Trabalho realizado na Sysadvance, Sistemas de Engenharia, SA, orientado pelo Doutor Patrick Bárcia / Tese de Mestrado Integrado. Engenharia Química. Universidade do Porto. Faculdade de Engenharia.. 2011

Serial prostate-specific antigen (PSA) measurements (PSA velocity) as an additional instrument to detect prostatic cancer was introduced in 1992. It has previously been reported that PSA increase per year differed in the last 5 years prior to diagnosis in patients with benign prostatic hyperplasia (0.18 ng/ml/year), locally confined (0.75 ng/ml/year) and metastasized (4.4 ng/ml/year) cancer of the prostate (CaP) in contrast to healthy men (0.04 ng/ml/year). The ability of PSA velocity to detect organ-confined CaP in patients with intermediate PSA serum values depends therefore on a reliable and reproducible PSA result. The present study comprised 85 men with PSA values between 3 and 8 ng/ml (Abbott IMx). PSA measurements were repeated with Abbott IMx (n = 85 patients) and Hybritech Tandem-E (n = 59 patients) assays. The PSA serum values differed from one examination to the other from 0.02 to 2.74 ng/ml with the Abbott IMx. Standard deviation amounted to 0.35 ng/ml with the Abbott IMx PSA assay. Using the Hybritech Tandem-E assay, mean standard deviation was 1.15 ng/ml and therefore higher than with the Abbott IMx assay. The difference from one test to the other ranged from 0.05 to 4.05 ng/ml with the Hybritech Tandem-E. Using the Abbott IMx assay, 10.6% of all repeat measurements exceeded 1 ng/ml whereas in the Hybritech Tandem-E assay 62.7% of the second measurements differed >1 ng/ml from the first PSA result. An increase in PSA serum values may therefore be due to intratest variation, physiological day-to-day variation as well as prostatic disease. It is important to notice that the intra-assay variation may be greater than the PSA increase per year in a patient with CaP. Therefore, PSA velocity seems to be of limited value. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

This PhD aimed to elucidate the mechanisms by which polymorphisms may alter androgen-induced transactivation of androgen receptor (AR) target genes which may be important in prostate cancer aetiology. The second aspect of this PhD focused on identifying and characterising functional polymorphisms that may have utility as predictive risk indicators for prostate cancer and which may aid in earlier therapeutic intervention and better disease management. Analyses were carried out on the kallikrein-related peptidase 3 (KLK3), also known as the prostate specific antigen (PSA), gene and the kallikrein-related peptidase 4 (KLK4) gene. The PSA and KLK4 genes are part of the serine protease family that have trypsin or chymotrypsin like activity and are thought to play a role in the development of hormone-dependent cancers in tissues such as those in the prostate, breast, endometrium and ovaries. In the prostate, PSA is regulated by androgens and three androgen response elements (AREs) have been described in the promoter and upstream enhancer region. The PSA ARE I harbours a polymorphism at -158 bp from the transcription initiation site (TIS) that results in a G to A transition (G-158A). This PhD investigated the functional significance of the PSA G-158A polymorphism which has been reported to be associated with prostate cancer risk. Electromobility shift assays (EMSAs) investigating the interaction of ARE I variants with the AR DNA binding domain (AR-DBD) demonstrated that the A allele had a two-fold increased binding affinity for the AR-DBD when compared with the G allele. This was confirmed with endogenous AR in limited proteolysis-EMSA experiments. The limited proteolysis-EMSA experiments also demonstrated differential sensitivities of PSA ARE I alleles to trypsin digestion, which suggests that the G-158A polymorphism has an allosteric effect on the AR that alters AR/ARE I complex stability. Furthermore, Chromatin Immunoprecipitation (ChIP) assays suggest that the A allele more readily recruited the AR in vivo when compared with the G allele and is consistent with the in vitro binding data. Luciferase reporter assays carried out in both LNCaP and 22Rv1 prostate cancer cells, and using the natural (dihydrotestosterone; DHT) ligand demonstrated that the A allele was more responsive to androgens in LNCaP cells. Hence, this study has elucidated the potential mechanisms by which the G-158A polymorphism may differentially regulate PSA expression (of which up-regulation of PSA is thought to be important in prostate cancer development and progression). KLK4 has similar tissue-restricted expression as PSA and is up-regulated by steroid hormones in many endocrine cells including those in the prostate. A putative ARE (KLK4-pARE) located at -1,005 to -1019 relative to the more predominantly used transcription initiation site, TIS3, was initially found in supershift assays using AR antibodies to interact with endogenous AR. However, subsequent EMSA analysis using purified AR-DBD suggest that KLK4-pARE may be interacting with the AR indirectly. To investigate this hypothesis, a tandem construct of KLK4-pARE was cloned into the pGL3-Promoter vector for hormone-induced reporter assays. However, reporter assays did not demonstrate any responsiveness of KLK4-pARE to androgens, estradiol or progestins. Consequently, Real-Time PCR was carried out to reassess the hormonal regulation of KLK4 at the mRNA level. Consistent with the literature, data from this study suggests that KLK4 may be up-regulated by androgens, progestins and estradiol in a cyclical manner. Hormone-induced luciferase reporter assays were then carried out on seven promoter constructs that span 2.8 kb of the KLK4 promoter from TIS3. However, none of the seven promoter constructs demonstrated any significant responsiveness to androgens, estradiol or progestins. This study suggests that hormone response elements (HREs) that may drive the hormonal regulation of KLK4 in prostate cancer may be located further upstream from the promoter region investigated in this PhD, or alternatively, may lie 3' of TIS3. The characterisation of KLK4 promoter polymorphisms and their flanking sequences were also carried out in parallel to the functional work with the intent to assess the functional significance of any polymorphisms that may be located within HREs. In total 19 polymorphisms were identified from the public databases and from direct sequencing within 2.8 kb of the KLK4 promoter from TIS3. However, the functional and clinical significance of these 19 polymorphisms were not further pursued given the negative findings from the functional work. The PSA AR enhancer region was also assessed for potential polymorphisms that may be associated with prostate cancer risk. A total of 12 polymorphisms were identified in the PSA enhancer of which two (A-4643G and T-5412C) have been reported to alter functionality of the enhancer region and thus, prioritised for further analysis. Association analysis for prostate cancer risk was then carried out on these PSA enhancer polymorphisms as none of the KLK4 promoter polymorphisms were found in functional HREs. No significant association for either the A-4643G or T-5412C polymorphism with prostate cancer risk was found at the P = 0.05 level. However, under an age-adjusted dominant model a 1.22- (95% CI = 1.16-1.26) and 1.23-fold (95% CI = 1.17-1.29) increased risk for prostate cancer was found for the A-4643G or T-5412C polymorphisms, respectively. Both polymorphisms were also assessed for association with tumour grade and stage and PSA levels. Genotypes were significantly different for the A-4643G and T-5412C polymorphisms with tumour stage and PSA levels, respectively. However, these results are likely to be biased by the case population which consist primarily of men who presented with incidental (pT1) and organ-confined (pT2) tumours. To summarise, the A-4643G and T-5412C polymorphisms are unlikely to be associated with prostate cancer risk, PSA levels or stage/grade of disease. However, further analyses in a larger cohort is warranted given that these polymorphisms alter androgen responsiveness of the PSA enhancer and that elevated PSA levels are indicative of men with prostate cancer. To summarise, this PhD has elucidated the functional significance of the PSA G-158A polymorphism in prostate cancer and which may be important in prostate cancer patho-physiology. This PhD has also furthered the understanding of the hormonal regulation of KLK4 in prostate cancer cells. Finally, this PhD has carried out a pilot study on two functional PSA enhancer polymorphisms (A-4643G and T-5412C) with prostate cancer risk.

Made available in DSpace on 2016-06-02T19:33:09Z (GMT). No. of bitstreams: 1 KAWAICHI_Vanessa_2012.pdf: 2539108 bytes, checksum: a9216ce702fc0aba843b8b6a772550de (MD5) Previous issue date: 2012-03-30 / The present study aims to estimate the supply of environmental services Sorocaba Rivers Basin and Middle Tietê, located in the central-southeastern state of Sao Paulo. The focus of the environmental service is given to the hydrological services, which are responsible for providing farmers in the region. Thus, from the provision of environmental services in the Basin, we estimated the value of PES to farmers based on the opportunity cost of changing agricultural practices on farms in the region. The methodology is based on the opportunity cost of changing a conventional practice for a conservation practice. The opportunity cost was obtained from the comparison between the yield of each crop analyzed by adopting the conventional practice and practice conservation. Thus, we analyzed the yields of major crops Basin (coffee, corn, beans, potatoes, sugar cane, orange, tomato, onion, grapes, rice and cassava), comparing the difference between the yields for both practices. The classification for conservation practice and conventional practice followed at the discretion of the forest area of the basin, considering the municipalities with an area of forest above average as conservation and below average, as conventional. Thus, the forest area was used as a proxy for measuring the environmental service in question, knowing that the preservation of vegetation provides a number of benefits, including conservation of water resources. The results show that the estimated values of PES for the main crops of the Basin can range from R$100/ha/year to R$30.000/ha/year. In relation to the cultures analyzed in this study, we can say that with the highest potential for implementation of programs for PES were the crops of rice and cassava, which, besides having more profitable return for the conservation practice, have a initial offering of high environmental services and the lowest values for PES. Cultures orange, tomato, onions and grape also have an initial supply of high environmental services, however, PES values are estimated quite high in comparison with other cultures, thereby implementing programs to increase the supply PES environmental services in this case should consider how much the payment would compensate for the provision of environmental services provided by these cultures. / O presente trabalho tem como objetivo estimar a oferta dos serviços ambientais na Bacia Hidrográfica dos Rios Sorocaba e Médio Tietê, localizada na região centrosudeste do Estado de São Paulo. O enfoque de serviço ambiental é dado aos serviços hidrológicos, cujos responsáveis pela oferta são os produtores rurais da região. Assim, a partir da oferta de serviços ambientais da Bacia, estimou-se o valor do PSA aos produtores rurais baseado no custo de oportunidade da mudança de práticas agrícolas nas propriedades rurais da região. A metodologia adotada é baseada no custo de oportunidade da mudança de uma prática convencional para uma prática conservacionista. O custo de oportunidade foi obtido a partir da comparação entre o rendimento de cada cultura analisada, adotando-se a prática que gera menos serviços ambientais e e a prática que gera mais serviços ambientais. Dessa forma, foram analisados os rendimentos das principais culturas da Bacia (café, milho, feijão, batata, cana, laranja, tomate, cebola, uva, arroz e mandioca), comparando-se a diferença entre os rendimentos para ambas as práticas. A classificação das práticas obedeceu ao critério de área de mata da Bacia, considerando-se os municípios com área de mata acima da média como B e, abaixo da média, como A. Dessa forma, a área de mata foi utilizada como proxy para a mensuração do serviço ambiental em questão, sabendo-se que a preservação da cobertura vegetal proporciona uma série de benefícios, inclusive, a conservação dos recursos hidrológicos. Os resultados mostram que os valores estimados do PSA para as principais culturas da Bacia podem variar de R$100/ha/ano a R$30.000/ha/ano. Em relação às culturas analisadas neste estudo, pode-se dizer que as que apresentaram maior potencial para a implementação de programas de PSA foram as culturas do arroz e da mandioca, as quais, além de apresentarem retorno mais rentável para a prática conservacionista, apresentam uma oferta inicial de serviços ambientais elevada e os menores valores para o PSA. As culturas laranja, tomate, cebola e uva também apresentam uma oferta inicial de serviços ambientais elevada, no entanto, os valores de PSA estimados são bem elevados em comparação com as demais culturas, dessa forma, a implementação de programas de PSA para aumentar a oferta de serviços ambientais neste caso deve considerar o quanto o pagamento compensaria a oferta de serviços ambientais proporcionados por essas culturas.

The content of this dissertation is the life and œuvre of Johann Georg de Hamilton, a relatively obscure painter of hunts, portraits of horses, hunting still-lifes and hunting scenes. Johann Georg de Hamilton (1672-1737), a painter belonging to a famous Scottish family, was influenced in his creative work considerably by the 17th -century Flemish painters of still-lifes and hunting scenes. He was active predominantly in Vienna and in South Bohemia, in service of Adam František, Prince of Schwarzenberg, as well as Karl VI of House Habsburg. He created hunting-themed paintings and portraits of horses to members of both the secular and the ecclesiastic aristocracy of the lands of the Austrian Empire, such as the Houses of Liechtenstein, Serényi, Althan and others. This dissertation is the very first attempt at a monographic analysis of the life and œuvre of this painter. In addition to new bibliographic data, it offers an in-depth insight into the relationship between the person who commissioned his work, Adam František, Prince of Schwarzenberg, and the painter Johann Georg de Hamilton on the basis of extant correspondence, and also attempts to present the painter's œuvre in a cultural-historical and artistic context. The dissertation mentions the first exhibition of a collection of Hamilton's work,...

The aim of this thesis is to estimate the value of the company PCA Slovakia, s.r.o. to the 31st December 2015. The thesis is divided into two parts, in the concrete the theoretical and practical parts. The theoretical part describes the basic concepts necessary for the valuation of the company and it is kind of the point for the practical part. On the other hand, the practical part is focused on the introduction of the company and the application of strategic analysis and financial analysis, the prognosis of revenue and other value drivers, financial plan and finally the actual valuation of the company. There was used the DCF method of valuation in the term of FCFF and EVA for the valuation of the company.

Different sampling devices for paper spray - mass spectrometry (PS - MS) were investigated to improve the assay’s simplicity and sensitivity over traditional approaches. In the first one, pressure-sensitive adhesive paper was used as both sampling tool to collect drug residues on surfaces and paper substrate in PS - MS analysis. This method showed a significant improvement in drug collection on surfaces leading to low nano-gram level detection limits. Other sampling device being investigated was snap-in solid-phase extraction column, which demonstrated the ability to detect trace amounts of drugs in plasma while allowed easy transportation and the use of PS - MS automated system.

Serial prostate-specific antigen (PSA) measurements (PSA velocity) as an additional instrument to detect prostatic cancer was introduced in 1992. It has previously been reported that PSA increase per year differed in the last 5 years prior to diagnosis in patients with benign prostatic hyperplasia (0.18 ng/ml/year), locally confined (0.75 ng/ml/year) and metastasized (4.4 ng/ml/year) cancer of the prostate (CaP) in contrast to healthy men (0.04 ng/ml/year). The ability of PSA velocity to detect organ-confined CaP in patients with intermediate PSA serum values depends therefore on a reliable and reproducible PSA result. The present study comprised 85 men with PSA values between 3 and 8 ng/ml (Abbott IMx). PSA measurements were repeated with Abbott IMx (n = 85 patients) and Hybritech Tandem-E (n = 59 patients) assays. The PSA serum values differed from one examination to the other from 0.02 to 2.74 ng/ml with the Abbott IMx. Standard deviation amounted to 0.35 ng/ml with the Abbott IMx PSA assay. Using the Hybritech Tandem-E assay, mean standard deviation was 1.15 ng/ml and therefore higher than with the Abbott IMx assay. The difference from one test to the other ranged from 0.05 to 4.05 ng/ml with the Hybritech Tandem-E. Using the Abbott IMx assay, 10.6% of all repeat measurements exceeded 1 ng/ml whereas in the Hybritech Tandem-E assay 62.7% of the second measurements differed >1 ng/ml from the first PSA result. An increase in PSA serum values may therefore be due to intratest variation, physiological day-to-day variation as well as prostatic disease. It is important to notice that the intra-assay variation may be greater than the PSA increase per year in a patient with CaP. Therefore, PSA velocity seems to be of limited value. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

There is a growing demand for studies applying quantitative methods to large-scale data sets for the purpose of evaluating the effects of educational reforms (UVK, 2010). In this thesis the statistical method, Propensity Score Analysis (PSA), is presented and explored in the evaluating context of an extensive educational initiative within science and technology education; the Science and Technology for All-program (NTA). The research question put forward reads; under what conditions are PSA-analyses a useful method when measuring the effects from a school improvement initiative in S &amp; T? The study considers the use of PSA when looking for long-term effects that could be measured, what to take into consideration to be able to measure this, and how this could be done. The baseline references (outcome variables) used in order to measure/evaluate the long-term effects from the studied program is students’ achievements in the national test (score and grades) and their grades in year 9. Some findings revealed regarding the object of study (long-term effects from using NTA) are also presented. The PSA method is found to be a useful tool that makes it possible to create artificial control groups when experimental studies are impossible or inappropriate; which is often the case in school education research. The method opens up for making use of the rich source of registry data gathered by authorities. PSA proves reliable and relatively insensitive to the effects of covariates and heterogeneous effecter if the number of samples is large enough. The use of PSA (or other statistical methods) also makes it possible to measure outcomes several years after treatment. There are issues of concern when using PSA. One is the obvious demand for organized collection of measurement data. Another issue of concern is the choice of outcome variables. In this study the chosen outcome variables (pupils’ score and grading in national tests and grades in year 9) open up for discussions regarding aspects that might not be reflected/measured in national tests and/or teachers’ grading. Findings regarding the long-term effects from using NTA) show significantly positive effects in physics on test scores (average increase 16.5%) and test grades, but not in biology and chemistry. In this study no significant effects are found for course grades. PSA approach has proved to be a reliable method. There is however a limitation in terms of the method's ability to capture more subtle aspects of learning. A combination of quantitative and qualitative approach when studying long-term effects from educational intervention is therefore suggested. / <p>QC 20131120</p>

Prostate cancer diagnostic algorithm Aim: The aim of the study is to implement the latest scientific knowledge in the diagnosis of prostate cancer (PC). We focused on tumor markers, imaging methods, prostate biopsy methodology and we created a diagnostic algorithm based on a review of current literature in combination with our own experience. Material and methods: The algorithm is divided into several branches, which have been individually subjected to clinical studies. Due to the low sensitivity and specificity of PSA, prostate health index (PHI) was added to the first line of patient stratification. 787 patients were primarily examined and these subsequently underwent radical prostatectomy. PHI levels were compared with definitive staging and grading. Cut-off values for PC detection and high-risk stratification, including locally advanced PC were determined. Next, 320 patients underwent prostate biopsy followed by radical prostatectomy. The cohort was further divided into two subgroups, patients with GS = 6 and patients with GS > 6. The ability of PHI to distinguish between insignificant and significant prostate cancer was evaluated. In a multicentric study with 395 patients, PHI with additional markers (tPSA, PSAD) and multiparametric magnetic resonance imaging of prostate (mpMRI) was assessed....