Widespread epidemiological data support the notion that high isoflavoves intake is safe and provides health benefits similar to estrogen. Isoflavones have been reported to regulate genes that are involved in several cellular events, such as control of cell proliferation, cell cycle, cell apoptosis, transcription regulation, cancer invasion and metastasis and so on. Previously, soy isoflavones have been demonstrated to have a tumor suppressor effect on bladder cancer. We earlier identified 71 candidate bladder tumor suppressors by using the isoflavones treatment vs. non-treatment in RT4 (Stage I, TMN) cells in conjunction with the strategy of suppression subtractive cDNA libraries. In a preliminary screening, epithelial membrane protein 2 (EMP2) mRNA was downregulated in higher TMN-grade bladder tumor cells. In this study, we aim at the mechanism of isoflavones upregulation of the candidate bladder tumor suppressor: EMP2. In order to clarify the regulatory mechanism, we have constructed a series of 5¡¦ deletion fragment of EMP2 promoter (-1,420 to +268 bp) into a luciferase reporter plasmid pGL3-baisc and analyzed the promoter activity. The rapidly loss of the promoter activities when the region between pH (-44 to +268) and pI (-24 to +268) is deleted, suggesting that at least one critical transcriptional factor binding site may exist between -44 to -24 bp region. By bioinformatics, the transcription factor CREB1 binding site is predicted in the critical region. The ChIP assay and site-directed mutagenesis of the CREB site reveal its critical role for the EMP2 transcription. The EMP2 mRNA is downregulated after knock-down of CREB1 protein by shCREB1 in J82 cells. It can be evidence for EMP2 transcription is regulated by transcription factor CREB1. In investigations of genistein (one component of isoflavones) up-regulated EMP2 transcripts, we find there are consistent results for genistein upregulated of EMP2 promoter region. In conclusion, transcription factor CREB1 plays an important role in the regulation of EMP2 transcription.

Isoflavonoids are a class of plant secondary metabolites which have multiple biological roles in plants as pest feeding deterrents, phytoalexins and signals to rhizobial microbes. Some isoflavonoids, or their breakdown products, are estrogenic when ingested by animals, and pastures with high levels of the isoflavonoid formononetin can cause sterility in ewes. White clover has low levels of isoflavonoids and is susceptible to pests like the clover root weevil. The overall aim of this project was to test whether isoflavonoids could be manipulated in white clover through metabolic engineering. The genes of the key isoflavonoid biosynthesis enzymes have been cloned from a range of legumes and three major genes, chalone reductase (CHR), isoflavone synthase (IFS) and isoflavonoid O-methyltransferase (IOMT), were cloned from white clover in this study. The white clover IFS2_12 gene was expressed in transgenic tobacco. Genistein, an isoflavonoid that is not naturally present in tobacco, was detected in the IFS-expressing tobacco, thus confirming the functionality of the IFS2_12 gene. Tobacco plants were transformed with ANT1, a transcription factor that induces the production of anthocyanins that share precursors with the isoflavonoid biosynthesis pathway. When IFS was expressed in red tobacco leaves, where anthocyanin biosynthesis was occurring, the levels of genistein were greater than in anthocyanin-free green leaves. White clover was transformed to overexpress the cloned IFS2_12 gene and some transformants had greater levels of IFS gene expression, up to 12.9 times the average wild type level. However, these transformants did not produce formononetin levels greater than the wild-type. A gene fusion of alfalfa chalcone isomerase (CHI), which produces the precursors naringenin and liquiritigenin, and soybean IFS, which converts the precursors to genistein and daidzein, respectively, was received from the Noble Foundation. Transgenic white clover plants expressing IFS/CHI were produced using a novel method that also regenerated wild-type clones of the transgenic plants. When compared with their wild-type clones, two IFS/CHI transformants produced higher levels of formononetin, thus supporting the suggestion that isoflavonoid levels can be increased in white clover through overexpression of isoflavonoid biosynthesis genes.

Techn. Universiẗat, Diss., 2004--Berlin.

Zugl.: Braunschweig, Techn. Univ., Diss., 2008

Edamame's (Glycine max L. Merrill) consumption in the US has also been growing due to purported health benefits. Edamame grows well around the US, but few have measured the growth and yield in the mid-Atlantic region. The objective of these studies were to determine the potential yield of edamame, determine how yield components change with planting date and cultivar, and to measure total protein, lipids, antioxidant activity and isoflavone concentrations at harvest.. The five cultivars (BeSweet 292, BeSweet 2015, BeSweet 2001, Midori Giant and Sunrise) used in the cultivar evaluation trial and for nutritional constituents analysis were grown in Painter, Virginia in 2008 and 2009. The cultivar evaluation trial yielded between 5,600 and 8,400 kg per ha. Percent marketable pods ranged from 74.3 and 85.6% in the cultivar evaluation trial. There were significant differences in average seed size across cultivars in both years. Cultivar lipid content followed the same patterns in both years with 2009 having lower overall concentrations than 2008. Protein contents were similar in 2008 and 2009. Both years, "BeSweet 2015" and "BeSweet 2001" had high radical scavenging ability and Midori Giant had the lowest. In 2008, there were no significant differences in the ORAC assay. "BeSweet 292" had significantly more reducing activity in the DPPH assay than Sunrise in 2009. Isoflavones were measured in 2008 and 2009 at harvest and temporally in 2009. Of all isoflavones, Malonyl genistin had the highest concentration. Edamame is a suitable crop for cultivation in Virginia, and is high in nutritional quality. / Master of Science

A menopausa é conseqüência da falência ovariana, caracterizada pela queda na produção de estrógenos, podendo causar na mulher vários sintomas e condições patológicas como vaginite atrófica, osteoporose e doençsa cardiovasculares. A terapia estroprogestiva é largamente prescrita na pós-menopausa, no entanto existem freqüentes efeitos colaterais e contra-indicações, inclusive possibilidade de aumento do câncer de mama. Devido a toda esta polêmica as terapias alternativas para combaterem os sintomas da menopausa estão largamente sendo exploradas, neste contexto cita-se os fitohormônios. A isoflavona é um deles obtido de várias plantas e mais usualmente da soja, que possui na sua molécula uma estrutura fenólica que se liga aos receptores de estrogênio e exerce um efeito estrogênico. Neste estudo enfocando a isoflavona, extraída da soja Glycine max, para o tratamento dos sintomas da menopausa, avaliou-se o efeito desse composto sobre a maturação do epitélio cérvico-vaginal e microbiota vaginal. Para isto propô-se um estudo randomizado, duplo cego, placebo controlado do qual participaram 49 voluntárias. Um grupo de 23 mulheres recebeu 1 cápsula/dia de 40 mg de caseína de leite, constituindo o grupo placebo e o outro grupo de 26 mulheres tratadas com 1 cápsula/dia de 40 mg de isoflavona. Amostras de urina e secreção vaginal para colpocitograma, colpocitograma com coleta úmida, bacterioscopia foram colhidas antes do início e ao término do tratamento. Foram determinados índice de maturação (I.M.) e valor de maturação (V.M.) pela técnica da citologia hormonal. O estudo da microbiota foi avaliado por exame direto a fresco e bacterioscopia de Gram. As mulheres participantes do estudo forneceram dados através de entrevistas mensais, antes do início e após o término do estudo, respondendo a questionários sobre queixas, sintomas, história da vida sexual, percepção da menopausa, estado de saúde e questões sócio demográficas. A avaliação destes dados aparece no índice de Kuppermann, nas maiores queixas e melhoras mais evidentes das mulheres tratadas com isoflavona. A avaliação do I.M. resultou no grupo tratado com isoflavona em redução significativa das células parabasais (P=0,003) no colpocitograma tradicional, em aumento significativo das células superficiais cianofílicas (P=0,006) e tendência de aumento das células superficiais eosinofílicas (P=0,18). No colpocitograma obtido por coleta úmida houve diminuição significativa das células parabasais (P=0,004), aumento significativo na porcentagem de células superficiais cianofílicas (P=0,008) e tendência de aumento das células superficiais eosinofílicas (P=0,18). No urocitograma houve redução significativa de células parabasais (P=0,008), aumento significativo na porcentagem de células superficiais cianofílicas (P=0,013) e aumento das células superficiais eosinofílicas (P=0,002). No grupo placebo não se observou variação significativa nos índices acima mencionados. O V.M. no grupo tratado com isoflavona apresentou aumento significativo (P=0,007) no colpocitograma tradicional, colpocitograma, obtido por coleta úmida (P=0,004); e no urocitograma (P=0,0008). No grupo placebo houve diminuição significativa (P=0,02) para este índice. Após o tratamento pela isoflavona houve aumento significativo de bacilos de Döederlein (P=0,005), diminuição significativa de 19,3% nos casos de vaginose bacteriana (índice de Nugent) e diminuição de outros agentes microbiológicos patogênicos. Houve diminuição dos sintomas da menopausa após o tratamento com isoflavona, também se observou diminuição das queixas após o tratamento com placebo. / The lack of estrogens characterizes postmenopausal condition and is associated with pathological conditions and symptoms, including atrophic vaginites, osteoporosis and cardiovascular disease. Hormone replacement therapy is recommended for postmenopausal women but there are contraindications and side effects. There are many alternative therapy researches in order to alleviate these symptoms. Soy isoflavone has hormonal effects in postmenopausal women, by binding a common phenolic structure to estrogens receptors. The aim of this study focusing isoflavone, extracted from soy beans Glycine max, was to evaluate the effects of isofavone on vaginal cytology, vaginal maturation index (M.I.), maturation value (M.V.), vaginal microbiota and menopausal symptoms. This study involved 49 postmenopausal women, a randomized double blind placebo controlled. The first group of 23 women was treated with 40 mg/day capsule of placebo (caseyne) and the second group of 26 women was treated with 40 mg/day capsule of isoflavone. Urine and vaginal samples for 3 slides: colpocytogram, wet sample colpocytogram, Gram-stained smears was taken at baseline and after treatment in order to determine M.I., M.V., vaginal microbiological findings and colpocytologic findings. Study participants answered an in-person interview with questions in order to determine menopausal symptoms, complains, menopause perception, sexual life, general health to determine, for instance the Kuppermann menopausal index (KMI) at baseline, every month and after six-months of treatment. The M.I. from traditional colpocytogram reduced significantly the parabasal cells (P=0,003), increased significantly the percentage of Superficial Cianophilic cells (P=0,006), there was a tendence of increasing the Superficial Eosinofilic cells (P=0,18). From wet sample colpocytogram, similar results were obtained: reduced significantly the parabasal cells (P=0,004), increased significantly the percentage of Superficial Cianophilic cells (P=0,008) there was a tendence of increasing the Superficial Eosinofilic cells (P=0,18).. In urocytogram, the results obtained were: reduced significantly the parabasal cells (P=0,008), increased significantly the percentage of Superficial Cianophilic cells (P=0,013), increase of Superficial Eosinofilic cells (P=0,002). For placebo group there were no significative changes. The Maturation Value, M.V. increased significtively (P=0,007) in traditional colpocytogram; in wet sample colpocytogram (P=0,004) and in urocytogram (P=0,0008), comparing with placebo was observed significant decrease (P=0,02) for M.V. After treatment with isoflavone there was significative increase of Döederlein bacillus (P=0,005), decrease of 19,3% in cases of bacterial vaginosis (Nugent score) and decrease of others pathogenic microbial agents. The symptoms decreased significatively in woman treated with placebo and decreased significatively in women treated with isoflavone.

Made available in DSpace on 2017-07-10T14:17:19Z (GMT). No. of bitstreams: 1 CAROLINA BOAMORTE.pdf: 1397771 bytes, checksum: 77780c428457cc0480306357b2380e44 (MD5) Previous issue date: 2016-06-16 / The ratio of osteoporosis in postmenopausal women is controversial, but may be a risk factor for periodontal disease, as well as having a direct or indirect role in bone due to estrogen action, since this can inhibit production of proinflammatory cytokines -inflammatory such as TNF-alpha and IL-6 that favor bone resorption. One of the alternatives in hormone replacement therapy is the isoflavone, a phyto-hormone that has established itself as replacement therapy for having a similar action to estrogen, but with protective effects against cancers, osteoporosis, among others. The objective was to evaluate the effect of isoflavone on alveolar bone of ovariectomized rats with induced periodontitis. Sixty (60) Female rats with 8 weeks of age were randomly divided into 6 groups: 1) control group (CON); 2) Bandage Group (LIG); 3) Group Ovariectomy (OVX); 4) Ovariectomia and Phytoestrogen group (OVX + FIT); 5) Ovariectomia and ligature group (OVX + LIG); 6) Ovariectomia Group, Phytoestrogen and Ligation (OVX + FIT + LIG) .All the animals received standard diet and water at will. After 70 days of life, the animals were anesthetized (xylazine 10 mg / kg and ketamine 75 mg / kg) and were placed in proper operating table, which allowed the maintenance of the mouth opening of the rats facilitating access to the teeth of the posterior region jaw. With the aid of a modified forceps and an explorer, it was placed a cotton yarn number 40 around the mandibular first molar right and left. After this procedure, after 79 days of life, the rats were submitted to anesthesia and ovariectomy surgery to mimic the absence of female sex hormones of menopause in women. Ligation served as irritating gums for 30 days and favored the accumulation of plaque, while hormone replacement therapy (isoflavone) was held for 21 days (0.25 g / kg / day sc) in the above-mentioned groups. At 100 days of life, the rats were euthanized. Macroscopic, microscopic and X-ray analyzes were performed with the left and right jaws of each rat as well with samples of gingival tissue to assess the periodontal tissues and their effects induced by hormone replacement. As a result of analysis, it was observed that the groups of phytoestrogen showed less bone loss than the groups that received no therapy isoflavone (p <0.05) and the treated groups had a higher phytoestrogen osteoblasts and osteocytes osteoclasts and lower than the groups not receiving hormone replacement therapy (p <0.05). Therefore, the isoflavone may confer a protective effect on bone loss in periodontal disease progression, compounded by osteoporosis. / A relação da osteoporose na pós-menopausa é controversa, mas pode ser um fator de risco para doença periodontal, bem como possuir uma atuação direta e indireta no tecido ósseo devido a ação do estrógeno, uma vez que este pode inibir a produção de citocinas pró-inflamatórias como o TNF-alfa e a IL-6 que favorecem a reabsorção óssea.Uma das alternativas na terapia de reposição hormonal atualmente é a isoflavona, um fito-hormônio que vem se estabelecendo como terapia substitutiva por ter sua ação similar ao estrógeno, porém com efeitos protetores contra as doenças cancerígenas, osteoporose, entre outras. Assim, o objetivo foi avaliar a ação da isoflavona no tecido ósseo alveolar de ratas ovariectomizadas com peridontite induzida. Sessenta (60) Ratas fêmeas com 8 semanas de vida foram divididas aleatoriamente em 6 grupos: 1) Grupo Controle (CON); 2) Grupo Ligadura(LIG); 3) Grupo Ovariectomia (OVX+DMSO); 4) Grupo Ovariectomia e Fitoestrógeno (OVX+FIT); 5) Grupo Ovariectomia e Ligadura (OVX+LIG+DMSO); 6) Grupo Ovariectomia, Fitoestrógeno e Ligadura (OVX+FIT+LIG).Todos os animais receberam dieta padrão e água a vontade. Aos 70 dias de vida, os animais foram anestesiados (xilazina 10 mg/Kg e quetamina 75 mg/Kg) e foram posicionados em mesa operatória apropriada, a qual permitia a manutenção da abertura bucal das ratas facilitando o acesso aos dentes da região posterior da mandíbula. Com o auxílio de uma pinça modificada e de uma sonda exploradora, foi colocado um fio de algodão número 40 ao redor do primeiro molar inferior direito e esquerdo. Após este procedimento, aos 79 dias de vida, as ratas também foram submetidas à anestesia e à cirurgia de ovariectomia para mimetizar a ausência de hormônios sexuais femininos da menopausa em mulheres. A ligadura atuou como irritante gengival por 30 dias e favoreceu o acúmulo de placa bacteriana, enquanto a reposição hormonal (isoflavona) se realizou por 21 dias (0,25g/kg/dia s.c.) nos grupos anteriormente citados. Aos 100 dias de vida, as ratas foram eutanasiadas. Foram realizadas análises macroscópica, microscópica e radiográfica com as mandíbulas esquerda e direita de cada rata, bem como com amostras do tecido gengival para avaliar os tecidos periodontais e seus efeitos induzidos pela reposição hormonal. Como resultado das análises, foi observado que os grupos com fitoestrógeno apresentaram menor perda óssea alveolar do que os grupos que não receberam terapia com isoflavona (p< 0,05), bem como os grupos tratados com fitoestrógeno apresentaram uma maior quantidade de osteoblastos e osteócitos e menor de osteoclastos do que os grupos que não receberam terapia de reposição hormonal (p<0,05). Logo, a isoflavona pode conferir um efeito protetor à perda óssea alveolar, na progressão da doença periodontal, agravada pela osteoporose.

In wissenschaftlichen wie auch in öffentlichen Kreisen wird kontrovers diskutiert, ob durch die Aufnahme von hormonell aktiven Substanzen aus Nahrungs‐ oder Nahrungsergänzungsmitteln mit positiven oder negativen Einflüssen auf die menschliche Gesundheit und Lebensqualität zu rechnen ist. Das Interesse an solchen Substanzen ist in den letzten Jahren permanent gewachsen, und der Markt erhielt einen zusätzlichen Zulauf, als Studien zeigten, dass die klassische Hormonersatztherapie für postmenopausale Frauen aus Östrogen‐Progestagen‐Kombinationen mit einem erhöhten Risiko für hormonabhängige Krebsarten wie Brust‐ oder Endometriumskarzinomen verbunden ist. Demgegenüber implizieren epidemiologische Beobachtungen aus ostasiatischen Ländern, dass die Aufnahme sojareicher Nahrung mit einer verringerten Inzidenz für diese hormonabhängigen Krebsarten korreliert zu sein scheint. Stofflich betrachtet stehen in diesem Zusammenhang Isoflavone wie Genistein und Daidzein, die in großen Mengen z. B. in Sojabohnen vorkommen, im Fokus des wissenschaftlichen Interesses. Eine Vielzahl von in vitro und in vivo Studien ergab für Isoflavone ein heterogenes Wirkspektrum. Neben hormonellen Wirkungen wie schwach östrogenen bzw. antiöstrogenen Eigenschaften, sind ebenfalls eine Reihe nicht‐hormoneller Wirkungen, wie beispielsweise antioxidative Eigenschaften oder die Fähigkeit zur Hemmung von Tyrosinkinasen, beschrieben. Trotz der hohen Anzahl an Studien zu Isoflavonen ist immer noch unklar wie valide die Beobachtungen beim Menschen sind, da ihre Langzeitwirkungen experimentell kaum untersucht sind. Daher war es das Ziel dieser Arbeit, zur Aufklärung der Langzeitwirkung von Isoflavonen als Nahrungsbestandteile und damit zum Verständnis ihrer Wirkmechanismen beizutragen, um anschließend mit den erzielten Resultaten die Risikobewertung dieser Substanzklasse zu optimieren. Zu diesem Zweck wurden in einem generationsübergreifenden Fütterungsexperiment die Einflüsse eines sojabasierten Futters mit denen eines, mit isoliertem Genistein supplementierten, Futters verglichen und anschließend denen eines phytoöstrogenfreien Futters gegenüber gestellt. Sowohl in juvenilen als auch adulten Ratten wurden die gewebespezifischen, physiologischen und molekularen Antworten des Uterus evaluiert. In einer zusätzlichen Studie sollte zudem geklärt werden, inwieweit Gestagene die genistein‐ bzw. östrogenregulierten Prozesse im Uterus adulter ovariektomierter Ratten zu modulieren vermögen. Anhand etablierter molekularbiologischer Methoden zur Gen‐ (qPCR) und Proteinexpression (Western Blot), aber auch durch neu zu etablierende immunohistochemische Analysen wurden diverse physiologische (z. B. Körpergewicht, Futteraufnahme, spezifische Organgewichte) und molekulare Marker (z. B. uterine Markergene für Östrogenität oder Proliferation) analysiert und untereinander verglichen. Bei der Betrachtung der Ergebnisse wird deutlich, dass die lebenslange, kontinuierliche Isoflavonexposition der Mütter während fetaler und pränataler Entwicklungsphasen unterschiedliche gewebespezifische Effekte in juvenilen und adulten Weibchen der Tochtergeneration auslöst. Während auf der einen Seite die bereits in utero begonnene Exposition mit Genistein in juvenilen Tieren in einer relativ starken Uterotrophie sowie östrogenen Effekten hinsichtlich der Expressionsmuster einiger Markergene resultiert, zeigt Genistein in den gleichen Tieren andererseits auch Eigenschaften die als antiproliferativ und antiöstrogen zu bewerten sind. Interessanterweise sind diese Effekte im adulten Organismus anschließend nicht mehr nachweisbar. Des Weiteren zeigen unsere Untersuchungen, dass die Östrogenresponsivität des juvenilen Uterus nicht durch diätetische Isoflavone modifiziert wird. Genistein wirkt in diesen Tieren als Östrogenrezeptor β Partialagonist, wobei das antiproliferative sowie antiöstrogene Potenzial erst in hohen Konzentrationen erkennbar wird. Im Gegensatz zum juvenilen zeigt sich im adulten Uterus eine erheblich gesteigerte Östrogenresponsivität infolge der lebenslangen, kontinuierlichen und bereits in utero begonnenen Isoflavon‐ bzw. Genisteinexposition. Diese Tiere reagieren mit dramatisch erhöhten Uterusfeuchtgewichten auf Östrogenrezeptor α vermittelte östrogene Stimuli. Bemerkenswerterweise sind diese stark erhöhten Uterusfeuchtgewichte jedoch nicht auf vermehrte Proliferation, sondern auf eine gesteigerte Einlagerung von Luminalflüssigkeit zurückzuführen. Analysen zur Genexpression von Aquaporinen deuten zwar auf östrogenabhängige Mechanismen hin, jedoch konnte kein direkter Bezug zur chronischen Isoflavon‐ bzw. Genisteinexposition nachgewiesen werden. Wie schon in den Untersuchungen am juvenilen Uterus beobachtet, vermittelt Genistein auch im adulten Uterus seine molekularen Wirkungen als tendenzieller Östrogenrezeptor β‐selektiver Partialagonist. Unsere Analysen der Expression spezifischer uteriner Markergene legen darüber hinaus die Vermutung nahe, dass Genistein seine molekularen Wirkungen präferenziell über den klassischen ERE‐abhängigen Signalweg vermittelt, da durch Genistein besonders Gene reguliert werden, die diese Transkriptionsfaktor Bindungsstellen in der Promotorregion besitzen. Die Rolle von Gestagenen bei genisteinregulierten Prozessen im Uterus ist nicht ganz klar. Während unsere Ergebnisse zeigen, dass Progesteron in einem kurzfristigen dreitägigen experimentellen Design, unabhängig von der Ab‐ bzw. Anwesenheit von 17β‐Östradiol, kaum einen Einfluss auf genisteinregulierte Prozesse hat, legen die Ergebnisse unserer längerfristigen Experimente hingegen die Vermutung nahe, dass Gestagene die chronischen Effekte von Isoflavonen modulieren können. Schlussfolgernd kann anhand unserer Studien festgehalten werden, dass die molekularen Wirkungen von Genistein stark von Faktoren wie der Expositionsdauer, der Expositionskonzentration sowie dem Alter des Organismus abhängen. Unsere Daten zeigen eindeutig, dass eine traditionelle sojabasierte ostasiatische Diät in keiner Weise mit einer, durch isolierte Isoflavone supplementierten, westlichen Diät gleichzusetzen ist. Des Weiteren konnten wir im Rahmen dieser Arbeit zeigen, dass die Exposition mit diätetischen Isoflavonen während der Embryogenese einen lebenslangen Einfluss auf die Nachkommen hat, indem die östrogenresponsivität dieser Individuen massiv beeinflusst wird.

A menopausa é conseqüência da falência ovariana, caracterizada pela queda na produção de estrógenos, podendo causar na mulher vários sintomas e condições patológicas como vaginite atrófica, osteoporose e doençsa cardiovasculares. A terapia estroprogestiva é largamente prescrita na pós-menopausa, no entanto existem freqüentes efeitos colaterais e contra-indicações, inclusive possibilidade de aumento do câncer de mama. Devido a toda esta polêmica as terapias alternativas para combaterem os sintomas da menopausa estão largamente sendo exploradas, neste contexto cita-se os fitohormônios. A isoflavona é um deles obtido de várias plantas e mais usualmente da soja, que possui na sua molécula uma estrutura fenólica que se liga aos receptores de estrogênio e exerce um efeito estrogênico. Neste estudo enfocando a isoflavona, extraída da soja Glycine max, para o tratamento dos sintomas da menopausa, avaliou-se o efeito desse composto sobre a maturação do epitélio cérvico-vaginal e microbiota vaginal. Para isto propô-se um estudo randomizado, duplo cego, placebo controlado do qual participaram 49 voluntárias. Um grupo de 23 mulheres recebeu 1 cápsula/dia de 40 mg de caseína de leite, constituindo o grupo placebo e o outro grupo de 26 mulheres tratadas com 1 cápsula/dia de 40 mg de isoflavona. Amostras de urina e secreção vaginal para colpocitograma, colpocitograma com coleta úmida, bacterioscopia foram colhidas antes do início e ao término do tratamento. Foram determinados índice de maturação (I.M.) e valor de maturação (V.M.) pela técnica da citologia hormonal. O estudo da microbiota foi avaliado por exame direto a fresco e bacterioscopia de Gram. As mulheres participantes do estudo forneceram dados através de entrevistas mensais, antes do início e após o término do estudo, respondendo a questionários sobre queixas, sintomas, história da vida sexual, percepção da menopausa, estado de saúde e questões sócio demográficas. A avaliação destes dados aparece no índice de Kuppermann, nas maiores queixas e melhoras mais evidentes das mulheres tratadas com isoflavona. A avaliação do I.M. resultou no grupo tratado com isoflavona em redução significativa das células parabasais (P=0,003) no colpocitograma tradicional, em aumento significativo das células superficiais cianofílicas (P=0,006) e tendência de aumento das células superficiais eosinofílicas (P=0,18). No colpocitograma obtido por coleta úmida houve diminuição significativa das células parabasais (P=0,004), aumento significativo na porcentagem de células superficiais cianofílicas (P=0,008) e tendência de aumento das células superficiais eosinofílicas (P=0,18). No urocitograma houve redução significativa de células parabasais (P=0,008), aumento significativo na porcentagem de células superficiais cianofílicas (P=0,013) e aumento das células superficiais eosinofílicas (P=0,002). No grupo placebo não se observou variação significativa nos índices acima mencionados. O V.M. no grupo tratado com isoflavona apresentou aumento significativo (P=0,007) no colpocitograma tradicional, colpocitograma, obtido por coleta úmida (P=0,004); e no urocitograma (P=0,0008). No grupo placebo houve diminuição significativa (P=0,02) para este índice. Após o tratamento pela isoflavona houve aumento significativo de bacilos de Döederlein (P=0,005), diminuição significativa de 19,3% nos casos de vaginose bacteriana (índice de Nugent) e diminuição de outros agentes microbiológicos patogênicos. Houve diminuição dos sintomas da menopausa após o tratamento com isoflavona, também se observou diminuição das queixas após o tratamento com placebo. / The lack of estrogens characterizes postmenopausal condition and is associated with pathological conditions and symptoms, including atrophic vaginites, osteoporosis and cardiovascular disease. Hormone replacement therapy is recommended for postmenopausal women but there are contraindications and side effects. There are many alternative therapy researches in order to alleviate these symptoms. Soy isoflavone has hormonal effects in postmenopausal women, by binding a common phenolic structure to estrogens receptors. The aim of this study focusing isoflavone, extracted from soy beans Glycine max, was to evaluate the effects of isofavone on vaginal cytology, vaginal maturation index (M.I.), maturation value (M.V.), vaginal microbiota and menopausal symptoms. This study involved 49 postmenopausal women, a randomized double blind placebo controlled. The first group of 23 women was treated with 40 mg/day capsule of placebo (caseyne) and the second group of 26 women was treated with 40 mg/day capsule of isoflavone. Urine and vaginal samples for 3 slides: colpocytogram, wet sample colpocytogram, Gram-stained smears was taken at baseline and after treatment in order to determine M.I., M.V., vaginal microbiological findings and colpocytologic findings. Study participants answered an in-person interview with questions in order to determine menopausal symptoms, complains, menopause perception, sexual life, general health to determine, for instance the Kuppermann menopausal index (KMI) at baseline, every month and after six-months of treatment. The M.I. from traditional colpocytogram reduced significantly the parabasal cells (P=0,003), increased significantly the percentage of Superficial Cianophilic cells (P=0,006), there was a tendence of increasing the Superficial Eosinofilic cells (P=0,18). From wet sample colpocytogram, similar results were obtained: reduced significantly the parabasal cells (P=0,004), increased significantly the percentage of Superficial Cianophilic cells (P=0,008) there was a tendence of increasing the Superficial Eosinofilic cells (P=0,18).. In urocytogram, the results obtained were: reduced significantly the parabasal cells (P=0,008), increased significantly the percentage of Superficial Cianophilic cells (P=0,013), increase of Superficial Eosinofilic cells (P=0,002). For placebo group there were no significative changes. The Maturation Value, M.V. increased significtively (P=0,007) in traditional colpocytogram; in wet sample colpocytogram (P=0,004) and in urocytogram (P=0,0008), comparing with placebo was observed significant decrease (P=0,02) for M.V. After treatment with isoflavone there was significative increase of Döederlein bacillus (P=0,005), decrease of 19,3% in cases of bacterial vaginosis (Nugent score) and decrease of others pathogenic microbial agents. The symptoms decreased significatively in woman treated with placebo and decreased significatively in women treated with isoflavone.

Orientador: Cristiano Soleo de Funari / Resumo: Aproximadamente 3,7 x 109 toneladas de resíduos agrícolas são produzidos anualmente no mundo. Por ser uma das grandes potências agrícolas atualmente, grandes quantidades de resíduos agrícolas são geradas constantemente no Brasil. Uma das principais culturas produzidas no país é a soja. Além de seu uso na pecuária, utilizada na alimentação animal, assim como para alimentação humana, a soja desperta grande interesse do ponto de vista farmacológico, por ser uma das principais fontes de isoflavonas, uma forma de fitoestrogênio, sendo umas das espécies listadas na RENISUS. Este trabalho teve como objetivo investigar o perfil químico dos resíduos provenientes da cultura da soja (caules, folhas e vagens) por meio de uma abordagem analítica verde. Os extratos foram obtidos em triplicata por maceração dinâmica a 30 °C, com acetona e etanol, com e sem adição de ácido. O solvente acetonitrila foi utilizado como solvente de referência, pois é comumente empregado tanto para a extração como para a separação de isoflavonas presentes em grãos de soja. Os extratos foram analisados por cromatografia líquida de alta eficiência acoplada a um espectrofotômetro de untravioleta/visível (HPLC-PAD/UV) e a comparação da eficiência de extração entre os solventes testados foi feita com base em número total de picos, área total sob os picos e rendimento da extração. Os solventes verdes etanol e acetona mostraram-se mais eficientes do que o solvente de referência acetonitrila. Posteriormente, uma estratég... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Approximately 3.7 x 109 tonnes of agricultural waste is produced annually in the world. Because it is one of the great agricultural powers today, large amounts of agricultural residues are constantly generated in Brazil. One of the main crops produced in the country is soy. In addition to its use in livestock and human nutrition, the soybean awakes great interest from the pharmacological point of view, being one of the main sources of isoflavones, a form of phytoestrogen, being one of the species listed in RENISUS. The objective of this work was to investigate the chemical profile of residues from soybean (shoots, leaves and pods) using a green analytical approach. The extracts were obtained in triplicate by dynamic maceration at 30 ° C, with acetone and ethanol, with and without acid addition. The solvent acetonitrile was used as reference solvent, since it is commonly used both for the extraction and the separation of isoflavones present in soybean grains. The extracts were analyzed by high performance liquid chromatography coupled to an untraviolet/visible spectrophotometer (HPLC-PAD/UV) and the comparison of the extraction efficiency among the solvents tested was done based on total number of peaks, total area under peaks and extraction yield. The green solvents ethanol and acetone were more efficient than the reference solvent acetonitrile. Subsequently, a strategy for the identification of compounds in mixture, based on data obtained by high performance liquid chromatog... (Complete abstract click electronic access below) / Mestre