Differential expression of CDC25 phosphatases splice variants in human breast cancer cells

Background: CDC25 phosphatases control cell cycle progression by activating cyclin dependent kinases. The three CDC25 isoforms encoding genes are submitted to alternative splicing events which generate at least two variants for CDC25A and five for both CDC25B and CDC25C. An over-expression of CDC25 was reported in several types of cancer, including breast cancer, and is often associated with a poor prognosis. Nevertheless, most of the previous studies did not address the expression of CDC25 splice variants. Here, we evaluated CDC25 spliced transcripts expression in anti-cancerous drug-sensitive and resistant breast cancer cell lines in order to identify potential breast cancer biomarkers. Methods: CDC25 splice variants mRNA levels were evaluated by semi-quantitative RT-PCR and by an original real-time RT-PCR assay. Results: CDC25 spliced transcripts are differentially expres-sed in the breast cancer cell lines studied. An up-regulation of CDC25A2 variant and an increase of the CDC25C5/C1 ratio are associated to the multidrug-resistance in VCREMS and DOXOR breast cancer cells, compared to their sensitive counterpart cell line MCF-7. Additionally, CDC25B2 tran-script is exclusively over-expressed in VCREMS resistant cells and could therefore be involved in the development of certain type of drug resistance. Conclusions: CDC25 splice variants could represent interesting potential breast cancer prognostic biomarkers.

The D1822V APC polymorphism interacts with fat, calcium, and fiber intakes in modulating the risk of colorectal cancer in Portuguese persons

Background - Both genetic and environmental factors affect the risk of colorectal cancer (CRC). Objective - We aimed to examine the interaction between the D1822V polymorphism of the APC gene and dietary intake in persons with CRC. Design - Persons with CRC (n = 196) and 200 healthy volunteers, matched for age and sex in a case-control study, were evaluated with respect to nutritional status and lifestyle factors and for the D1822V polymorphism. Results - No significant differences were observed in energy and macronutrient intakes. Cases had significantly (P < 0.05) lower intakes of carotenes, vitamins C and E, folate, and calcium than did controls. Fiber intake was significantly (P = 0.004) lower in cases than in controls, whereas alcohol consumption was associated with a 2-fold risk of CRC. In addition, cases were significantly (P = 0.001) more likely than were controls to be sedentary. The homozygous variant for the APC gene (VV) was found in 4.6% of cases and in 3.5% of controls. Examination of the potential interactions between diet and genotype found that a high cholesterol intake was associated with a greater risk of colorectal cancer only in noncarriers (DD) of the D1822V APC allele (odds ratio: 1.66; 95% CI: 1.00, 2.76). In contrast, high fiber and calcium intakes were more markedly associated with a lower risk of CRC in patients carrying the polymorphic allele (DV/VV) (odds ratio: 0.50; 95% CI: 0.27, 0.94 for fiber; odds ratio: 0.51; 95% CI: 0.28, 0.93 for calcium) than in those without that allele. Conclusion - These results suggest a significant interaction between the D1822V polymorphism and the dietary intakes of cholesterol, calcium, and fiber for CRC risk.

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.

Mod��les animaux pour l�����valuation pr��clinique des dispositifs m��dicaux en radiologie interventionnelle au CR21 AP-HP INRA

Disserta����o de Mestrado, Engenharia Zoot��cnica, 14 de Maio de 2015, Universidade dos A��ores.

Implementa����o e valida����o de um m��todo de avalia����o do perfil de ��cidos gordos e do teor de gordura em alimentos por cromatografia gasosa

Nos dias de hoje �� necess��rio criar h��bitos de vida mais saud��veis que contribuam para o bem-estar da popula����o. Adoptar medidas e pr��ticas de modo regular e disciplinado, pode diminuir o risco do aparecimento de determinadas doen��as, como a obesidade, as doen��as cardiovasculares, a hipertens��o, a diabetes, alguns tipos de cancro e tantas outras. �� tamb��m importante salientar que, uma alimenta����o cuidada d�� sa��de e aumenta a esperan��a m��dia de vida. Em Portugal, nos ��ltimos anos, os costumes alimentares da popula����o t��m vindo a alterar-se significativamente. As refei����es caseiras confeccionadas com produtos frescos d��o lugar �� designada ���cultura do fast food���. Em contrapartida, os consumidores s��o cada vez mais exigentes, estando em permanente alerta no que se refere ao estado dos alimentos. A rotulagem de um produto, para al��m da fun����o publicit��ria, tem vindo a ser objecto de legisla����o espec��fica de forma a fornecer informa����o simples e clara, correspondente �� composi����o, qualidade, quantidade, validade ou outras caracter��sticas do produto. Estas informa����es devem ser acess��veis a qualquer tipo de p��blico, com mais ou menos forma����o e de qualquer estrato social. A qualidade e seguran��a dos produtos deve basear-se na garantia de que todos os ingredientes, materiais de embalagem e processos produtivos s��o adequados �� produ����o de produtos seguros, saud��veis e saborosos. A Silliker Portugal, S.A. �� uma empresa independente de presta����o de servi��os para o sector agro-alimentar, l��der mundial na presta����o de servi��os para a melhoria da qualidade e seguran��a alimentar. A Silliker dedica-se a ajudar as empresas a encontrar solu����es para os desafios actuais do sector, oferecendo uma ampla gama de servi��os, onde se inclui o servi��o de an��lises microbiol��gicas, qu��micas e sensorial; consultadoria em seguran��a alimentar e desenvolvimento; auditorias; rotulagem e legisla����o. A actualiza����o permanente de procedimentos na procura de uma melhoria cont��nua �� um dos objectivos da empresa. Para responder a um dos desafios colocados �� Silliker, surgiu este trabalho, que consistiu no desenvolvimento de um novo m��todo para determina����o de ��cidos gordos e da gordura total em diferentes tipos de alimentos e compara����o dos resultados, com os obtidos com o m��todo anal��tico at�� ent��o adoptado. Se a gordura �� um elemento de grande import��ncia na alimenta����o, devido ��s suas propriedades nutricionais e organol��ticas, recentemente, os investigadores t��m focado a sua aten����o nos mais diversos ��cidos gordos (saturados, monoinsaturados e polinsaturados), em particular nos ��cidos gordos essenciais e nos is��meros do ��cido linoleico conjugado (CLA), uma mistura de is��meros posicionais e geom��tricos do ��cido linoleico com actividade biol��gica importante. A t��cnica usada nas determina����es foi a cromatografia gasosa com ioniza����o de chama, GC-FID, tendo as amostras sido previamente tratadas e extra��das de acordo com o tipo de matriz. A metodologia anal��tica desenvolvida permitiu a correcta avalia����o do perfil em ��cidos gordos, tendo-se para isso usado uma mistura de 37 ��steres met��licos, em que o ��cido gordo C13:0 foi usado como padr��o interno. A identifica����o baseou-se nos tempos de reten����o de cada ��cido gordo da mistura e para a quantifica����o usaram-se os factores de resposta. A valida����o do m��todo implementado foi baseada nos resultados obtidos no estudo de tr��s matrizes relativas a materiais certificados pela BIPEA (Bureau Interprofessionnel des Etudes Analytiques), para o que foram efectuadas doze r��plicas de cada matriz. Para cada r��plica efectuada foi calculado o teor de mat��ria gorda, sendo posteriormente o resultado comparado com o emitido pela entidade certificada. Ap��s an��lise de cada constituinte foi tamb��m poss��vel calcular o teor em ��cidos gordos saturados, monoinsaturados e polinsaturados. A determina����o do perfil em ��cidos gordos dos materiais certificados foi aceit��vel atendendo aos valores obtidos, os quais se encontravam no intervalo de valores admiss��veis indicados nos relat��rios. A quantifica����o da mat��ria gorda no que se refere �� matriz de ���Pat�� �� Tartinier��� apresentou um z-score de 4,3, o que de acordo com as exig��ncias internas da Silliker, n��o �� v��lido. Para as outras duas matrizes (���M��lange Nutritif��� e ���Plat cuisin�� �� base de viande���) os valores de z-score foram, respectivamente, 0,7 e -1,0, o que permite concluir a validade do m��todo. Para que o m��todo possa vir a ser adoptado como m��todo alternativo �� necess��rio um estudo mais alargado relativamente a amostras com diferentes composi����es. O m��todo foi aplicado na an��lise de amostras de fiambre, leite gordo, queijo, ovo com ��mega 3, amendoim e ��leo de girassol, e os resultados foram comparados com os obtidos pelo m��todo at�� ent��o adoptado.

Ecosapentanoic acid (EPA) does not affect cell kinetics in peripheral lymphocites from patients with Crohn's disease (CD) according to IL6 174G/C polymorphism

Rationale: Omega 3 fatty acids have been shown to be of potential benefit in patients with CD. The aim of the present study was to evaluate whether EPA can modulate the inflammatory response according to different genotypes of IL6G174G/C polymorphism. Methods: Peripheral blood cells were collected from CD patients with different genotypes for IL6 174G/C (GG, n = 16, GC, n = 8, CC, n = 7), and lymphocytes were established in culture media. Replicates with the addition of EPA (25 mM) were analysed in a period of 24h, 48h and 72h. Expression of IL6 e a PGE2 was assessed by ELISA. Apoptosis and cellular proliferation was determined by flow cytometry.

Osmolality of preterm formulas suplemented with monprotein energy supplements

Background: Addition of energy supplements to preterm formulas is an optional strategy to increase the energy intake in infants requiring fluid restriction, in conditions like bronchopulmonary dysplasia. This strategy may lead to an undesirable increase in osmolality of feeds, the maximum recommended safe limit being 400���mOsm/kg. The aim of the study was to measure the changes in osmolality of several commercialized preterm formulas after addition of glucose polymers and medium-chain triglycerides. Methods: Osmolality was measured by the freezing point depression method. Six powdered formulas with concentrations of 14���g/100���ml and 16���g/100���ml, and five ready-to-feed liquid formulas were analyzed. All formulas, were supplemented with 10% (low supplementation) or 20% (high supplementation) of additional calories, respectively, in the form of glucose polymers and medium chain triglycerides, maintaining a 1:1 glucose:lipid calorie ratio. Inter-analysis and intra-analysis coefficients of variation of the measurements were always < 3.9%. Results: The mean osmolality (mOsm/kg) of the non-supplemented formulas varied between 268.5 and 315.3���mOsm/kg, increasing by 3���5% in low supplemented formulas, and by 6���10% in high supplemented formulas. None of the formulas analyzed exceeded 352.8���mOsm/kg. Conclusion: The supplementation of preterm formulas with nonprotein energy supplements with up to 20% additional calories did not exceed the maximum recommended osmolality for neonatal feedings.

Reading performance in children with visual function anomalies

Aims - To compare reading performance in children with and without visual function anomalies and identify the influence of abnormal visual function and other variables in reading ability. Methods - A cross-sectional study was carried in 110 children of school age (6-11 years) with Abnormal Visual Function (AVF) and 562 children with Normal Visual Function (NVF). An orthoptic assessment (visual acuity, ocular alignment, near point of convergence and accommodation, stereopsis and vergences) and autorefraction was carried out. Oral reading was analyzed (list of 34 words). Number of errors, accuracy (percentage of success) and reading speed (words per minute - wpm) were used as reading indicators. Sociodemographic information from parents (n=670) and teachers (n=34) was obtained. Results - Children with AVF had a higher number of errors (AVF=3.00 errors; NVF=1.00 errors; p<0.001), a lower accuracy (AVF=91.18%; NVF=97.06%; p<0.001) and reading speed (AVF=24.71 wpm; NVF=27.39 wpm; p=0.007). Reading speed in the 3rd school grade was not statistically different between the two groups (AVF=31.41 wpm; NVF=32.54 wpm; p=0.113). Children with uncorrected hyperopia (p=0.003) and astigmatism (p=0.019) had worst reading performance. Children in 2nd, 3rd, or 4th grades presented a lower risk of having reading impairment when compared with the 1st grade. Conclusion - Children with AVF had reading impairment in the first school grade. It seems that reading abilities have a wide variation and this disparity lessens in older children. The slow reading characteristics of the children with AVF are similar to dyslexic children, which suggest the need for an eye evaluation before classifying the children as dyslexic.

Interac����o de prote��nas com superf��cies nanoestruturadas e nanopart��culas de metais nobres

Disserta����o apresentada para obten����o do Grau de Doutor em Qu��mica Inorg��nica pela Universidade Nova de Lisboa, Faculdade de Ci��ncias e Tecnologia

Qualidade dos sistemas de informa����o no servi��o de radiologia do Hospital de Curry Cabral: um estudo de caso

Mestrado em Interven����o S��cio-Organizacional na Sa��de - ��rea de especializa����o: Pol��ticas de Administra����o e Gest��o de Servi��os de Sa��de

Serum trace elements in dysphagic gastrostomy candidates before endoscopic gastrostomy for long term enteral feeding

Background & aims - Patients who underwent endoscopic gastrostomy (PEG) present protein-energy malnutrition, but little is known about Trace Elements (TE), Zinc (Zn), Copper (Cu), Selenium (Se), Iron (Fe), Chromium (Cr). Our aim was the evaluation of serum TE in patients who underwent PEG and its relationship with serum proteins, BMI and nature of underlying disorder. Methods - A prospective observational study was performed collecting: patient's age, gender, underlying disorder, NRS-2002, BMI, serum albumin, transferrin and TE concentration. We used ferrozine colorimetric method for Fe; Inductively Coupled Plasma-Atomic Emission Spectroscopy for Zn/Cu; Furnace Atomic Absorption Spectroscopy for Se/Cr. The patients were divided into head and neck cancer (HNC) and neurological dysphagia (ND). Results - 146 patients (89 males), 21���95 years: HNC-56; ND-90. Low BMI in 78. Low values mostly for Zn (n = 122) and Fe (n = 69), but less for Se (n = 31), Cu (n = 16), Cr (n = 7); low albumin in 77, low transferrin in 94 and 66 with both proteins low. Significant differences between the groups of underlying disease only for Zn (t140.326 = ���2,642, p < 0.01) and a correlation between proteins and TE respectively albumin and Zn (r = 0.197, p = 0.025), and albumin and Fe (r = 0.415, p = 0.000). Conclusions - When gastrostomy was performed, patients display low serum TE namely Zn, but also Fe, less striking regarding others TE. It was related with prolonged fasting, whatever the underlying disease. Low proteins were associated with low TE. Teams taking care of PEG-patients should use Zn supplementation and include other TE evaluation as part of the nutritional assessment of PEG candidates.

Preditores da ades��o �� medica����o na epilepsia: estudo longitudinal

Introdu����o: A n��o ades��o �� medica����o na epilepsia �� prevalente, pelo que a compreens��o dos fatores associados deve ser promovida. Objetivos: Analisar a capacidade preditiva das estrat��gias de coping e da espiritualidade em rela����o �� ades��o �� medica����o ao longo do tempo. Metodologia: Foram avaliados 60 indiv��duos atrav��s de um Question��rio Sociodemogr��fico e Cl��nico, a Medida de Ades��o aos Tratamentos, o COPER e a Escala de Avalia����o de Espiritualidade em Contextos de Sa��de, em dois momentos. Resultados: A espiritualidade Momento 1 n��o se relacionou com ades��o �� medica����o no Momento 3, e as estrat��gias de coping: Desinvestimento Comportamental e Aceita����o no Momento 1 predizem negativamente a ades��o �� medica����o no Momento 2. Conclus��o: A rela����o entre a ades��o �� medica����o, estrat��gias de coping e espiritualidade varia em fun����o do tempo, o que deve ser considerado ao n��vel da interven����o.

Pr��ticas de gest��o de recursos humanos em PME���s portuguesas

O presente estudo tem como principal objectivo caracterizar as pr��ticas de Gest��o de Recursos Humanos mais desenvolvidas em pequenas e m��dias empresas (PME���s) portuguesas. Embora parte integrante de uma investiga����o mais ampla, a qual resultou numa disserta����o de mestrado que pretendeu estudar as pr��ticas de Gest��o de Recursos Humanos (GRH) nas empresas de m��dia dimens��o do distrito de Aveiro, este estudo pretende apresentar uma das suas principais dimens��es de an��lise. De referir que a investiga����o inicial abrangia o conceito de m��dia empresa (de 50 a 500 trabalhadores). No entanto, e para conferir originalidade ao estudo que agora se apresenta publicamente, a realidade emp��rica em estudo foi circunscrita ��s denominadas PME���s (50 a 249 trabalhadores). Para melhor contextualizar a realidade objecto de estudo, tomou-se como ponto de partida a abordagem te��rica sobre as diferentes fases evolutivas do conceito e conte��do da GRH e, consequentemente, a sua caracteriza����o no nosso pa��s. Adicionalmente �� analisado o papel que o departamento de recursos humanos assume no desenvolvimento da GRH no interior das PME���s atrav��s da dissemina����o que faz das suas diferentes pr��ticas, previamente identificadas a partir da revis��o de literatura analisada. A partir da literatura revista foi identificado um conjunto de 21 pr��ticas de GRH como as mais comummente desenvolvidas em contexto organizacional. Dado ser objectivo principal perceber se a GRH das PME���s portuguesas acompanha a tend��ncia evolutiva desta disciplina de gest��o a n��vel internacional, fez-se a diferencia����o entre aquelas que s��o consideradas pr��ticas tradicionais e pr��ticas estrat��gicas de GRH. Delimitado ao distrito de Aveiro, de um universo constitu��do por 613 empresas, foi aplicado um inqu��rito por question��rio, via postal, tendo sido recolhidos 111 question��rios v��lidos. A an��lise dos resultados permite concluir que n��o obstante predominarem em Portugal as PME���s, estas apresentam pouca intensidade no desenvolvimento de pr��ticas de GRH, ao mesmo tempo que se apresentam como limitadoras do grau de interven����o do departamento de RH, no desenvolvimento dessas pr��ticas. H��, portanto, motivos para propor alguns desafios de mudan��a �� GRH vigente neste grupo de empresas, ainda muito dependente dos processos burocr��tico-administrativos na gest��o do recurso mais din��mico das organiza����es: as pessoas. �� ainda longo o caminho a percorrer para que se possa falar numa efectiva gest��o estrat��gica de recursos humanos nas PME���s portuguesas. Contudo, os primeiros passos est��o dados. Este estudo permitiu n��o s�� uma primeira aproxima����o a um conjunto de PME���s portuguesas bem como a obten����o de maior conhecimento da fun����o recursos humanos das PME���s estudadas. As principais conclus��es extra��das deste estudo pretendem ser um importante input que auxilie as empresas portuguesas com esta tipologia a melhorarem a sua efici��ncia no dom��nio da GRH.

Medication regimen complexity in institutionalized elderly people in an aging society

Background: Complex medication regimens may adversely affect compliance and treatment outcomes. Complexity can be assessed with the medication regimen complexity index (MRCI), which has proved to be a valid, reliable tool, with potential uses in both practice and research. Objective: To use the MRCI to assess medication regimen complexity in institutionalized elderly people. Setting: Five nursing homes in mainland Portugal. Methods: A descriptive, cross-sectional study of institutionalized elderly people (n = 415) was performed from March to June 2009, including all inpatients aged 65 and over taking at least one medication per day. Main outcome measure: Medication regimen complexity index. Results: The mean age of the sample was 83.9 years (��6.6 years), and 60.2 % were women. The elderly patients were taking a large number of drugs, with 76.6 % taking more than five medications per day. The average medication regimen complexity was 18.2 (��SD = 9.6), and was higher in the females (p < 0.001). The most decisive factors contributing to the complexity were the number of drugs and dosage frequency. In regimens with the same number of medications, schedule was the most relevant factor in the final score (r = 0.922), followed by pharmaceutical forms (r = 0.768) and additional instructions (r = 0.742). Conclusion: Medication regimen complexity proved to be high. There is certainly potential for the pharmacist's intervention to reduce it as part as the medication review routine in all the patients.

In Situ Near-Infrared (NIR) Versus High-Throughput Mid- Infrared (MIR) Spectroscopy to Monitor Biopharmaceutical Production

The development of biopharmaceutical manufacturing processes presents critical constraints, with the major constraint being that living cells synthesize these molecules, presenting inherent behavior variability due to their high sensitivity to small fluctuations in the cultivation environment. To speed up the development process and to control this critical manufacturing step, it is relevant to develop high-throughput and in situ monitoring techniques, respectively. Here, high-throughput mid-infrared (MIR) spectral analysis of dehydrated cell pellets and in situ near-infrared (NIR) spectral analysis of the whole culture broth were compared to monitor plasmid production in recombinant Escherichia coil cultures. Good partial least squares (PLS) regression models were built, either based on MIR or NIR spectral data, yielding high coefficients of determination (R-2) and low predictive errors (root mean square error, or RMSE) to estimate host cell growth, plasmid production, carbon source consumption (glucose and glycerol), and by-product acetate production and consumption. The predictive errors for biomass, plasmid, glucose, glycerol, and acetate based on MIR data were 0.7 g/L, 9 mg/L, 0.3 g/L, 0.4 g/L, and 0.4 g/L, respectively, whereas for NIR data the predictive errors obtained were 0.4 g/L, 8 mg/L, 0.3 g/L, 0.2 g/L, and 0.4 g/L, respectively. The models obtained are robust as they are valid for cultivations conducted with different media compositions and with different cultivation strategies (batch and fed-batch). Besides being conducted in situ with a sterilized fiber optic probe, NIR spectroscopy allows building PLS models for estimating plasmid, glucose, and acetate that are as accurate as those obtained from the high-throughput MIR setup, and better models for estimating biomass and glycerol, yielding a decrease in 57 and 50% of the RMSE, respectively, compared to the MIR setup. However, MIR spectroscopy could be a valid alternative in the case of optimization protocols, due to possible space constraints or high costs associated with the use of multi-fiber optic probes for multi-bioreactors. In this case, MIR could be conducted in a high-throughput manner, analyzing hundreds of culture samples in a rapid and automatic mode.