A number of investigations during the last two decades describe adverse trends in male reproductive health, which have been proposed to be caused by environmental factors with endocrine disrupting properties. In contrast to many other toxicants, endocrine disruptors often do not show linear dose-response relationships typical of those found in traditional toxicological studies. For many compounds, low-dose exposure causes effects opposite to the ones seen after high-dose exposure. In addition, the timing of exposure has been found to be critical. Hence, to correctly assess the impact of endocrine disruptors on reproductive health requires in-depth knowledge of their mechanisms of action. This thesis aimed at identifying the mechanisms underlying the effects of cadmium (Cd), phthalates and phytoestrogens on testicular steroidogenesis. For this purpose, in vitro as well as in vivo models were used. Cd was found to inhibit testosterone synthesis in vivo by down-regulating LH receptor gene expression and reducing the testicular levels of cAMP and StAR protein. In addition, Cd caused a pronounced increase in testicular prostaglandin F2ɑ (PGF2ɑ), suggesting that Cd exerts its suppressive effect on steroidogenesis also by inducing the inhibitory PKC pathway. Pre-treatment with zinc (Zn) protected completely against Cd-induced effects on testosterone and PGF2ɑ. Furthermore, we observed that Cd exposure increased glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA expression in the testis. GAPDH is a potent coactivator of androgen receptor-mediated transcription and the up-regulation found in our study is probably a compensatory response to reduced testosterone concentrations. This finding is interesting since GAPDH has been proposed to have an important role in the regulation of apoptosis as well as sperm motility. We discovered that mono-(2-ethylhexyl) phthalate (MEHP), the active metabolite of the frequently used phthalate di-(2-ethylhexyl) phthalate (DEHP), stimulates Leydig cell steroidogenesis in vitro, by a cAMP- and StAR-independent mechanism. MEHP exposure caused a similar effect in granulosa cells. Gene expression analysis revealed that MEHP is likely to stimulate steroidogenesis by increasing the amount of cholesterol available for steroid synthesis. In the last investigation, we examined the effects of low-dose phytoestrogen exposure on testosterone synthesis during puberty in male goats. Isoflavones present in clover increased plasma concentrations of testosterone and free as well as total triiodothyronine (T3). T3 has previously been shown to induce testosterone synthesis and it is possible that an elevated T3 secretion underlies the increased plasma testosterone levels. Reduced fertility and reproductive tract malformations affect both the individual and the society. Hence, a sound knowledge of reproductive toxicants is of crucial importance. The findings presented in this thesis provide new insights into the reproductive toxicology of endocrine disruptors and may be valuable for risk assessment purposes.

No description available.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 4 uppsatser.

Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Pavlína Zlochová Supervisor: PharmDr. Marie Kašparová, Ph.D. Title of diploma thesis: Secondary metabolites of plant cultures in vitro I. Key words: Suspension cultures, Trifolium pratense, phytoestrogens, flavonoids, isoflavonoids Explant cultures are source of plant secondary metabolites. Nevertheless the production of secondary metabolites is usually low in explant cultures. This production can be increased by elicitation metods. Suitable elicitor is added into the cultivate medium and leads to gene expression and production of secondary metabolites. The aim of this study was to observe the influence of sodium chloride on the production of flavonoids and isoflavonoids by the Trifolium pratense L. suspension culture (Tempus variety). The culture was cultivated on Gamborg nutriet medium with addition of 2 mg.l-1 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 6-benzylaminopurine. Cultivation proceed in 25řC temperature and 16 hours light/8 hours dark period. Then determination of flavonoids in according to Czech Pharmacopoeia 2009 and the determination of isoflavonoids by HPLC method was performed. The best elicitation effect on the production of flavonoids was 175 mmol.l-1 of sodium chloride after a 3...

Orientador: Adriana Orcesi Pedro / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-27T01:25:39Z (GMT). No. of bitstreams: 1 Carmignani_LucioOmar_D.pdf: 1313710 bytes, checksum: e5a2ce3ebfaf92d14e105ee597ca752a (MD5) Previous issue date: 2015 / Resumo: Objetivos: Comparar os efeitos da ingestão diária de um suplemento alimentar à base de isoflavona de soja, terapia hormonal (TH) de baixa dosagem e placebo sobre a qualidade de vida (QV) em mulheres sintomáticas na pós-menopausa e avaliar a correlação entre a melhora dos sintomas da menopausa e a QV de acordo com o tipo de tratamento e comparar os efeitos de cada uma das intervenções sobre o sistema urogenital da mulher na pós-menopausa. Métodos: Ensaio clínico randomizado, duplo-cego e controlado envolvendo 60 mulheres na pós-menopausa, com idade entre 40 e 60 anos. Foram selecionadas e randomizadas em três grupos: um grupo recebeu um suplemento alimentar à base de soja (isoflavona 90mg/dia), outro grupo recebeu terapia hormonal de baixa dose (estradiol 1mg e acetato de noretisterona 0,5mg) e um grupo-controle que recebeu placebo, por um período de 16 semanas. O Menopause Rating Scale (MRS) foi utilizado para avaliar as mudanças nos sintomas climatéricos. A QV foi avaliada através da Versão Abreviada do Instrumento de Avaliação de Qualidade de Vida da Organização Mundial de Saúde (WHOQOL-BREF). Os instrumentos foram aplicados no início e após 16 semanas de tratamento. As queixas urinárias, vaginais e sexuais foram avaliadas através da subescala urogenital do MRS. A avaliação dos efeitos terapêuticos no sistema urogenital foi realizada através da medida do pH vaginal e calculo do valor de maturação vaginal. A espessura endometrial foi mensurada através da ultrassonografia transvaginal. A análise estatística foi realizada usando-se o teste do qui-quadrado, teste exato de Fisher, teste t de Student pareado, teste de Kruskal-Wallis, teste não paramétrico de Kruskal-Wallis e análise de variância (ANOVA). Para a análise intergrupo dos escores do MRS e da QV foi utilizado ANOVA, teste de Kruskal-Wallis e Kruskal-Wallis seguido de Mann-Whitney. Para a análise de correlação usou-se o coeficiente de correlação de Spearman. Para a comparação intergrupo das subescalas urogenitais do MRS utilizou-se o teste não paramétrico de Kruskal-Wallis seguido de Mann-Whitney. Resultados: A média de idade das participantes foi de 52,4 (±3,9) anos com tempo médio desde a menopausa de 4,1 (±3,1) anos. Os escores de QV aumentaram significativamente no domínio físico da QV nas usuárias da TH e em menor escala no grupo da soja (233,1% x 39,7%; p=0,02). Apenas no grupo que utilizou TH houve correlação significativa entre a melhora dos sintomas climatéricos e a melhora nos domínios físico (p=0,01) e psicológico (p=0,01) da QV e na questão da saúde geral (p<0,01). A avaliação do sistema urogenital mostrou que houve melhora da secura vaginal nos grupos da soja e da TH (p=0,04). As queixas sexuais e urinárias não se alteraram ao longo do tratamento. Apenas no grupo que usou TH houve um aumento significativo no índice de maturação vaginal (p<0,01) e uma diminuição no pH vaginal (p<0,01). Não houve mudança na espessura endometrial e os efeitos colaterais foram semelhantes nos três grupos estudados. Conclusões: O uso da terapia hormonal mostrou-se eficaz na melhora nos escores do domínio físico da qualidade de vida, isto também foi observado com o uso da isoflavona em uma escala muito menor, mas superior ao uso do placebo, porém apenas o grupo que utilizou a terapia hormonal apresentou uma correlação significativa entre a melhora dos sintomas climatéricos e a melhora na qualidade de vida. O uso de suplemento alimentar à base de soja contendo isoflavonas mostrou eficácia comparável à da terapia hormonal na melhora do ressecamento vaginal, sem exercer ação estrogênica no trato urogenital, em mulheres na pós-menopausa, e superior ao placebo / Abstract: Objectives: To compare the effects of a soy-based dietary supplement, low-dose hormone therapy (HT) and placebo on climacteric symptoms and on quality of life (QOL) of symptomatic postmenopausal women, and to evaluate the correlation between the improvement in menopausal symptoms and QOL according to treatment type, and also assess the effects of each intervention on the urogenital system in postmenopausal women. Methods: This was a double-blind, randomized and placebo-controlled clinical trial. Sixty healthy postmenopausal women, aged 40-60, were recruited and randomly assigned to three groups: a soy dietary supplement group (isoflavone 90mg/day), a low-dose HT group (estradiol 1mg plus noretisterone acetate 0.5mg) and a placebo group. Menopausal symptoms changes were evaluated through Menopause Rating Scale (MRS). QOL was measured by the abbreviated version of the World Health Organization¿s Quality of Life instrument (WHOQOL-BREF). These instruments were applied at baseline and after 16 weeks of treatment. Urinary, vaginal and sexual complaints were evaluated by using the urogenital subscale of the MRS. Evaluation of therapeutic effects on urogenital system was performed by vaginal pH measurement and maturation value calculation. Transvaginal sonography was performed to evaluate endometrial thickness. Statistical analysis were performed using the chi-square test, Fisher's exact test, paired Student¿s t-test, Kruskal-Wallis test, Kruskal-Wallis nonparametric test, and analysis of variance (ANOVA). For MRS and QOL scores intergroup analysis were used ANOVA, Kruskal-Wallis test, and Kruskal-Wallis followed Mann-Whitney test. Correlation analysis was performed using the Correlation Spearman Coefficient. For Intergroup comparisons related to MRS urogenital subscale, the Kruskal-Wallis nonparametric test was used, followed by the Mann-Whitney test. Results: The mean age of the patients was 52.4 (±3.9) years, with 4.1 (±3.1) years mean time since menopause. QOL scores increased significantly in the physical health domain of QOL in the users of HT and in a much lesser extent in the soy group (233,1% x 39,7%; p=0,02). It was observed that only in the HT group there was a statistically significant correlation between the improvement of symptoms of total MRS and improvement in the QOL physical (p=0.01) and psychological (p=0.01) domains and also on general health assessment (p<0.01). Urogenital system evaluation showed a significantly improvement in vaginal dryness in the soy group and HT group (p=0.04). Urinary and sexual symptoms did not change with treatment in the three groups. After 16 weeks of treatment, there was a significant increase in maturation value only in the HT group (p<0.01). Vaginal pH decreased only in this group (p<0.01). There were no statistically significant differences in endometrial thickness between the three groups and the adverse effects evaluated were similar. Conclusions: Hormone therapy was effective in improving the physical health domain of QOL, it was also observed with the use of isoflavones on a much lesser extent, but superior to placebo. However, only the HT group showed a significant correlation between the improvement of climacteric symptoms and the improvement in QOL. The use of soy-based dietary supplement containing isoflavones showed an efficacy similar to that of HT in improving vaginal dryness, and greater than placebo, without exerting estrogen action on urogenital tract in postmenopausal women / Doutorado / Fisiopatologia Ginecológica / Doutor em Ciências da Saúde

Os riscos provocados pela Terapia de Reposição Hormonal, levaram à busca de novas terapias, como os fitoestrógenos. São substâncias com ação estrogênica e propriedades que podem retardar a formação de placas ateroscleróticas. Isoflavonas são os fitoestrógenos mais estudados e são encontradas na soja, no red clover e em outras plantas. Avaliamos os efeitos dos fitoestrógenos extraídos da soja Glycine max: genisteína, formononetina, biocanina A e daidzeína; a mistura entre eles (Mix1); o extrato padronizado de red clover (Menoflavon 40mg) e uma segunda mistura com os fitoestrógenos extraídos da Glycine max nas concentrações encontradas no Menoflavon (Mix2), na expressão de moléculas de adesão de leucócitos, VCAM-1, ICAM-1 e E-selectina, em cultura de células endoteliais de cordão umbilical humano (HUVEC), assim como na linhagem modificada de célula endotelial, ECV304, estimuladas com LPS. Resultados: foram padronizados os tempos e concentrações de exposição ao LPS no cultivo de HUVEC de 1ug durante 12 horas de exposição para as três moléculas de adesão; e no cultivo de ECV304 para a expressão de VCAM-1, de 500ng durante 12 horas, para ICAM-1 de 1ug durante 18 horas, para E-selectina 100ng durante 18 horas na superfície celular e 200ng durante 24 horas no sobrenadante de culturas de ECV304, permitindo que este tipo celular seja utilizado como modelo de inflamação. Os fitoestrógenos reduziram VCAM-1, ICAM-1 e E-selectina na superfície celular assim como as formas solúveis dessas moléculas, tanto em ECV304 como em HUVEC, sendo efetivos como agentes preventivos e também para tratamento da aterosclerose. A mistura entre os fitoestrógenos não apresentou maior eficiência na redução das moléculas de adesão na superfície celular, mas apresentou diferenças significativas na produção das formas solúveis. Tanto em ECV304, quanto em HUVEC os fitoestrógenos extraídos do red clover e os extraídos da soja Glycine max reduziram as moléculas de adesão na superfície celular e no sobrenadante, sendo que o Menoflavon, apresentou maior efetividade na redução das moléculas de adesão que os fitoestrógenos extraídos da soja Glycine max, em HUVEC. Ocorreram interações entre os fitoestrógenos e o 17 estradiol, tanto em ECV304 quanto em HUVEC, principalmente quando este se encontrava em baixas concentrações, sugerindo proteção para mulheres na menopausa. Esses efeitos dos fitoestrógenos ocorreram via receptor de estrógeno, como demonstrado pela inibição de suas ações por ICI. Conclusão: tanto os fitoestrógenos extraídos da soja Glycine max quanto os extraídos do red clover apresentaram efeitos anti-aterogênicos, podendo atuar como cardioprotetores para mulheres pós-menopausa. / The risks of hormone replacement therapy have led to a search for new alternatives such as the use of phytoestrogens, plant compounds with estrogen-like biological activity. Isoflavones are the phytoestrogens most extensively studied and can be found in soy, red clover and other plants. Due this estrogen-like activity phytoestrogens can have some effect on atherosclerosis. We evaluated the effects of the phytoestrogens extracts from Glycine max soy: genistein, formononetin, biocanin A and daidzein; a Mix between them (Mix1); a standardized red clover extracts (Menoflavon 40mg) and a second Mix using phytoestrogens from Glycine max with same Menoflavon concentrations (Mix2) on adhesion molecules expression, VCAM-1, ICAM-1 and E-selectin by endothelial cell HUVEC, and by endothelial cell line ECV304, stimulated with LPS. Results: were standardized time and concentration to LPS exposure, being 1ug during 12 hours for the three adhesion molecules expression on HUVEC, and 500ng during 12 hours for VCAM-1 expression, 1ug during 18 hours for ICAM-1 expression and 100ng during 18 hours for E-selectin expression on cell surface as well as 200ng during 24 hours to E-selectin increase on culture supernadant, on ECV 304 cell line. The phytoestrogens decreased VCAM-1, ICAM-1 and E-selectin levels on cell surface and on culture supernadant in HUVEC and ECV304, being useful as preventive agents as well as treatment agents. Mix1 were not most effective than isolated phytoestrogens on cell surface, but presented decreased results on soluble forms. Menoflavon presented more effectiveness than Glycine max on HUVEC. Phytoestrogens interacted with 17 oestradiol mainly, in low concentrations (10pg), showing protection for post menopausal women. These phytoestrogens effects happened by oestrogen receptor activation, this was demonstrated through phytoestrogens inhibition by ICI. Conclusions: the phytoestrogens from Glycine max as well as phytoestrogens from red clover presented antiatherogenic effects, mainly when 17 estradiol is low, being usefull for postmenopausal women.

Phytoestrogens are very intensively studied in recent years because they show different pharmacological effects on human health. For example, it was demonstrated that phytoestrogens reduce the risk of certain cancers (breast, prostate or colon), act as antioxidants in the body and affect the activity of certain enzymes. In addition, also it was demonstrated number of positive effects of soybean phytoestrogens on the health status of women in the menopause, such as reducing the risk of atherosclerotic cardiovascular disease or osteoporosis. Various technologically processed soy beans are important part of the feed rations mainly for high-lactating dairy cows, because they are a valuable source of the energy and high-quality protein, despite their negative effect on reproductive performance. Phytoestrogens consumed from the feed rations are hydrolyzed in the rumen of dairy cow and together with their metabolites are absorbed in the small intestine into the blood. They can be excreted from the body in urine or milk. From this perspective, cow's milk may become a source of phytoestrogens substances, especially equol in the human nutrition. On the other hand, some research results suggest possible potential risks associated with phytoestrogens intake in the diet on human health. At the some age (infants and young children) and group (vegetarians) categories may be dietary intake of phytoestrogens undiserable. According to most experts, a beneficial effect on human health prevail the potential risks. However, the risks associated with phytoestrogens intake in food can not be ignored and therefore phytoestrogens are still actual problem in the field of scientific research.

This study investigated potential apoptotic and anti-proliferative effects of the phytochemicals, genistein and anthocyanin extract, as single and combined treatments in MCF-7 human breast cancer cells. Cells were exposed to single and combined treatments with the phytochemiclas for 48 and 72 hours. Cell viability was assessed using the MTT bioassay. Apoptosis induction was assessed using acridine orange ethidium bromide and rhodamine 123 ethidium bromide fluorescence assays. Both singe and combination treatments induced dose- and time-dependent apoptotic cell death in MCF-7 cells. The percentage of apoptosis was higher in combination treatments than single treatments with either phytochemical, although the difference was not statistically significant. The combination of genistein and anthocyanin extract peaked in efficacy at 48 hours of treatment, to exhibit significantly greater (P<. O5) dose- and time-dependent cell cytotoxicity than single treatments. This study reveals potential chemopreventive implications for the complementary effects of genistein and anthocyanin extract. / by Corine M. Stinson. / Thesis (M.S.)--Florida Atlantic University, 2011. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2011. Mode of access: World Wide Web.

A Solanum lycocarpum é uma planta encontrada em quase todos os estados do Brasil, podendo ser ingerida por animais em idade fértil e em condições naturais. Os frutos desta planta possuem glicoalcalóides esteroidais com propriedades semelhantes aos esteróides. Neste estudo investigou-se os efeitos na esfera reprodutiva causados pela ingestão de S. lycocarpum durante a puberdade de ratos. Os frutos secos foram administrados na ração (10%) desde o desmame até a idade adulta (70dias). Foram avaliados o peso corpóreo e de órgãos, o comportamento e a orientação sexual, os níveis de hormônios sexuais nas fezes, a bioquímica sérica, os níveis de neurotransmissores centrais e os achados anatomopatológico, tanto de ratos machos como de fêmeas. Observou-se os seguintes resultados: 1) Redução no peso corpóreo dos machos e das fêmeas; 2) Em machos, o peso relativo aumentou nos epidídimos, vesícula seminal e testículos; 3) Não houve diferenças no hemograma entre os grupos controle e experimental de ambos os sexos; 4) Alterações na bioquímica sangüínea sugerem leve toxicidade em células musculares e hepática; 5) Em fêmeas, não constatou-se nenhuma alteração no ciclo estral e no comportamento sexual; enquanto nos machos houve melhora do comportamento sexual e 6) Os níveis de neurotransmissores do estriado foi modificado mas não os do hipotálamo. Esses achados indicam que a exposição a S. lycocarpum durante a puberdade prejudicou parâmetros da esfera reprodutiva, em particular, dos machos. / Solanum lycocarpum St. Hil, a toxic plant common in almost all states of Brazil, may be consumed by animals in fertile age and in natural condition. The fruits of this plant contain steroidal glycoalkaloids with steroid-like properties. Presently the reproductive effects of S. lycocarpum fruit ingestion during puberty were investigated. The dried fruits were administered in the diet (10%) from the weaning up to adult age (70 days). The body and organs weights, sexual behavior as well as sexual orientation, the feces hormonal levels, hemogram and biochemical blood evaluations, hypothalamic and striatal neurotransmitter levels, and anatomopathological studies of male and female rats were performed. The results showed: 1) a reduced body weight in male and female; 2) in male, an increased epydidimis, seminal vesicle and testis relative weights; 3) no differences of controls and experimental groups of both sex in the hemogram; 4) blood biochemical alterations meaning slight toxicity in muscular and hepatic cells; 5) in female, no interference was detected in estrous cycle and sexual behavior; an improvement of male sexual behavior was observed; 6) that striatal neurotransmitter levels were modified but not in the hypothalamus. These results indicate that exposure to S. lycocarpum during puberty prejudiced the reproductive system, particularly in male rats.

A Solanum lycocarpum é uma planta encontrada em quase todos os estados do Brasil, podendo ser ingerida por animais em idade fértil e em condições naturais. Os frutos desta planta possuem glicoalcalóides esteroidais com propriedades semelhantes aos esteróides. Neste estudo investigou-se os efeitos na esfera reprodutiva causados pela ingestão de S. lycocarpum durante a puberdade de ratos. Os frutos secos foram administrados na ração (10%) desde o desmame até a idade adulta (70dias). Foram avaliados o peso corpóreo e de órgãos, o comportamento e a orientação sexual, os níveis de hormônios sexuais nas fezes, a bioquímica sérica, os níveis de neurotransmissores centrais e os achados anatomopatológico, tanto de ratos machos como de fêmeas. Observou-se os seguintes resultados: 1) Redução no peso corpóreo dos machos e das fêmeas; 2) Em machos, o peso relativo aumentou nos epidídimos, vesícula seminal e testículos; 3) Não houve diferenças no hemograma entre os grupos controle e experimental de ambos os sexos; 4) Alterações na bioquímica sangüínea sugerem leve toxicidade em células musculares e hepática; 5) Em fêmeas, não constatou-se nenhuma alteração no ciclo estral e no comportamento sexual; enquanto nos machos houve melhora do comportamento sexual e 6) Os níveis de neurotransmissores do estriado foi modificado mas não os do hipotálamo. Esses achados indicam que a exposição a S. lycocarpum durante a puberdade prejudicou parâmetros da esfera reprodutiva, em particular, dos machos. / Solanum lycocarpum St. Hil, a toxic plant common in almost all states of Brazil, may be consumed by animals in fertile age and in natural condition. The fruits of this plant contain steroidal glycoalkaloids with steroid-like properties. Presently the reproductive effects of S. lycocarpum fruit ingestion during puberty were investigated. The dried fruits were administered in the diet (10%) from the weaning up to adult age (70 days). The body and organs weights, sexual behavior as well as sexual orientation, the feces hormonal levels, hemogram and biochemical blood evaluations, hypothalamic and striatal neurotransmitter levels, and anatomopathological studies of male and female rats were performed. The results showed: 1) a reduced body weight in male and female; 2) in male, an increased epydidimis, seminal vesicle and testis relative weights; 3) no differences of controls and experimental groups of both sex in the hemogram; 4) blood biochemical alterations meaning slight toxicity in muscular and hepatic cells; 5) in female, no interference was detected in estrous cycle and sexual behavior; an improvement of male sexual behavior was observed; 6) that striatal neurotransmitter levels were modified but not in the hypothalamus. These results indicate that exposure to S. lycocarpum during puberty prejudiced the reproductive system, particularly in male rats.