Immunohistochemical Evaluation of TNF Expression in Canine Cystic Endometrial Hyperplasia

Tumour Necrosis Factor (TNF) has been identified in the uterus of several species, and altered TNF expression is reported in some pathological conditions. This study sought to evaluate TNF expression in canine cystic endometrial hyperplasia (CEH; n=20) and compare it with expression in postpartum samples (PP; n=5).This work was supported by Portuguese Foundation for Science and Technology, Center for Studies in Education, Technologies and by Health and by strategic project PEst-OE/CED/UI4016/201

Pre-selection of fibroblasts subsets prompt prevascularization of tissue engineered skin analogues

The papillary and reticular dermis harbors phenotypically distinct fibroblasts, whose functions such as maintenance of skin's microvasculature are also distinct. Thus, we hypothesized that pre-selection of the subpopulations of fibroblasts would benefit the generation of skin tissue engineered (TE) constructs, promoting their prevascularization in vitro. We first isolated papillary and reticular fibroblasts using fluorescence-activated cell sorting and studied the effect of their secretome and extracellular matrix (ECM) on human dermal microvascular endothelial cell (hDMEC) organization. Subsequently, we developed a bilayered 3D polymeric structure with distinct layer-associated features to house the subpopulations of fibroblasts, to generate a skin analogue. Both papillary and reticular fibroblasts were able to stimulate capillary-like network formation in a Matrigel assay. However, the secretome of the two subpopulations was substantially different, being enriched in VEGF, IGF-1, and Angio-1 in the case of papillary fibroblasts and in HGF and FGF-2 for the reticular subset. In addition, the fibroblast subpopulations deposited varied levels of ECM proteins, more collagen I and laminin was produced by the reticular subset, but these differences did not impact hDMEC organization. Vessel-like structures with lumens were observed earlier in the 3D skin analogue prepared with the sorted fibroblasts, although ECM deposition was not affected by the cell's pre-selection. Moreover, a more differentiated epidermal layer was obtained in the skin analogue formed by the sorted fibroblasts, confirming that its whole structure was not affected. Overall, we provide evidence that pre-selection of papillary and reticular fibroblasts is relevant for promoting the in vitro prevascularization of skin TE constructs.The authors would like to acknowledge the financial support from the Consolidator Grant “ECM_INK” (ERC-2016- COG-726061), to the FSE/POCH (Fundo Social Europeu através do Programa Operacional do Capital Humano) under the scope of the PD/169/2013, NORTE-08-5369-FSE-000037 (H.R. M.)

Alginate-lysozyme nanofibers hydrogels with improved rheological behavior, printability and biological properties for 3D bioprinting applications

In this study, alginate nanocomposite hydrogel bioinks reinforced with lysozyme nanofibers (LNFs) were developed. Alginate-LNF (A-LNF) suspensions with different LNF contents (1, 5 and 10 wt.%) were prepared and pre-crosslinked with 0.5% (w/v) CaCl2 to formulate A-LNF inks. These inks exhibit proper shear-thinning behavior and good recovery properties (~90%), with the pre-crosslinking step playing a crucial role. A-LNF fully crosslinked hydrogels (with 2% (w/v) CaCl2) that mimic 3D printing scaffolds were prepared, and it was observed that the addition of LNFs improved several properties of the hydrogels, such as the morphology, swelling and degradation profiles, and mechanical properties. All formulations are also noncytotoxic towards HaCaT cells. The printing parameters and 3D scaffold model were then optimized, with A-LNF inks showing improved printability. Selected A-LNF inks (A-LNF0 and A-LNF5) were loaded with HaCaT cells (cell density 2 × 106 cells mL-1), and the cell viability within the bioprinted scaffolds was evaluated for 1, 3 and 7 days, with scaffolds printed with the A-LNF5 bioink showing the highest values for 7 days (87.99 ± 1.28%). Hence, A-LNF bioinks exhibited improved rheological performance, printability and biological properties representing a good strategy to overcome the main limitations of alginate-based bioinks.publishe

High resolution melting : improvements in the genetic diagnosis of hypertrophic cardiomyopathy in a Portuguese cohort

© 2012 Susana et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Background: Hypertrophic Cardiomyopathy (HCM) is a complex myocardial disorder with a recognized genetic heterogeneity. The elevated number of genes and mutations involved in HCM limits a gene-based diagnosis that should be considered of most importance for basic research and clinical medicine. Methodology: In this report, we evaluated High Resolution Melting (HRM) robustness, regarding HCM genetic testing, by means of analyzing 28 HCM-associated genes, including the most frequent 4 HCM-associated sarcomere genes, as well as 24 genes with lower reported HCM-phenotype association. We analyzed 80 Portuguese individuals with clinical phenotype of HCM allowing simultaneously a better characterization of this disease in the Portuguese population. Results: HRM technology allowed us to identify 60 mutated alleles in 72 HCM patients: 49 missense mutations, 3 nonsense mutations, one 1-bp deletion, one 5-bp deletion, one in frame 3-bp deletion, one insertion/deletion, 3 splice mutations, one 5’UTR mutation in MYH7, MYBPC3, TNNT2, TNNI3, CSRP3, MYH6 and MYL2 genes. Significantly 22 are novel gene mutations. Conclusions: HRM was proven to be a technique with high sensitivity and a low false positive ratio allowing a rapid, innovative and low cost genotyping of HCM. In a short return, HRM as a gene scanning technique could be a cost-effective gene-based diagnosis for an accurate HCM genetic diagnosis and hopefully providing new insights into genotype/phenotype correlations.info:eu-repo/semantics/publishedVersio

Aplicação das técnicas de deteção de apoptose em ensaios clínicos

A aplicação de metodologias capazes de identificar células apoptóticas constitui uma valiosa ferramenta em vários estudos biomédicos. A hemorragia grave continua a ser a principal causa de morbilidade e mortalidade em animais de companhia vítimas de trauma, situação também comum em humanos, sobretudo jovens vítimas de acidentes de viação. A reperfusão é fundamental, nestas situações, para fomentar a redistribuição do fluxo sanguíneo, repor a disponibilidade de oxigénio e retomar a síntese de ATP, para reconstruir as reservas de energia perdidas e prevenir a morte celular, incluindo a apoptose. Este estudo tem como objectivo principal avaliar os niveis de apoptose no intestino delgado e rim, num ensaio clínico que reproduz a situação de hemorragia pós-trauma, comparando a reperfusão feita com hidroxietilamido 130/0,4 (HES), um coloide, e o Lactato de Ringer (LR), um cristaloide. Dezoito porcos da raça Large White foram submetidos a anestesia total intravenosa (TIVA) com propofol e remifentanil. Nos grupos 1 e 2, os animais foram submetidos a hemorragia controlada e a reposição de volume foi feita usando solução de LR no grupo 1 e HES 130/0.4 no grupo 2. O grupo 3 (grupo controlo), foi apenas submetido a TIVA, sem nenhum outro procedimento. Uma hora após a reposição do volume, os animais foram eutanasiados

An antibody-based survey of Toxoplasma gondii and Neospora caninum Infection in client-owned cats from Portugal

Toxoplasma gondii and Neospora caninum are intracellular parasites with a great impact on human and animal health, respectively. This work aims to investigate the presence of antibodies against T. gondii and N. caninum in client-owned cats from Portugal and to identify risk factors. A total of 183 domestic cats were sampled and their owners answered an online questionnaire designed to obtain background information. The overall anti-T. gondii and anti-N. caninum seroprevalences were 13.1% and 3.8%, respectively. An indoor lifestyle was identified as a significant protection factor against T. gondii infection, while the presence of a chronic disease and the presence of antibodies against N. caninum were identified as significant risk factors to T. gondii seroprevalence. To the best of our knowledge, this is the first serosurvey on N. caninum seroprevalence in cats from Portugal.A91F-E8B8-FA62 | Teresa Susana Letra MateusN/

REASE – Rede regional de sensibilização dos serviços prestados pelos ecossistemas das pradarias de ervas marinhas e sapais na região do Algarve.

Projeto financiado pelo Fundo Ambiental da República PortuguesaA Rede de Educação Ambiental para os Serviços Ambientais (REASE) é uma iniciativa recentemente criada no Algarve, por instituições de pesquisa científica, educação, divulgação científica e uma ONG ambiental. O REASE tem como objetivo desenvolver projetos de educação ambiental na área dos Serviços Ecossistémicos (SE), especialmente aqueles gerados pela vegetação costeira, incluindo sapais e pradarias marítimas. Atualmente, a rede está trabalhando em: 1) capacitação de professores e gestores ambientais, a fim de promover o conhecimento científico sobre os serviços prestados pelos ecossistemas costeiros; 2) a criação de uma “incubadora de projetos” para poder desenvolver em escolas locais projetos SE inovadores de educação formal; 3) sensibilização dos estudantes e do público em geral sobre o papel dos ecossistemas para o bem-estar humano através de exposições e palestras em centros de divulgação científica; 4) publicação de um livro para crianças sobre SE de ervas marinhas, num processo participativo de base comunitária (ilustrações feitas por crianças nas escolas); 5) o lançamento de um projeto de Ciência Cidadã, para avaliar a produção de Carbono Azul no Algarve, envolvendo escolas que recebem kits de campo e de laboratório e respetivo apoio científico; 6) o desenvolvimento de uma aplicação móvel para fazer upload diretamente das medições de campo e dados do carbono azul no site do projeto, onde os dados são cientificamente validados e analisados.SFRH/BGCT/52704/2014 - REFORÇO DA INTERAÇÃO COM ENTIDADES EXTERNAS E RENOVAÇÃO DE ACTIVIDADES E DE CONTEÚDOS (SFRH/BGCT/52704/2014)info:eu-repo/semantics/publishedVersio

Effect of Cr(V) on reproductive organ morphology and sperm parameters: An experimental study in mice

BACKGROUND: Cr(V) species are formed during the intracellular reduction of Cr(VI), a ubiquitous environmental pollutant. In this study, the acute toxicity of a physiologically stable Cr(V) compound, [Cr(V)-BT](2- )(BT = bis(hydroxyethyl)aminotris(hydroxymethyl)methane) was investigated in the male reproductive system of sexually mature 60-day-old male ICR-CD1 mice. METHODS: Eight-week-old animals were subcutaneously injected daily with a dose of ca 8 μmol of Cr/mouse, during 5 days. The control group was injected with 0.5 mL of BT buffer. Testis and epididymis morphology was evaluated using light and transmission electron microscopy. Epididymal sperm counts, motility and acrosome integrity were also assayed using standard methods. RESULTS: Seminiferous epithelium abnormalities were detected in the Cr(V)-BT experimental group, including intraepithelial vacuolation, and remarkable degeneration of Sertoli cells, spermatocytes and spermatids. The premature release of germ cells into the tubular lumen was also evident. Histological evaluation of epididymal compartments revealed apparently normal features. However, the epididymal epithelium presented vacuolation. [Cr(V)-BT](2- )induced a reduction in sperm acrosome integrity. However, sperm motility and density were not significantly affected. CONCLUSION: This in vivo study using a Cr(V) compound, provides evidence for the potential reproductive hazards caused on male reproductive system by species containing chromium in intermediate oxidation states