Consequences of Lmna Exon 4 Mutations in Myoblast Function.

Laminopathies are causally associated with mutations on the Lamin A/C gene (LMNA). To date, more than 400 mutations in LMNA have been reported in patients. These mutations are widely distributed throughout the entire gene and are associated with a wide range of phenotypes. Unfortunately, little is known about the mechanisms underlying the effect of the majority of these mutations. This is the case of more than 40 mutations that are located at exon 4. Using CRISPR/Cas9 technology, we generated a collection of Lmna exon 4 mutants in mouse C2C12 myoblasts. These cell models included different types of exon 4 deletions and the presence of R249W mutation, one of the human variants associated with a severe type of laminopathy, LMNA-associated congenital muscular dystrophy (L-CMD). We characterized these clones by measuring their nuclear circularity, myogenic differentiation capacity in 2D and 3D conditions, DNA damage, and levels of p-ERK and p-AKT (phosphorylated Mitogen-Activated Protein Kinase 1/3 and AKT serine/threonine kinase 1). Our results indicated that Lmna exon 4 mutants showed abnormal nuclear morphology. In addition, levels and/or subcellular localization of different members of the lamin and LINC (LInker of Nucleoskeleton and Cytoskeleton) complex were altered in all these mutants. Whereas no significant differences were observed for ERK and AKT activities, the accumulation of DNA damage was associated to the Lmna p.R249W mutant myoblasts. Finally, significant myogenic differentiation defects were detected in the Lmna exon 4 mutants. These results have key implications in the development of future therapeutic strategies for the treatment of laminopathies.

A pessoa com problemas ligados ao álcool - interpretação e intervenção sistémica / The person with alcohol-related problems - interpretation and systemic intervention

Os cuidados de Enfermagem na Comunidade englobam um vasto leque de intervenções, no entanto o aprofundamento em determinadas áreas assume uma importância elevada. O presente trabalho pretende demonstrar a evidência científica no tratamento das pessoas com Problemas Ligados ao Álcool. A dependência alcoólica é uma doença caracterizada pela cronicidade, comorbilidade patológica e consequentes alterações ao nível orgânico, psicológico e social. A intervenção da Enfermagem nestas pessoas implica uma abordagem específica e sistémica, com o objetivo de reduzir a probabilidade de recaídas e promover estilos de vida saudáveis. Foram realizadas diversas intervenções individuais e de grupo assentes nos princípios da capacitação e do empowerment, as quais resultaram de uma contextualização teórica sistémica e enquadramento na unidade de tratamento especializada. Daqui decorreu a construção de um modelo original de interpretação sistémica da problemática. Com a finalidade de demonstrar a prática baseada na evidência foi também realizada uma Revisão Sistemática da Literatura

Community Nursing care cover a wide range of interventions, however scoop out knowledge in certain areas is very important. This document aims to demonstrate the scientific evidence in the treatment of people with Alcohol Related Problems. Alcohol dependence is a disease characterized by chronicity, pathological comorbidity and consequential amendments to organic, psychological and social level. The intervention of Nursing in these persons implies a specific and systemic approach, aiming to reduce the chance of relapse and promote healthy lifestyles. There have been made several individual and group interventions based on the principles of empowerment, wich resulted from a systemic theoretical context and framed in the specialized care unit. Here took place the building of an original model of systemic interpretation. In order to demonstrate evidence-based practice, a Systematic Literature Review was also performed