A síndrome dos ovários policísticos (SOP) é um distúrbio endócrino caracterizado por anovulação crônica e hiperandrogenismo. Modelos animais em ratas são usados para estudar processos complexos da SOP. As procineticinas (PROKs) são proteínas relacionadas a apoptose, proliferação vascular e regulação do sistema reprodutor. No entanto, seu padrão de expressão e função não são bem conhecidos no ovário com SOP. Este estudo propõe-se a identificar e localizar as PROKs e seus receptores nos ovários de ratas com SOP experimental. Foram utilizadas 33 ratas Wistar divididas em 3 grupos que receberam, entre o 1º e o 3º dia de vida, uma única injeção por via subcutânea de: propionato de testosterona (1,25 mg/0,1 mL, GT, n=12); benzoato de estradiol (0,5 mg/0,1 mL, GE, n=11) e óleo de oliva (0,1 mL, GC, n=10). Aos 90-95 dias de idade, os animais foram eutanaziados e os ovários removidos para avaliação da expressão gênica e proteica das procineticinas 1 e 2 e seus receptores por qRT-PCR e imunoistoquímica. A expressão dos genes Prok1 e Prok2 nos ovários de ratas do GT foi maior quando comparado ao GC (p=0,0157 e p=0,0354). Houve maior expressão da proteína PROK2 em células da teca interna (p=0,0049) e nas células intersticiais (p=0,0068) de folículos antrais nos ovários de ratas do GT em relação ao GC. PROK2 foi mais expressa em células da granulosa dos folículos pré-antrais comparado aos folículos antrais no GT (p=0,0098). Concluímos que as procineticinas estão expressas no ovário do modelo estudado em diferentes padrões, e que a PROK2 parece exibir maior expressão no grupo testosterona. Por apresentar papéis relevantes no controle do eixo hipotálamo-hipófise-gonadal, acreditamos que esses resultados podem abrir uma linha de investigação sobre o papel dessa proteína na fisiopatologia da síndrome / Polycystic ovary syndrome (PCOS) is an endocrine disorder characterized by chronic anovulation and hyperandrogenism. Animal models have been used to study PCOS pathophysiology. Prokineticins (PROKs) are proteins with functions related to apoptosis, vascular proliferation and reproductive physiology. However, their expression patterns and functions are unknown in the PCOS ovary. The aim of this study is to identify and localize the PROKs and their receptors in PCOS rat models induced by testosterone or estradiol. Thirty-three female Wistar rats aged between 1-3 days were sorted into three groups according to the compounds injected subcutaneously: Testosterone propionate (1.25 mg / 0.1 mL, TG, n = 12); Estradiol benzoate (0.5 mg / 0.1 mL, EG, n = 11) and olive oil (0.1 mL, CG, n = 10). At 90-95 days of age, the animals were euthanized and the ovaries removed for evaluation of prokineticins 1 and 2 and their receptors by qRT-PCR and immunohistochemistry. The expression of the genes Prok1 and Prok2 in the ovaries was higher in TG compared to CG (p=0.0157 and p=0.0345). There was higher expression of PROK2 in theca interna (p=0.0049) and interstitial cells (p=0.0068) of antral follicles in the ovaries of TG vs CG; PROK2 was higher expressed in the granulosa cells of the preantral follicles compared to the antral follicles in the TG (p=0.0098). We conclude that prokineticins are expressed in the ovary of the animal model studied and they present different patterns, PROK2 seems to exhibit higher expression in the testosterone group. Due important roles in the control of the hypothalamicpituitary- gonadal axis, we believe that these results may open a line of investigation about the role of this protein in the pathophysiology of the syndrome

A qualidade do ar nas áreas urbanas tende a apresentar concentrações indesejáveis de contaminantes provenientes da queima de biomassa, não havendo um sistema abrangente de monitoramento. Achados in vivo mostram que o diesel pode conter compostos que modulam a atividade do estrógeno. Estudos in vitro apoiam essa ideia, como o estudo realizado com cloreto de metileno extraído do diesel que comprova atuar como ativadores de ligação para os receptores de estrógeno. Esse trabalho tem por objetivo avaliar o efeito da exposição à metais presentes nas partículas de exaustao de diesel (PED) na morfologia do ovaário através da histologia, e análise do padrão de expressão dos receptores de estrógeno (ERs) no ovário de zebrafish (Danio rerio) usando a metodologia de PCR em tempo real. Fêmeas adultas de zebrafish foram expostas à concentrações ambientais de combinado de metais (Ni = 0,181 mg/L ; Fe = 0,07455 mg/L ; Pb= 0,05 mg/ L; Cd = 0,029 mg/ L; Cr = 0,161 mg/L ; Cu = 0,017 mg / L) por 3 dias. Em relação a contagem de ovócitos atrésicos a média observada nos animais expostos foi de 1,5 ± 0,4 e nos controles foi de 1,34 ± 0,3. Contudo, não houve diferença significativa entre os grupos (p > 0,05). Na expressão dos três genes receptores de estrógeno (esr1, esr2a, esr2b) as médias foram: 0,68; 1,32; 0,54; respectivamente. No entanto, não houve alteração significativa na expressão desses genes em relação aos controles. Os elementos traços presentes nas PED em concentração ambiental e em exposição aguda na fase adulta não alteram a expressão dos ERs o também não afetam o número de folículos atrésicos nos ovários / The air quality in urban areas tends to have undesirable concentrations of contaminants from biomass combustion, without a monitoring system. In vivo findings show that diesel may contain compounds that modulate the activity of estrogen. In vitro studies support this idea, as the study with methylene chloride extract from diesel proves that this substance act as activators binding to the estrogen receptors (ERs). This study aims to evaluate the effect of exposure to metals present in diesel exhaust particles (PED) in ovary morphology through histology, and analysis of expression pattern of ERs in ovary of zebrafish (Danio rerio) using real time PCR metodology. Adult females were exposed to ambient concentrations of combined metals (Ni = 0.181 mg / L, Fe = 0.07455 mg / L Pb = 0,05 mg / L; Cd = 0.029 mg / L; = 0.161 mg Cr / L, Cu = 0.017 mg / L) for 3 days. The mean of atretic oocytes counting observed in animals exposed was 1.5 ± 0.4 and the controls was 1.34 ± 0.3. However, no significant difference between groups was observed (p> 0.05). The expression of the three genes estrogen receptor (esr1, esr2a, esr2b) mean were: 0.68, 1.32, 0.54, respectively. However, no significant change in the expression of these genes in relation to controls was observed. In conclusion, the trace elements present in the PED concentration in environmental and acute exposure in adulthood not alter the expression of ERs and does not affect the number of atretic follicles in the ovary

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O conhecimento da morfologia e ultraestrutura dos helmintos permite a correta classificação destes organismos, bem como fornece subsídios que poderão ser utilizados para diagnóstico e controle. A microscopia laser confocal é uma ferramenta para estudar a organização estrutural de várias espécies de helmintos, possibilitando acesso a detalhes morfológicos não evidenciados pela microscopia óptica. Echinostoma paraensei é um trematódeo, digenético, hermafrodita parasito de numerosos hospedeiros vertebrados. Neste trabalho foi investigado o desenvolvimento dos órgãos reprodutivo e a morfometria de E. paraensei, desde a fase jovem até a adulta, como contribuição ao conhecimento do desenvolvimento reprodutivo desta espécie. Os trematódeos foram recuperados aos 3, 4, 5, 6, 7, 10, 14 e 21 dias posterior à infecção (dpi) experimental em hamsters. Estes foram corados em carmim clorídrico, desidratados em série alcoólica e montados em lâmina permanente em bálsamo do Canadá, fotografados e medidos usando microscopia de luz de campo claro (MCC) e microscopia de varredura laser confocal (MVLC). Entre 3 e 4 dpi, os primórdios genitais estavam presentes e nenhuma organização do sistema reprodutivo foi visualizada por MCC e MVLC. Os primórdio do ovário, dos testículos e da bolsa do cirro foram visualizados por MCC aos 5 e 6 dpi, no entanto, MVLC dos helmintos aos 5dpi mostra que estes primórdios, o ootipo e o útero estavam presentes, como estruturas individualizadas. A bolsa do cirro apresenta metratermo e o ovário com primórdio do oótipo adjacente aos 7dpi por MVLC. A vesícula seminal, receptáculo seminal, células diferenciadas nos testículos, ducto e reservatório vitelínico e oviducto foram visualizados após 10 dias, enquanto os espermatozóides na vesícula seminal, ovos e oócitos, células vitelínicas, poro e canal de Laurer aos 14 dias. A morfometria evidencia um acelerado crescimento dos órgãos reprodutores a partir do 7 dia. Os testículos apresentam aumento significativo no comprimento do 7 ao 21 dia e o ovário durante o período de 7 à 10 dpi. Aos 21 dpi, todos os helmintos apresentaram glândulas vitelínicas, útero contendo ovos e espermatozóides no oviducto enquanto outros ovos estão sendo formados. As mudanças morfológicas acentuadas durante a gametogênese consistem no aumento do comprimento do helminto, maturação das gônadas, desenvolvimento e maturação das glândulas vitelínicas. O desenvolvimento do helminto como um todo está relacionado à maturação dos órgãos reprodutivo masculino e feminino indicando o investimento deste trematódeo em garantir a produção e eliminação dos ovos ao meio exterior. / The knowledge about morphology and ultra structure of helminthes are great importance in correct classification these organisms. The Scanning Laser Microscopy (LSM) is an important tool to study the structural organization of several helminthes species. Echinostoma paraensei is a trematode, digenetic, hermaphroditic parasite of several hosts. In this study, the development of reproductive organs and the morphometry of E. paraensei from young stage to adult worm were investigated, to contribute knowledge of the reproductive development of this specie. The trematodes were recovered on 3, 4, 5, 6, 7, 10, 14 and 21 days post infection (dpi) from experimental hamsters. It were dehydrated in alcohol series, stained with hydrochloric carmine, mounted on permanent slide using Canada balsam, photographed and measured using light microscopy (LM) and Scanning Laser Microscopy (LSM). Between 3 and 4 dpi the genital anlage were present and were not observed reproductive system organization by either LM and LSM. The anlage of ovary, testes and Cirruss sac were seen at 5 and 6dpi by LM, however LSM from 5dpi image shows theses anlage, ootype and uterus are present as individualized structure. Cirrus sac showed metraterm and ootype adjacent to primordial ovary were seen at 7 dpi by LSM. The seminal vesicle, seminal receptacle, differentiated cells in the testes, viteline ducts and oviduct were visualized after 10 days infection, while sperms in seminal vesicle, eggs, oocyte, Laurer canal and pore from 14 dpi by LSM. The morphometry shows a rapid growth of reproductive organs from 7th day. The testes have significantly increased length from 7 until 21dpi and ovary from 7 until 10dpi. All helminthes showed vitellines glands, uterus contained eggs and sperm in oviduct while another eggs were forming at 21 dpi. The marked morphologic changes during gametogenesis are increase of body length of helminthes, gonad maturation and development and maturation of vitelline glands. The development of helminthes as a whole is related to maturation of female and male organs of reproductive system showing the investment this trematode taken to ensuring the production, maintenance and delivery the eggs to external environment.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2018-09-24T15:13:44Z No. of bitstreams: 1 2017 - Enely Maris da Silveira Firmiano.pdf: 12307016 bytes, checksum: d58bee7e5ddc0113fa072ab4e9aabc29 (MD5) / Made available in DSpace on 2018-09-24T15:13:44Z (GMT). No. of bitstreams: 1 2017 - Enely Maris da Silveira Firmiano.pdf: 12307016 bytes, checksum: d58bee7e5ddc0113fa072ab4e9aabc29 (MD5) Previous issue date: 2017-08-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Cap. I The aim of the current study is to investigate the histological and histochemical description, as well as the presence and immunohistochemical distribution of cytoskeleton proteins such as alpha smooth muscle actin, desmin and vimentin in the ovaries, testes and epididymis of Enyalius bilineatus, Enyalius perditus and Urostrophus vautieri. These species are representatives of family Leiosauridae, in order to help developing a database on the reproductive biology of these lizards, which could be compared to that of other reptile species and vertebrates. The herein studied specimens are deposited in the Herpetological Collection of Federal University of Juiz de Fora. The animals? ovaries, epididymis and testes were removed and sent to the Histology and Embryology Laboratory (UFRRJ), where they were fixed, processed and then subjected to histological, histochemical and immunohistochemical techniques. The ovaries of these leiosauridae are functionally-active paired organs filled with ovarian follicles at different follicular development stages. These organs are covered by a simple cubic epithelium, which presents an underlying connective tissue layer called tunica albuginea. The testes are covered by the tunica albuginea and hold septa forming lobes where the seminiferous tubules are located in. The epididymis of these lizards are convolute ducts internally coated by an epithelium, whose shape ranges from simple cylindrical to simple cubic with stereocilia. The immunohistochemical analysis applied to the ovaries of the herein studied lizards showed moderate to strong reaction to the alpha smooth muscle actin in the teak layer and in endothelial cells of blood vessels. With respect to the species? testes, this cytoskeleton protein showed strong reaction in the tunica albuginea, interstitial tissue, and in endothelial cells of blood vessels. As for the epididymis, the alpha-actin showed immunohistochemical reaction in the capsule and in the interstitial tissue of the lizards. The antibody ?desmin? showed strong immunostaining in the albuginea and ovarian stroma of E. perditus, only; however, the endothelial cells in the blood vessels of the three herein analyzed species showed positive reaction to this intermediate filament. Desmin showed immunoreactivity in the testicular albuginea and in the endothelial cells of the blood vessels in the herein analyzed leiosauridae testes; however, only E. bilineatus showed immunostaining in the interstitial region. Desmin showed positive immunoreaction in the epididymis of E. perditus and E. bilineatus, only. Vimentin did not show immunostaining in the ovaries, testes and epididymis of the three analyzed lizards. The consistent results found in the current study provide additional data on the reproductive biology of the investigated species. These data may be used for phylogenetic and behavioral comparisons between the herein studied species and other species belonging to family Leiosauridae or other reptiles Cap. II The aim of the current study is to investigate the histological description, as well as the presence and immunohistochemical distribution of alpha smooth muscle actin, desmin and vimentin in the testes and epididymis of Enyalius bilineatus, Enyalius perditus and Urostrophus vautieri, which are representatives of family Leiosauridae, in order to help developing a database on the reproductive biology of these lizards, which could be compared to that of other reptile species and vertebrates. The herein studied specimens are deposited in the Herpetological Collection of Federal University of Juiz de Fora. The animals? epididymis and testes were removed and sent to the Histology and Embryology Laboratory (UFRRJ), where they were fixed, processed and then subjected to histological and immunohistochemical techniques. The shape of the testes varies reasonably among the studied leiosauridae. These organs are covered by the tunica albuginea and hold septa forming lobes, where the seminiferous tubules are located in. The epididymis of these lizards are convolute ducts internally coated by an epithelium, whose shape ranges from simple cylindrical to simple cubic with stereocilia. The immunohistochemical analysis applied to the testes of the herein studied lizards showed strong positive reaction to the alpha smooth muscle actin in the tunica albuginea, interstitial tissue, and in endothelial cells of blood vessels. Desmin showed strong immunoreactivity in the testicular albuginea of E. bilineatus and moderate immunoreactivity in E. perditus and U. vautieri; however, only E. bilineatus showed immunostaining in the interstitial tissue region. The endothelial cells of blood vessels showed positive immunoreaction to desmin in the three herein analyzed species. The epididymis showed moderate immunohistochemical reaction to alpha smooth muscle actin in the capsule and in the interstitial tissue, whereas desmin showed positive immunoreaction in the epididymis of E. perditus and E. bilineatus, only. Vimentin did not show immunostaining in the testes and epididymis of the three analyzed lizards. The consistent results found in the current study provide additional data on the reproductive biology of the investigated species. These data may be used for phylogenetic and behavioral comparisons between the herein studied species and other species belonging to family Leiosauridae or other reptiles. / Cap. I Este trabalho investiga, al?m da descri??o histol?gica e histoqu?mica, a presen?a e distribui??o imuno-histoqu?mica de alfa-actina de m?sculo liso, desmina e vimentina nos ov?rios, test?culos e epid?dimos de Enyalius bilineatus, Enyalius perditus e Urostrophus vautieri, representantes da fam?lia Leiosauridae, para ajudar a formar uma base de dados sobre a biologia reprodutiva destes lagartos que possam ser comparados com outras esp?cies de r?pteis, bem como com outros vertebrados. Os esp?cimes estudados encontram-se depositados na Cole??o Herpetol?gica da UFJF. Os ov?rios, epid?dimos e test?culos dos animais foram removidos e encaminhados ao Laborat?rio de Histologia e Embriologia (UFRRJ), onde foram fixados, processados e ent?o submetidos ?s t?cnicas histol?gicas, histoqu?micas e imuno-histoqu?micas. Os ov?rios destes leiosaur?deos s?o ?rg?os pareados, repletos de fol?culos ovarianos, em distintos est?gios de desenvolvimento folicular. Estes s?o revestidos por epit?lio c?bico simples, com uma camada de tecido conjuntivo subjacente, a t?nica albug?nea. Os test?culos encontram-se envolvidos pela t?nica albug?nea e em seu interior cont?m septos que formam l?bulos, nos quais se localizam os t?bulos semin?feros. Os epid?dimos destes lagartos s?o ductos convolutos, revestidos internamente por um epit?lio que varia de cil?ndrico simples a c?bico simples com estereoc?lios. A imuno-histoqu?mica, nos ov?rios dos lagartos estudados, mostra uma rea??o moderada a acentuada para alfa-actina de m?sculo liso na camada da teca e em c?lulas endoteliais dos vasos sangu?neos, enquanto nos test?culos destas esp?cies nota-se uma acentuada rea??o para esta prote?na do citoesqueleto, na t?nica albug?nea, no tecido intersticial e nas c?lulas endoteliais dos vasos sangu?neos. Nos epid?dimos observa-se imunomarca??o para alfa-actina na c?psula e no tecido intersticial destes lagartos. Quanto ao anticorpo desmina, apenas em E. perditus, foi poss?vel imunorrea??o na albug?nea e no estroma ovariano, no entanto, nas tr?s esp?cies analisadas, as c?lulas endoteliais dos vasos sangu?neos demonstraram rea??o positiva a este filamento intermedi?rio. Nos test?culos dos leiosaur?deos estudados, a desmina foi imunolocalizada na albug?nea testicular e nas c?lulas endoteliais dos vasos sangu?neos, entretanto, apenas em E. bilineatus ocorreu imunomarca??o, na regi?o do interst?cio. Nos epid?dimos, a desmina demonstrou imunorrea??o positiva apenas em E. perditus e E. bilineatus. N?o houve rea??o imuno-histoqu?mica para vimentina nos ov?rios, test?culos e epid?dimos dos tr?s lagartos analisados. Este estudo obteve resultados consistentes, fornecendo assim mais dados sobre a biologia reprodutiva das esp?cies aqui investigadas, que poder?o ser utilizados para compara??es filogen?ticas e comportamentais com outras esp?cies da fam?lia Leiosauridae e outros r?pteis. Cap. II Este trabalho investiga, al?m da descri??o histol?gica, a presen?a e distribui??o imuno-histoqu?mica de alfa-actina de m?sculo liso, desmina e vimentina nos test?culos e epid?dimos de Enyalius bilineatus, Enyalius perditus e Urostrophus vautieri, representantes da fam?lia Leiosauridae, para ajudar a formar uma base de dados sobre a biologia reprodutiva destes lagartos que possam ser comparados com outras esp?cies de r?pteis, bem como com outros vertebrados. Os esp?cimes estudados encontram-se depositados na Cole??o Herpetol?gica da Universidade Federal de Juiz de Fora. Os epid?dimos e test?culos dos animais foram removidos e encaminhados ao Laborat?rio de Histologia e Embriologia (UFRRJ), onde foram fixados, processados e ent?o submetidos ?s t?cnicas histol?gicas e imuno-histoqu?micas. O formato dos test?culos varia razoavelmente entre os leiosaur?deos estudados. Estes ?rg?os encontram-se envoltos pela t?nica albug?nea e no seu interior cont?m septos que formam l?bulos incompletos, nos quais se localizam os t?bulos semin?feros. Os epid?dimos destes lagartos s?o ductos convolutos. Estes ductos s?o revestidos internamente por um epit?lio que varia de cil?ndrico simples a c?bico simples com estereoc?lios. A imuno-histoqu?mica revela uma acentuada rea??o positiva para alfa-actina de m?sculo liso nos test?culos dos leiosaur?deos analisados, na t?nica albug?nea e no tecido intersticial, assim como, nas c?lulas endoteliais dos vasos sangu?neos. A desmina apresentou acentuada imunorreatividade na albug?nea testicular em E. bilineatus e moderada em E. perditus e U. vautieri, entretanto, na regi?o do tecido intersticial, apenas em E. bilineatus ocorreu imunomarca??o. Nas tr?s esp?cies analisadas, as c?lulas endoteliais dos vasos sangu?neos demostraram imunorrea??o positiva para desmina. Nos epid?dimos observou-se rea??o imuno-histoqu?mica moderada para alfa-actina de m?sculo liso na c?psula e no tecido intersticial, enquanto a desmina demonstrou imunorrea??o positiva apenas em E. perditus e E. bilineatus. N?o houve imunomarca??o para vimentina nos test?culos e epid?dimos dos tr?s lagartos analisados. Este estudo obteve resultados consistentes, fornecendo assim mais dados sobre a biologia reprodutiva das esp?cies aqui investigadas, que poder?o ser utilizados para compara??es filogen?ticas e comportamentais com outras esp?cies da fam?lia Leiosauridae e outros r?pteis.

<p>Endocrine&ndash / disrupting compounds (EDCs) are exogenous compounds/chemicals which interfere with, or have adverse effects on the production, distribution and function of natural hormones, thereby affecting normal endocrine activity, health and quality of life of both humans and wildlife. The reproductive system is highly susceptible to EDCs due to it being controlled by an array of hormonal signals. The effects of EDCs on the male reproductive system include infertility, decreased sperm count, function and morphology, abnormal development of secondary sex characteristics, reproductive function and sexual behaviour as well as decreased libido. There are various sources by which EDCs enter the environment which include effluents from several industries (mining, agriculture, smelting, hazardous waste sites, manufacturing industries, etc.), sewage treatment effluents, urban and agricultural runoff and effluents which include natural and pharmaceutical chemicals excreted in the urine of humans and domestic livestock, pesticides, polychlorinated biphenyls, dioxins, plasticizers, surfactants, etc. Humans and animals can also be affected by EDCs by consuming food containing endocrine active substances. The growing concern regarding adverse effects due to EDC exposure of humans and wildlife, as well as the increased incidence of EDC contamination has prompted extensive research into the development and validation of screening tests to detect and monitor known EDCs and new substances with endocrine-disrupting capability. These screening tests involve assessing the effect of known and potential EDCs on reproductive function and development as well as&nbsp / hormone production. To assess the effect of EDCs on the reproductive system different methods are employed which include in vitro, in vivo and ex vivo methods. In vitro methods have been suggested as a suitable screening tool for EDC monitoring due to low costs, reduced animal usage, the use of standard and basic equipment as well as the ability to screen a large number of samples with multiple endpoints. Of the available in vitro methods, the minced testes method has been suggested as the most suitable method for screening EDCs and for this reason has been employed in this study. The aim of this study was thus to employ a minced testes method to screen samples for male reproductive toxicity using cell viability and hormone production (testosterone and estradiol) as endpoints.The first objective of this study was to optimize an in vitro testicular cell culture assay by determining both optimal luteinizing hormone (LH)&nbsp / concentration and incubation time needed for testosterone production. Testicular cell cultures were prepared and cells were treated with varying concentrations of LH (10, 1, 0.1, 0.01 and 0 mu/ml) and incubated for 4 hours and 20 hours. Testosterone production was evaluated for each incubation period. Testosterone production was significantly increased for both incubation periods at all LH concentrations tested as compared to the control. For both incubation periods, there was no significant difference in testosterone production between the different LH concentrations tested. From the data obtained, the 4 hour incubation period as well as the LH concentration of 10 mu/ml were selected as optimal for the testicular cell culture assay. The second objective of this study was to determine the effect of Tulbaghia violacea Harv. on the male reproductive system. T. violacea is a plant species indigenous to southern Africa and is used locally as a herbal remedy/medicine to treat several ailments. Cells were treated with varying concentrations of the T. violacea ethanol extract (with/without LH-treatment) and incubated for 4 hours. Hormone production and cell viability were evaluated. The results obtained from this pilot in vitro study demonstrated that the ethanol extract of T.violacea has androgenic properties by significantly increasing LH-induced testosterone production in mouse testes with no significant change in cell viability. The third objective of this study was to assess the effect of Sutherlandia frutescens(L.) R.Br and Artemisia afra Jacq. Ex Willd. on the male reproductive system. S. frutescens and A. afra are also plant species indigenous to southern Africa and used locally as a herbal remedy/medicine to treat several ailments. Ethanol extracts of each plant was prepared and cells were treated with varying concentrations of each extract (0, 156.25, 312.5, 625, 1250,2500 and 5000 &mu / g/ml) with or without LH-treatment and incubated for 4 hours. Cytotoxicity by LDH measurement and hormone production (testosterone and estradiol) were endpoints that were evaluated. The results obtained showed that the ethanol extracts of both plants are not cytotoxic to testicular cells and that A. afra decreases testosterone production at high concentrations. The fourth and final objective of this study was to assess the acute effect of four heavy metals, namely manganese, copper, cadmium and magnesium on the male reproductive system. These heavy metals are used extensively in manufacturing and mining industries. Cells were treated with varying concentrations of each metal salt (200, 100, 50, 25, 12.5, and 6.25&nbsp / &mu / M) with or without LH-treatment and incubated for 4 hours. Endpoints evaluated included cell viability, testosterone and estradiol production. The results obtained showed that manganese, cadmium and copper are highly toxic to testicular cells in vitro and therefore may potentially cause reproductive toxicity.</p>

The effect of case based learning on 10th grade students&rsquo / academic achievement in the unit of human reproductive system and their perceived motivation (Intrinsic Goal Orientation, Extrinsic Goal Orientation and Task Value) was investigated in this study. The study was conducted during 2007-2008 spring semester in a private high school in Ankara, Turkey. A total of 80 (48 males and 32 females) tenth grade students from four biology classes of two teachers were involved in the study. One class of each individual teacher was assigned as control group and the other class of the same teacher was assigned as experimental group to sustain randomization. Two of the classes assigned as experimental group were instructed with case based learning, while two classes of the control group were received traditionally designed biology instruction. In the experimental group, two cases that are divided into several parts were assigned in an interrupted manner. Students learned the human reproductive system via cases in experimental group without any additional method. The cases include incomplete data and students were needed to search for and discuss to answer the questions posed in the cases and by themselves and to complete the data. When the students completed a part of the case, next part was distributed. While dealing with the dilemma presented in cases, students did independent study as well as group work. In the control group, instruction was based on teacher explanations and web based notes prepared by the biology department of the high school. Human Reproductive System Achievement Test, Motivated Strategies for Learning Questionnaire were administered as pre-test and post-test to the students in both groups to measure the students&rsquo / academic achievement in the unit of human reproductive system / and their perceived motivation / specifically, task value, intrinsic goal orientation, and extrinsic goal orientation. Multivariate Analysis of Variance (MANOVA) was used to investigate the effect of case based learning on the students&rsquo / academic achievement in the unit of human reproductive system and their perceived motivation. Results revealed that case based learning improved students&rsquo / academic achievement and task value.

<p>Endocrine&ndash / disrupting compounds (EDCs) are exogenous compounds/chemicals which interfere with, or have adverse effects on the production, distribution and function of natural hormones, thereby affecting normal endocrine activity, health and quality of life of both humans and wildlife. The reproductive system is highly susceptible to EDCs due to it being controlled by an array of hormonal signals. The effects of EDCs on the male reproductive system include infertility, decreased sperm count, function and morphology, abnormal development of secondary sex characteristics, reproductive function and sexual behaviour as well as decreased libido. There are various sources by which EDCs enter the environment which include effluents from several industries (mining, agriculture, smelting, hazardous waste sites, manufacturing industries, etc.), sewage treatment effluents, urban and agricultural runoff and effluents which include natural and pharmaceutical chemicals excreted in the urine of humans and domestic livestock, pesticides, polychlorinated biphenyls, dioxins, plasticizers, surfactants, etc. Humans and animals can also be affected by EDCs by consuming food containing endocrine active substances. The growing concern regarding adverse effects due to EDC exposure of humans and wildlife, as well as the increased incidence of EDC contamination has prompted extensive research into the development and validation of screening tests to detect and monitor known EDCs and new substances with endocrine-disrupting capability. These screening tests involve assessing the effect of known and potential EDCs on reproductive function and development as well as&nbsp / hormone production. To assess the effect of EDCs on the reproductive system different methods are employed which include in vitro, in vivo and ex vivo methods. In vitro methods have been suggested as a suitable screening tool for EDC monitoring due to low costs, reduced animal usage, the use of standard and basic equipment as well as the ability to screen a large number of samples with multiple endpoints. Of the available in vitro methods, the minced testes method has been suggested as the most suitable method for screening EDCs and for this reason has been employed in this study. The aim of this study was thus to employ a minced testes method to screen samples for male reproductive toxicity using cell viability and hormone production (testosterone and estradiol) as endpoints.The first objective of this study was to optimize an in vitro testicular cell culture assay by determining both optimal luteinizing hormone (LH)&nbsp / concentration and incubation time needed for testosterone production. Testicular cell cultures were prepared and cells were treated with varying concentrations of LH (10, 1, 0.1, 0.01 and 0 mu/ml) and incubated for 4 hours and 20 hours. Testosterone production was evaluated for each incubation period. Testosterone production was significantly increased for both incubation periods at all LH concentrations tested as compared to the control. For both incubation periods, there was no significant difference in testosterone production between the different LH concentrations tested. From the data obtained, the 4 hour incubation period as well as the LH concentration of 10 mu/ml were selected as optimal for the testicular cell culture assay. The second objective of this study was to determine the effect of Tulbaghia violacea Harv. on the male reproductive system. T. violacea is a plant species indigenous to southern Africa and is used locally as a herbal remedy/medicine to treat several ailments. Cells were treated with varying concentrations of the T. violacea ethanol extract (with/without LH-treatment) and incubated for 4 hours. Hormone production and cell viability were evaluated. The results obtained from this pilot in vitro study demonstrated that the ethanol extract of T.violacea has androgenic properties by significantly increasing LH-induced testosterone production in mouse testes with no significant change in cell viability. The third objective of this study was to assess the effect of Sutherlandia frutescens(L.) R.Br and Artemisia afra Jacq. Ex Willd. on the male reproductive system. S. frutescens and A. afra are also plant species indigenous to southern Africa and used locally as a herbal remedy/medicine to treat several ailments. Ethanol extracts of each plant was prepared and cells were treated with varying concentrations of each extract (0, 156.25, 312.5, 625, 1250,2500 and 5000 &mu / g/ml) with or without LH-treatment and incubated for 4 hours. Cytotoxicity by LDH measurement and hormone production (testosterone and estradiol) were endpoints that were evaluated. The results obtained showed that the ethanol extracts of both plants are not cytotoxic to testicular cells and that A. afra decreases testosterone production at high concentrations. The fourth and final objective of this study was to assess the acute effect of four heavy metals, namely manganese, copper, cadmium and magnesium on the male reproductive system. These heavy metals are used extensively in manufacturing and mining industries. Cells were treated with varying concentrations of each metal salt (200, 100, 50, 25, 12.5, and 6.25&nbsp / &mu / M) with or without LH-treatment and incubated for 4 hours. Endpoints evaluated included cell viability, testosterone and estradiol production. The results obtained showed that manganese, cadmium and copper are highly toxic to testicular cells in vitro and therefore may potentially cause reproductive toxicity.</p>

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-05-03T13:11:41Z No. of bitstreams: 1 AndreMV_TESE.pdf: 15675614 bytes, checksum: fe55971bec8a35c1310c8ede4fbbf2f5 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-05-04T16:04:13Z (GMT) No. of bitstreams: 1 AndreMV_TESE.pdf: 15675614 bytes, checksum: fe55971bec8a35c1310c8ede4fbbf2f5 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-05-04T16:04:48Z (GMT) No. of bitstreams: 1 AndreMV_TESE.pdf: 15675614 bytes, checksum: fe55971bec8a35c1310c8ede4fbbf2f5 (MD5) / Made available in DSpace on 2017-05-04T16:05:05Z (GMT). No. of bitstreams: 1 AndreMV_TESE.pdf: 15675614 bytes, checksum: fe55971bec8a35c1310c8ede4fbbf2f5 (MD5) Previous issue date: 2017-02-17 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The gestation period, the characteristics of the reproductive cycle, including the selection of the influence of the male effect, the description of the female reproductive system, placentation, embryonic and fetal development – added of the morphometric analysis - and the expression of sulfated glycosaminoglycans (GAGs) in the reproductive and placental organs were the ways to contribute with the conservation of the species. For this, 45 non-pregnant females were used in the following experiments: 15 females were distributed in three experimental groups, followed by the addition of one male in each group, so that after the copulation and fertilization the females were separeted in accordance with the compatible dates of begining (days 5, 10 and 15), middle (days 20, 25, 30 and 35) and end (days 40, 45, 50 and 55) of gestation. This condition was obtained to obtain fragments of gestational sacs, placentas and organs of the female reproductive system, which were fixed in 4% paraformaldehyde and submitted to light microscopy techniques. The samples fixed in glutaraldehyde were performed in scanning and transmission electron microscopy techniques. In addition, prozima was used for GAG extraction, followed by agarose gel electrophoresis and subsequent quantification by densitometry. The second experiment consisted of the use of five females, housed in a room that contained a male to monitor the duration of gestation. The third experiment aimed to characterize the estrous cycle in a group containing five females plus one male, which was found trapped in a cage and another five, in a box exempt from the latter. The fourth experiment was performed by means of the morphometric analysis of embryos with 20, 25, 30 and 35 days plus fetuses on days 40, 45, 50, 55, 60 and newborns. The results revealed the presence of an incomplete double uterus, formed by two horns, a septate body and a cervix. The organs of the reproductive system presented macroscopic and histological characteristics similar to those reported for other rodents. The duration of gestation showed an average of 59 ± 2,7487 days and the sexual cycle was classified as continuous polystyric, with a mean period of 14.8 ± 0.73 days for females submitted to the male effect and 14.6 ± 0.75 days for those of the other group. In addition, the male presence significantly influenced (D <0.05) the duration of the diestrus, making it longer. The yolk sac inversion occurred on the 14th day of gestation and the visceral endoderm was placed in apposition to the uterine tissues. The chorioallantoic placenta presented a discoidal shape and had well defined regions of spongiotrophoblast, labyrinth and subplacenta from the intermediate gestation. Embryonic development was similar to that observed in histricognatis rodents, being the transition to the fetal phase, occurring at 40 days of gestation. Morphometric data allowed the establishment of correlations with gestational age. Dermatan sulfate was the predominant GAG in samples of uterine tubes, vaginas and placentas, whereas heparam expressed more in the body of the uterus at the beginning of gestation. There were also high concentrations of GAGs in the pregnant uterus and still in the reproductive system in the estrogenic phase when compared to the diestrus. It is concluded that the reproductive events presented characteristics of the species correlated with others of the order Rodentia, fact that indicates a set of evolutionary strategies related to its maintenance / O período de gestação, características do ciclo reprodutivo, incluindo verificação da influência do efeito macho, descrição do sistema reprodutor feminino, placentação, desenvolvimento embrionário e fetal – acrescido de análises morfométricas - e expressão dos glicosaminoglicanos (GAGs) sulfatados nos órgãos reprodutivos e placentários foram caracterizados em fêmeas de preás como forma de contribuir para a conservação da espécie. Para tanto, utilizaram-se 45 fêmeas não gestantes nos seguintes experimentos: 15 fêmeas foram distribuídas em três grupos experimentais, seguido da adição de um macho em cada grupo de modo que após a cópula e fecundação, as fêmeas eram separadas segundo datas compatíveis com início (dias cinco, 10 e 15), meio (dias 20, 25, 30 e 35) e fim (dias 40, 45, 50 e 55) da gestação. Tal condição visou à obtenção de fragmentos dos sacos gestacionais, placentas e órgãos do sistema reprodutor feminino, os quais foram fixados em paraformaldeído 4% e submetidos às técnicas de microscopia de luz. As amostras fixadas em glutaraldeído foram destinadas à realização de técnicas de microscopia eletrônica de varredura e transmissão. Além disto, usou-se a prozima para extração dos GAGs, seguida da eletroforese em gel de agarose e posterior quantificação por densitometria. O segundo experimento consistiu na utilização de cinco fêmeas, alocadas em recinto que continha um macho para o acompanhamento da duração da gestação. O terceiro experimento objetivou a caracterização do ciclo estral em um grupo contendo cinco fêmeas mais um macho, o qual encontrou-se preso em gaiola e outras cinco, em box isento deste último. O quarto experimento foi realizado por meio da análise morfométrica de embriões com 20, 25, 30 e 35 dias além de fetos nos dias 40, 45, 50, 55, 60 e recém-nascidos. Os resultados revelaram a presença de um útero duplo incompleto, formado por dois cornos, um corpo septado e uma cérvice. Os órgãos do sistema reprodutor apresentaram características macroscópicas e histológicas semelhantes as relatadas para outros roedores. A duração da gestação demonstrou média de 59 ± 2,7487 dias e o ciclo sexual foi classificado como poliéstrico contínuo, com período médio de 14,8 ± 0,73 dias para as fêmeas submetidas ao efeito macho e 14,6 ± 0,75 dias para aquelas do outro grupo. Além disto, a presença do macho influenciou significativamente (P<0,05) a duração do diestro, tornando-o mais longo. A inversão do saco vitelino ocorreu no décimo quarto dia de gestação e o endoderma visceral situou-se em aposição aos tecidos uterinos. A placenta corioalantoide apresentou forma discoidal e possuiu regiões bem definidas de espongiotrofoblasto, labirinto e subplacenta a partir da gestação intermediária. O desenvolvimento embrionário foi semelhante ao observado em roedores histricognatis, sendo a transição para a fase fetal, ocorrida aos 40 dias de gestação. Os dados morfométricos possibilitaram o estabelecimento de correlações com a idade gestacional. O dermatam sulfato foi o GAG predominante em amostras de tubas uterinas, vaginas e placentas, ao passo que o heparam expressou-se mais no corpo do útero, no início da gestação. Houve ainda elevadas concentrações de GAGs no útero gestante e no sistema reprodutor na fase estrogênica quando comparada ao diestro. Conclui-se que os eventos reprodutivos apresentaram características da espécie correlacionáveis com outras da ordem Rodentia, fato este que indica um conjunto de estratégias evolutivas relacionadas com a sua manutenção / 2017-05-03

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O conhecimento da morfologia e ultraestrutura dos helmintos permite a correta classificação destes organismos, bem como fornece subsídios que poderão ser utilizados para diagnóstico e controle. A microscopia laser confocal é uma ferramenta para estudar a organização estrutural de várias espécies de helmintos, possibilitando acesso a detalhes morfológicos não evidenciados pela microscopia óptica. Echinostoma paraensei é um trematódeo, digenético, hermafrodita parasito de numerosos hospedeiros vertebrados. Neste trabalho foi investigado o desenvolvimento dos órgãos reprodutivo e a morfometria de E. paraensei, desde a fase jovem até a adulta, como contribuição ao conhecimento do desenvolvimento reprodutivo desta espécie. Os trematódeos foram recuperados aos 3, 4, 5, 6, 7, 10, 14 e 21 dias posterior à infecção (dpi) experimental em hamsters. Estes foram corados em carmim clorídrico, desidratados em série alcoólica e montados em lâmina permanente em bálsamo do Canadá, fotografados e medidos usando microscopia de luz de campo claro (MCC) e microscopia de varredura laser confocal (MVLC). Entre 3 e 4 dpi, os primórdios genitais estavam presentes e nenhuma organização do sistema reprodutivo foi visualizada por MCC e MVLC. Os primórdio do ovário, dos testículos e da bolsa do cirro foram visualizados por MCC aos 5 e 6 dpi, no entanto, MVLC dos helmintos aos 5dpi mostra que estes primórdios, o ootipo e o útero estavam presentes, como estruturas individualizadas. A bolsa do cirro apresenta metratermo e o ovário com primórdio do oótipo adjacente aos 7dpi por MVLC. A vesícula seminal, receptáculo seminal, células diferenciadas nos testículos, ducto e reservatório vitelínico e oviducto foram visualizados após 10 dias, enquanto os espermatozóides na vesícula seminal, ovos e oócitos, células vitelínicas, poro e canal de Laurer aos 14 dias. A morfometria evidencia um acelerado crescimento dos órgãos reprodutores a partir do 7 dia. Os testículos apresentam aumento significativo no comprimento do 7 ao 21 dia e o ovário durante o período de 7 à 10 dpi. Aos 21 dpi, todos os helmintos apresentaram glândulas vitelínicas, útero contendo ovos e espermatozóides no oviducto enquanto outros ovos estão sendo formados. As mudanças morfológicas acentuadas durante a gametogênese consistem no aumento do comprimento do helminto, maturação das gônadas, desenvolvimento e maturação das glândulas vitelínicas. O desenvolvimento do helminto como um todo está relacionado à maturação dos órgãos reprodutivo masculino e feminino indicando o investimento deste trematódeo em garantir a produção e eliminação dos ovos ao meio exterior. / The knowledge about morphology and ultra structure of helminthes are great importance in correct classification these organisms. The Scanning Laser Microscopy (LSM) is an important tool to study the structural organization of several helminthes species. Echinostoma paraensei is a trematode, digenetic, hermaphroditic parasite of several hosts. In this study, the development of reproductive organs and the morphometry of E. paraensei from young stage to adult worm were investigated, to contribute knowledge of the reproductive development of this specie. The trematodes were recovered on 3, 4, 5, 6, 7, 10, 14 and 21 days post infection (dpi) from experimental hamsters. It were dehydrated in alcohol series, stained with hydrochloric carmine, mounted on permanent slide using Canada balsam, photographed and measured using light microscopy (LM) and Scanning Laser Microscopy (LSM). Between 3 and 4 dpi the genital anlage were present and were not observed reproductive system organization by either LM and LSM. The anlage of ovary, testes and Cirruss sac were seen at 5 and 6dpi by LM, however LSM from 5dpi image shows theses anlage, ootype and uterus are present as individualized structure. Cirrus sac showed metraterm and ootype adjacent to primordial ovary were seen at 7 dpi by LSM. The seminal vesicle, seminal receptacle, differentiated cells in the testes, viteline ducts and oviduct were visualized after 10 days infection, while sperms in seminal vesicle, eggs, oocyte, Laurer canal and pore from 14 dpi by LSM. The morphometry shows a rapid growth of reproductive organs from 7th day. The testes have significantly increased length from 7 until 21dpi and ovary from 7 until 10dpi. All helminthes showed vitellines glands, uterus contained eggs and sperm in oviduct while another eggs were forming at 21 dpi. The marked morphologic changes during gametogenesis are increase of body length of helminthes, gonad maturation and development and maturation of vitelline glands. The development of helminthes as a whole is related to maturation of female and male organs of reproductive system showing the investment this trematode taken to ensuring the production, maintenance and delivery the eggs to external environment.

A qualidade do ar nas áreas urbanas tende a apresentar concentrações indesejáveis de contaminantes provenientes da queima de biomassa, não havendo um sistema abrangente de monitoramento. Achados in vivo mostram que o diesel pode conter compostos que modulam a atividade do estrógeno. Estudos in vitro apoiam essa ideia, como o estudo realizado com cloreto de metileno extraído do diesel que comprova atuar como ativadores de ligação para os receptores de estrógeno. Esse trabalho tem por objetivo avaliar o efeito da exposição à metais presentes nas partículas de exaustao de diesel (PED) na morfologia do ovaário através da histologia, e análise do padrão de expressão dos receptores de estrógeno (ERs) no ovário de zebrafish (Danio rerio) usando a metodologia de PCR em tempo real. Fêmeas adultas de zebrafish foram expostas à concentrações ambientais de combinado de metais (Ni = 0,181 mg/L ; Fe = 0,07455 mg/L ; Pb= 0,05 mg/ L; Cd = 0,029 mg/ L; Cr = 0,161 mg/L ; Cu = 0,017 mg / L) por 3 dias. Em relação a contagem de ovócitos atrésicos a média observada nos animais expostos foi de 1,5 ± 0,4 e nos controles foi de 1,34 ± 0,3. Contudo, não houve diferença significativa entre os grupos (p > 0,05). Na expressão dos três genes receptores de estrógeno (esr1, esr2a, esr2b) as médias foram: 0,68; 1,32; 0,54; respectivamente. No entanto, não houve alteração significativa na expressão desses genes em relação aos controles. Os elementos traços presentes nas PED em concentração ambiental e em exposição aguda na fase adulta não alteram a expressão dos ERs o também não afetam o número de folículos atrésicos nos ovários / The air quality in urban areas tends to have undesirable concentrations of contaminants from biomass combustion, without a monitoring system. In vivo findings show that diesel may contain compounds that modulate the activity of estrogen. In vitro studies support this idea, as the study with methylene chloride extract from diesel proves that this substance act as activators binding to the estrogen receptors (ERs). This study aims to evaluate the effect of exposure to metals present in diesel exhaust particles (PED) in ovary morphology through histology, and analysis of expression pattern of ERs in ovary of zebrafish (Danio rerio) using real time PCR metodology. Adult females were exposed to ambient concentrations of combined metals (Ni = 0.181 mg / L, Fe = 0.07455 mg / L Pb = 0,05 mg / L; Cd = 0.029 mg / L; = 0.161 mg Cr / L, Cu = 0.017 mg / L) for 3 days. The mean of atretic oocytes counting observed in animals exposed was 1.5 ± 0.4 and the controls was 1.34 ± 0.3. However, no significant difference between groups was observed (p> 0.05). The expression of the three genes estrogen receptor (esr1, esr2a, esr2b) mean were: 0.68, 1.32, 0.54, respectively. However, no significant change in the expression of these genes in relation to controls was observed. In conclusion, the trace elements present in the PED concentration in environmental and acute exposure in adulthood not alter the expression of ERs and does not affect the number of atretic follicles in the ovary