Felicidade no trabalho e interação familiar: projeto de investigação com enfermeiros de um Agrupamento de Centros de Saúde

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Defects of splicing in antithrombin deficiency

Background: There is increasing evidence supporting the relevance of aberrant splicing in multiple disorders. In antithrombin deficiency only 22 intronic mutations affecting splicing sites (7% of SERPINC1 mutations) are considered as splicing mutations. Methods: SERPINC1 was analyzed by Sanger sequencing and MLPA in 141 unrelated cases with antithrombin deficiency. Plasma antithrombin was studied by functional and western blot assays, purified by FPLC and characterized by proteomic analysis. In silico predictions on splicing was done with the Human Splicing Finder software. Results: We detected 89 different SERPINC1 defects, 13 with potential effect on splicing. Ten cases presented 9 mutations disturbing splicing sites, 5 new. Three gross or small gene defects also disturbed a correct splicing. Interestingly, the first duplication of a single exon ever described (c.1154-13_1218+115dup), caused mild deficiency (75%). A deeper intronic mutation (c.1154-14G>A), identified in three unrelated patients with traces of disulphide dimers of antithrombin in plasma, created a cryptic splicing site that might generate a variant with 4 additional in frame residues according to in silico predictions. This aberrant splicing was confirmed by proteomic analysis of the dimer purified from plasma. Conclusions: A high proportion of cases with antithrombin deficiency (up to 13%) may be explained by an aberrant splicing. Up to 15% of mutations in SERPINC1: splicing site variations, gross gene defects and deep intronic mutations, may affect a correct splicing with three potential consequences type I, type II, and even moderate antithrombin deficiency

Interacção familiar e felicidade no trabalho: estudo comparativo com enfermeiros em pandemia COVID-19

Objetivo: comparar os níveis de felicidade e interação familiar em enfermeiros portugueses que exercem em contexto hospitalar e de cuidados de saúde primários, em plena vivencia de pandemia por COVID-19. Metodologia: estudo descritivo, correlacional e transversal, realizado com uma amostra de conveniência de 468 enfermeiros portugueses, a exercer funções em contexto hospitalar e de cuidados de saúde primários. Entre março e abril de 2021, em contexto de pandemia COVID-19, aplicou-se um questionário sociodemográfico e profissional e a versão portuguesa da Survey Work-Home Interaction e da Shorted Happiness at Work Scale. Os dados foram colhidos por via digital e tratados com recurso ao programa IBM-SPSS® versão 26.0. Foi obtida aprovação da Comissão de Ética para a saúde e a autorização das unidades de saúde envolvidas. Resultados: identificaram-se scores médios tendencialmente mais elevados de interação familiar positiva, na relação família-trabalho, nos enfermeiros que exercem no hospital e, de felicidade no trabalho, nos enfermeiros dos cuidados de saúde primários, ainda que sem relevância estatística entre os dois grupos de participantes. Em ambas as amostras, as dimensões engagement e compromisso organizacional afetivo evidenciaram médias superiores à da satisfação com o trabalho. Conclusão: o impacto que a pandemia por COVID-19 exerceu em enfermeiros, fez-se manifestar tendencialmente no seu equilíbrio de interação trabalho-família. Os valores superiores de engagement e compromisso organizacional afetivo, comparativamente à da satisfação com o trabalho, obtidos nesta amostra de enfermeiros, poderá ter estado associado à dedicação, atividade e energia destes profissionais, quer pela ligação com a sua profissão, quer pelo compromisso para fazer face aos desafios da pandemia.info:eu-repo/semantics/publishedVersio

Lentiviral gene therapy reverts GPIX expression and phenotype in Bernard-Soulier syndrome type C

Bernard-Soulier syndrome (BSS) is a rare congenital disease characterized by macrothrombocytopenia and frequent bleeding. It is caused by pathogenic variants in three genes (GP1BA, GP1BB, or GP9) that encode for the GPIbα, GPIbβ, and GPIX subunits of the GPIb-V-IX complex, the main platelet surface receptor for von Willebrand factor, being essential for platelet adhesion and aggregation. According to the affected gene, we distinguish BSS type A1 (GP1BA), type B (GP1BB), or type C (GP9). Pathogenic variants in these genes cause absent, incomplete, or dysfunctional GPIb-V-IX receptor and, consequently, a hemorrhagic phenotype. Using gene-editing tools, we generated knockout (KO) human cellular models that helped us to better understand GPIb-V-IX complex assembly. Furthermore, we developed novel lentiviral vectors capable of correcting GPIX expression, localization, and functionality in human GP9-KO megakaryoblastic cell lines. Generated GP9-KO induced pluripotent stem cells produced platelets that recapitulated the BSS phenotype: absence of GPIX on the membrane surface and large size. Importantly, gene therapy tools reverted both characteristics. Finally, hematopoietic stem cells from two unrelated BSS type C patients were transduced with the gene therapy vectors and differentiated to produce GPIX-expressing megakaryocytes and platelets with a reduced size. These results demonstrate the potential of lentiviral-based gene therapy to rescue BSS type C

Introducing high-throughput sequencing into mainstream genetic diagnosis practice in inherited platelet disorders

Inherited platelet disorders are a heterogeneous group of rare diseases, caused by inherited defects in platelet production and/or function. Their genetic diagnosis would benefit clinical care, prognosis and preventative treatments. Until recently, this diagnosis has usually been performed via Sanger sequencing of a limited number of candidate genes. High-throughput sequencing is revolutionizing the genetic diagnosis of diseases, including bleeding disorders. We have designed a novel high-throughput sequencing platform to investigate the unknown molecular pathology in a cohort of 82 patients with inherited platelet disorders. Thirty-four (41.5%) patients presented with a phenotype strongly indicative of a particular type of platelet disorder. The other patients had clinical bleeding indicative of platelet dysfunction, but with no identifiable features. The high-throughput sequencing test enabled a molecular diagnosis in 70% of these patients. This sensitivity increased to 90% among patients suspected of having a defined platelet disorder. We found 57 different candidate variants in 28 genes, of which 70% had not previously been described. Following consensus guidelines, we qualified 68.4% and 26.3% of the candidate variants as being pathogenic and likely pathogenic, respectively. In addition to establishing definitive diagnoses of well-known inherited platelet disorders, high-throughput sequencing also identified rarer disorders such as sitosterolemia, filamin and actinin deficiencies, and G protein-coupled receptor defects. This included disease-causing variants in DIAPH1 (n=2) and RASGRP2 (n=3). Our study reinforces the feasibility of introducing high-throughput sequencing technology into the mainstream laboratory for the genetic diagnostic practice in inherited platelet disorders.This study was supported by research grants from the Gerencia Regional de Salud (GRS 1370/A/16), ISCIII & Feder (PI14/01956), CIBERER CB15/00055, Fundación Séneca (19873/GERM/15) and Sociedad Española de Trombosis y Hemostasia (SETH). SPW holds a British Heart Foundation chair.Peer Reviewe

Happiness at work and family interaction: A study with nurses from a grouping of health centers

A promoção de ambientes de trabalho positivos e a Interação familiar em Enfermeiros, têm implicação na Felicidade no trabalho, representando um verdadeiro desafio para o Enfermeiro Gestor. Inserido no projeto Internacional de Saúde Ocupacional (INT-SO): dos contextos de trabalho à saúde ocupacional dos profissionais de Enfermagem, um estudo comparativo entre Portugal, Brasil e Espanha, este estudo, visa identificar: os níveis de Felicidade no trabalho e de Interação familiar, a sua relação em função de variáveis sociodemográficas e profissionais e, analisar a variação entre Felicidade no trabalho e a Interação familiar. Para a sua concretização desenvolveu-se um estudo quantitativo, descritivo, correlacional e transversal com 105 Enfermeiros de um Agrupamento de Centros de Saúde (ACeS) da região norte de Portugal, selecionados através de uma amostra de conveniência. O instrumento de colheita de dados contemplou um questionário sociodemográfico e profissional, a Shorted Happiness at Work Scale (SHAW) (Salas-Vallina & Alegre, 2018a; Queirós et al., 2020), e a Survey WorkHome Interaction Nijmegen (SWING, Geurts et al., 2005; Pereira et al., 2014), tendo a análise de dados sido efetuada através de estatística descritiva e inferencial. Os Enfermeiros apresentaram níveis moderados de Felicidade no trabalho, sendo estes mais elevados na dimensão do engagement e, níveis moderados a baixos de Interação familiar, com o score médio mais elevado na Interação positiva família-trabalho. Os Enfermeiros mais jovens e os que exerciam funções no Centro de Saúde 2 evidenciaram níveis superiores de engagement, os Enfermeiros Especialistas apresentaram níveis mais elevados de compromisso organizacional afetivo e, os que percecionam o trabalho stressante, níveis mais baixos de Felicidade no trabalho na escala total e em todas as suas dimensões. Os Enfermeiros do sexo feminino evidenciaram um nível mais elevado de Interação familiar e de influência negativa família-trabalho. Os participantes com filhos obtiveram níveis mais elevados de Interação negativa trabalho-família, os que percecionam trabalho stressante demonstraram níveis mais elevados de Interação familiar e de Interação negativa trabalhofamília e Interação positiva família-trabalho e, os que praticam atividades de lazer revelaram níveis médios mais baixos de Interação familiar (total e subescalas). Verificou-se, ainda, uma correlação negativa entre a Interação negativa trabalho-família e a escala total da Felicidade no trabalho, assim como na dimensão de engagement. Em suma, avaliar o modo como os Enfermeiros percecionam a sua Felicidade no trabalho e os níveis de Interação familiar, torna-se essencial para que os Enfermeiros gestores possam definir estratégias que potenciem ambientes de trabalho saudáveis.The promotion of positive work environments and family interactions in nurses has implications on work happiness, presenting as a challenge to the Nurse Manager. This work, incorporated in International Occupational Health Project (INT-SO): link between work environments and occupational health in nursing, a comparative study between Portugal, Brazil and Spain, aims to identify the levels of work happiness and family interactions, and their connection in relation to sociodemographic and professional variables. For this, a quantitative, descriptive, comparative and transverse study was developed, analysing a sample of 105 nurses, gathered through convenience sampling, from an Agrupamento de Centros de Saúde – Grouping of Health Centers (ACeS) of the North region of Portugal. The methods used for data collection were based on sociodemographic and professional surveys, such as a Shorted Happiness at Work Scale (SHAW) (Salas-Vallina & Alegre, 2018a; Queirós et al., 2020) and a Survey Work-Home Interaction Nijmegen (SWING, Geurts et al., 2005; Pereira et al., 2014). The data collected was then processed through descriptive and inferential statistical analysis. Overall, the nurses presented moderate levels of Work happiness, being higher in the engagement component; moderate to low levels of Family interaction, having obtained the highest average score on positive interaction family-work. Younger nurses and those working on Health Center 2 presented higher levels of engagement, Specialist Nurses presented higher levels of emotional organizational compromise and the lowest levels of work happiness on all its components. Female Nurses had generally higher levels of Family Interaction and of negative influence family-work. Participants with children had the highest levels of negative influence workfamily; those who view work as stressful had higher levels of family interaction, negative influence work-family and of positive influence family-work; those who enrol on hobbies or other free-time activities presented medium to low levels of Family Interaction (total and components). It was also verified a negative correlation between Negative interaction workfamily and overall work happiness as with the engagement component. Therefore, evaluating how nurses view their work happiness and family interactions becomes essential to Nurse Managers for the creation and planning of strategies that boost healthy work environment

Familial thrombotic risk based on the genetic background of Protein C Deficiency in a Portuguese Study

INTRODUCTION: Inherited protein C (PC) deficiency is a well-known risk factor for venous thrombosis (VT). Plasma PC levels are reliable in moderate to severe deficiencies; however, in mildly deficient individuals, the levels may overlap with those considered normal. Genetic studies of PROC, which encodes PC, could help identify carriers; genome-wide association studies (GWAS) have shown that approximately 50% of phenotypic variation in PC deficiency is caused by the cumulative effects of mutations in several other loci, namely in the PROCR. PATIENTS AND METHODS: With the main objective of determining the genotype/phenotype correlation in 59 Portuguese individuals from 26 unrelated families with history of thrombosis and repeatedly low/borderline PC plasma levels, we conducted a molecular study by direct sequencing of PROC; PROC promoter haplotypes and PROCR c.4600A>G polymorphism (rs867186), which are known to influence plasma PC concentrations, were also screened. RESULTS: Twelve different PROC mutations were identified, one of them not previously reported, p.Cys105Arg. The mutation types and locations as well as haplotype combinations correlated with the phenotypic severity. The most frequent mutation, p.Arg199X, correlated with the CGTC haplotype and was identified in nine families containing patients with higher numbers of VT episodes. This mutation in homozygous individuals for the CGTC haplotype is a significant risk factor for VT in Portuguese. CONCLUSION: These genetic family studies allowed the identification of the unknown carriers and individuals at a higher thrombotic risk within each family, thus permitting the evaluation of the need for prophylactic measures, particularly in at-risk situations.info:eu-repo/semantics/publishedVersio

Cerebral Venous Sinus Thrombosis in a Child with Idiopathic Nephrotic Syndrome: a case report

ABSTRACT Complications are rare in pediatric cases of idiopathic nephrotic syndrome (NS). Thromboembolism ranks among the most uncommon and difficult complications to diagnose, particularly in the first episode of NS, since clinical signs might be unspecific. This report describes the case of a 5-year-old girl with NS for the first time presenting with severe hypoalbuminemia (< 2g/dL). The patient responded poorly to therapy with corticosteroids. On day 8 of hospitalization she started having headaches and vomiting; she did not present hemodynamic alterations, fever or exanthems, and her neurological parameters were normal. The patient was suspected for intracranial hypertension, and computed tomography scans revealed she had cerebral venous sinus thrombosis (CVST). She was started on anticoagulants and showed clinical signs of improvement. The patient had no evident prothrombotic risk factors. She had three other episodes since she was diagnosed, one in which her plasma antithrombin level was low. Although antithrombin levels were normal in her first episode, she was tested after the resolution of proteinuria. The low levels of antithrombin seen in the first recurrence might have mirrored the initial drop in plasma antithrombin levels, an idea supported by the severe hypoalbuminemia she had when diagnosed. This severe manifestation of acquired thrombophilia might be in the origin of CVST. This report presents a rare case of thromboembolic complication in a pediatric patient with NS. The patient progressed well since she was started on anticoagulants. Although she did not present any evident risk factors at first, the development of her case indicated that severe acquired thrombophilia might have worked as the pathophysiological mechanism leading to CVST

Functional and molecular characterization of inherited platelet disorders in the Iberian Peninsula: results from a collaborative study

Abstract Background: The diagnostic evaluation of inherited platelet disorders (IPDs) is complicated and time-consuming, resulting in a relevant number of undiagnosed and incorrectly classified patients. In order to evaluate the spectrum of IPDs in individuals with clinical suspicion of these disorders, and to provide a diagnostic tool to centers not having access to specific platelets studies, we established the project &quot;Functional and Molecular Characterization of Patients with Inherited Platelet Disorders&quot; under the scientific sponsorship of the Spanish Society of Thrombosis and Haemostasis