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THE ROLE OF SERINE/THREONINE PHOSPHATASES IN SPERMFUNCTIONJoudeh, Nidaa M. 01 May 2017 (has links)
No description available.
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Evaluation of standard and development of new sperm function tests in selected primate speciesPrag, Farren Chelsea January 2017 (has links)
Magister Scientiae - MSc / Male infertility in humans has increased in the last few decades and could be as high as 40%, while up to 50% of these men have ''unexplained'' (idiopathic) infertility. Although newly developed molecular techniques have great value in detecting subtle causes of male infertility, more detailed sperm functional tests are required to identify compromised fertility, especially in a clinical set-up. Since ethical constraints often preclude the pursuit of many basic research questions in humans, non-human primates (NHPs) have been identified as key models in human-related studies. NHPs are often used in studies on male fertility/infertility, IVF or assisted reproductive technology (ART) procedures, male contraception and reproductive toxicology. However, comparing results of NHP and human studies require that techniques used for assessment must be objective, standardized and sensitive to recognize compromised sperm function. The aim of this study was to evaluate standard sperm functional tests and develop new functional tests using NHP sperm, specifically from vervet monkeys (Chlorocebus aethiops), chacma baboons (Papio ursinus) and rhesus monkeys (Macaca mulatta), for application in human and NHP studies and to ultimately develop a basic primate model. The sperm functions investigated included sperm motility, longevity, vitality, DNA integrity, acrosome reaction, and hyperactivation. The sperm functional tests evaluated were: CASA motility analysis; Sperm Longevity test; Eosin-Nigrosin and Hoechst and Propidium Iodide staining, as well as the use of WST-1 cytotoxicity assay for vitality; the TUNEL assay for DNA integrity; Acrosome Intactness Test; and induction of hyperactivation via stimulants. The validity of each test was investigated by inhibiting sperm function through the use of copper sulphate and cadmium chloride. All functional tests were successfully performed across all three species, except the TUNEL assay for DNA integrity, and was further used for validation testing. Validation testing proved that all sperm functional parameters were significantly affected by the highest concentrations of the chemicals (250 µg/ml CuSO4 and 500 µg/ml CdCl2) and if not significant, trends of reduction were seen. The tests employed were therefore sensitive to the inhibitory effect of the metals. By evaluating these established sperm functional tests we found that primates would serve as good models for research study. Furthermore, we optimized and modified techniques for sperm and functional analysis in these three primate species and this study will standardize protocols for use in future studies on male infertility. Additionally, comparing human and NHP sperm function can possibly reveal or explain the high infertility rates in humans.
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Comparação entre os critérios de análise seminal da OMS de 1999 e de 2010 e contagem total de espermatozoides móveisZorzi, Patricia de Moraes de January 2016 (has links)
Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática. / Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.
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Comparação entre os critérios de análise seminal da OMS de 1999 e de 2010 e contagem total de espermatozoides móveisZorzi, Patricia de Moraes de January 2016 (has links)
Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática. / Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.
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Comparação entre os critérios de análise seminal da OMS de 1999 e de 2010 e contagem total de espermatozoides móveisZorzi, Patricia de Moraes de January 2016 (has links)
Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática. / Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.
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AnÃlise proteÃmica do plasma seminal de carneiros Santa InÃs adulto / Proteomic analysis of seminal plasma Santa InÃs rams adultJoÃo Paulo Arcelino do RÃgo 16 March 2010 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O Nordeste brasileiro à detentor de um expressivo rebanho ovino, e, dentre as raÃas deslanadas existentes na regiÃo, a Santa InÃs se destaca por apresentar bom desenvolvimento corporal, ganho de peso e adaptabilidade Ãs condiÃÃes tropicais. Recentemente, demonstrou-se que o perfil protÃico do plasma seminal de carneiros Santa InÃs passa por alteraÃÃes significativas durante o desenvolvimento reprodutivo, simultaneamente a mudanÃas nos parÃmetros de motilidade e concentraÃÃo espermÃtica. Contudo, ainda nÃo se conhece a identidade de todas essas proteÃnas. Dessa forma, o presente trabalho foi conduzido com o objetivo de identificar as proteÃnas do plasma seminal de carneiros Santa InÃs maduros, utilizando as tÃcnicas de eletroforese bidimensional associada à espectrometria de massa. Amostras de sÃmen foram coletadas de oito carneiros adultos e o plasma seminal obtido atravÃs de centrifugaÃÃo e submetido à eletroforese bidimensional. Os gÃis foram corados com Coomassie coloidal, digitalizados e analisados no aplicativo PDQuest. Os spots foram cortados individualmente dos gÃis, digeridos com tripsina, e submetidos a identificaÃÃo por espectrometria de massa (MALDI-ToF/ToF). A sequÃncia peptÃdica das proteÃnas mais abundantes e a distribuiÃÃo dos domÃnios foram representadas graficamente utilizando o aplicativo Caititu. Foram detectados, em mÃdia, 302 spots por gel. Destes, 143 estavam presentes em todos os gÃis. Trinta e nove spots foram identificados, correspondendo a 26 diferentes proteÃnas. As proteÃnas mais abundantes foram as RSVPs 14 e 22 kDa, pertencentes à famÃlia das binder of sperm proteins, e as Bodesinas-1 e 2, pertencentes à famÃlia das espermadesinas. Outras proteÃnas foram identificas no estudo, incluindo albumina, clusterina, peroxiredoxina, lactotransferrina, metaloproteinase de matriz 2 (MMP-2), beta galactosidase e heat shock proteins. Dessa forma, a identidade dessas proteÃnas sugere que o plasma seminal de carneiros Santa InÃs contÃm componentes que participam de diversos processos fisiolÃgicos, incluindo proteÃÃo do espermatozÃide, modulaÃÃo da motilidade e capacitaÃÃo espermÃtica, modificaÃÃo da membrana do espermatozÃide e interaÃÃo entre gametas. O conhecimento dessas proteÃnas contribuirà para uma melhor compreensÃo dos mecanismos de regulaÃÃo do plasma seminal sobre a funÃÃo espermÃtica em ovinos / Northeastern Brazil has a significant population of sheep and, among the
native hairy breeds, Santa InÃs is known for its good performance and adaptability to
tropical environments. Recently, it has been shown that the protein profile of the
seminal plasma of Santa InÃs rams changes during sexual development, in concert
with changes in semen parameters, such as sperm motility, morphology and
concentration. However, the identity of seminal plasma proteins from these hairy
rams is still unknown. Therefore, we pursued the identification of seminal plasma
proteins from Santa InÃs rams using a proteomic approach. Semen samples were
collected from eight mature rams, and seminal plasma saved after centrifugation.
Seminal plasma protein maps, obtained by two-dimensional electrophoresis, were
stained with colloidal Coomassie. The gels were scanned and analyzed using
PDQuest software. Protein spots were individually cut, in-gel digested with trypsin
and identified after tandem mass spectrometry and database search. On average , we
detected 302 spots per gel, from which 143 were present on every member of the
match set generated by PDQuest. Thirty-nine spots were positively identified by
mass spectrometry, corresponding to 26 different proteins. The major proteins
present in the maps were the BSP-like RSVP14 and RSVP22 and the spermadhesins
bodhesin 1 and 2. Other proteins detected in the maps included albumin, clusterin,
peroxiredoxin, lactoferrin, transferrin, matrix metalloproteinase 2, beta galactosidase
and heat shock proteins. The Identity of these proteins suggest their participation on
several physiological events, such as sperm protection, regulation of sperm motility
and capacitation, modification of sperm membrane and fertilization. The knowledge
of the proteome of the ovine seminal plasma is a necessary step towards the understanding of the mechanisms underlying the regulation of sperm function by
seminal plasma components in rams
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IMPORTANCE OF THE MAMMAL - SPECIFIC PROTEIN PHOSPHATASE ISOFORM, PPP1CC2, IN SUPPORTING SPERMATOGENESIS AND SPERM FUNCTIONSen, Sabyasachi 25 November 2014 (has links)
No description available.
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Effect of tea and herbal infusions on mammalian reproduction and fertilityOpuwari, Chinyerum Sylvia January 2013 (has links)
<p>Camellia sinensis (tea) and Aspalathus linearis (rooibos) may improve reproductive function owing to their antioxidant properties. To test this<br />
hypothesis, male and female rats were given 2% and 5% green tea (Gt), black tea (Bt), unfermented rooibos (Ur) or fermented rooibos (Fr) as sole source of drinking for 52 and 21 days respectively. Control rats received tap water. In addition, TM3 Leydig cells were exposed to 0.025, 0.05, 0.1 and 0.5 % aqueous extracts of green tea, black tea, unfermented and fermented rooibos for 24h. In vitro analysis of tea and the herbal infusion revealed the phenolic property and antioxidant capacity (FRAP) in the order Gt > / Bt > / Ur > / Fr. Camellia sinensis and Aspalathus linearis revealed no significant effect on serum antioxidant capacity (p > / 0.05) and lipid peroxidation (MDA) in the kidney or liver in both male and female rats and in the testes of the male rats (p > / 0.05). In addition, the antioxidant levels were maintained in the testes, liver and kidneys in both the male and female rats. In the male rats, no significant alterations were observed in body weight gain, liver and reproductive organs weight, and serum testosterone (p > / 0.05). Only, 5% green tea significantly increased testosterone level (p < / 0.05). Seminiferous tubules displayed complete spermatogenesis with abundant sperm in the lumen in all treated groups. However, a significant decrease in diameter and germinal epithelial height of these tubules were observed (p < / 0.05). In the epididymides, epithelial height of caput region showed a significant increase (p < / 0.01), while the cauda region was increased by Camellia sinensis but decreased by Aspalathus linearis. Sperm concentration improved significantly by green tea and unfermented rooibos (p < / 0.05), while black tea and fermented rooibos produced a non significant effect (p > / 0.05). Sperm viability was enhanced in all treatment groups (p < / 0.05). Furthermore, green tea, black tea and unfermented rooibos significantly improved the motility of rat sperm (p < / 0.05) / fermented rooibos tended to improve it (p > / 0.05). In addition, green tea, black tea and fermented rooibos enhanced acrosome reaction (p < / 0.05). Creatinine activity was significantly higher in rats treated with black tea, unfermented rooibos or fermented rooibos (p < / 0.05), green tea tended to increase it (p > / 0.05) reflecting the significant increased kidney weight in the treatment groups at high concentrations. Liver markers, ALT and AST, decreased significantly in all treated groups (p < / 0.05), except in 5% fermented rooibos where a significant increase in AST level was observed (p < / 0.01). In the female rats, the body weight gain, and reproductive organs weight was no affected (p > / 0.05). However, 5% fermented rooibos reduced the ovarian weight (p < / 0.05), while 5% unfermented rooibos significantly increased the uterine weight (p < / 0.05). Liver weight increased significantly by black tea and unfermented rooibos (p < / 0.05) while the kidney weight increased significantly by 5% black tea (p < / 0.05). No significant effect was observed in the level of FSH produced, on the other hand, Camellia sinensis significantly lowered the level of LH (p < / 0.05), while Aspalathus linearis had no effect (p > / 0.05). Creatinine activity was enhanced significantly only by 5% fermented rooibos (p < / 0.05). Liver markers, ALT and AST were reduced in most treated groups except in fermented rooibos where an increase was observed. In addition, histological sections revealed no obvious alteration in the ovaries, uteri, kidneys and liver of all treated female rats. Camellia sinensis and Aspalathus linearis significantly reduced the level of testosterone produced in TM3 Leydig cells under stimulated conditions in vitro (p< / 0.05). Furthermore, both plants maintained the viability and morphology of the cells. However, at 0.5% of either plant extracts, a significant decrease in the viability (p < / 0.05) and altered morphology of the TM3 Leydig cells was observed. In conclusion, Camellia sinensis and Aspalathus linearis significantly improved certain sperm function which might be attributed to their high level of antioxidant activity. However, the prolonged exposure of both plant extracts might result in subtle structural changes in the male reproductive system and impair kidney function. In addition, fermented rooibos at high concentration may also impair the functions of the liver. In vitro, both plants were shown to possess anti-androgenic property on TM3 Leydig cells. Furthermore, both Camellia sinensis and Aspalathus linearis may be classified as weak phytoestrogens due to the changes in the weight of the uterus and ovaries observed.</p>
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Effect of tea and herbal infusions on mammalian reproduction and fertilityOpuwari, Chinyerum Sylvia January 2013 (has links)
<p>Camellia sinensis (tea) and Aspalathus linearis (rooibos) may improve reproductive function owing to their antioxidant properties. To test this<br />
hypothesis, male and female rats were given 2% and 5% green tea (Gt), black tea (Bt), unfermented rooibos (Ur) or fermented rooibos (Fr) as sole source of drinking for 52 and 21 days respectively. Control rats received tap water. In addition, TM3 Leydig cells were exposed to 0.025, 0.05, 0.1 and 0.5 % aqueous extracts of green tea, black tea, unfermented and fermented rooibos for 24h. In vitro analysis of tea and the herbal infusion revealed the phenolic property and antioxidant capacity (FRAP) in the order Gt > / Bt > / Ur > / Fr. Camellia sinensis and Aspalathus linearis revealed no significant effect on serum antioxidant capacity (p > / 0.05) and lipid peroxidation (MDA) in the kidney or liver in both male and female rats and in the testes of the male rats (p > / 0.05). In addition, the antioxidant levels were maintained in the testes, liver and kidneys in both the male and female rats. In the male rats, no significant alterations were observed in body weight gain, liver and reproductive organs weight, and serum testosterone (p > / 0.05). Only, 5% green tea significantly increased testosterone level (p < / 0.05). Seminiferous tubules displayed complete spermatogenesis with abundant sperm in the lumen in all treated groups. However, a significant decrease in diameter and germinal epithelial height of these tubules were observed (p < / 0.05). In the epididymides, epithelial height of caput region showed a significant increase (p < / 0.01), while the cauda region was increased by Camellia sinensis but decreased by Aspalathus linearis. Sperm concentration improved significantly by green tea and unfermented rooibos (p < / 0.05), while black tea and fermented rooibos produced a non significant effect (p > / 0.05). Sperm viability was enhanced in all treatment groups (p < / 0.05). Furthermore, green tea, black tea and unfermented rooibos significantly improved the motility of rat sperm (p < / 0.05) / fermented rooibos tended to improve it (p > / 0.05). In addition, green tea, black tea and fermented rooibos enhanced acrosome reaction (p < / 0.05). Creatinine activity was significantly higher in rats treated with black tea, unfermented rooibos or fermented rooibos (p < / 0.05), green tea tended to increase it (p > / 0.05) reflecting the significant increased kidney weight in the treatment groups at high concentrations. Liver markers, ALT and AST, decreased significantly in all treated groups (p < / 0.05), except in 5% fermented rooibos where a significant increase in AST level was observed (p < / 0.01). In the female rats, the body weight gain, and reproductive organs weight was no affected (p > / 0.05). However, 5% fermented rooibos reduced the ovarian weight (p < / 0.05), while 5% unfermented rooibos significantly increased the uterine weight (p < / 0.05). Liver weight increased significantly by black tea and unfermented rooibos (p < / 0.05) while the kidney weight increased significantly by 5% black tea (p < / 0.05). No significant effect was observed in the level of FSH produced, on the other hand, Camellia sinensis significantly lowered the level of LH (p < / 0.05), while Aspalathus linearis had no effect (p > / 0.05). Creatinine activity was enhanced significantly only by 5% fermented rooibos (p < / 0.05). Liver markers, ALT and AST were reduced in most treated groups except in fermented rooibos where an increase was observed. In addition, histological sections revealed no obvious alteration in the ovaries, uteri, kidneys and liver of all treated female rats. Camellia sinensis and Aspalathus linearis significantly reduced the level of testosterone produced in TM3 Leydig cells under stimulated conditions in vitro (p< / 0.05). Furthermore, both plants maintained the viability and morphology of the cells. However, at 0.5% of either plant extracts, a significant decrease in the viability (p < / 0.05) and altered morphology of the TM3 Leydig cells was observed. In conclusion, Camellia sinensis and Aspalathus linearis significantly improved certain sperm function which might be attributed to their high level of antioxidant activity. However, the prolonged exposure of both plant extracts might result in subtle structural changes in the male reproductive system and impair kidney function. In addition, fermented rooibos at high concentration may also impair the functions of the liver. In vitro, both plants were shown to possess anti-androgenic property on TM3 Leydig cells. Furthermore, both Camellia sinensis and Aspalathus linearis may be classified as weak phytoestrogens due to the changes in the weight of the uterus and ovaries observed.</p>
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Análise proteômica do plasma seminal de carneiros Santa Inês adulto / Proteomic analysis of seminal plasma Santa Inês rams adultRêgo, João Paulo Arcelino do January 2010 (has links)
RÊGO, João Paulo Arcelino do. Análise proteômica do plasma seminal de carneiros Santa Inês adulto. 2010. 107 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Zootecnia, Fortaleza-CE, 2010 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-29T14:08:18Z
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Previous issue date: 2010 / Northeastern Brazil has a significant population of sheep and, among the native hairy breeds, Santa Inês is known for its good performance and adaptability to tropical environments. Recently, it has been shown that the protein profile of the seminal plasma of Santa Inês rams changes during sexual development, in concert with changes in semen parameters, such as sperm motility, morphology and concentration. However, the identity of seminal plasma proteins from these hairy rams is still unknown. Therefore, we pursued the identification of seminal plasma proteins from Santa Inês rams using a proteomic approach. Semen samples were collected from eight mature rams, and seminal plasma saved after centrifugation. Seminal plasma protein maps, obtained by two-dimensional electrophoresis, were stained with colloidal Coomassie. The gels were scanned and analyzed using PDQuest software. Protein spots were individually cut, in-gel digested with trypsin and identified after tandem mass spectrometry and database search. On average , we detected 302 spots per gel, from which 143 were present on every member of the match set generated by PDQuest. Thirty-nine spots were positively identified by mass spectrometry, corresponding to 26 different proteins. The major proteins present in the maps were the BSP-like RSVP14 and RSVP22 and the spermadhesins bodhesin 1 and 2. Other proteins detected in the maps included albumin, clusterin, peroxiredoxin, lactoferrin, transferrin, matrix metalloproteinase 2, beta galactosidase and heat shock proteins. The Identity of these proteins suggest their participation on several physiological events, such as sperm protection, regulation of sperm motility and capacitation, modification of sperm membrane and fertilization. The knowledge of the proteome of the ovine seminal plasma is a necessary step towards the understanding of the mechanisms underlying the regulation of sperm function by seminal plasma components in rams / O Nordeste brasileiro é detentor de um expressivo rebanho ovino, e, dentre as raças deslanadas existentes na região, a Santa Inês se destaca por apresentar bom desenvolvimento corporal, ganho de peso e adaptabilidade às condições tropicais. Recentemente, demonstrou-se que o perfil protéico do plasma seminal de carneiros Santa Inês passa por alterações significativas durante o desenvolvimento reprodutivo, simultaneamente a mudanças nos parâmetros de motilidade e concentração espermática. Contudo, ainda não se conhece a identidade de todas essas proteínas. Dessa forma, o presente trabalho foi conduzido com o objetivo de identificar as proteínas do plasma seminal de carneiros Santa Inês maduros, utilizando as técnicas de eletroforese bidimensional associada à espectrometria de massa. Amostras de sêmen foram coletadas de oito carneiros adultos e o plasma seminal obtido através de centrifugação e submetido à eletroforese bidimensional. Os géis foram corados com Coomassie coloidal, digitalizados e analisados no aplicativo PDQuest. Os spots foram cortados individualmente dos géis, digeridos com tripsina, e submetidos a identificação por espectrometria de massa (MALDI-ToF/ToF). A sequência peptídica das proteínas mais abundantes e a distribuição dos domínios foram representadas graficamente utilizando o aplicativo Caititu. Foram detectados, em média, 302 spots por gel. Destes, 143 estavam presentes em todos os géis. Trinta e nove spots foram identificados, correspondendo a 26 diferentes proteínas. As proteínas mais abundantes foram as RSVPs 14 e 22 kDa, pertencentes à família das binder of sperm proteins, e as Bodesinas-1 e 2, pertencentes à família das espermadesinas. Outras proteínas foram identificas no estudo, incluindo albumina, clusterina, peroxiredoxina, lactotransferrina, metaloproteinase de matriz 2 (MMP-2), beta galactosidase e heat shock proteins. Dessa forma, a identidade dessas proteínas sugere que o plasma seminal de carneiros Santa Inês contém componentes que participam de diversos processos fisiológicos, incluindo proteção do espermatozóide, modulação da motilidade e capacitação espermática, modificação da membrana do espermatozóide e interação entre gametas. O conhecimento dessas proteínas contribuirá para uma melhor compreensão dos mecanismos de regulação do plasma seminal sobre a função espermática em ovinos
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