Introdução: Isoflavonóides são compostos encontrados em vários vegetais e apresentam diversos efeitos farmacológicos. Dentre estes compostos, encontramos os fitoestrógenos, assim chamados por possuírem ações que mimetizam o efeito do estrógeno natural sobre as células. A soja (Glycine max), um dos vegetais ricos nos fitoestrógenos daidzeína e genisteína, tem sido indicada pela literatura como terapia alternativa para a menopausa pela atividade estrogênica que apresenta, visto que a terapia estroprogestiva para tratar os sintomas desta fase aponta um aumento da incidência de câncer de mama. Objetivo: Avaliar a promoção de apoptose e/ou necrose por um Extrato de Soja (Glycine max) Biotransformado pelo fungo Aspergillus awamori (ESBF) em células de linhagem de adenocarcinoma mamário estrógeno-dependentes (MCF-7) e estrógeno-independentes (SK-BR-3). Materiais e métodos: o ESBF foi produzido na Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP/USP), com concentração determinada de daidzeína (D) e genisteína (G) por CLAE e avaliado em dois modelos de células de adenocarcinoma mamário: estrógeno-dependentes (MCF-7) e estrógeno-independentes (SK-BR-3). Nestes modelos experimentais, foram avaliados também, o Extrato de Soja (ES) e os padrões comerciais de daidzeína (D) e genisteína (G) isoladas ou em combinação (D+G). Neste estudo avaliamos estes compostos perante os parâmetros: a) citotoxicidade pelo método de MTT; b) necrose e apoptose celular pelo ensaio de marcação por iodeto de propídio (IP) e anexina-V e IP; c) a atividade da caspase-3 por western blotting. Resultados: o ESBF nas linhagem MCF-7 e SK-BR-3 apresentou citotoxicidade dose-dependente a partir de 2,184 mg/mL; o ES apresentou aumento na viabilidade celular em todas as concentrações estudadas; os padrões D e G nas concentrações 1,3 e 1,5 µM respectivamente aumentou a viabilidade celular apenas para a linhagem MCF-7; resultado este não observado nas células SK-BR-3. Quanto aos ensaios de necrose e apoptose, encontramos que as duas linhagens celulares apresentaram marcação pelo IP a partir da concentração de 1,638 mg/mL do ESBF, enquanto que o ES e D+G não apresentaram marcação nas concentrações testadas. Somente para a linhagem MCF-7 encontramos no teste de anexina-V + IP apoptose precoce a partir da concentração 0,819 mg/mL e apoptose tardia/necrose a partir da concentração de 2,717 mg/mL frente ao ESBF, enquanto que frente ao ES e aos padrões D+G este resultado não foi observado. Utilizando apenas a linhagem MCF-7, com relação a detecção da caspase-3 íntegra, não foi possível visualizar sua presença a partir da concentração de 1,638 mg/mL do ESBF. Conclusão: Com este estudo verificamos que o ESBF favorece a indução a morte celular das linhagens MCF-7 e SK-BR-3, não acontecendo o mesmo com o ES e os padrões D+G. Nossos achados sugerem que componentes do fungo são os responsáveis por este efeito biológico, e não os metabólitos da soja, visto que os padrões de daidzeína e genisteína, bem como o ES, não apresentaram os resultados de morte celular evidenciados aqui. / Introduction: Isoflavones are compounds found in various vegetables and have different pharmacological effects. Among these compounds there are phytoestrogens, so called because they have actions that mimic the effects of natural estrogen on cells. Soybean (Glycine max), a plant rich in phytoestrogens genistein and daidzein, have been cited in the literature as an alternative therapy for menopause because this plant has estrogen activity. Since oestroprogestative therapy to treat the symptoms of this phase, has many collateral effects, like increased incidence of breast cancer. Objective: To evaluate the promotion of apoptosis and/or necrosis caused by an extract of soybean (Glycine max) biotransformed by the fungus Aspergillus awamori (ESBF) by cell lineage of estrogen-dependent (MCF-7) and estrogen-independent (SK- BR-3) breast adenocarcinoma. Materials and methods: ESBF was produced at the Faculty of Pharmaceutical Sciences of Ribeirão Preto (FCFRP / USP), with known concentration of daidzein (D) and genistein (G) by HPLC and subjected to two models of breast adenocarcinoma cells: estrogen- dependent (MCF-7) and estrogen-independent (SK-BR-3). In these experimental models were also evaluated, Soy Extract (ES) and the commercial standards of daidzein (D) and genistein (G) alone or in combination (D+G). In this study we evaluated all these compounds the following parameters: a) cytotoxicity by MTT method; b) necrosis and apoptosis assay by dialing propidium iodide (PI) and annexin-V + PI; c) the activity of caspase-3 by western blotting. Results: ESBF in cell line MCF-7 and SK-BR-3 showed dose-dependent cytotoxicity starting from 2.184 mg/mL, the ES showed an increase in cell viability at all concentrations studed, D and G standards at concentrations of 1, 3 and 1.5 mM respectively increased cell viability only in line MCF-7, this result not observed in SK-BR-3. For the tests of necrosis and apoptosis, we found that that two cell lines presented labeling IP from the concentration of 1.638 mg/mL of ESBF, while the ES and D + G showed no labeling at all concentrations tested. Only line MCF-7 in the test of annexin-V + PI early apoptosis from the concentration 0.819 mg / mL and late apoptosis or necrosis from the concentration of 2.717 mg / mL against the ESBF, while facing the ES and D+G standards this result was not observed. Using only the cell line MCF-7 in assay to detection of caspase-3 intact, we could not see his presence from the concentration of 1.638 mg/mL ESBF. Conclusion: This study verified that the ESBF favors the induction of cell death in cell line MCF-7 and SK-BR-3, the same not happening with the ES and D+G standards. Our findings suggest that components of the fungus are responsible for this biological effect and not the soy metabolites, since the standards of daidzein and genistein, as well as the ES, the results showed no cell death.

O presente estudo visou avaliar os efeitos da administração de isoflavonas da soja (ISOs) sobre parâmetros bioquímicos e atividade imunológica em ratas ovariectomizadas. As ISOs foram administradas na apresentação não isolada nas doses de 100, 300 e 900 mg/kg, por gavage, durante 28 dias. Ao longo do tratamento foram avaliados o peso e o consumo de ração que evidenciaram efeito moderador na ingesta de alimentos e perda de peso com as maiores doses das ISOs. Foram avaliadas também a bioquímica sérica e urinária, que revelaram alteração do perfil lipídico (⇑ HDL) de forma dose-dependente, aumento de glicemia e diminuição de glicosúria, embora esta última não diferente estatisticamente do controle. No grupo de 900 mg/kg, o estresse oxidativo foi maior e caracterizado pelo aumento de glutationa oxidada (GSSG). No sistema imune foram avaliados diversos parâmetros, a saber: peso relativo de baço e timo e suas celularidades; celularidade de medula óssea; atividades de neutrófilos circulantes e macrófagos peritoneais; resposta imune do tipo tardia (DTH); fenotipagem de linfócitos T e B no sangue e baço e proliferação de linfócitos esplênicos em resposta a ConA e LPS. As maiores doses de ISOs causaram diminuição do peso absoluto do timo e aumento do peso relativo do baço, além de aumento da intensidade de fagocitose de macrófagos na dose de 900 mg/kg. Com a dose de 300 mg/kg de ISOs foi observado menor intensidade na fagocitose de neutrófilos. Assim, pode-se concluir que a utilização de ISOs, na apresentação não isolada, não induziu melhora no perfil lipídico, na proteção ao estresse oxidativo e nem alterou a capacidade de resposta do sistema imune de ratas ovariectomizadas, e, ainda, que a exposição a doses mais elevadas de ISOs apresenta potente efeito sobre a ingesta de alimentos, piora do perfil lipídico, aumento de glicemia e alteração na sinalização redox das células, portanto, futuros experimentos são necessários para melhor caracterizar os efeitos destas agliconas sobre o metabolismo dos lipídeos e também na proteção ao estresse oxidativo. / The present study aimed to evaluate the effects of administration of soybean isoflavones (ISOs) on biochemical and immunological parameters in ovariectomized rats. For this, the ISOs were administered in non-isolated presentation at doses of 100; 300 and 900 mg/kg by gavage for 28 days. Throughout the treatment, the body weight gain and feed intake were evaluated and higher doses of ISOs showed a moderating effect on food intake and weight loss. It were also evaluated the serum and urine biochemistry, which showed altered lipid profile (⇑ HDL), increase in blood glucose and decreased glycosuria in a dose-dependent fashion, although the latter is not statistically different from the control. In the group of 900 mg/kg, the oxidative stress was higher, characterized by increased in oxidized glutathione (GSSG) levels. In the immune system several parameters were evaluated, relative weight of thymus and spleen and their cellularity, bone marrow cellularity, neutrophils and peritoneal macrophages activity, delayed type immune response (DTH), T and B lymphocytes phenotyping in the blood and spleen and splenic lymphocyte proliferation in response to ConA and LPS. Higher doses of ISOs caused decreased in the absolute thymus weight and increase in relative spleen weight. In addition, the dose of 900 mg/kg increased the intensity of phagocytosis of macrophages. On the other hand, it was observed lower phagocytic activity of the neutrophils in the group of 300 mg/kg. Thus, it can be concluded that the use of ISOs in a non-isolated presentation did not induce an improvement in the lipid profile, in the cellular protection to oxidative stress and did not alter the immune response of ovariectomized rats; furthermore, the exposure to higher doses of ISOs has potent effect on food intake, worsening lipid profile, increased blood glucose and changes in redox signaling cells, so further experiments are needed to better characterize the effects of these aglycones on the metabolism of lipids and also in protecting the oxidative stress.

Decreased testosterone levels (hypogonadism) in middle-aged men (aged 45-64) has been associated with increased levels of depression. Studies have suggested that increases in anxiety and/or attention problems may also be associated with hypogonadism but have not provided empirical evidence to support these suggestions. The purpose of this quantitative study was to examine depression, anxiety, and attention problems in middle-aged men using a psychological self-report inventory. The theoretical model used in this study was the biomedical model, which combined pharmacological treatment with psychological self-report inventories to determine if there was an association or relationship between the testosterone levels in men and the psychological distress experienced by men who have become hypogonadal. A total of 179 males were recruited through local physicians. There was a statistically significant difference and a small size effect in the level of depression, anxiety, and/or attention issues experienced by those who were receiving TRT versus those who were not. This study may provide some guidance to medical clinicians, such as psychiatrists, primary-care physicians, and endocrinologists, as well as clinical psychologists who see middle-aged men in their practice settings.

One of the most critical elements in empowering the patient, and ensuring concordance, is communication of the possible benefits and harms of different actions in health care. Risk assessment is a complex task due both to the different interpretations of the concept of risk, and the common lack of hard facts. Hormone, or hormone replacement, therapy (HT) is used by many women in, and after, the menopause. The benefits and possible harms associated with short and long term treatment with HT have been extensively discussed the last decade and the use of HT has decreased dramatically internationally the last few years. The aims of this thesis were to study the interaction between patient and physician when discussing risks and benefits of different treatment alternatives, and to suggest strategies to improve risk communication in clinical practice. The studies have focused on how risks and benefits with HT were communicated between women and physicians during firsttime consultations in 1999- 2000 on this subject (20 women, 5 gynaecologists), and through questionnaires how attitudes towards HT have changed between 1999 (n=1,760) and 2003 (n=1,733) among women entering the menopause (53-54 years). Through a qualitative analysis of the risk communication in the consultations a system was constructed to classify how risk is communicated in relation to benefits. This was used to assess and present differences in risk communication in the consultations. Different rhetorical strategies by the physicians were identified and the dominating tendency was a move from the woman’s current problems to the long-term effects of HT. The questionnaires showed a marked difference in attitudes towards HT between the years. In 2003 women perceived HT to be associated with higher risk and less benefits than in 1999. This correlated to a drastic reduction in the use of HT over the same period. Media was the most frequent source of information about HT during the last twelve months before the questionnaire in 2003. Possible explanations for the different attitudes towards HT between women entering the menopause and gynaecologist; how this difference might have influenced the results; and how they may have implications for future communication strategies are discussed. This thesis illustrates the importance of a deeper understanding in health care of the concept of risk in order to achieve an adequate communication of risk. This is important both in consultations and in campaigns to educate and inform the public. / Reprinted figure 1 on page 32 with permission from Science Ref # 05-17260-Revised. Copyright 2006 AAAS.

Background: Numerous studies have shown that estrogen replacement therapy (ERT) is an effective treatment for vasomotor symptoms, insomnia and vaginal dryness. Beneficial effects have also been shown on lipid patterns and on the incidence of osteoporotic fractures. As ERT increases the risk of endometrial adenocarcinoma, combinations with various progestogens have been developed in order to protect the endometrium. However, the addition of progestogens tends to reduce the beneficial effects of estrogens on mood, cognition and lipid metabolism. The added progestogen often causes side effects such as irritability and depression. There is evidence that the effect on wellbeing varies between women and with the type of progestogen used. Women who prefer to avoid withdrawal bleedings can be given continuous combined hormone replacement therapy (HRT). Unfortunately, irregular bleedings are common at the beginning of treatment and reduces compliance. Recently, several studies have reported an increased risk of breast cancer and venous thrombosis, and therefore long-term treatment with HRT for women without climacteric symptoms is no longer recommended. The ongoing debate has, for the time being, resulted in a recommendation that improving quality of life (QoL) by treatment of climacteric symptoms should be the only indication for prescribing HRT. Aims and methods: The aims of the study were to investigate bleeding patterns, changes in wellbeing at onset and during long-term treatment, and lipid and lipoprotein profiles with two different types of continuous combined HRT. In addition, women starting, and women switching from mainly sequential HRT were compared. The design was a randomised, double-blind, one year, prospective, multicentre study including 249 healthy postmenopausal women who were given continuous daily oral treatment with either combined 0.625mg conjugated estrogen (CE) and 5mg medroxyprogesterone acetate (MPA) or combined 2mg 17β - estradiol (E2) and 1mg norethisterone acetate (NETA). Bleedings, if any, were recorded daily throughout the study. The main outcome measures (changes in wellbeing and climacteric symptoms) consisted of daily ratings of 12 items on a validated symptom scale. Serum concentrations of lipids and lipoproteins were measured at baseline and after one year of treatment. Results and conclusions: The majority of drop-outs were confined to the first three months, and the main reasons were bleedings and/or decreased wellbeing. Drop-outs were three times more common in the E2/NETA group. During the first month, 67% of the women reported irregular bleedings. The number of bleeding days decreased on both treatments during the first four months. Treatment with CE/MPA resulted in less irregular bleedings and a shorter time to amenorrhoea compared to E2/NETA. As expected, "starters" experienced more sweats than "switchers" at the onset of treatment, but both groups improved significantly. Side effects such as breast tenderness, swelling, depression and irritability appeared during the first treatment week in both groups. The side effects of HRT appeared much more quickly than the benefits and were more frequent in women with a history of premenstrual syndrome (PMS). Breast tenderness was more common in the E2/NETA group throughout the whole study period. Apart from that, there were no differences between the two treatment regimens as regards effects on well-being at the end of the study. Lipoprotein(a) levels, an important risk factor for cardiovascular disease, decreased in both treatment groups. Triglyceride levels increased in women treated with CE/MPA, and levels of total cholesterol, high density lipoprotein and low density lipoprotein fell in the E2/NETA group. In conclusion, treatment with E2/NETA caused more bleeding problems than treatment with CE/MPA. CE/MPA was better tolerated than E2/NETA at the beginning of the study, but among the women remaining in the study there was no difference in QoL between the two treatment groups. HRT counselling should take into account that a history of PMS increases the likelihood of side effects and that these may precede any beneficial effects. Both treatments produced beneficial effects on lipid and lipoprotein levels, and neither of the regimens was superior in this respect.

Hormone replacement therapy (HRT) has been studied as a protective factor for cognitive decline and dementia. However, study findings have been inconsistent. Variation in study findings may be due to differences in study designs, small sample size, exposure ascertainment, diagnostic procedures, and inclusion of relevant risk and confounding factors. Moreover, there may be significant differences between the characteristics of women choosing to use HRT and those opting not to use the therapy. Using a large-scale, population-based, cohort study, we examined the relationship between HRT and cognition while paying particular attention to moderating and confounding factors. The main outcomes of interest were to assess differences in risk for cognitive impairments and dementia between HRT user and never user groups; examine HRTs impact on age of onset of dementia; and explore the relationship between duration of HRT and cognitive decline. Logistic regression and Cox Proportional Hazards models were used to test HRT as a predictor for cognitive impairments, Alzheimers disease and vascular dementia, as well as to assess the effect of duration. Linear regression was used to consider the putative relationship between age at onset of dementia and HRT status. HRT use was found to be a statistically significant predictor for Alzheimers disease and vascular dementia. Overall, HRT use did not significantly predict for milder cognitive impairments, although significant interaction effects indicate that HRT may be protective at least for specific sub-groups of women. No durational effect was found for any of the outcomes. Neither did HRT appear to predict for age at onset of dementia. Notably, a large proportion of women in the current study reported using estrogen-only hormone supplements, and therefore generalizations regarding the findings are likely limited to estrogen-only preparations, not combination estrogen-progestin therapies. These findings must be considered within the context of the other known and potential risks and benefits that HRT may afford.

Hormone replacement therapy (HRT) has been studied as a protective factor for cognitive decline and dementia. However, study findings have been inconsistent. Variation in study findings may be due to differences in study designs, small sample size, exposure ascertainment, diagnostic procedures, and inclusion of relevant risk and confounding factors. Moreover, there may be significant differences between the characteristics of women choosing to use HRT and those opting not to use the therapy. Using a large-scale, population-based, cohort study, we examined the relationship between HRT and cognition while paying particular attention to moderating and confounding factors. The main outcomes of interest were to assess differences in risk for cognitive impairments and dementia between HRT user and never user groups; examine HRTs impact on age of onset of dementia; and explore the relationship between duration of HRT and cognitive decline. Logistic regression and Cox Proportional Hazards models were used to test HRT as a predictor for cognitive impairments, Alzheimers disease and vascular dementia, as well as to assess the effect of duration. Linear regression was used to consider the putative relationship between age at onset of dementia and HRT status. HRT use was found to be a statistically significant predictor for Alzheimers disease and vascular dementia. Overall, HRT use did not significantly predict for milder cognitive impairments, although significant interaction effects indicate that HRT may be protective at least for specific sub-groups of women. No durational effect was found for any of the outcomes. Neither did HRT appear to predict for age at onset of dementia. Notably, a large proportion of women in the current study reported using estrogen-only hormone supplements, and therefore generalizations regarding the findings are likely limited to estrogen-only preparations, not combination estrogen-progestin therapies. These findings must be considered within the context of the other known and potential risks and benefits that HRT may afford.

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 117-132).

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 54-58).

The purpose of this study was to determine the factors that influence women's decisions to use hormone replacement therapy (HRT) during menopause. Using the Theory of Planned Behavior and constructs of risk perception, this study was able to explore the beliefs and attitudes of women about HRT use during menopause. The study was also able to explore how risk perception of HRT and the conditions that affect women during midlife impact their decision to use HRT during that time. Focus groups were conducted to develop the questionnaire used in the larger study. A community-based sample of women from Houston area churches participated in the study. The major theme that emerged from the focus groups was the weighing of cancer risks and the protection benefits of HRT. The most interesting factor that emerged as a barrier to HRT use was "negative publicity" and myths toward taking HRT. Results from the larger study demonstrated that the construct of attitude was the predominate predictor of intention when direct measures were used in a model to predict intention. Meaning that women's attitudes towards HRT use during menopause (whether they are safe, wise to use, good or bad, beneficial, risky, pleasant, or valuable) played a significant role in their intention. In contrast when the belief-based measures were used in the model, subjective norm and perceived behavioral control were significant predictors of intention. Meaning that the intention to use HRT was based more on the influence of their husbands, physicians, and families. This also meant that the dosage form, cost, negative publicity, family history of cancer, personal fear of developing cancer, and education about HRT would affect their intention to use HRT during menopause more than the advantages and disadvantages of using HRT (advantages such as protection from osteoporosis, relief from hot flashes; or, disadvantages such as risk of breast cancer). Finally, women's perception of risk with regards to HRT was highest for breast cancer followed by heart disease, endometrial cancer, and osteoporosis. This study found that there are many factors that may affect the decision to use HRT during menopause and that overall these factors affect women’s attitudes towards HRT and their intention to use it. / text