GJB2: Frequency of the less common variants in a sample of the Portuguese population

Introduction: Sequence variants in the GJB2 gene account for up to 50% of cases of non-syndromic sensorineural hearing loss in the Caucasian population. In this study, we report the frequency of the less common variants of the GJB2 gene in a Portuguese sample and compare these frequencies with those of a group of hearing-impaired patients. Material and Methods: In order to select the less common GJB2 variants, 147 hearing-impaired patients followed in Centro Hospitalar Universitário de São João were evaluated. Afterwards, the presence of those variants was tested in 360 individuals from Generation 21. Results: The patient assessment enabled the selection of 11 GJB2 variants. Of those, 10 were investigated in Generation 21 participants, with only four being detected, in heterozygosity: p.Phe83Leu, p.Arg127His, p.Val153Ile and p.Asn206Ser, with the allelic frequencies (95% confidence interval) of 0.14% (0.01% - 0.87%), 0.28% (0.01% - 1.08%), 0.97% (0.43% - 2.04%) and 0.14% (0.01% - 0.88%), respectively. Two variants, p.Val37Ile and p.Val95Met, were more frequent in the patients’ group with statistical significance. Discussion: Our results allow for the p.Arg127His and p.Val153Ile variants to comply with polymorphism criteria and support the pathogenicity of p.Val37Ile and p.Val95Met variants. Moreover, two cases of moderate hearing loss were explained by the p.Val37Ile/p. Asn206Ser genotype, substantiating both the pathogenicity of such variants and the hypothesis that compound heterozygosity with p.Ans206Ser is associated with mild-moderate genotypes. Conclusion: Understanding the role of the variants is essential in order to provide genetic counselling to patients and their families. We explored a set of uncommon GJB2 variants that comprised 12% of the hearing-impaired patients in this study, supporting the relevance of their description.Generation XXI was funded by Programa Operacional de Saúde – Saúde XXI, Quadro Comunitário de Apoio III and Administração Regional de Saúde Norte (Regional Department of Ministry of Health). This study was funded by FEDER through the Operational Programme Competitiveness and Internationalization and national funding from the Foundation for Science and Technology – FCT (Portuguese Ministry of Science, Technology and Higher Education) by the Unidade de Investigação em Epidemiologia – Instituto de Saúde Pública da Universidade do Porto (EPIUnit) (POCI-01-0145-FED-ER-006862; Ref. UID/DTP/ 04750/2013), and the Calouste Gulbenkian Foundation. Ana Cristina Santos holds an FCT Investigator contract IF/ 01060/2015. (POCI-01-0145-FEDER-016837)

Direct isolation of single cells from living brains of Drosophila melanogaster without dissociation for transcriptome analysis

Here, we describe a protocol to remove single identified cells directly from Drosophila living brains and analyze their transcriptome. We detail the steps to harvest fluorescent cells using a capillary under epifluorescence and transmitted light to avoid contamination. We then outline the procedure to obtain the transcriptome by reverse transcription and amplification. The process from cell harvesting to the initiation of reverse transcription only takes 2 min, thus avoiding transcriptional activation of cell damage response or cell death genes. For complete details on the use and execution of this protocol, please refer to Barros and Bossing (2021), Bossing et al. (2012), Gil-Ranedo et al. (2019), and Liu and Bossing (2016)

Queixas musculoesqueléticas em uma Unidade Básica de Saúde: implicações para o planejamento das ações em saúde e fisioterapia

OBJETIVO: O objetivo deste estudo foi analisar a prevalência de queixas musculoesqueléticas em adultos em uma Unidade Básica de Saúde. MÉTODO: Foram avaliados os usuários atendidos na recepção espontânea no período de março de 2010 a maio de 2011. Ao todo, foram estudados 1.023 indivíduos. A caracterização das queixas foi realizada por meio de questionário com dados sociodemográficos e motivo da procura por atendimento. RESULTADOS: Os dados mostraram que a maioria dos usuários pertence ao sexo feminino (71,2%), está na faixa etária de 31 a 60 anos (50,0%), é solteira (31,6%), aposentada (14,2%) e apresenta queixas em vários sistemas (77,1%). O sistema musculoesquelético é o mais acometido (14,4%), representando o segundo motivo de procura por atendimento (31,0%). Analisando as razões de chance de ocorrência de queixas musculoesqueléticas com relação às variáveis estudadas, verificou-se que pessoas com idade entre 40 e 59 anos apresentaram 3,49 (IC95% 2,17-5,57) vezes mais chances de associação com essas dores do que as demais. Não houve associação entre outros sistemas e variáveis. CONCLUSÃO: A alta prevalência de queixas musculoesqueléticas requer um novo olhar de gestores em saúde para o atendimento destas demandas, pensando em incluir o fisioterapeuta na atenção básica para tratamento de dores de menor complexidade

Limit on the Radiative Neutrinoless Double Electron Capture of 36^{36}Ar from GERDA Phase I

Neutrinoless double electron capture is a process that, if detected, would give evidence of lepton number violation and the Majorana nature of neutrinos. A search for neutrinoless double electron capture of $^{36}$Ar has been performed with germanium detectors installed in liquid argon using data from Phase I of the GERmanium Detector Array (GERDA) experiment at the Gran Sasso Laboratory of INFN, Italy. No signal was observed and an experimental lower limit on the half-life of the radiative neutrinoless double electron capture of $^{36}$Ar was established: $T_{1/2} >$ 3.6 $\times$ 10$^{21}$ yr at 90 % C.I.Comment: 7 pages, 3 figure