutational mechanism for DAB1 (ATTTC) n insertion in SCA37: ATTTT repeat lengthening and nucleotide substitution

Dynamic mutations by microsatellite instability are the molecular basis of a growing number of neuromuscular and neurodegenerative diseases. Repetitive stretches in the human genome may drive pathogenicity, either by expansion above a given threshold, or by insertion of abnormal tracts in nonpathogenic polymorphic repetitive regions, as is the case in spinocerebellar ataxia type 37 (SCA37). We have recently established that this neurodegenerative disease is caused by an (ATTTC)n insertion within an (ATTTT)n in a noncoding region of DAB1. We now investigated the mutational mechanism that originated the (ATTTC)n insertion within an ancestral (ATTTT)n . Approximately 3% of nonpathogenic (ATTTT)n alleles are interspersed by AT-rich motifs, contrarily to mutant alleles that are composed of pure (ATTTT)n and (ATTTC)n stretches. Haplotype studies in unaffected chromosomes suggested that the primary mutational mechanism, leading to the (ATTTC)n insertion, was likely one or more T>C substitutions in an (ATTTT)n pure allele of approximately 200 repeats. Then, the (ATTTC)n expanded in size, originating a deleterious allele in DAB1 that leads to SCA37. This is likely the mutational mechanism in three similar (TTTCA)n insertions responsible for familial myoclonic epilepsy. Because (ATTTT)n tracts are frequent in the human genome, many loci could be at risk for this mutational process.We are grateful to the families and individuals who participated in this work. We thank Patricia Ribeiro for technical assistance. This study was financed by Fundo Europeu de Desenvolvimento Regional (FEDER), through the COMPETE 2020 Operational Pro- gram for Competitiveness and Internationalization (POCI) of Portugal 2020, and by the Fundacão para a Ciência e a Tecnologia (FCT) and Ministério da Ciência, Tecnologia e Ensino Superior (Portugal), in the framework of the project POCI-01-0145-FEDER-029255; (PTDC/MED-GEN/29255/2017) to I.S. J.R.L. and C.L.O. were sup- ported by scholarships from PEst-C/SAU/LA0002/2013. S.M. is funded by the project IF/00930/2013/ CP1184/CT0002 from FCT. This work was also funded by the Porto Neurosciences and Neurologic Disease Research Initiative at the Instituto de Investigação e Inovação em Saúde (Norte-01-0145-FEDER- 000008), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTU- GAL 2020 Partnership Agreement with FEDER

Challenges of university teaching practice in distance education.

O artigo busca compreender a pr?tica pedag?gica de professores rec?m-doutores durante suas primeiras experi?ncias na educa??o a dist?ncia, tendo como foco principal a forma??o docente no contexto universit?rio. Buscamos refletir sobre os desafios enfrentados pelos sujeitos investigados analisando suas narrativas escritas ap?s as primeiras experi?ncias na doc?ncia na EaD. Os estudos sobre narrativas (auto)biogr?ficas balizam metodologicamente a coleta e an?lise de dados. Ao revisitarem suas trajet?rias de inser??o na EaD, os sujeitos t?m a oportunidade de constru?rem e reconstru?rem seus saberes sobre a doc?ncia no contexto universit?rio. Sabemos que as tecnologias t?m favorecido novas concep??es sobre o que ? ensinar, foco da Did?tica. Os resultados apontam a necessidade e a urg?ncia de forma??o docente no que se refere ? educa??o a dist?ncia universit?ria.The present paper aims to understand the pedagogical practice of newly doctoral graduated teachers during their first experiences in distance learning, focusing mainly on teacher training in the university context. We seek to pounder on the challenges faced by the investigated subjects analyzing their written narratives after the first experiences teaching in distance education. Studies on (auto)biography narratives methodologically guide the collection and analysis of data. When revisiting their paths of insertion in distance education, the subjects have the opportunity to build and reconstruct their knowledge about teaching in the university context. We do know that technologies have favored new conceptions of what it is to teach, which is what Didactics is focused on. The results point out the necessity and urgency for teacher training regarding university distance educatio

Allosteric communication in class A β-lactamases occurs via cooperative coupling of loop dynamics

Understanding allostery in enzymes and tools to identify it, offer promising alternative strategies to inhibitor development. Through a combination of equilibrium and nonequilibrium molecular dynamics simulations, we identify allosteric effects and communication pathways in two prototypical class A β-lactamases, TEM-1 and KPC-2, which are important determinants of antibiotic resistance. The nonequilibrium simulations reveal pathways of communication operating over distances of 30 Å or more. Propagation of the signal occurs through cooperative coupling of loop dynamics. Notably, 50% or more of clinically relevant amino acid substitutions map onto the identified signal transduction pathways. This suggests that clinically important variation may affect, or be driven by, differences in allosteric behavior, providing a mechanism by which amino acid substitutions may affect the relationship between spectrum of activity, catalytic turnover and potential allosteric behavior in this clinically important enzyme family. Simulations of the type presented here will help in identifying and analyzing such differences

Effect of maternal Schistosoma mansoni infection and praziquantel treatment during pregnancy on Schistosoma mansoni infection and immune responsiveness among offspring at age five years.

INTRODUCTION: Offspring of Schistosoma mansoni-infected women in schistosomiasis-endemic areas may be sensitised in-utero. This may influence their immune responsiveness to schistosome infection and schistosomiasis-associated morbidity. Effects of praziquantel treatment of S. mansoni during pregnancy on risk of S. mansoni infection among offspring, and on their immune responsiveness when they become exposed to S. mansoni, are unknown. Here we examined effects of praziquantel treatment of S. mansoni during pregnancy on prevalence of S. mansoni and immune responsiveness among offspring at age five years. METHODS: In a trial in Uganda (ISRCTN32849447, http://www.controlled-trials.com/ISRCTN32849447/elliott), offspring of women treated with praziquantel or placebo during pregnancy were examined for S. mansoni infection and for cytokine and antibody responses to SWA and SEA, as well as for T cell expression of FoxP3, at age five years. RESULTS: Of the 1343 children examined, 32 (2.4%) had S. mansoni infection at age five years based on a single stool sample. Infection prevalence did not differ between children of treated or untreated mothers. Cytokine (IFNγ, IL-5, IL-10 and IL-13) and antibody (IgG1, Ig4 and IgE) responses to SWA and SEA, and FoxP3 expression, were higher among infected than uninfected children. Praziquantel treatment of S. mansoni during pregnancy had no effect on immune responses, with the exception of IL-10 responses to SWA, which was higher in offspring of women that received praziquantel during pregnancy than those who did not. CONCLUSION: We found no evidence that maternal S. mansoni infection and its treatment during pregnancy influence prevalence and intensity of S. mansoni infection or effector immune response to S. mansoni infection among offspring at age five years, but the observed effects on IL-10 responses to SWA suggest that maternal S. mansoni and its treatment during pregnancy may affect immunoregulatory responsiveness in childhood schistosomiasis. This might have implications for pathogenesis of the disease