Identificação de uma nova mutação na síndrome de insensibilidade aos androgénios

Introdução: A Síndrome de Insensibilidade aos Androgénios (SIA), uma doença rara de hereditariedade recessiva ligada ao X, é causada por mutações no gene que codifica o recetor dos androgénios (AR). Esta disfunção leva à resistência dos tecidos-alvo aos androgénios, impedindo a masculinização e virilização normal num indivíduo com a constituição cromossómica 46,XY. O diagnóstico pode ser feito in utero ou na infância, mas muitos casos passam despercebidos até à puberdade, quando a amenorreia primária com desenvolvimento mamário normal e pêlos púbicos e axilares escassos levam ao estudo dessas jovens. A presença de uma mutação no gene AR é o principal critério diagnóstico de SIA. O objetivo deste trabalho foi estudar geneticamente uma paciente adulta de fenótipo feminino e com cariótipo 46,XY. Materiais e Métodos: A paciente em estudo apresentava aparência feminina normal, mas com cariótipo XY e ausência de útero e ovários mas com gónada à esquerda em ecografia. Extraiuse ADN leucocitário da paciente e amplificaram-se os exões 4-8 do gene AR, que foram sequenciados. Analisou-se a estrutura do ARN mensageiro do AR através da realização de RTPCR a partir de ARN leucocitário e sequenciação do produto resultante. Resultados: Foi encontrada uma mutação c.2173+2T>C, localizada na zona doadora de splicing do intrão 4, com a substituição de uma timina por uma citosina. A sequenciação do produto de RT-PCR demonstrou uma deleção de 123 nucleótidos do exão 4, com justaposição do exão 5 ao restante exão 4. A tradução deste transcrito aberrante produz uma proteína AR com deleção in-frame de 41 aminoácidos, correspondentes aos resíduos 674-714, do domínio de ligação da testosterona. Discussão: Confirmou-se o diagnóstico de síndrome de insensibilidade aos androgénios nesta paciente. A mutação encontrada é a primeira mutação reportada na posição +2 de um intrão do gene AR. Esta mutação inativou o local de doação de splicing do intrão 4 e iniciou um mecanismo de splicing alternativo na posição c.2049 do exão 4, e parte do exão foi eliminada. No recetor dos androgénios, a região deletada corresponde a grande parte da hélice 3, cuja presença é crítica para a mudança conformacional do recetor que facilita a entrada dos androgénios no núcleo. A inexistência da hélice 3 explica a ausência de ligação fisiológica da testosterona ao seu recetor nesta paciente. Após o diagnóstico de SIA, a abordagem terapêutica assenta no tratamento da densidade mineral óssea mais baixa, na exérese cirúrgica das gónadas masculinas, e no apoio psicológico e aconselhamento genético. Palavras-chave: Síndrome de Insensibilidade aos Androgénios; amenorreia primária; infertilidade; recetor dos androgénios; genética; mutação; testosterona; endocrinologiaIntroduction: Androgen Insensitivity Syndrome (AIS), a rare X-linked recessive disorder, is caused by mutations in the gene coding the androgen receptor (AR). Its dysfunction leads to target-tissue resistance to androgens, preventing a normal masculinization and virilization of a 46,XY individual. Diagnosis can be made in utero or during infancy, but many cases go unnoticed until puberty, when primary amenorrhea with normal breast development and scarce axillary and pubic hair lead to assessment of these girls. Demonstration of a mutation in the AR gene is the main diagnostic criteria for AIS. The aim of this study was to genetically study an adult patient with a female phenotype and 46,XY karyotype. Materials and Methods: The patient presented a normal female appearance, but with an XY karyotype and absence of a uterus and ovaries, but with a gonad on the left, demonstrated by ultrasound. Leukocyte DNA was extracted from the patient and exons 4-8 of the AR gene were amplified and then sequenced. The structure of the messenger RNA was analyzed using RTPCR with leukocyte RNA with subsequent sequencing of the resulting product. Results: A c.2173+2T>C mutation was found, located at the splice donor site of intron 4, with the substitution of a thymine for a cytosine. Sequencing of the RT-PCR product demonstrated a 123-nucleotide deletion in exon 4, with exon 5 being spliced directly to the remaining exon 4. Translation of this aberrant transcript produces an AR protein with an in-frame deletion of 41 aminoacids, corresponding to residues 674-714, in the testosterone-binding domain. Discussion: The diagnosis of AIS was confirmed in this patient. This is the first reported mutation at the +2 position of an intron in the AR gene. This mutation inactivated the donor splice region of intron 4 and initiated an alternative splicing at position c.2049 of exon 4, and part of the exon was deleted. In the androgen receptor, the deleted region corresponds to the majority of helix 3, and its presence is critical to the conformational change of the receptor that enables the entry of androgens into the nucleus. The inexistence of helix 3 explains the absence of physiological binding of testosterone to its receptor in this patient. After the diagnosis of AIS, the therapeutic approach is based on the treatment of the lower bone mineral density, in the surgical removal of the male gonads, and in psychological support and genetic counseling

Por uma família participativa: estratégias e propostas de atividade para o envolvimento parental

Relatório da Prática Profissional Supervisionada Mestrado em Educação Pré-EscolarEste relatório foi realizado no âmbito da Prática Profissional supervisionada para a obtenção de grau Mestre em Educação Pré-Escolar. O presente relatório apresenta o percurso realizado durante o estágio em creche e jardim-de-infância, tendo como objectivo promover a participação dos pais através da criação e desenvolvimento de estratégias e propostas para o envolvimento parental. Neste sentido, este trabalho apresenta e explicita as diferentes estratégias utilizadas para melhorar a e favorecer a participação dos pais no contexto de jardim-de-infância. Neste processo de intervenção educacional procurei compreender e conhecer as relações que se estabelecem entre Família Jardim de Infância e a influência que o envolvimento dos pais exerce sobre as crianças. O processo permitiu extrair algumas conclusões. Ao longo do processo e nas fases finais revelou-se a importância de apoiar e suportar oportunidades para criar interações de qualidade entre todos os elementos da comunidade educativa. Por sua vez, os pais puderam compreender que o jardim-de-infância é um contexto que promove a aprendizagem ativa e a interação social e puderam conhecer o trabalho desenvolvido pela educadora e pela Instituição.Abstract This report was fulfilled in the domain of the supervised professional practice, in order to obtain the master’s degree in Preschool Education. The present report presents the course taken during the nursery and kindergarten’s internship, having as an objective promoting parental participation through the implementation and development of strategies and proposals for parental involvement. In this sense, this assignment presents and expresses the different strategies used to ameliorate and favour the participation of parents in the kindergarten context. In this process of educational intervention, I sought to know and understand the relations established between the families and the kindergarten, and the influence that parental involvement exerts on the children. The process allowed drawing some conclusions. Throughout the process and in the final stages the importance of aiding and supporting opportunities that could create quality interaction between all of the educational community members. In turn, parents could comprehend that the kindergarten is an active learning and social interaction promoter and, in addition, could also get to know the work developed by the kindergarten teacher and by the institution

Case report: VEXAS syndrome: an atypical indolent presentation as sacroiliitis with molecular response to azacitidine

VEXAS syndrome is a recently described autoinflammatory syndrome caused by the somatic acquisition of UBA1 mutations in myeloid precursors and is frequently associated with hematologic malignancies, chiefly myelodysplastic syndromes. Disease presentation can mimic several rheumatologic disorders, delaying the diagnosis. We describe a case of atypical presentation resembling late-onset axial spondylarthritis, later progressing to a systemic inflammatory syndrome with chondritis, cutaneous vasculitis, and transfusion-dependent anemia, requiring high doses of steroids. Ruxolitinib was used as the first steroid-sparing strategy without response. However, azacitidine showed activity in controlling both inflammation and the mutant clone. This case raises the question of whether azacitidine’s anti-inflammatory effects are dependent on or independent of clonal control. We discuss the potential relevance of molecular remission in VEXAS syndrome and highlight the importance of a multidisciplinary team for the care of such complex patients

Score CTo-aBCDE : um novo score preditor de sucesso nas CTOs

© 2020 Sociedade Portuguesa de Cardiologia. Published by Elsevier España, S.L.U. This is an open access article under the CC-BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).Introduction: Patient selection for percutaneous coronary intervention (PCI) in chronic total occlusions (CTOs) is crucial to procedural success. Our aim was to identify independent predictors of success in CTO PCI in order to create an accurate score. Methods: In a single-center observational registry of CTO PCI, demographic and clinical data and anatomical characteristics of coronary lesions were recorded. Linear and logistic regression analysis were used to identify predictors of success. A score to predict success was created and its accuracy was measured by receiver operating curve analysis. Results: A total of 377 interventions were performed (334 patients, age 68±11 years, 75% male). The success rate was 65% per patient and 60% per procedure. Predictors of success in univariate analysis were absence of active smoking (OR 2.02, 95% CI 1.243-3.29; p=0.005), presence of tapered stump (OR 5.2, 95% CI 2.7-10.2; p8 with high probability (95%). Conclusion: In our sample only anatomical characteristics were predictors of success. The creation of a score to predict success, with good accuracy, may enable selection of cases that can be treated by any operator, those in which a dedicated operator will be desirable, and those with an extremely low probability of success, which should be considered individually for conservative management, surgical revascularization or PCI by a team experienced in CTO.info:eu-repo/semantics/publishedVersio

Covid-19 e o tempo mundial : a crise iniciadora de uma nova realidade?

info:eu-repo/semantics/publishedVersio

Tissue Oxygenation in Response to Different Relative Levels of Blood-Flow Restricted Exercise

Blood flow restrictive (BFR) exercise elicits a localized hypoxic environment compatible with greater metabolic stress. We intended to compare the acute changes in muscle microvascular oxygenation following low-intensity knee extension exercise, combined with different levels of BFR. Thirteen active young men (age: 23.8 ± 5.4 years) were tested for unilateral knee extension exercise (30 + 15 + 15 + 15 reps at 20% one repetition maximum) on four different conditions: no-BFR (NOBFR), 40, 60, and 80% of arterial occlusion pressure (AOP). Deoxyhemoglobin+myoglobin concentration Deoxy[Hb+Mb], total hemoglobin [T(H+Mb)] and tissue oxygen saturation [TOI] were measured on the vastus lateralis muscle using near-infrared spectroscopy (NIMO, Nirox srl, Brescia, Italy). The magnitude of change in Deoxy[Hb+Mb]during exercise was similar between 60 and 80% AOP. Overall, compared to that seen during 60 and 80% AOP, NOBFR as well as 40% AOP resulted in a lower magnitude of change in Deoxy[Hb+Mb] (p < 0.05). While the oxygen extraction decreased during each inter-set resting interval in NOBFR and 40% AOP, this was not the case for 60 or 80% AOP. Additionally, TOI values obtained during recovery from each set of exercise were similarly affected by all conditions. Finally, our data also show that, when performed at higher restrictive values (60 and 80%), BFR exercise increases total Deoxy[Hb+Mb] extraction (p < 0.05). Taken together, we provide evidence that BFR is effective for increasing deoxygenation and reducing tissue oxygenation during low-intensity exercise. We also showed that when using low loads, a relative pressure above 40% of the AOP at rest is required to elicit changes in microvascular oxygenation compared with the same exercise with unrestricted conditions

Differential Temporal Dynamics of Axial and Appendicular Ataxia in SCA3

Background: Disease severity in spinocerebellar ataxia type 3 (SCA3) is commonly defined by the Scale for the Assessment and Rating of Ataxia (SARA) sum score, but little is known about the contributions and progression patterns of individual items. Objectives: To investigate the temporal dynamics of SARA item scores in SCA3 patients and evaluate if clinical and demographic factors are differentially associated with evolution of axial and appendicular ataxia. Methods: In a prospective, multinational cohort study involving 11 European and 2 US sites, SARA scores were determined longitudinally in 223 SCA3 patients with a follow-up assessment after 1 year. Results: An increase in SARA score from 10 to 20 points was mainly driven by axial and speech items, with a markedly smaller contribution of appendicular items. Finger chase and nose-finger test scores not only showed the lowest variability at baseline, but also the least deterioration at follow-up. Compared with the full set of SARA items, omission of both tests would result in lower sample size requirements for therapeutic trials. Sex was associated with change in SARA sum score and appendicular, but not axial, subscore, with a significantly faster progression in men. Despite considerable interindividual variability, the average annual progression rate of SARA score was approximately three times higher in subjects with a disease duration over 10 years than in those within 10 years from onset. Conclusion: Our findings provide evidence for a difference in temporal dynamics between axial and appendicular ataxia in SCA3 patients, which will help inform the design of clinical trials and development of new (etiology-specific) outcome measures. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.Funding agencies: This publication is an outcome of ESMI, an EU Joint Programme — Neurodegenerative Disease Research (JPND) Project (www.jpnd.eu). The project is supported through the following funding organizations under the aegis of JPND: Germany, Federal Ministry of Education and Research (BMBF; funding codes 01ED1602A/B); Netherlands, The Netherlands Organization for Health Research and Development; Portugal, Foundation for Science and Technology and Regional Fund for Science and Technology of the Azores; United Kingdom, Medical Research Council. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement no. 643417. At the United States sites this work was in part supported by the National Ataxia Foundation.Spinocerebellar ataxia type 3Natural historyScale for the Assessment and Rating of AtaxiaDisease progressio

A standardized protocol for blood and cerebrospinal fluid collection and processing for biomarker research in ataxia

The European Spinocerebellar Ataxia Type 3/Machado-Joseph Disease Initiative (ESMI) is a consortium established with the ambition to set up the largest European longitudinal trial-ready cohort of Spinocerebellar Ataxia Type 3/Machado Joseph disease (SCA3/MJD), the most common autosomal dominantly inherited ataxia worldwide. A major focus of ESMI has been the identification of SCA3/MJD biomarkers to enable future interventional studies. As biosample collection and processing variables significantly impact the outcomes of biomarkers studies, biosampling procedures standardization was done previously to study visit initiation. Here, we describe the ESMI consensus biosampling protocol, developed within the scope of ESMI, that ultimately might be translated to other neurodegenerative disorders, particularly ataxias, being the first step to protocol harmonization in the field

Autoantibody screening in Guillain-Barré syndrome

Background: Guillain?Barré syndrome (GBS) is an acute inflammatory neuropathy with a heterogeneous presentation. Although some evidences support the role of autoantibodies in its pathogenesis, the target antigens remain unknown in a substantial proportion of GBS patients. The objective of this study is to screen for autoantibodies targeting peripheral nerve components in Guillain-Barré syndrome. Methods: Autoantibody screening was performed in serum samples from all GBS patients included in the International GBS Outcome study by 11 different Spanish centres. The screening included testing for anti-ganglioside antibodies, anti-nodo/paranodal antibodies, immunocytochemistry on neuroblastoma-derived human motor neurons and murine dorsal root ganglia (DRG) neurons, and immunohistochemistry on monkey peripheral nerve sections. We analysed the staining patterns of patients and controls. The prognostic value of anti-ganglioside antibodies was also analysed. Results: None of the GBS patients (n = 100) reacted against the nodo/paranodal proteins tested, and 61 (61%) were positive for, at least, one anti-ganglioside antibody. GBS sera reacted strongly against DRG neurons more frequently than controls both with IgG (6% vs 0%; p = 0.03) and IgM (11% vs 2.2%; p = 0.02) immunodetection. No differences were observed in the proportion of patients reacting against neuroblastoma-derived human motor neurons. Reactivity against monkey nerve tissue was frequently detected both in patients and controls, but specific patterns were only detected in GBS patients: IgG from 13 (13%) patients reacted strongly against Schwann cells. Finally, we confirmed that IgG anti-GM1 antibodies are associated with poorer outcomes independently of other known prognostic factor

Autoantibody screening in Guillain-Barré syndrome

Background: Guillain-Barré syndrome (GBS) is an acute inflammatory neuropathy with a heterogeneous presentation. Although some evidences support the role of autoantibodies in its pathogenesis, the target antigens remain unknown in a substantial proportion of GBS patients. The objective of this study is to screen for autoantibodies targeting peripheral nerve components in Guillain-Barré syndrome. Methods: Autoantibody screening was performed in serum samples from all GBS patients included in the International GBS Outcome study by 11 different Spanish centres. The screening included testing for anti-ganglioside antibodies, anti-nodo/paranodal antibodies, immunocytochemistry on neuroblastoma-derived human motor neurons and murine dorsal root ganglia (DRG) neurons, and immunohistochemistry on monkey peripheral nerve sections. We analysed the staining patterns of patients and controls. The prognostic value of anti-ganglioside antibodies was also analysed. Results: None of the GBS patients (n = 100) reacted against the nodo/paranodal proteins tested, and 61 (61%) were positive for, at least, one anti-ganglioside antibody. GBS sera reacted strongly against DRG neurons more frequently than controls both with IgG (6% vs 0%; p = 0.03) and IgM (11% vs 2.2%; p = 0.02) immunodetection. No differences were observed in the proportion of patients reacting against neuroblastoma-derived human motor neurons. Reactivity against monkey nerve tissue was frequently detected both in patients and controls, but specific patterns were only detected in GBS patients: IgG from 13 (13%) patients reacted strongly against Schwann cells. Finally, we confirmed that IgG anti-GM1 antibodies are associated with poorer outcomes independently of other known prognostic factors. Conclusion: Our study confirms that (1) GBS patients display a heterogeneous repertoire of autoantibodies targeting nerve cells and structures; (2) gangliosides are the most frequent antigens in GBS patients and have a prognostic value; (3) further antigen-discovery experiments may elucidate other potential antigens in GBS