Estrogen receptor dependent genetic and epigenetic factors of tamoxifen resistance

Resumo: A decisão da terapêutica hormonal no tratamento do cancro da mama baseiase na determinação do receptor de estrogénio alfa por imunohistoquímica (IHC). Contudo, a presença deste receptor não prediz a resposta em todas as situações, em parte devido a limitações do método IHC. Investigámos se a expressão dos genes ESR1 e ESR2, bem como a metilação dos respectivos promotores, pode estar relacionada com a evolução desfavorável de uma proporção de doentes tratados com tamoxifeno assim como com a perda dos receptores de estrogénio alfa (ERα) e beta (ERß). Amostras de 211 doentes com cancro da mama diagnosticado entre 1988 e 2004, fixadas em formalina e preservadas em parafina, foram utilizadas para a determinação por IHC da presença dos receptores ERα e ERß. O mRNA total do gene ESR1 e os níveis específicos do transcrito derivado do promotor C (ESR1_C), bem como dos transcritos ESR2_ß1, ESR2_ß2/cx, and ESR2_ß5 foram avaliados por Real-time PCR. Os promotores A e C do gene ESR1 e os promotores 0K e 0N do gene ESR2 foram investigados por análise de metilação dos dinucleotidos CpG usando bisulfite-PCR para análise com enzimas de restrição, ou para methylation specific PCR. Atendendo aos resultados promissores relacionados com a metilação do promotor do gene ESR1, complementamos o estudo com um método quantitativo por matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) suportado pelo software Epityper para a medição da metilação nos promotores A e C. Fez-se a avaliação da estabilidade do mRNA nas linhas celulares de cancro da mama MCF-7 e MDA-MB-231 tratadas com actinomicina D. Baixos níveis do transcrito ESR1_C associaram-se a uma melhor sobrevivência global (p = 0.017). Níveis elevados do transcrito ESR1_C associaram-se a uma resposta inferior ao tamoxifeno (HR = 2.48; CI 95% 1.24-4.99), um efeito mais pronunciado em doentes com tumores de fenótipo ERα/PgR duplamente positivo (HR = 3.41; CI 95% 1.45-8.04). A isoforma ESR1_C mostrou ter uma semi-vida prolongada, bem como uma estrutura secundária da região 5’UTR muito mais relaxada em comparação com a isoforma ESR1_A. A análise por Western-blot mostrou que ao nível da 21 proteína, a selectividade de promotores é indistinguivel. Não se detectou qualquer correlação entre os níveis das isoformas do gene ESR2 ou entre a metilação dos promotores do gene ESR2, e a detecção da proteína ERß. A metilação do promotor C do gene ESR1, e não do promotor A, foi responsável pela perda do receptor ERα. Estes resultados sugerem que os níveis do transcrito ESR1_C sejam usados como um novo potencial marcador para o prognóstico e predição de resposta ao tratamento com tamoxifeno em doentes com cancro da mama. Abstract: The decision of endocrine breast cancer treatment relies on ERα IHC-based assessment. However, ER positivity does not predict response in all cases in part due to IHC methodological limitations. We investigated whether ESR1 and ESR2 gene expression and respective promoter methylation may be related to non-favorable outcome of a proportion of tamoxifen treated patients as well as to ERα and ERß loss. Formalin-fixed paraffin-embedded breast cancer samples from 211 patients diagnosed between 1988 and 2004 were submitted to IHC-based ERα and ERß protein determination. ESR1 whole mRNA and promoter C specific transcript levels, as well as ESR2_ß1, ESR2_ß2/cx, and ESR2_ß5 transcripts were assessed by real-time PCR. ESR1 promoters A and C, and ESR2 promoters 0N and 0K were investigated by CpG methylation analysis using bisulfite-PCR for restriction analysis, or methylation specific PCR. Due to the promising results related to ESR1 promoter methylation, we have used a quantification method by matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDITOF MS) together with Epityper software to measure methylation at promoters A and C. mRNA stability was assessed in actinomycin D treated MCF-7 and MDA-MB-231 cells. ERα protein was quantified using transiently transfected breast cancer cells. Low ESR1_C transcript levels were associated with better overall survival (p = 0.017). High levels of ESR1_C transcript were associated with non-favorable response in tamoxifen treated patients (HR = 2.48; CI 95% 1.24-4.99), an effect that was more pronounced in patients with ERα/PgR double-positive tumors (HR = 3.41; CI 95% 1.45-8.04). The ESR1_C isoform had a prolonged mRNA half-life and a more relaxed 5’UTR structure compared to ESR1_A isoform. Western-blot analysis showed that at protein level, the promoter selectivity is undistinguishable. There was no correlation between levels of ESR2 isoforms or ESR2 promoter methylation and ERß protein staining. ESR1 promoter C CpG methylation and not promoter A was responsible for ERα loss. We propose ESR1_C levels as a putative novel marker for breast cancer prognosis and prediction of tamoxifen response.

Quality of Life Predictors and Normative Data

This study identifies predictors and normative data for quality of life (QOL) in a sample of Portuguese adults from general population. A cross-sectional correlational study was undertaken with two hundred and fifty-five (N = 255) individuals from Portuguese general population (mean age 43 years, range 25–84 years; 148 females, 107 males). Participants completed the European Portuguese version of the World Health Organization Quality of Life short-form instrument and the European Portuguese version of the Center for Epidemiologic Studies Depression Scale. Demographic information was also collected. Portuguese adults reported their QOL as good. The physical, psychological and environmental domains predicted 44 % of the variance of QOL. The strongest predictor was the physical domain and the weakest was social relationships. Age, educational level, socioeconomic status and emotional status were significantly correlated with QOL and explained 25 % of the variance of QOL. The strongest predictor of QOL was emotional status followed by education and age. QOL was significantly different according to: marital status; living place (mainland or islands); type of cohabitants; occupation; health. The sample of adults from general Portuguese population reported high levels of QOL. The life domain that better explained QOL was the physical domain. Among other variables, emotional status best predicted QOL. Further variables influenced overall QOL. These findings inform our understanding on adults from Portuguese general population QOL and can be helpful for researchers and practitioners using this assessment tool to compare their results with normative data

Multidimensional Frailty and Pain in Community Dwelling Elderly

Artigo científico disponível actualmente em Early View (Online Version of Record published before inclusion in an issue)

Life-cycle performance of structures: combining expert judgment and results of inspection

Life-Cycle Civil Engineering – Biondini & Frangopol

Improving discovery in the life sciences using semantic Web technologies and linked data: design principles for life sciences knowledge organization systems

Dissertation presented to obtain the Ph.D degree in Bioinformatics

An Exercise Program Improves Health-Related Quality of Life of Workers

Low back problems are associated with decreased quality of life. Specific exercises can improve quality of life, resulting in better professional performance and functionality. The purpose of this study was to evaluate the effect of following a 21-month exercise program on the quality of life of warehouse workers. The population included 557 male warehouse workers from a food distribution company in Oporto, Portugal. Upon application of the selection criteria, 249 workers were deemed eligible, which were randomized into two groups (125 in the intervention group and 124 in the control group). Then, subjects were asked to volunteer for the study, the sample being formed by 229 workers (112 in the intervention group and 117 in the control group). All subjects completed the SF-36 questionnaire prior to beginning the program and on the 11th and 21st months following it. The exercises were executed in the company facilities once a day for 8 min. Data were analyzed using SPSS® 17.0 for Windows®. After 11 months of following the exercise program, there was an increase in all scores for the experimental group, with statistically significant differences in the dimensions physical functioning (0.019), bodily pain (0.010), general health (0.004), and rolephysical (0.037). The results obtained at the end of the study (21 months) showed significant improvements in the dimensions physical functioning (p = 0.002), rolephysical (p = 0.007), bodily pain (p = 0.001), social functioning (p = 0.015), role-emotional (p = 0.011), and mental health (p = 0.001). In the control group all dimensions showed a decrease in mean scores. It can be concluded that the implementation of a low back specific exercise program has changed positively the quality of life of warehouse workers.

Limit Cycle Prediction of Systems with Fractional Controllers and Backlash

This article investigates the limit cycle (LC) prediction of systems with backlash by means of the describing function (DF) when using discrete fractional-order (FO) algorithms. The DF is an approximate method that gives good estimates of LCs. The implementation of FO controllers requires the use of rational approximations, but such realizations produce distinct dynamic types of behavior. This study analyzes the accuracy in the prediction of LCs, namely their amplitude and frequency, when using several different algorithms. To illustrate this problem we use FO-PID algorithms in the control of systems with backlash.

Fatigue life assessment of exhaust systems for naval gas turbines

Dissertação para obtenção do Grau de Mestre em Engenharia Mecânica

Using data mining with time series data in short-term stocks prediction

Data Mining (DM) methods are being increasingly used in prediction with time series data, in addition to traditional statistical approaches. This paper presents a literature review of the use of DM with time series data, focusing on short- time stocks prediction. This is an area that has been attracting a great deal of attention from researchers in the field. The main contribution of this paper is to provide an outline of the use of DM with time series data, using mainly examples related with short-term stocks prediction. This is important to a better understanding of the field. Some of the main trends and open issues will also be introduced.

Development and validation of a risk model for prediction of hazardous alcohol consumption in general practice attendees : the PredictAL study

Background: Little is known about the risk of progression to hazardous alcohol use in people currently drinking at safe limits. We aimed to develop a prediction model (predictAL) for the development of hazardous drinking in safe drinkers. Methods: A prospective cohort study of adult general practice attendees in six European countries and Chile followed up over 6 months. We recruited 10,045 attendees between April 2003 to February 2005. 6193 European and 2462 Chilean attendees recorded AUDIT scores below 8 in men and 5 in women at recruitment and were used in modelling risk. 38 risk factors were measured to construct a risk model for the development of hazardous drinking using stepwise logistic regression. The model was corrected for over fitting and tested in an external population. The main outcome was hazardous drinking defined by an AUDIT score >= 8 in men and >= 5 in women. Results: 69.0% of attendees were recruited, of whom 89.5% participated again after six months. The risk factors in the final predictAL model were sex, age, country, baseline AUDIT score, panic syndrome and lifetime alcohol problem. The predictAL model's average c-index across all six European countries was 0.839 (95% CI 0.805, 0.873). The Hedge's g effect size for the difference in log odds of predicted probability between safe drinkers in Europe who subsequently developed hazardous alcohol use and those who did not was 1.38 (95% CI 1.25, 1.51). External validation of the algorithm in Chilean safe drinkers resulted in a c-index of 0.781 (95% CI 0.717, 0.846) and Hedge's g of 0.68 (95% CI 0.57, 0.78). Conclusions: The predictAL risk model for development of hazardous consumption in safe drinkers compares favourably with risk algorithms for disorders in other medical settings and can be a useful first step in prevention of alcohol misuse.

Life Cycle Assessment of the Hybrid Vehicle – Toyota Prius

Die Luftverschmutzung, die globale Erwärmung sowie die Verknappung der endlichen Ressourcen sind die größten Bedenken der vergangenen Jahrzehnte. Die Nachfrage nach jeglicher Mobilität steigt rapide. Dementsprechend bemüht ist die Automobilindustrie Lösungen für Mobilität unter dem Aspekt der Nachhaltigkeit und dem Umweltschutz anzubieten. Die Elektrifizierung hat sich hierbei als der beste Weg herausgestellt, um die Umweltprobleme sowie die Abhängigkeit von fossilen Brennstoffen zu lösen. Diese Arbeit soll einen Einblick über die Umweltauswirkungen des Hybridfahrzeuges Toyota Prius geben. Hierbei findet eine Gliederung in vier verschiedene Lebensphasen statt. Im Anschluss bietet die Sachbilanz die Möglichkeit die Umweltauswirkungen mit verschiedenen Antriebsmöglichkeiten und Brennstoffen zu vergleichen. Das Modell hat gezeigt, dass der Toyota Prius während der Nutzung einen hohen Einfluss auf das Treibhauspotenzial aufweist. Durch die Nutzung anderer Brennstoffe, wie beispielsweise Ethanol oder Methanol lassen sich die Auswirkungen am Treibhauspotenzial sowie der Verbrauch an abiotischen Ressourcen reduzieren. Vergleicht man die Elektromobilität mit der konventionellen, so ist festzustellen, dass diese Art der Mobilität die derzeit beste Möglichkeit zur Reduzierung der Umweltbelastungen bietet. Die Auswirkungen der Elektromobilität sind im hohen Maße abhängig von der Art des verwendeten Strommixes.

A Clinical Support System Based on Quality of Life Estimation

Quality of life is a concept influenced by social, economic, psychological, spiritual or medical state factors. More specifically, the perceived quality of an individual's daily life is an assessment of their well-being or lack of it. In this context, information technologies may help on the management of services for healthcare of chronic patients such as estimating the patient quality of life and helping the medical staff to take appropriate measures to increase each patient quality of life. This paper describes a Quality of Life estimation system developed using information technologies and the application of data mining algorithms to access the information of clinical data of patients with cancer from Otorhinolaryngology and Head and Neck services of an oncology institution. The system was evaluated with a sample composed of 3013 patients. The results achieved show that there are variables that may be significant predictors for the Quality of Life of the patient: years of smoking (p value 0.049) and size of the tumor (p value < 0.001). In order to assign the variables to the classification of the quality of life the best accuracy was obtained by applying the John Platt's sequential minimal optimization algorithm for training a support vector classifier. In conclusion data mining techniques allow having access to patients additional information helping the physicians to be able to know the quality of life and produce a well-informed clinical decision.

Adhesive selection for single lap bonded joints: Experimentation and advanced techniques for strength prediction

The integrity of multi-component structures is usually determined by their unions. Adhesive-bonding is often used over traditional methods because of the reduction of stress concentrations, reduced weight penalty, and easy manufacturing. Commercial adhesives range from strong and brittle (e.g., Araldite® AV138) to less strong and ductile (e.g., Araldite® 2015). A new family of polyurethane adhesives combines high strength and ductility (e.g., Sikaforce® 7888). In this work, the performance of the three above-mentioned adhesives was tested in single lap joints with varying values of overlap length (LO). The experimental work carried out is accompanied by a detailed numerical analysis by finite elements, either based on cohesive zone models (CZM) or the extended finite element method (XFEM). This procedure enabled detailing the performance of these predictive techniques applied to bonded joints. Moreover, it was possible to evaluate which family of adhesives is more suited for each joint geometry. CZM revealed to be highly accurate, except for largely ductile adhesives, although this could be circumvented with a different cohesive law. XFEM is not the most suited technique for mixed-mode damage growth, but a rough prediction was achieved.