Magister Scientiae (Medical Bioscience) - MSc(MBS) / The prostate is prone to three pathological processes that include inflammation, benign prostate hyperplasia (BPH) and tumors. According to the center for Disease and Control 1999-2012 report, prostate cancer is the second leading cause of death in the United States. Scientific evidence suggests that up to 30% of men in the general population aged from 50 years and above, irrespective of geographic origin, have foci of prostate neoplastic growth. Unbalanced ROS production and a dysregulated antioxidant defence system have been implicated in prostate cancer development. The transformation of a normal cell into cancer takes a very long period. This observation provides the advantage of using nutraceuticals to prevent, arrest or reverse the cellular and molecular processes of carcinogenesis. Based on scientifically observed positive health roles of green tea (Cameli sinensis) and rooibos (Aspalathus linearis) on major diseases like atherosclerosis, hepatitis and certain types of cancer, this thesis evaluated the effects of these two teas on benign (RPWE 1) and malignant (LNCaP) prostate cells. This was done through the quantification of reactive oxygen species (ROS) using a fluorescence dye 5,6 CM-H2DCFDA, total prostate specific antigen (PSA) levels using a PSA ELISA kit, cell viability using the MTT assay, apoptosis using Tali annexin V stain and cell imaging studies using a Zeiss axiovert 200M inverted fluorescence microscope. Statistical analysis was done using graphpad prism. The findings of this study show that aqueous extracts of green and black tea, fermented and unfermented rooibos and their active compounds epigallocatechin gallate (EGCG) and aspalatin, respectively, are cytotoxic in malignant (LNCaP) prostate cells but exert protective effects in benign (RPWE 1) prostate cells. This thesis implicates the pro-oxidant and anti-oxidant properties of the plant extracts, respectively, for the above mentioned effects. In this regard, tea and rooibos promoted ROS production in malignant (LNCaP) prostate cells, which subsequently promoted cell death of the malignant cells through apoptosis and necrosis. Further to this, tea and rooibos used in this thesis, protected normal prostate cells from the adverse effects of ROS. In this regard, fluorescence microscope photographs showed RPWE 1 cells with low DCF fluorescence compared to the malignant prostate cells. Low magnification light microscope photographs showed RPWE 1 cells with flat polygonal shapes and increased adherence both at low and high concentrations of tea and rooibos. On the contrary, high concentrations of tea and rooibos on malignant (LNCaP) prostate cells induced stress, which made the cells attain irregular shapes and as the stress levels increased, cells became detached and appeared dead. Flow cytometry confirmed the presence of apoptotic and necrotic cell in malignant (LNCaP) prostate cells. In this thesis, EGCG and aspalathin were responsible for the high rates of apoptosis observed whereas green tea and unfermented rooibos induced the highest rate of necrosis. Further to this, tea and rooibos and the main active compounds EGCG and aspalathin, respectively, significantly promoted the reduction of total serum prostate specific antigen (PSA) in malignant prostate cells. In normal prostate cells, these plant extracts maintained the total serum PSA at its basal physiological level. In this thesis, to the best of our knowledge, we report for the first time the cell-specific effects of fermented rooibos, unfermented rooibos and their main active component aspalathin, on prostate cancer cells. We showed that rooibos and aspalathin exert pro-oxidant effects on malignant LNCaP cells and anti-oxidant effects on benign RPWE 1 cells. In conclusion, tea (C. sinensis) and rooibos (A. linearis) and their respective main active compounds, epigallocatechin gallate and aspalathin, are cytotoxic to malignant prostate cells whereas in normal prostate cells, they have protective effects against ROS induced stress. The pro-oxidant and anti-oxidant effects are responsible for the aforementioned effects respectively. The decrease in total serum PSA demonstrate the strong therapeutic effects that tea and rooibos have on malignant (LNCaP) prostate cells. / Malawi Government: Department of Human Resources Development and Management

1. INTRODUCTION 1.1. Work actuality In recent years the number of males with urination disorders mostly induced by prostate pathology has significantly increased [Gas et al., 1998; Khan, Khan et al., 2005; Amaral, Coeli et al., 2004]. Benign prostate hyperplasia (BPH) affects males at a much younger age making them complain of urination disorders of different types and intensity. It is quite a common pathology among males (which is) conditioned by life style and nutrition [Saga & Sugimura, 2004; Shabbir & Mumtaz, 2004; Cambell, 2005]. Clinical symptoms of BPH are not restricted only to urination complaints. With an increase of age of males, the balance of androgens and estrogens undergoes changes in the male organism, affecting the power of libido. According to C.M.Porth, 2005, two theories related to senility have been created, trying to explain biological processes occurring with age: The first theory is related to the so called genetically programmed changes. This theory affirms that changes brought on by aging, are genetically predetermined. Another theory is called the stochastic theory which says that all changes are of accidental nature, i.e. the body undergoes accidental changes. There is one more theory, the so-called neuroendocrinic theory of senility. This theory involves three basic factors of the process of aging: 1) facilitated degradation of hormones, 2) decreased synthesis and secretion of hormones, 3) diminished sensitivity of “target” receptors to... [to full text]

Resumo: O aumento da glândula prostática canina com o envelhecimento pode ser decorrente de uma proliferação excessiva do estroma e epitélio acinar, uma diminuição na taxa de apoptose, entre outros. Esses processos celulares são mediados por hormônios, fatores de crescimento e proteínas, dentre elas podemos citar as proteínas CYR61 e p-ERK1/2 e os fatores de crescimento EGF, TGF- e EGFr. Próstatas originadas de 29 cães, com pesos variando entre 6 a 40 kg, foram retiradas, lavadas e pesadas e o volume medido, os valores variaram em 3,8 a 130 g e 2,4 a 90 cm3 respectivamente. Dos fragmentos avaliados 55,17% apresentaram diagnóstico de HPB-ep e 5,75% apresentaram diagnóstico normal. Exames de IHQ foram realizados e para a proteína CYR61 houve uma forte imunorreatividade nas FML, MLA e núcleo das células epiteliais dos fragmentos avaliados. Houve diferenças estatísticas entre grupos de HPB quanto a fração da área de marcação, no exame de W.B. a proteína teve como peso molecular médio 37,72 kDa. Quanto ao EGF houve uma predominância de marcação na FML e núcleo das células epiteliais, os pesos moleculares médio deste fator de crescimento foi de 47,03 e 41,65 kDa nos fragmentos prostáticos avaliados. O TGF- teve imunorreatividade nas FML e citoplasma das células epiteliais, e no exame de W.B. houve 2 bandas de pesos moleculares médios de 52,23 e 38,88 kDa. O receptor de membrana EGFr apresentou imunorreatividade nas FML e núcleo das células epiteliais, no W.B. houve apenas uma banda visível de peso molecular médio de 48,49 kDa. A proteína p-ERK1/2 esteve predominante nas FML e núcleo das células e no W.B. apresentou 3 bandas de pesos moleculares médios de 42,61; 39,56 e 37,26 kDa. Houve correlação positiva entre as proteínas e os fatores de crescimento. / Abstract: The canine prostate gland enlargement also aging could be resulting of excessive stroma and acinar epithelium proliferation, reduced apoptosis, among other. These cellular mechanisms are mediated by hormones, growth factors and proteins, like the proteins CYR61 and p-ERK1/2 and the growth factors EGF, TGF- e EGFr. Prostate from 29 dogs, weighing 6 to 40 kg, were collected, washed, wheighed and volume was measured, with average from 3,8 to 130 g and 2,4 to 90 cm3 respectively. Approximately 55,17% of samples were diagnosed with HPB-ep and 5,75% were normal. IHQ exams were accomplished and high CYR61 immunoreactivity in SMF, ASM and epithelial cells nucleus was observed. There were statistical differences between BHP groups related to immunoreactivity area, by means of W. B. and the mean molecular weight was 37,72 kDa. By analyzing EGF, immunoreactivity in SMF epithelial cells nucleus was prevalent, the mean molecular weights was 47,03 41,65 kDa. TGF- presented immunoreactivity in SMF and epithelial cells cytoplasm and W.B. showed two bands with mean weight 52,23 and 38,88 kDa. The EGFr membrane receptor presented immunoreactivity in SMF and epithelial cells nucleus, W.B. showed only one visible band with mean molecular weight 48,49 kDa. The p-ERK1/2 protein was prevalent in SMF and cell nucleus, W.B. showed three bands with mean molecular weights 42, 61; 39,56 and 37,26 kDa. There was positive correlation between proteins and growth factors. / Orientador: Gilson Hélio Toniollo / Coorientadora: Renée Laufer Amorin / Coorientador: José Félix Pérez Gutiérrez / Banca: Rosangela Zacarias Machado / Banca: Mirela Tinucci Costa / Banca: Eugênio Gonçalves de Araújo / Banca: Sebastião Roberto Taboga / Doutor

Made available in DSpace on 2014-06-11T19:35:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-06Bitstream added on 2014-06-13T19:24:33Z : No. of bitstreams: 1 oliveira_ks_dr_jabo.pdf: 9693100 bytes, checksum: cbce02641faf86a0db97373d5c457eff (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O aumento da glândula prostática canina com o envelhecimento pode ser decorrente de uma proliferação excessiva do estroma e epitélio acinar, uma diminuição na taxa de apoptose, entre outros. Esses processos celulares são mediados por hormônios, fatores de crescimento e proteínas, dentre elas podemos citar as proteínas CYR61 e p-ERK1/2 e os fatores de crescimento EGF, TGF- e EGFr. Próstatas originadas de 29 cães, com pesos variando entre 6 a 40 kg, foram retiradas, lavadas e pesadas e o volume medido, os valores variaram em 3,8 a 130 g e 2,4 a 90 cm3 respectivamente. Dos fragmentos avaliados 55,17% apresentaram diagnóstico de HPB-ep e 5,75% apresentaram diagnóstico normal. Exames de IHQ foram realizados e para a proteína CYR61 houve uma forte imunorreatividade nas FML, MLA e núcleo das células epiteliais dos fragmentos avaliados. Houve diferenças estatísticas entre grupos de HPB quanto a fração da área de marcação, no exame de W.B. a proteína teve como peso molecular médio 37,72 kDa. Quanto ao EGF houve uma predominância de marcação na FML e núcleo das células epiteliais, os pesos moleculares médio deste fator de crescimento foi de 47,03 e 41,65 kDa nos fragmentos prostáticos avaliados. O TGF- teve imunorreatividade nas FML e citoplasma das células epiteliais, e no exame de W.B. houve 2 bandas de pesos moleculares médios de 52,23 e 38,88 kDa. O receptor de membrana EGFr apresentou imunorreatividade nas FML e núcleo das células epiteliais, no W.B. houve apenas uma banda visível de peso molecular médio de 48,49 kDa. A proteína p-ERK1/2 esteve predominante nas FML e núcleo das células e no W.B. apresentou 3 bandas de pesos moleculares médios de 42,61; 39,56 e 37,26 kDa. Houve correlação positiva entre as proteínas e os fatores de crescimento. / The canine prostate gland enlargement also aging could be resulting of excessive stroma and acinar epithelium proliferation, reduced apoptosis, among other. These cellular mechanisms are mediated by hormones, growth factors and proteins, like the proteins CYR61 and p-ERK1/2 and the growth factors EGF, TGF- e EGFr. Prostate from 29 dogs, weighing 6 to 40 kg, were collected, washed, wheighed and volume was measured, with average from 3,8 to 130 g and 2,4 to 90 cm3 respectively. Approximately 55,17% of samples were diagnosed with HPB-ep and 5,75% were normal. IHQ exams were accomplished and high CYR61 immunoreactivity in SMF, ASM and epithelial cells nucleus was observed. There were statistical differences between BHP groups related to immunoreactivity area, by means of W. B. and the mean molecular weight was 37,72 kDa. By analyzing EGF, immunoreactivity in SMF epithelial cells nucleus was prevalent, the mean molecular weights was 47,03 41,65 kDa. TGF- presented immunoreactivity in SMF and epithelial cells cytoplasm and W.B. showed two bands with mean weight 52,23 and 38,88 kDa. The EGFr membrane receptor presented immunoreactivity in SMF and epithelial cells nucleus, W.B. showed only one visible band with mean molecular weight 48,49 kDa. The p-ERK1/2 protein was prevalent in SMF and cell nucleus, W.B. showed three bands with mean molecular weights 42, 61; 39,56 and 37,26 kDa. There was positive correlation between proteins and growth factors.

A hiperplasia prostática benigna (HPB) é a doença na qual a próstata demonstra um crescimento anormal e sua prevalência aumenta com o envelhecimento. Leptina, a adipocina mais notável, tem um importante papel na regulação do sistema reprodutivo. Esse trabalho tem por fim avaliar o papel da leptina no tecido prostático humano, utilizando a cultura de tecido in vitro, avaliando a proliferação celular e a expressão dos genes do fator de crescimento do fibroblasto 2 (FGF2), da enzima aromatase e dos genes apoptóticos. De 2009 a 2011, amostras de tecido hiperplásico humano foram obtidas pela prostatectomia transvesical em quinze pacientes com próstatas de volume aumentadas. Cada amostra foi dividida em quatro partes simétricas, mantidas no meio RPMI suplementado com soro fetal bovino a 10%, e 1ng/mL de gentamicina, adicionados a 16 ng/mL de leptina (Leptina) ou não (Controle). Após três horas a expressão dos genes de FGF2, aromatase, Bax, Bcl-x e Bcl-2 foram avaliados por RT-PCR em tempo real. A proliferação celular foi avaliada por imunohistoquímica para PCNA. O tratamento com leptina levou a um aumento na expressão de Bax (C=0.40.1; L=0.90.2; p<0.05), enquanto as expressões de Bcl-2 (C=19.95.6; L=5.61.8; p< 0.05) e Bcl-x (C=0.20.06; L=0.070.02; p<0.05) foram significativamente reduzidas. Não houve alteração significativa na expressão de FGF2, enquanto a expressão da aromatase foi significativamente (C=1.90.6; L=0.40.1; p<0.04) reduzida. A leptina também levou a um aumento na proliferação celular (C=21.80.5; L=64.80.9; p<0.0001). Dessa forma, concluímos que a leptina tem um importante papel na manutenção do crescimento fisiológico da próstata, desde que estimula tanto a proliferação celular como a apoptose, com diminuição na expressão do gene da aromatase. / The benign prostate hyperplasia (BPH) is the disease in which the prostate shows an abnormal growth and its prevalence rise up with aging. Leptin, the most well characterized adipokine, has an important role regulating the reproductive system. This paper aimed to evaluate the leptin role in the human prostate tissue, using an in vitro tissue culture system, on the cellular proliferation and the expression of fibroblast growth factor 2, aromatase enzyme and apoptotic genes. From 2009 to 2011, hyperplasic prostate tissue samples obtained by transvesical prostatectomy in fifteen patients with enlarged prostates were used. Each sample was divided in four symmetric parts which were maintained in RPMI medium supplemented with 10% fetal bovine serum, 1 ng/mL of gentamicin and added with 16 ng/mL leptin (L) or not (C). After 3 hours gene expression of FGF2, aromatase, Bax, Bcl-x and Bcl-2 were evaluated by Real Time RT-PCR. Cellular proliferation was evaluated by imunohistochemistry for PCNA. The leptin treatment led to an increase in the expression of Bax (C=0.40.1; L=0.90.2; p<0.05) while Bcl-2 (C=19.95.6; L=5.61.8; p< 0.05) and Bcl-x (C=0.20.06; L=0.070.02; p<0.05) expressions were significantly reduced. There was no significant alteration in the FGF2 expression, while the aromatase expression was significantly (C=1.90.6; L=0.40.1; p<0.04) reduced. Leptin also resulted in an increase in cellular proliferation (C=21.80.5; L=64.80.9; p<0.0001). The conclusion is that leptin has an important role in maintaining the physiological growth of the prostate since it stimulates both cellular proliferation and apoptosis, with the decrement in the aromatase gene expression

Background: To compare the clinical value of the measurement of complex and total PSA in the discrimination between benign prostatic hyperplasia (BPH) and prostate cancer. Methods: In serum samples collected from 166 men with histopathologically proven clinically localized prostate cancer and of 97 men with BPH, total prostate-specific antigen (PSA), complexed PSA and the free to total PSA ratio were determined. The statistical analysis was done by the comparison of the receiver operator characteristic (ROC) curves. Results: The areas under the ROC curves were 0.776 for total PSA, 0.799 for complexed PSA (total PSA vs. cPSA: p < 0.0001) and 0.812 for the free to total PSA ratio. With a cut-off of 3.0 ng/ml for complexed PSA, the sensitivity was 90%, the specificity 58%, the positive and the negative predictive values 79 and 78%, respectively. With a cut-off of 4.0 ng/ml for total PSA, the sensitivity was 87%, the specificity 59%, the positive and the negative predictive values were 78 and 72%, respectively. Conclusions: There was a statistically significant advantage for complexed PSA compared to total PSA in the discrimination between BPH and prostate cancer. The difference was, however, small and its clinical relevance is questionable. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

A hiperplasia prostática benigna (HPB) é a doença na qual a próstata demonstra um crescimento anormal e sua prevalência aumenta com o envelhecimento. Leptina, a adipocina mais notável, tem um importante papel na regulação do sistema reprodutivo. Esse trabalho tem por fim avaliar o papel da leptina no tecido prostático humano, utilizando a cultura de tecido in vitro, avaliando a proliferação celular e a expressão dos genes do fator de crescimento do fibroblasto 2 (FGF2), da enzima aromatase e dos genes apoptóticos. De 2009 a 2011, amostras de tecido hiperplásico humano foram obtidas pela prostatectomia transvesical em quinze pacientes com próstatas de volume aumentadas. Cada amostra foi dividida em quatro partes simétricas, mantidas no meio RPMI suplementado com soro fetal bovino a 10%, e 1ng/mL de gentamicina, adicionados a 16 ng/mL de leptina (Leptina) ou não (Controle). Após três horas a expressão dos genes de FGF2, aromatase, Bax, Bcl-x e Bcl-2 foram avaliados por RT-PCR em tempo real. A proliferação celular foi avaliada por imunohistoquímica para PCNA. O tratamento com leptina levou a um aumento na expressão de Bax (C=0.40.1; L=0.90.2; p<0.05), enquanto as expressões de Bcl-2 (C=19.95.6; L=5.61.8; p< 0.05) e Bcl-x (C=0.20.06; L=0.070.02; p<0.05) foram significativamente reduzidas. Não houve alteração significativa na expressão de FGF2, enquanto a expressão da aromatase foi significativamente (C=1.90.6; L=0.40.1; p<0.04) reduzida. A leptina também levou a um aumento na proliferação celular (C=21.80.5; L=64.80.9; p<0.0001). Dessa forma, concluímos que a leptina tem um importante papel na manutenção do crescimento fisiológico da próstata, desde que estimula tanto a proliferação celular como a apoptose, com diminuição na expressão do gene da aromatase. / The benign prostate hyperplasia (BPH) is the disease in which the prostate shows an abnormal growth and its prevalence rise up with aging. Leptin, the most well characterized adipokine, has an important role regulating the reproductive system. This paper aimed to evaluate the leptin role in the human prostate tissue, using an in vitro tissue culture system, on the cellular proliferation and the expression of fibroblast growth factor 2, aromatase enzyme and apoptotic genes. From 2009 to 2011, hyperplasic prostate tissue samples obtained by transvesical prostatectomy in fifteen patients with enlarged prostates were used. Each sample was divided in four symmetric parts which were maintained in RPMI medium supplemented with 10% fetal bovine serum, 1 ng/mL of gentamicin and added with 16 ng/mL leptin (L) or not (C). After 3 hours gene expression of FGF2, aromatase, Bax, Bcl-x and Bcl-2 were evaluated by Real Time RT-PCR. Cellular proliferation was evaluated by imunohistochemistry for PCNA. The leptin treatment led to an increase in the expression of Bax (C=0.40.1; L=0.90.2; p<0.05) while Bcl-2 (C=19.95.6; L=5.61.8; p< 0.05) and Bcl-x (C=0.20.06; L=0.070.02; p<0.05) expressions were significantly reduced. There was no significant alteration in the FGF2 expression, while the aromatase expression was significantly (C=1.90.6; L=0.40.1; p<0.04) reduced. Leptin also resulted in an increase in cellular proliferation (C=21.80.5; L=64.80.9; p<0.0001). The conclusion is that leptin has an important role in maintaining the physiological growth of the prostate since it stimulates both cellular proliferation and apoptosis, with the decrement in the aromatase gene expression

Background: To compare the clinical value of the measurement of complex and total PSA in the discrimination between benign prostatic hyperplasia (BPH) and prostate cancer. Methods: In serum samples collected from 166 men with histopathologically proven clinically localized prostate cancer and of 97 men with BPH, total prostate-specific antigen (PSA), complexed PSA and the free to total PSA ratio were determined. The statistical analysis was done by the comparison of the receiver operator characteristic (ROC) curves. Results: The areas under the ROC curves were 0.776 for total PSA, 0.799 for complexed PSA (total PSA vs. cPSA: p < 0.0001) and 0.812 for the free to total PSA ratio. With a cut-off of 3.0 ng/ml for complexed PSA, the sensitivity was 90%, the specificity 58%, the positive and the negative predictive values 79 and 78%, respectively. With a cut-off of 4.0 ng/ml for total PSA, the sensitivity was 87%, the specificity 59%, the positive and the negative predictive values were 78 and 72%, respectively. Conclusions: There was a statistically significant advantage for complexed PSA compared to total PSA in the discrimination between BPH and prostate cancer. The difference was, however, small and its clinical relevance is questionable. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

The theoretical part of my thesis at first introduces the most common benign neoplasm in men over fifty, a non cancerous prostate gland enlargement called the benign prostate hyperplasia (BPH). The main focus of this section will be on BPH's anatomy, etiology, symptomatology, complications, diagnosis and therapy. I will also describe four cost analyses used in health services: CMA, CEA, CUA and CBA. In the practical section of my thesis I will analyze the therapy effectiveness of transurethral resection of the prostate and photoselective vaporization of the prostate with the use of cost-effectiveness analysis.

No description available.