Tubulin cofactor A gene silencing in mammalian cells induces changes in microtubule cytoskeleton, cell cycle arrest and cell death

Microtubules are polymers of alpha/beta-tubulin participating in essential cell functions. A multistep process involving distinct molecular chaperones and cofactors produces new tubulin heterodimers competent to polymerise. In vitro cofactor A (TBCA) interacts with beta-tubulin in a quasi-native state behaving as a molecular chaperone. We have used siRNA to silence TBCA expression in HeLa and MCF-7 mammalian cell lines. TBCA is essential for cell viability and its knockdown produces a decrease in the amount of soluble tubulin, modifications in microtubules and G1 cell cycle arrest. In MCF-7 cells, cell death was preceded by a change in cell shape resembling differentiation.

Sequencing of a 9.9 kb segment on the right arm of yeast chromosome VII reveals four open reading frames, including PFK1, the gene coding for succinyl-CoA synthetase (beta-chain) and two ORFs sharing homology with ORFs of the yeast chromosome VIII

A 9.9 kb DNA fragment from the right arm of chromosome VII of Saccharomyces cerevisiae has been sequenced and analysed. The sequence contains four open reading frames (ORFs) longer than 100 amino acids. One gene, PFK1, has already been cloned and sequenced and the other one is the probable yeast gene coding for the beta-subunit of the succinyl-CoA synthetase. The two remaining ORFs share homology with the deduced amino acid sequence (and their physical arrangement is similar to that) of the YHR161c and YHR162w ORFs from chromosome VIII.

Sequencing of a 17.6 kb segment on the right arm of yeast chromosome VII reveals 12 ORFs, including CCT, ADE3 and TR-I genes, homologues of the yeast PMT and EF1G genes, of the human and bacterial electron-transferring flavoproteins (beta-chain) and of the Escherichia coli phosphoserine phosphohydrolase, and five new ORFs.

A 17.6 kb DNA fragment from the right arm of chromosome VII of Saccharomyces cerevisiae has been sequenced and analysed. The sequence contains twelve open reading frames (ORFs) longer than 100 amino acids. Three genes had already been cloned and sequenced: CCT, ADE3 and TR-I. Two ORFs are similar to other yeast genes: G7722 with the YAL023 (PMT2) and PMT1 genes, encoding two integral membrane proteins, and G7727 with the first half of the genes encoding elongation factors 1gamma, TEF3 and TEF4. Two other ORFs, G7742 and G7744, are most probably yeast orthologues of the human and Paracoccus denitrificans electron-transferring flavoproteins (beta chain) and of the Escherichia coli phosphoserine phosphohydrolase. The five remaining identified ORFs do not show detectable homology with other protein sequences deposited in data banks. The sequence has been deposited in the EMBL data library under Accession Number Z49133.

The complete sequence of a 9000 bp fragment of the right arm of Saccharomyces cerevisiae chromosome VII contains four previously unknown open reading frames

We report the sequence of a 9000 bp fragment from the right arm of Saccharomyces cerevisiae chromosome VII. Analysis of the sequence revealed four complete previously unknown open reading frames, which were named G7587, G7589, G7591 and G7594 following standard rules for provisional nomenclature. Outstanding features of some of these proteins were the homology of the putative protein coded by G7589 with proteins involved in transcription regulation and the transmembrane domains predicted in the putative protein coded by G7591.

Identification of tsunami-induced deposits using numerical modeling and rock magnetism techniques: A study case of the 1755 Lisbon tsunami in Algarve, Portugal

Storm- and tsunami-deposits are generated by similar depositional mechanisms making their discrimination hard to establish using classic sedimentologic methods. Here we propose an original approach to identify tsunami-induced deposits by combining numerical simulation and rock magnetism. To test our method, we investigate the tsunami deposit of the Boca do Rio estuary generated by the 1755 earthquake in Lisbon which is well described in the literature. We first test the 1755 tsunami scenario using a numerical inundation model to provide physical parameters for the tsunami wave. Then we use concentration (MS. SIRM) and grain size (chi(ARM), ARM, B1/2, ARM/SIRM) sensitive magnetic proxies coupled with SEM microscopy to unravel the magnetic mineralogy of the tsunami-induced deposit and its associated depositional mechanisms. In order to study the connection between the tsunami deposit and the different sedimentologic units present in the estuary, magnetic data were processed by multivariate statistical analyses. Our numerical simulation show a large inundation of the estuary with flow depths varying from 0.5 to 6 m and run up of similar to 7 m. Magnetic data show a dominance of paramagnetic minerals (quartz) mixed with lesser amount of ferromagnetic minerals, namely titanomagnetite and titanohematite both of a detrital origin and reworked from the underlying units. Multivariate statistical analyses indicate a better connection between the tsunami-induced deposit and a mixture of Units C and D. All these results point to a scenario where the energy released by the tsunami wave was strong enough to overtop and erode important amount of sand from the littoral dune and mixed it with reworked materials from underlying layers at least 1 m in depth. The method tested here represents an original and promising tool to identify tsunami-induced deposits in similar embayed beach environments.

Conversor estático de potência tolerante a falhas

O trabalho apresentado nesta dissertação refere-se à concepção, projecto e realização experimental de um conversor estático de potência tolerante a falhas. Foram analisados trabalhos de investigação sobre modos de falha de conversores electrónicos de potência, topologias de conversores tolerantes a falhas, métodos de detecção de falhas, entre outros. Com vista à concepção de uma solução, foram nomeados e analisados os principais modos de falhas para três soluções propostas de conversores com topologias tolerantes a falhas onde existem elementos redundantes em modo de espera. Foram analisados os vários aspectos de natureza técnica dos circuitos de potência e guiamento de sinais onde se salientam a necessidade de tempos mortos entre os sinais de disparo de IGBT do mesmo ramo, o isolamento galvânico entre os vários andares de disparo, a necessidade de minimizar as auto-induções entre o condensador DC e os braços do conversor de potência. Com vista a melhorar a fiabilidade e segurança de funcionamento do conversor estático de potência tolerante a falhas, foi concebido um circuito electrónico permitindo a aceleração da actuação normal de contactores e outro circuito responsável pelo encaminhamento e inibição dos sinais de disparo. Para a aplicação do conversor estático de potência tolerante a falhas desenvolvido num accionamento com um motor de corrente contínua, foi implementado um algoritmo de controlo numa placa de processamento digital de sinais (DSP), sendo a supervisão e actuação do sistema realizados em tempo-real, para a detecção de falhas e actuação de contactores e controlo de corrente e velocidade do motor utilizando uma estratégia de comando PWM. Foram realizados ensaios que, mediante uma detecção adequada de falhas, realiza a comutação entre blocos de conversores de potência. São apresentados e discutidos resultados experimentais, obtidos usando o protótipo laboratorial.

Identification of tsunami-induced deposits using numerical modeling and rock magnetism techniques: A study case of the 1755 Lisbon tsunami in Algarve, Portugal

Storm- and tsunami-deposits are generated by similar depositional mechanisms making their discrimination hard to establish using classic sedimentologic methods. Here we propose an original approach to identify tsunami-induced deposits by combining numerical simulation and rock magnetism. To test our method, we investigate the tsunami deposit of the Boca do Rio estuary generated by the 1755 earthquake in Lisbon which is well described in the literature. We first test the 1755 tsunami scenario using a numerical inundation model to provide physical parameters for the tsunami wave. Then we use concentration (MS. SIRM) and grain size (chi(ARM), ARM, B1/2, ARM/SIRM) sensitive magnetic proxies coupled with SEM microscopy to unravel the magnetic mineralogy of the tsunami-induced deposit and its associated depositional mechanisms. In order to study the connection between the tsunami deposit and the different sedimentologic units present in the estuary, magnetic data were processed by multivariate statistical analyses. Our numerical simulation show a large inundation of the estuary with flow depths varying from 0.5 to 6 m and run up of similar to 7 m. Magnetic data show a dominance of paramagnetic minerals (quartz) mixed with lesser amount of ferromagnetic minerals, namely titanomagnetite and titanohematite both of a detrital origin and reworked from the underlying units. Multivariate statistical analyses indicate a better connection between the tsunami-induced deposit and a mixture of Units C and D. All these results point to a scenario where the energy released by the tsunami wave was strong enough to overtop and erode important amount of sand from the littoral dune and mixed it with reworked materials from underlying layers at least 1 m in depth. The method tested here represents an original and promising tool to identify tsunami-induced deposits in similar embayed beach environments.

Validação do dispositivo automático de medição da pressão arterial, OMRON® M6 Comfort, segundo o Protocolo Internacional da Sociedade Europeia de Hipertensão (2010)

Introdução – A medição da pressão arterial (PA), utilizando dispositivos automáticos, é frequentemente realizada na prática clínica e na automedição, permitindo adquirir informação fiável para o diagnóstico, controlo e tratamento da hipertensão arterial. Porém, muitos dos dispositivos automáticos disponíveis no mercado não estão validados segundo protocolos existentes para o efeito. O objetivo do estudo foi confirmar a validação do dispositivo de medição automática da PA, OMRON® M6 Comfort, segundo o Protocolo Internacional da European Society of Hypertension (ESH), de 2010, para a validação de dispositivos de medição automática da PA em adultos. Metodologia – Foram estudados 33 indivíduos, aos quais foram realizadas 9 medições sequenciais da PA, no braço esquerdo, com um esfignomanómetro aneróide alternando com o dispositivo automático. Seguidamente avaliaram-se as diferenças entre os valores obtidos pelos dispositivos para a pressão arterial sistólica (PAS) e diastólica (PAD), classificando-as em três níveis (≤ 5, ≤ 10 ou ≤ 15 mmHg). O número das diferenças em cada nível foi comparado ao requerido pelo Protocolo (fase 1.1). Para cada sujeito foi ainda determinado o número de diferenças com valores ≤ 5 mmHg. Pelo menos 24 dos 33 indivíduos devem ter 2 ou 3 diferenças com valores ≤ 5 mmHg e no máximo 3 dos 33 indivíduos podem apresentar as 3 diferenças com valores > 5 mmHg (fase 1.2). Resultados – O dispositivo OMRON® M6 Comfort foi aprovado nas fases 1.1 e 1.2 para a PAS e PAD. A média das diferenças entre as medições da PA, determinada pelos dispositivos automático e manual, foi de -0,82 ± 5,62 mmHg para a PAS e 2,14 ± 5,15 mmHg para a PAD. Considerações Finais – O dispositivo OMRON® M6 Comfort é válido para a medição da PA em adultos, de acordo com o Protocolo Internacional da ESH, de 2010. - ABSTRACT - Introduction – The measurement of blood pressure (BP) using automatic devices is often performed in clinical practice and self-measurement allowing the acquisition of reliable information for the diagnosis, monitoring and treatment of hypertension. However not all of the automated devices available in the market are validated in accordance with the existing protocols for this purpose. The purpose of this study was to confirm the validation of the automatic measuring device of the BP, OMRON® M6 Comfort, according to the “European Society of Hypertension International Protocol revision 2010 for the validation of blood pressure measuring devices in adults”. Methodology – The study involved 33 subjects, in each one of them, 9 sequential measurements of BP were performed, in the left arm, with the aneroid sphygmomanometer alternating with the automatic device. Afterwards, the differences on the values obtained by the different devices were evaluated, for systolic blood pressure (SBP) and diastolic (DBP), and these differences were then classified into three levels (≤ 5, ≤ 10 or ≤ 15 mmHg). The number of differences at each level was compared to the number required by the protocol (phase 1.1). For each subject the number of differences with values ≤ 5 mmHg was also determined. At least 24 of the 33 subjects should have 2 or 3 differences with values ≤ 5 mmHg and a maximum of 3 of the 33 subjects may have all differences with values > 5 mmHg (phase 1.2). Results – The device OMRON M6 Comfort ® was approved in phases 1.1 and 1.2 for SBP and DBP. The average difference between measurements of BP, as determined by automatic and manual devices, was -0.82 ± 5.62 mmHg for SBP and 2.14 ± 5.15 mmHg for DBP. Conclusion – The device OMRON M6 Comfort® is valid for measuring BP in adults, according to the ESH International Protocol of 2010.

Identification of tsunami-induced deposits using numerical modeling and rock magnetism techniques: Astudy case of the 1755 Lisbon tsunami in Algarve, Portugal (vol 182, pg 187, 2010)

Storm- and tsunami-deposits are generated by similar depositional mechanisms making their discrimination hard to establish using classic sedimentologic methods. Here we propose an original approach to identify tsunami-induced deposits by combining numerical simulation and rock magnetism. To test our method, we investigate the tsunami deposit of the Boca do Rio estuary generated by the 1755 earthquake in Lisbon which is well described in the literature. We first test the 1755 tsunami scenario using a numerical inundation model to provide physical parameters for the tsunami wave. Then we use concentration (MS. SIRM) and grain size (chi(ARM), ARM, B1/2, ARM/SIRM) sensitive magnetic proxies coupled with SEM microscopy to unravel the magnetic mineralogy of the tsunami-induced deposit and its associated depositional mechanisms. In order to study the connection between the tsunami deposit and the different sedimentologic units present in the estuary, magnetic data were processed by multivariate statistical analyses. Our numerical simulation show a large inundation of the estuary with flow depths varying from 0.5 to 6 m and run up of similar to 7 m. Magnetic data show a dominance of paramagnetic minerals (quartz) mixed with lesser amount of ferromagnetic minerals, namely titanomagnetite and titanohematite both of a detrital origin and reworked from the underlying units. Multivariate statistical analyses indicate a better connection between the tsunami-induced deposit and a mixture of Units C and D. All these results point to a scenario where the energy released by the tsunami wave was strong enough to overtop and erode important amount of sand from the littoral dune and mixed it with reworked materials from underlying layers at least 1 m in depth. The method tested here represents an original and promising tool to identify tsunami-induced deposits in similar embayed beach environments.

Neonatal anthropometry: a tool to evaluate the nutritional status and predict early and late risks

Neonatal anthropometry is an inexpensive, noninvasive and convenient tool for bedside evaluation, especially in sick and fragile neonates. Anthropometry can be used in neonates as a tool for several purposes: diagnosis of foetal malnutrition and prediction of early postnatal complications; postnatal assessment of growth, body composition and nutritional status; prediction of long-term complications including metabolic syndrome; assessment of dysmorphology; and estimation of body surface. However, in this age group anthropometry has been notorious for its inaccuracy and the main concern is to make validated indices available. Direct measurements, such as body weight, length and body circumferences are the most commonly used measurements for nutritional assessment in clinical practice and in field studies. Body weight is the most reliable anthropometric measurement and therefore is often used alone in the assessment of the nutritional status, despite not reflecting body composition. Derived indices from direct measurements have been proposed to improve the accuracy of anthropometry. Equations based on body weight and length, mid-arm circumference/head circumference ratio, and upper-arm cross-sectional areas are among the most used derived indices to assess nutritional status and body proportionality, even though these indices require further validation for the estimation of body composition in neonates.

Influência do tratamento de radioterapia na qualidade de vida dos doentes com cancro de mama

RESUMO: Introdução – A evolução tecnológica permitiu uma deteção precoce do cancro e, consequentemente, uma abordagem terapêutica mais eficaz e efetiva, resultando no aumento da sobrevida dos doentes. Assim, pretende-se avaliar a influência da radioterapia na qualidade de vida dos doentes com cancro da mama. Metodologia – 47 participantes que efetuaram radioterapia no Centro Hospitalar Barreiro-Montijo preencheram os dois questionários, o EORTC QLQ-C30 e o QLQ-BR23, antes de iniciar radioterapia e no último dia do tratamento. Os resultados foram analisados através do programa informático SPSS, versão 19.0. Resultados – Observou-se um ligeiro decréscimo na qualidade da saúde global no segundo momento de avaliação. Verificou-se uma diminuição nas escalas sintomática e funcional no segundo momento de avaliação. Em relação ao questionário QLQ-BR23, na escala sintomática, observou-se uma diminuição nos efeitos secundários resultantes da radioterapia no último momento de avaliação. Discussão dos Resultados – Os resultados obtidos no questionário QLQ-C30 revelam que a avaliação global da saúde dos doentes não é influenciada pelo tratamento de radioterapia. Em relação ao questionário QLQ-BR23, os sintomas relacionados com a mama e o braço tiveram um aumento ligeiramente significativo no segundo momento de avaliação, sendo estes associados à radioterapia. De acordo com os resultados obtidos, a radioterapia não influencia a qualidade de vida em doentes com cancro da mama.

In vivo and in vitro effects of RAD001 on bladder cancer

Objective: To evaluate the influence of Everolimus (RAD001) on chemically induced urothelial lesions in mice and its influence on in vitro human bladder cancer cell lines. Methods: ICR male mice were given N-butyl-N-(4-hydroxybutyl) nitrosamine in drinking water for a period of 12 weeks. Subsequently, RAD001 was administered via oral gavage, for 6 weeks. At the end of the experiment, all the animals were sacrificed and tumor development was determined by means of histopathologic evaluation; mammalian target of rapamycin (mTOR) expressivity was evaluated by immunohistochemistry. Three human bladder cancer cell lines (T24, HT1376, and 5637) were treated using a range of RAD001 concentrations. MTT assay, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and flow cytometry were used to assess cell proliferation, apoptosis index, and cell cycle analysis, respectively. Immunoblotting analysis of 3 cell line extracts using mTOR and Akt antibodies was performed in order to study the expression of Akt and mTOR proteins and their phosphorylated forms. Results: The incidence of urothelial lesions in animals treated with RAD001 was similar to those animals not treated. RAD001 did not block T24 and HT1376 cell proliferation or induce apoptosis. A reduction in cell proliferation rate and therefore G0/G1 phase arrest, as well as a statistically significant induction of apoptosis (P 0.001), was only observed in the 5637 cell line. Conclusion: RAD001 seems not to have a significant effect on chemically induced murine bladder tumors. The effect of RAD001 on tumor proliferation and apoptosis was achieved only in superficial derived bladder cancer cell line, no effect was observed in invasive cell lines.

Efetividade do Pnf-Chi® na funcionalidade do membro superior e na qualidade de vida, em mulheres submetidas a tratamento cirúrgico por cancro da mama

Mestrado em Fisioterapia.

Alterações da curvatura da coluna vertebral: influência da fisioterapia, a nível neuromuscular

Mestrado em Fisioterapia

Modulation of translation factor's gene expression by histone deacetylase inhibitors in breast cancer cells

The histone deacetylase inhibitors sodium butyrate (NaBu) and trichostatin A (TSA) exhibit anti-proliferative activity by causing cell cycle arrest and apoptosis. The mechanisms by which NaBu and TSA cause apoptosis and cell cycle arrest are not yet completely clarified, although these agents are known to modulate the expression of several genes including cell-cycle- and apoptosis-related genes. The enzymes involved in the process of translation have important roles in controlling cell growth and apoptosis, and several of these translation factors have been described as having a causal role in the development of cancer. The expression patterns of the translation mechanism, namely of the elongation factors eEF1A1 and eEF1A2, and of the termination factors eRF1 and eRF3, were studied in the breast cancer cell line MCF-7 by real-time quantitative reverse transcription-polymerase chain reaction after a 24-h treatment with NaBu and TSA. NaBu induced inhibition of translation factors' transcription, whereas TSA caused an increase in mRNA levels. Thus, these two agents may modulate the expression of translation factors through different pathways. We propose that the inhibition caused by NaBu may, in part, be responsible for the cell cycle arrest and apoptosis induced by this agent in MCF-7 cells.