Targeted Next-Generation Sequencing Study in Familial ALS-FTD Portuguese Patients Negative for C9orf72 HRE

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with clinical and etiological heterogeneity and a complex genetic contribution. Clinical, neuropathological, and genetic evidence revealed that ALS and frontotemporal dementia (FTD) are in part of a single disease continuum. Genetic causes have been identified in sporadic (SALS) and familial patients (FALS) and the recurrent genetic factor underlying ALS and FTD is the C9orf72 hexanucleotide repeat expansion (HRE). However, in our population, the concomitance of ALS and FTD cannot be explained by C9orf72 HRE in many FALS and SALS cases. Our aim is to further understand the genetic basis of ALS in Portuguese patients. 34 patients with FALS or SALS-FTD, negative for C9orf72 HRE, were screened for rare variants in a panel of 29 relevant genes by next-generation sequencing. We detected 15 variants in 11 genes, one classified as pathogenic in TARDBP, two as likely pathogenic in TARDBP and PRPH, and the others as variants of unknown significance (VUS). Gene variants, including VUS, were found in 41.2% FALS patients and 40% SALS-FTD. In most patients, no potential pathogenic variants were found. Our results emphasize the need to enhance the efforts to unravel the genetic architecture of ALS-FTD.info:eu-repo/semantics/publishedVersio

The Power of Non-Determinism in Higher-Order Implicit Complexity

We investigate the power of non-determinism in purely functional programming languages with higher-order types. Specifically, we consider cons-free programs of varying data orders, equipped with explicit non-deterministic choice. Cons-freeness roughly means that data constructors cannot occur in function bodies and all manipulation of storage space thus has to happen indirectly using the call stack. While cons-free programs have previously been used by several authors to characterise complexity classes, the work on non-deterministic programs has almost exclusively considered programs of data order 0. Previous work has shown that adding explicit non-determinism to cons-free programs taking data of order 0 does not increase expressivity; we prove that this - dramatically - is not the case for higher data orders: adding non-determinism to programs with data order at least 1 allows for a characterisation of the entire class of elementary-time decidable sets. Finally we show how, even with non-deterministic choice, the original hierarchy of characterisations is restored by imposing different restrictions.Comment: pre-edition version of a paper accepted for publication at ESOP'1

Endometrial Cancer Spheres Show Cancer Stem Cells Phenotype and Preference for Oxidative Metabolism

This study aimed to characterize endometrial cancer regarding cancer stem cells (CSC) markers, regulatory and differentiation pathways, tumorigenicity and glucose metabolism. Endometrial cancer cell line ECC1 was submitted to sphere forming protocols. The first spheres generation (ES1) was cultured in adherent conditions (G1). This procedure was repeated and was obtained generations of spheres (ES1, ES2 and ES3) and spheres-derived cells in adherent conditions (G1, G2 and G3). Populations were characterized regarding CD133, CD24, CD44, aldehyde dehydrogenase (ALDH), hormonal receptors, HER2, P53 and β-catenin, fluorine-18 fluorodeoxyglucose ([18F]FDG) uptake and metabolism by NMR spectroscopy. An heterotopic model evaluated differential tumor growth. The spheres self-renewal was higher in ES3. The putative CSC markers CD133, CD44 and ALDH expression were higher in spheres. The expression of estrogen receptor (ER)α and P53 decreased in spheres, ERβ and progesterone receptor had no significant changes and β-catenin showed a tendency to increase. There was a higher 18F-FDG uptake in spheres, which also showed a lower lactate production and an oxidative cytosol status. The tumorigenesis in vivo showed an earlier growth of tumours derived from ES3. Endometrial spheres presented self-renewal and differentiation capacity, expressed CSC markers and an undifferentiated phenotype, showing preference for oxidative metabolism.info:eu-repo/semantics/publishedVersio

Effects of mesenchymal stromal cells versus serum on tendon healing in a controlled experimental trial in an equine model

Abstract Background Mesenchymal stromal cells (MSC) have shown promising results in the treatment of tendinopathy in equine medicine, making this therapeutic approach seem favorable for translation to human medicine. Having demonstrated that MSC engraft within the tendon lesions after local injection in an equine model, we hypothesized that they would improve tendon healing superior to serum injection alone. Methods Quadrilateral tendon lesions were induced in six horses by mechanical tissue disruption combined with collagenase application 3 weeks before treatment. Adipose-derived MSC suspended in serum or serum alone were then injected intralesionally. Clinical examinations, ultrasound and magnetic resonance imaging were performed over 24 weeks. Tendon biopsies for histological assessment were taken from the hindlimbs 3 weeks after treatment. Horses were sacrificed after 24 weeks and forelimb tendons were subjected to macroscopic and histological examination as well as analysis of musculoskeletal marker expression. Results Tendons injected with MSC showed a transient increase in inflammation and lesion size, as indicated by clinical and imaging parameters between week 3 and 6 (p < 0.05). Thereafter, symptoms decreased in both groups and, except that in MSC-treated tendons, mean lesion signal intensity as seen in T2w magnetic resonance imaging and cellularity as seen in the histology (p < 0.05) were lower, no major differences could be found at week 24. Conclusions These data suggest that MSC have influenced the inflammatory reaction in a way not described in tendinopathy studies before. However, at the endpoint of the current study, 24 weeks after treatment, no distinct improvement was observed in MSC-treated tendons compared to the serum-injected controls. Future studies are necessary to elucidate whether and under which conditions MSC are beneficial for tendon healing before translation into human medicine

Balance of IL-10 and Interferon-γ plasma levels in human visceral leishmaniasis: Implications in the pathogenesis

BACKGROUND: Leishmaniasis remains a serious public health problem in several parts of the developing world. Effective prophylactic measurements are hampered by imprecise comprehension of different aspects of the disease, including its immunoregulation. A better comprehension of immunoregulation in human VL may be useful both for designing and evaluating immunoprophylaxis. METHODS: To explore immunoregulatory mechanisms, 20 visceral leishmaniasis (VL) patients were evaluated during active disease and at different periods up to one year after treatment determining their plasma cytokine levels, clinical parameters (palpable spleen and liver) and antibody levels. RESULTS: Elevated plasma levels of IFN-γ and of IL-12 p40 were observed during active disease, significantly decreasing after treatment whereas in vitro Leishmania antigen-stimulated IFN-γ production by PBMC exhibited an inverse pattern being low during disease and increasing steadily thereafter. Absence of IFN-γ activity is a hallmark of VL. The main candidate for blunting IFN-γ activity is IL-10, a cytokine highly elevated in plasma with sharp decrease after treatment. Activity of IL-10 is inferred by high levels of anti-Leishmania specific IgG1 and IgG3. TGF-β had elevated total, but not of active, levels lessening the likelihood of being the IFN-γ counterpart. Spleen or liver size presented a steady decrease but return to normal values at only 120 days after treatment. Anti-Leishmania IgG (total and subclasses) levels and DTH or Leishmania-stimulated lymphocyte proliferation conversion to positive also present a slow decrease after treatment. IL-6 plasma levels were elevated in only a few patients. CONCLUSION: Taken together our results suggest that IFN-γ and IL-10 are the molecules most likely involved in determining fate of disease. After treatment, there is a long delay before the immune profile returns to normal what precludes using plasma cytokine levels as criteria of cure as simpler clinical evaluations, as a palpable spleen or liver, can be used

DIAGNÓSTICOS DE ENFERMAGEM COMO INSTRUMENTOS NA FORMAÇÃO DO ENFERMEIRO: UMA REVISÃO DE LITERATURA

This research presents a review of articles on the topic of "Nursing Diagnosis," published during the years 2000-2007 and refers to a trend which has increasingly been mentioned in practice and in medical journals: The study is based on evidence. To this end, we used the methodology of a bibliographic search, using data sources such as the national nursing journals indexed in the LILACS database (Latin-American and Caribbean Health Sciences Literature), MEDLINE (International Science Literature Health) SciELO (Scientific Electronic Library Online). We chose to use the term "Nursing Diagnosis," that was found in 65 articles. For the analysis it was revealed the nature of the items, the assessment of the authors and how they evaluated the diagnosis teaching of nursing. It highlighted the main theoretical references, issues of proportionality and the years of the publication in question. It can be concluded that the production of nursing articles, within the theme of the diagnosis, has been made even in small quantities, which indicates the need of production and application of these articles in the fields of teaching and practice.  Esta investigación presenta una revisión bibliográfica de los artículos sobre el tema "Diagnóstico de Enfermería", publicados durante los años 2000-2007, y se refiere a una tendencia que se ha afirmado en la práctica y en las revistas médicas: el estudio basado en la evidencia. Con este fin, hemos utilizado la metodología de búsqueda bibliográfica, utilizando fuentes de datos como las revistas de enfermería nacional indexadas a la base de datos LILACS (Literatura Latino-Americana y del Caribe en Ciencias de la Salud), MEDLINE (Literatura Internacional en Ciencias Salud) y SCIELO (Scientific Electronic Library Online). Se optó por utilizar el término "Diagnóstico de Enfermería", donde fueron encontrados 65 artículos. Para el análisis se puso de manifiesto la naturaleza de los artículos, la valoración de los autores, y cómo estos evalúan el diagnóstico en el hacer/educar en enfermería. Se destacaron los principales teóricos referenciados, temas y proporcionalidad de los años de la publicación en cuestión. Se puede concluir que la producción de artículos de enfermería, dentro de la temática del diagnóstico, se ha hecho incluso en pequeñas cantidades, lo que indica la necesidad de la producción y aplicación de estos productos en las esferas de la enseñanza y la práctica, teniendo en cuenta la consideración de enfermería como ciencia.Esta pesquisa apresenta uma revisão de literatura dos artigos referentes ao tema “Diagnósticos de Enfermagem” publicados no período dos anos 2000-2007, e relaciona-os a uma tendência, a qual cada vez tem se afirmado na prática e nas publicações médicas: o estudo baseado em evidências. Para tal, utilizou-se a metodologia de pesquisa bibliográfica, utilizando-se como fontes de dados as Revistas de Enfermagem nacionais indexadas às base de dados LILACS (Literatura Latino-Americana e do Caribe em Ciências da Saúde), MEDLINE (Literatura Internacional em Ciências da Saúde) e Scielo (Scientific Electronic Library Online). Utilizando como descritor o termo “Diagnósticos de Enfermagem”, foram encontrados 65 artigos. Para a análise demonstramos a natureza dos artigos, a titulação dos autores, e como estes avaliam a visão diagnóstica dentro do fazer/ educar na Enfermagem, procuramos evidenciar os principais teóricos referenciados, temáticas e proporcionalidade dos anos de publicação em questão. Pode-se concluir que a produção de artigos na Enfermagem, dentro da temática diagnóstico, tem-se apresentado ainda em pequena quantidade, evidenciando a necessidade de produção e aplicabilidade destas produções, nos campos da docência e prática, visto a afirmação da Enfermagem como ciência.  

A randomized trial and novel SPR technique identifies altered lipoprotein-LDL receptor binding as a mechanism underlying elevated LDL-cholesterol in APOE4s

At a population level APOE4 carriers (~25% Caucasians) are at higher risk of cardiovascular diseases. The penetrance of genotype is however variable and influenced by dietary fat composition, with the APOE4 allele associated with greater LDL-cholesterol elevation in response to saturated fatty acids (SFA). The etiology of this greater responsiveness is unknown. Here a novel surface plasmon resonance technique (SPR) is developed and used, along with hepatocyte (with the liver being the main organ modulating lipoprotein metabolism and plasma lipid levels) uptake studies to establish the impact of dietary fatty acid composition on, lipoprotein-LDL receptor (LDLR) binding, and hepatocyte uptake, according to APOE genotype status. In men prospectively recruited according to APOE genotype (APOE3/3 common genotype, or APOE3/E4), triglyceride-rich lipoproteins (TRLs) were isolated at fasting and 4-6 h following test meals rich in SFA, unsaturated fat and SFA with fish oil. In APOE4s a greater LDLR binding affinity of postprandial TRL after SFA, and lower LDL binding and hepatocyte internalization, provide mechanisms for the greater LDL-cholesterol raising effect. The SPR technique developed may be used for the future study of the impact of genotype, and physiological and behavioral variables on lipoprotein metabolism