A dynamic role for HDAC7 in MEF2-mediated muscle differentiation

The overlapping expression profile of MEF2 and the class-II histone deacetylase, HDAC7, led us to investigate the functional interaction and relationship between these regulatory proteins. HDAC7 expression inhibits the activity of MEF2 (-A, -C, and -D), and in contrast MyoD and Myogenin activities are not affected. Glutathione S-transferase pulldown and immunoprecipitation demonstrate that the repression mechanism involves direct interactions between MEF2 proteins and HDAC7 and is associated with the ability of MEF2 to interact with the N-terminal 121 amino acids of HDAC7 that encode repression domain 1. The MADS domain of MEF2 mediates the direct interaction of MEF2 with HDAC7, MEF2 inhibition by HDAC7 is dependent on the N-terminal repression domain and surprisingly does not involve the C-terminal deacetylase domain. HDAC7 interacts with CtBP and other class-I and -II HDACs suggesting that silencing of MEF2 activity involves corepressor recruitment. Furthermore, we show that induction of muscle differentiation by serum withdrawal leads to the translocation of HDAC7 from the nucleus into the cytoplasm. This work demonstrates that HDAC7 regulates the function of MEF2 proteins and suggests that this class-II HDAC regulates this important transcriptional (and pathophysiological) target in heart and muscle tissue. The nucleocytoplasmic trafficking of HDAC7 and other class-II HDACs during myogenesis provides an ideal mechanism for the regulation of HDAC targets during mammalian development and differentiation.

Stimulation of protein kinase C-dependent and -independent signaling pathways by bistratene A in intestinal epithelial cells

The marine toxin bistratene A (BisA) potently induces cytostasis and differentiation in a variety of systems. Evidence that BisA is a selective activator of protein kinase C (PKC) delta implicates PKC delta signaling in the negative growth-regulatory effects of this agent. The current study further investigates the signaling pathways activated by BisA by comparing its effects with those of the PKC agonist phorbol 12-myristate 13-acetate (PMA) in the IEC-18 intestinal crypt cell line. Both BisA and PMA induced cell cycle arrest in these cells, albeit with different kinetics. While BisA produced sustained cell cycle arrest in G(o)/G(1) and G(2)/M, the effects of PMA were transient and involved mainly a G(o)/G(1), blockade. BisA also produced apoptosis in a proportion of the population, an effect not seen with PMA. Both agents induced membrane translocation/activation of PKC, with BisA translocating only PKC delta and PMA translocating PKC alpha, delta, and epsilon in these cells. Notably, while depletion of PKC alpha, delta, and epsilon abrogated the cell cycle-specific effects of PMA in IEC-18 cells, the absence of these PKC isozymes failed to inhibit BisA-induced G(o)/G(1), and G(2)/M arrest or apoptosis. The cell cycle inhibitory and apoptotic effects of BisA, therefore, appear to be PKC-independent in IEG-18 cells. On the other hand, BisA and PMA both promoted PKC-dependent activation of Erk 1 and 2 in this system. Thus, intestinal epithelial cells respond to BisA through activation of at least two signaling pathways: a PKC delta -dependent pathway, which leads to activation of mitogen-activated protein kinase and possibly cytostasis in the appropriate context, and a PKC-independent pathway, which induces both cell cycle arrest in G(o)/G(1) and G(2)/M and apoptosis through as yet unknown mechanisms. (C) 2001 Elsevier Science Inc. All rights reserved.

Mouse Sox8 is located between, not within, the t-complex deletions t(w18) and t(h20) on chromosome 17

The SOX family of developmental transcription factors is known to play critical roles in cell lineage specification, fate determination and differentiation during development in diverse phyla. Their importance is underscored by their involvement in a number of human diseases and mouse mutants, and by targeted mutation in mice. SOX8 is broadly expressed during development and is located on human chromosome 16p and within the t-complex on mouse chromosome 17, in the vicinity of two mutations t(w18) and t(h20). Here we analyse mutant genomic DNA to show that the Sox8 gene locus lies outside the deletion regions of both t(w18) and t(h20) and between these deletions. These data exclude Sox8 from contributing to the t(w18) and t(h20) phenotypes, and provide an additional marker for structural characterization of this complex genomic region. Copyright (C) 2001 S. Karger AG, Basel.

Coordinated expression of scleraxis and Sox9 genes during embryonic development of tendons and cartilage

Embryonic development of tendons is in close association with that of cartilage and bone. Although these tissues are derived from mesenchymal progenitor cells which also give rise to muscle and fat, their fates clearly diverse in early embryonic stages, Transcription factors may play pivotal roles in the process of determination and differentiation of tendon cells as well as other cells in the skeletal system. Scleraxis, a basic helix-loop-helix (bHLH) type transcription factor. is expressed in mesenchymal progenitors that later form connective tissues including tendons. Sox9 is an HMG-box containing transcription factor, which is expressed at high levels in chondrocytes. We hypothesized that the two transcription factors regulate the fate of cells that interact with each other at the interface between the two tissues during divergence of their differentiation pathways, To address this point, we investigated scleraxis and Sox9 rnRNA expression during mouse embyogenesis focusing on the coordinated development of tendons and skeletons, In the early stage of mesenchymal tissue development at 10.5 d.p.c., scleraxis and Sox9 transcripts were expressed in the mesenchymal progenitor cells in the appendicular and axial mesenchyme. At 11.5 d.p.c.. scleraxis transcripts were observed in the mesenchymal tissue surrounding skeletal primordia which express Sox9. From this stage, scleraxis expression was closely associated with, but distinct from, formation of skeletal primordia, At 13.5 d.p.c., scleraxis was expressed broadly in the interface between muscle and skeletal primordia while Sox9 expression is confined within the early skeletal primordia. Then. at 15.5 d.p.c., scleraxis transcripts were more restricted to tendons. These observations revealed the presence of temporal and spatial association of scleraxis expression during embryonic development of tendon precursor cells in close association with that of So,0 expression in chondrogenic cells in skeletal tissues. (C) 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.

Fibroblast growth factor receptor 4 (FGFR4) expression in newborn murine calvaria and primary osteoblast cultures

Fibroblast growth factor receptor (FGFR) signalling is important in the initiation and regulation of osteogenesis. Although mutations in FGFR1, 2 and 3 genes are known to cause skeletal deformities, the expression of FGFR4 in bony tissue remains unclear. We have investigated the expression pattern of FGFR4 in the neonatal mouse calvaria and compared it to the expression pattern in cultures of primary osteoblasts. Immunohistochemistry demonstrated that FGFR4 was highly expressed in rudimentary membranous bone and strictly localised to the cellular components (osteoblasts) between the periosteal and endosteal layers. Cells in close proximity to the newly formed osteoid (preosteoblasts) also expressed FGFR4 on both the endosteal and periosteal surfaces. Immunocytochemical analysis of primary osteoblast cultures taken from the same cranial region also revealed high levels of FGFR4 expression, suggesting a similar pattern of cellular expression in vivo and in vitro. RT-PCR and Western blotting for FGFR4 confirmed its presence in primary osteoblast cultures. These results suggest that FGFR4 may be an important regulator of osteogenesis with involvement in preosteoblast proliferation and differentiation as well as osteoblast functioning during intramembranous ossification. The consistent expression of FGFR4 in vivo and in vitro supports the use of primary osteoblast cultures for elucidating the role of FGFR4 during osteogenesis.

Internacionalização do vinho do Porto – estudo de caso Grupo Sogevinus

Dissertação apresentada ao Instituto Superior de Contabilidade para a obtenção do Grau de Mestre em Empreendedorismo e Internacionalização Orientada por Professor Doutor José Freitas Santos

Radiographic outcomes and evaluation of developmental dysplasia of the hip in children

The Developmental Dysplasia of the Hip (DDH), also know as Congenital Dislocation of the Hip, is common in infants and children and may persist into adulthood. The radiographic interpretation is highly conditioned by appropriate patient positioning and image quality criteria. The main goal of this study is to demonstrate the value of radiographic evaluation of DDH. Through the retrospective analysis of 65 radiographs of the hips, only 2 (3.1%) female patients with 1-2 years of age presented radiographic findings of DDH. The inappropriate field size and the improper placement and size of the gonadal shields, were the most common errors observed.

New mitochondrial and nuclear evidences support recent demographic expansion and an atypical phylogeographic pattern in the Spittlebug Philaenus spumarius (Hemiptera, Aphrophoridae)

Copyright: © 2014 Rodrigues et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

STIR, SPIR and SPAIR techniques in magnetic resonance of the breast: a comparative study

The amount of fat is a component that complicates the clinical evaluation and the differential diagnostic between benign and malign lesions in the breast MRI examinations. To overcome this problem, an effective erasing of the fat signal over the images acquisition process, is essentials. This study aims to compare three fat suppression techniques (STIR, SPIR, SPAIR) in the MR images of the breast and to evaluate the best image quality regarding its clinical usefulness. To mimic breast women, a breast phantom was constructed. First the exterior contour and, in second time, its content which was selected based on 7 samples with different components. Finally it was undergone to a MRI breast protocol with the three different fat saturation techniques. The examinations were performed on a 1.5 T MRI system (Philips®). A group of 5 experts evaluated 9 sequences, 3 of each with fat suppression techniques, in which the frequency offset and TI (Inversion Time) were the variables changed. This qualitative image analysis was performed according 4 parameters (saturation uniformity, saturation efficacy, detail of the anatomical structures and differentiation between the fibroglandular and adipose tissue), using a five-point Likert scale. The statistics analysis showed that anyone of the fat suppression techniques demonstrated significant differences compared to the others with (p > 0.05) and regarding each parameter independently. By Fleiss’ kappa coefficient there was a good agreement among observers P(e) = 0.68. When comparing STIR, SPIR and SPAIR techniques it was confirmed that all of them have advantages in the study of the breast MRI. For the studied parameters, the results through the Friedman Test showed that there are similar advantages applying anyone of these techniques.

Besnoitia besnoiti and Toxoplasma gondii: two apicomplexan strategies to manipulate the host cell centrosome and Golgi apparatus

Besnoitia besnoiti and Toxoplasma gondii are two closely related parasites that interact with the host cell microtubule cytoskeleton during host cell invasion. Here we studied the relationship between the ability of these parasites to invade and to recruit the host cell centrosome and the Golgi apparatus. We observed that T. gondii recruits the host cell centrosome towards the parasitophorous vacuole (PV), whereas B. besnoiti does not. Notably, both parasites recruit the host Golgi apparatus to the PV but its organization is affected in different ways. We also investigated the impact of depleting and over-expressing the host centrosomal protein TBCCD1, involved in centrosome positioning and Golgi apparatus integrity, on the ability of these parasites to invade and replicate. Toxoplasma gondii replication rate decreases in cells over-expressing TBCCD1 but not in TBCCD1-depleted cells; while for B. besnoiti no differences were found. However, B. besnoiti promotes a reorganization of the Golgi ribbon previously fragmented by TBCCD1 depletion. These results suggest that successful establishment of PVs in the host cell requires modulation of the Golgi apparatus which probably involves modifications in microtubule cytoskeleton organization and dynamics. These differences in how T. gondii and B. besnoiti interact with their host cells may indicate different evolutionary paths.

On the choice of posittioning and cluster size for fixed relay stations in a LTE network

The long term evolution (LTE) is one of the latest standards in the mobile communications market. To achieve its performance, LTE networks use several techniques, such as multi-carrier technique, multiple-input-multiple-output and cooperative communications. Inside cooperative communications, this paper focuses on the fixed relaying technique, presenting a way for determining the best position to deploy the relay station (RS), from a set of empirical good solutions, and also to quantify the associated performance gain using different cluster size configurations. The best RS position was obtained through realistic simulations, which set it as the middle of the cell's circumference arc. Additionally, it also confirmed that network's performance is improved when the number of RSs is increased. It was possible to conclude that, for each deployed RS, the percentage of area served by an RS increases about 10 %. Furthermore, the mean data rate in the cell has been increased by approximately 60 % through the use of RSs. Finally, a given scenario with a larger number of RSs, can experience the same performance as an equivalent scenario without RSs, but with higher reuse distance. This conduces to a compromise solution between RS installation and cluster size, in order to maximize capacity, as well as performance.

Characteristics of IEF, SDS and SAR-PAGE of Cuba EPO biosimilar

EPO is a glycoprotein produced in the kidney, which stimulates the division and differentiation of red cells in the bone marrow. Erythropoietin is available as a therapeutic agent produced by recombinant DNA technology in mammalian cell culture into which the human EPO gene has been transfected. Biosimilar Epoetins are mostly erythropoietins of the Epoetin alfa, beta or omega type, which are being produced at much lower cost due to expired patents. Recombinant human erythropoietin (rh-EPO) contains the identical amino acid sequence of natural EPO: 165 amino acids, with a molecular weight of 30,400 Da. Since glycosylation is not only dependent on the cell-line used for the expression of Epoetins but also on the entire biotechnological process the glycosylation patterns of biosimilars do not necessarily reflect the patterns of the originator compounds. Today biosimilar Epoetins are manufactured and distributed worldwide and under many different names. The use of recombinant EPOs for doping is prohibited because of its performance enhancing effect. The aim of the present study was to investigated whether biosimilar alpha r-HuEPO – ior®-EPOCIM, produced in Cuba and also available in other countries in all continents, could be differentiated from endogenous one by iso-electro-focusing plus double blotting, SDS-PAGE and SAR-PAGE for antidoping analysis.

Development of a system to monitor surfer’s balance and plantar pressure and surfboard’s movement

O uso da tecnologia tem crescido nas últimas décadas nas mais diversas áreas, seja na indústria ou no dia-a-dia, e é cada vez mais evidente os benefícios que traz. No desporto não é diferente. Cada dia surgem novos desenvolvimentos objetivando a melhoria do desempenho dos praticantes de atividades físicas, possibilitando atingir resultados nunca antes pensados. Além disto, a utilização da tecnologia no desporto permite a obtenção de dados biomecânicos que podem ser utilizados tanto no treinamento quando na melhoria da qualidade de vida dos atletas auxiliando na prevenção de lesões, por exemplo. Deste modo, o presente projeto se aplica na área do desporto, nomeadamente, na modalidade do surfe, onde a ausência de trabalhos científicos ainda é elevada, aliando a tecnologia eletrônica ao desporto para quantificar informações até então desconhecidas. Três fatores básicos de desempenho foram levantados, sendo eles: equilíbrio, posicionamento dos pés e movimentação da prancha de surfe. Estes fatores levaram ao desenvolvimento de um sistema capaz de medi-los dinamicamente através da medição das forças plantares e da rotação da prancha de surfe. Além da medição dos fatores, o sistema é capaz de armazenar os dados adquiridos localmente através de um cartão de memória, para posterior análise; e também enviá-los através de uma comunicação sem fio, permitindo a visualização do centro de pressões plantares; dos ângulos de rotação da prancha de surfe; e da ativação dos sensores; em tempo real. O dispositivo consiste em um sistema eletrônico embarcado composto por um microcontrolador ATMEGA1280; um circuito de aquisição e condicionamento de sinal analógico; uma central inercial; um módulo de comunicação sem fio RN131; e um conjunto de sensores de força Flexiforce. O firmware embarcado foi desenvolvido em linguagem C. O software Matlab foi utilizado para receção de dados e visualização em tempo real. Os testes realizados demostraram que o funcionamento do sistema atende aos requisitos propostos, fornecendo informação acerca do equilíbrio, através do centro de pressões; do posicionamento dos pés, através da distribuição das pressões plantares; e do movimento da prancha nos eixos pitch e roll, através da central inercial. O erro médio de medição de força verificado foi de -0.0012 ± 0.0064 N, enquanto a mínima distância alcançada na transmissão sem fios foi de 100 m. A potência medida do sistema foi de 330 mW.

Functional and therapeutical implications of ligand recognition by the scavenger-like lymphocyte receptors CD5 and CD6

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

A CSR communication strategy for Continente: Improving brand image and reinforcing local community bonds

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics