Thesis (M.A.)--University of Missouri-Columbia, 2004. / Typescript. Includes bibliographical references (leaves 51-67). Also available on the Internet.

Orientador: Alvimar José da Costa / Banca: Katia Denise Saraiva Bresciani / Banca: Anderson Barbosa Moura / Resumo: Oito reprodutores ovinos, isentos de Toxoplasma gondii e quaisquer outras doenças reprodutivas, foram distribuídos em três grupos para infecção com o respectivo protozoário: GI - três ovinos (2,0 x 105 oocistos da cepa P), GII - três ovinos (1,0 x 106 taquizoítos da cepa RH) e GIII - dois ovinos (controle). Parâmetros clínicos foram mensurados. Avaliações parasitêmicas e sorológicas (IFI) foram efetuadas. Qualidade espermática foi avaliada e a presença do parasito no sêmen foi investigada por meio das técnicas da bioprova e da PCR. Parasitismo tissular foi pesquisado pela bioprova, PCR e imunohistoquímica. Foi possível registrar alterações hipertermia e apatia nos ovinos infectados com oocistos de Toxoplasma gondii Parasitemia foi detectada em cinco ovinos. Todos os ovinos inoculados responderam ao estímulo antigênico, produzindo anticorpos contra T. gondii a partir do 5º dia pós-inoculação. Nos animais dos grupos I e II, os títulos observados foram de 1:4096 e 1:8192, respectivamente. Nos mesmos grupos, foram ainda, pela bioprova e PCR, constatada a presença do parasito no sêmen. Pela PCR detectou-se o DNA de T. gondii no sêmen dos animais 02 e 09 (do GI oocistos) e 07, 48 e 52 (do GII taquizoítos). Parasitismo tissular por T. gondii (bioprova e PCR) foi diagnosticada nos ovinos 09, 16 (oocistos) e 07 (taquizoítos). Vesícula seminal e próstata foram os tecidos de eleição da infecção por T. gondii (imunohistoquímica) no sistema reprodutor dos ovinos experimentalmente infectados. Em síntese, estes resultados sugerem a viabilidade da transmissão venérea deste coccídio. / Abstract: Eight sheep reproducers exempt of Toxoplasma gondii and any other reproductive illnesses, were distributed in three groups for infection with the respective protozoan: GI - three sheep (2.0 x 105 oocysts from P lineage), GII - three sheep (1.0 x 106 taquizoites from RH lineage) and GIII - two sheep (control). Clinical parameters were measured. Parasitemics and sorologicals evaluations (IFA) were effected. Spermetic quality was evaluated and the presence of the parasite in the semen was searched through techniques of bioassay and PCR. Tissue parasitism was researched through bioassay, PCR and immuneistochemistry. It was possible to registered hyperthermia and apathy alterations in sheep infected with oocysts from Toxoplasma gondii. Parasitemia was detected in five sheep. All inoculated sheep responded to antigenic stimulation, producing antibodies against T. gondii upon the 5th day post-inoculation. In animals from groups I and II, the titles observed were 1:4096 and 1:8192, respectively. The animals of group III (control), did not present antibodies against T. gondii during all experimental period. In the same groups, it still has been evidenced, through bioassay and PCR, the presence of the parasite in the semen. Through PCR, T. gondii DNA was detected in the semen of animals 02, 09 of GI (oocysts) and 07, 48 and 52 of GII (taquizoites). Tissue parasitism by T. gondii (biotest and PCR) was diagnosed in sheep 09, 16 (oocysts) and 07 (taquizoites). Seminal vesicle and prostate were the tissues elected for infection with T. gondii (immuneistochemistry) in the experimentally infected sheep reproductive system. In summery, these results suggests the viability of the venereal transmission for this coccidium. / Mestre

Made available in DSpace on 2014-06-11T19:27:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-01-31Bitstream added on 2014-06-13T18:55:50Z : No. of bitstreams: 1 lopes_wdz_me_jabo.pdf: 1609666 bytes, checksum: ef67fda0becfef41fe6aebcf7cfdad39 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Oito reprodutores ovinos, isentos de Toxoplasma gondii e quaisquer outras doenças reprodutivas, foram distribuídos em três grupos para infecção com o respectivo protozoário: GI - três ovinos (2,0 x 105 oocistos da cepa P), GII - três ovinos (1,0 x 106 taquizoítos da cepa RH) e GIII - dois ovinos (controle). Parâmetros clínicos foram mensurados. Avaliações parasitêmicas e sorológicas (IFI) foram efetuadas. Qualidade espermática foi avaliada e a presença do parasito no sêmen foi investigada por meio das técnicas da bioprova e da PCR. Parasitismo tissular foi pesquisado pela bioprova, PCR e imunohistoquímica. Foi possível registrar alterações hipertermia e apatia nos ovinos infectados com oocistos de Toxoplasma gondii Parasitemia foi detectada em cinco ovinos. Todos os ovinos inoculados responderam ao estímulo antigênico, produzindo anticorpos contra T. gondii a partir do 5º dia pós-inoculação. Nos animais dos grupos I e II, os títulos observados foram de 1:4096 e 1:8192, respectivamente. Nos mesmos grupos, foram ainda, pela bioprova e PCR, constatada a presença do parasito no sêmen. Pela PCR detectou-se o DNA de T. gondii no sêmen dos animais 02 e 09 (do GI oocistos) e 07, 48 e 52 (do GII taquizoítos). Parasitismo tissular por T. gondii (bioprova e PCR) foi diagnosticada nos ovinos 09, 16 (oocistos) e 07 (taquizoítos). Vesícula seminal e próstata foram os tecidos de eleição da infecção por T. gondii (imunohistoquímica) no sistema reprodutor dos ovinos experimentalmente infectados. Em síntese, estes resultados sugerem a viabilidade da transmissão venérea deste coccídio. / Eight sheep reproducers exempt of Toxoplasma gondii and any other reproductive illnesses, were distributed in three groups for infection with the respective protozoan: GI - three sheep (2.0 x 105 oocysts from P lineage), GII - three sheep (1.0 x 106 taquizoites from RH lineage) and GIII - two sheep (control). Clinical parameters were measured. Parasitemics and sorologicals evaluations (IFA) were effected. Spermetic quality was evaluated and the presence of the parasite in the semen was searched through techniques of bioassay and PCR. Tissue parasitism was researched through bioassay, PCR and immuneistochemistry. It was possible to registered hyperthermia and apathy alterations in sheep infected with oocysts from Toxoplasma gondii. Parasitemia was detected in five sheep. All inoculated sheep responded to antigenic stimulation, producing antibodies against T. gondii upon the 5th day post-inoculation. In animals from groups I and II, the titles observed were 1:4096 and 1:8192, respectively. The animals of group III (control), did not present antibodies against T. gondii during all experimental period. In the same groups, it still has been evidenced, through bioassay and PCR, the presence of the parasite in the semen. Through PCR, T. gondii DNA was detected in the semen of animals 02, 09 of GI (oocysts) and 07, 48 and 52 of GII (taquizoites). Tissue parasitism by T. gondii (biotest and PCR) was diagnosed in sheep 09, 16 (oocysts) and 07 (taquizoites). Seminal vesicle and prostate were the tissues elected for infection with T. gondii (immuneistochemistry) in the experimentally infected sheep reproductive system. In summery, these results suggests the viability of the venereal transmission for this coccidium.

Made available in DSpace on 2014-06-11T19:27:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-01-30Bitstream added on 2014-06-13T20:35:34Z : No. of bitstreams: 1 santana_lf_me_jabo.pdf: 793308 bytes, checksum: 144da7774a5d1aadcadce3f1534456c3 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Seis caprinos machos em idade reprodutiva foram selecionados e distribuídos em três grupos: GI dois caprinos mantidos como controle (placebo), GII dois caprinos inoculados com 1 x 106 taquizoítos de T. gondii (cepa RH) e GIII dois caprinos inoculados com 2 x 105 oocistos de T. gondii (cepa P). Periodicamente foram aferidos parâmetros clínicos, hematológicos e sorológicos, assim como avaliações parasitêmicas, além de exames andrológicos. A presença do parasito no sêmen e, também, nos tecidos do sistema reprodutor foi pesquisada por meio das técnicas de bioprova, PCR e imunohistoquímica. Surtos parasitêmicos foram detectados em todos os animais inoculados com taquizoítos ou com oocistos. A recíproca dos títulos sorológicos (RIFI) alcançou valores máximos de 4096 nos dois grupos de animais que receberam o Toxoplasma gondii. Pela técnica da bioprova foi possível revelar precocemente a presença do coccídio nas amostras seminais dos animais inoculados como taquizoítos (5°, 7°, 28°, 49°, 63° e 70° DPI) e um pouco mais tardiamente nas amostras seminais daqueles inoculados com oocistos (56° e 70° DPI). Pela PCR foi possível identificar, no sêmen, material genético de T. gondii, em cinco e em duas datas experimentais pós-inoculanção dos animais pertencentes aos grupos GII e GIII, respectivamente. Por esta mesma técnica foi possível, ainda, isolar material genético deste protozoário também em amostras teciduais (pool de próstata, testículo, vesícula seminal e epidídimo) dos caprinos 11 e 36 inoculados com taquizoítos e oocistos, respectivamente. Pela imunohistoquímica foi diagnosticado o T. gondii no epidídimo de todos os caprinos que receberam o protozoário. O isolamento de T. gondii, no sistema reprodutor de caprinos (sêmen e tecidos), sugere a possibilidade da transmissão sexual constituir uma importante via na disseminação desta zoonose tão difundida na caprinocultura mundial. / Six goat males in reproductive age had been selected and distributed in three groups: GI - two goat noninoculated (control), GII - two goat inoculated with 1 x 106 tachyzoites of T. gondii (RH strain) and GIII - two goat inoculated with 2 x 105 oocysts of T. gondii (P strain). Periodically had been measured clinical parameters, hematologics and sorologics evaluations, as well as parasitemics, beyond andrologics examinations. The presence of the parasite in the semen, and also in tissues of the reproductive system was searched by means of the techniques of bioassay, PCR and immunehistochemistry (only for tissues). Parasitemia was detected in the two animals inoculated with tachyzoites and in the inoculated others two with oocysts. The reciprocal one of the sorologicys titles (RIFI) reached maximal level of 4096 in the two groups of animals inoculated with the Toxoplasma gondii. Using biossay technique was possible precociously to disclose to the presence of the coccidy in the seminal samples of animals inoculated with tachyzoites (5°, 7°, 28°, 4 9°, 63° e 70° DPI) and a little more delayed in the seminal samples of it inoculated with oocysts (56° e 70° DPI). For the PCR it was possible to identify to the genetic material of the T. gondii, in the semen in five and two experimental dates after-inoculanção of the pertaining animals to groups GII and GIII respectively. For this same technique was possible still to isolate material genetic of this protozoario also in tissues samples (pool of prostate, testícule, vesicula seminal and epididim) of goat 11 and 36 inoculated with tachyzoites and oocysts, respectively. By the immunehistochemistry was diagnosised T. gondii in epididim of all goats ones that had received the protozoario. The isolation of T. gondii, in the reproductive system of goat (semen and tissues), suggests the possibility of the sexual transmission to constitute... (Complete abstract, click electronic access below)

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Modulation of the male reproductive system occurs as a result of exposure to endocrine disrupting compounds (EDCs) in different life stages. 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 behavior, as well as decreased libido. Phytochemicals are naturally occurring, biologically active chemical compounds in plants. They are divided into different groups. Isoflavonoids and lignans, are the two major groups of phytoestrogens. Phytoestrogens of teas, coffee and buchu have many beneficial effects on body systems such as antimutagenic, antidiabetic, anti-inflammatory, antibacterial and antiviral properties. They also elicit many adverse events, for example, heavy consumption of green and black tea may cause liver damage and added unwanted effects when combined with other herbal beverages. Chronic heavy consumption of coffee is positively correlated to acute myocardial infarction and can elevate serum cholesterol levels. Rooibos tea decreases steroidogenesis by steroid secreting cell lines.This study investigated the effects of these beverages on the male reproductive system, using a minced testes method for determination of cell viability and hormone (testosterone) production. The first objective of this study was to optimize protein supplement for in vitro testosterone production using human serum albumin (HSA) and foetal bovine serum (FBS). Testicular cultures were prepared and exposed overnight to different concentrations of both sera and then incubated for 4 hours with or without luteinizing hormone (LH). The results showed that addition of protein supplements (HSA or FBS) did not have a significant effect on testosterone production. The second objective of this study was to investigate the effects of green tea, black tea, rooibos tea, coffee and buchu on cell viability of testicular cultures. Cells were treated overnight with varying concentrations of the plant extracts followed by incubation with/without LH for 4 hours. The effects of the plant beverages on cellular protein production were determined by the Bradford assay. The results showed that treatment of cells with varying concentrations of the plant extracts (with/without LH-treatment) had no significant effect on total cellular protein. The third objective of this study was to investigate the effects of black, green and rooibos teas, coffee and buchu on testosterone production by testicular cultures. The results obtained from these experiments showed that rooibos tea and buchu did not affect testosterone production in the presence or absence of LH. The results also indicated that green tea, black tea and coffee inhibited testosterone production by mouse testis cultures in the presence of LH, but not in the absence of LH. Black tea was the most potent inhibitor of testosterone synthesis by mouse testis cultures (IC50= 48 μg/ml), followed by coffee (IC50= 64 μg/ml) and green tea (IC50= 173 μg/ml). Green tea, black tea and coffee inhibited LH-stimulated testosterone synthesis, suggesting that these beverages may impair testicular steroidogenesis in mice. Thus, in spite of their acclaimed beneficial effects, consumption of these beverages in high doses raises concerns for their inhibitory effects on male reproductive function. Further in vitro and in vivo studies are warranted to determine their exact mechanisms of action on the male reproductive system in general and testicular function in particular.

While many studies have shown harmful effects of space flight on many tissues and systems of the human body, few studies have been done on the effects of space flight on the reproductive system. While the microgravity conditions of space flight are common knowledge, there is another component of space flight, that being higher than ambient (on Earth) levels of radiation. The purpose of this study was to examine the effects of simulated space flight on follicular development in the ovaries of mice, and to determine which component of spaceflight, i.e. microgravity, radiation, or a combination of the two, might be responsible for any changes in this follicular development. To simulate the environment of space, mice were exposed to higher levels of radiation by the use of cobalt plates and to simulated microgravity using a technique known as hind limb unloading. Four groups of mice-were used in this study; a control or untreated group, a group exposed to higher levels of radiation, a group exposed to simulated microgravity, and a group treated in both high radiation and simulated microgravity. The mice were further subdivided within these groups based on the amount of time they were kept alive after treatment/exposure (one, four, and nine months). The ovarian tissues were then analyzed to see the effects of these simulated conditions on the development of follicles. In all three treatment groups, development of follicles was restricted compared to the control group. Follicles from the various treatment groups appeared to be in the early stages of their development. It should be noted that these are preliminary results as the study is still in progress. One of the overarching questions that has been put forth by NASA over the last few decades is, can an organism, in this case a mammalian organism, complete an entire life cycle in space? This study may help to answer some of that question. If any of the components of space flight proves to be harmful to the female reproductive tissues human colonization of space would be problematic. If the damage incurred during space flight is irreversible, colonization of other worlds would also be problematic.

The primary function of the testis is twofold: 1, it is responsible for production of testosterone and 2, it is responsible for spermatogenesis. Previous studies in alcohol fed mice have shown that chronic alcohol consumption causes reduced sperm counts and testicular lesions. CTRP3 is a novel adipokine which has been shown to promote follicular proliferation and reduce apoptosis in granulosa cells in the ovaries. Both folliculogenesis and spermatogenesis occur via the process of meiosis and therefore have some similarities. Since CTRP3 has been shown to be involved in folliculogenesis it would be reasonable to assume that it will play a role in spermatogenesis. CTRP3 has been shown to have protective properties in some organs in alcohol fed mice. This study was designed to determine if CTRP3 conveyed protective properties to the testicular tissue in chronic alcohol fed mice by comparing testicular morphology across 4 treatment groups: wild-type control mice, wild-type mice on a high alcohol diet, CTRP3 over expressing mice, and CTRP3 over expressing alcohol fed mice. To date this study indicates that alcohol did decrease germ cells due to apoptosis in the wild-type mice. Our study indicates that apoptosis of germ cells increased the intercellular space in seminiferous tubules and separated spermatogenic cells in the wild type mice. The CTRP3 mice do not show as aggressive results, indicating that CTRP3 may be playing a protective role. At this time, only a small number of tissues from the study have been analyzed so these results should be considered to be preliminary.

One of the major healthcare issues found almost worldwide, especially in the United States, is the obesity epidemic. Obesity is known to have deleterious effects on many body/organ systems. C1q TNF-related protein-3 (CTRP3) is effective at preventing high-fat diet-induced fatty liver. With these two factors taken into consideration, this study explores the possible effects of a high caloric diet on the muscle wall of the uterus, i.e., the myometrium, and how over-expression of CTRP3 may modify those effects. We hypothesize that consumption of excessive amounts of fat and sugar will have detrimental effects on the dual layers of the mouse myometrium. For this study, 17 mice were divided into 4 treatment groups: wild type/low fat diet, wild type/high fat diet, CTRP3-overexpressing/low fat diet, and CTRP3-overexpressing/high fat diet. The mice were placed on their respective diets at 7 weeks of age with a feeding duration of 12 weeks. At the conclusion of the feeding protocol, the female reproductive tissues were harvested and fixed in 4% paraformaldehyde and subsequently paraffin embedded. The uterine horns of each mouse were painstakingly paraffin embedded in a vertical position so that cross sections of the uterus could be obtained and measured. These 4µ sections were stained using standard H&E staining techniques and visualized under light microscopy. A randomization grid was utilized to determine measurement locations on the tissue. For each animal, 15 measurements were made of the outer longitudinal layer of the uterine horn, as well as 15 measurements of the inner circular layer, and 15 measurements of the thickness of the two layers combined. Two-way ANOVA was used to determine if any changes seen were statistically significant. At the time of the writing of this abstract, no appreciable differences have been found between the treatment groups, although there will be more data and final statistics completed before the presentation of our findings.

lin-28 was first characterized as a developmental timing regulator in Caenorhabditis elegans. Loss of lin-28 function (lin-28(lf)) mutants skip the hypodermal cell fates specific to the 2nd larval stage. Here, we studied two aspects of lin-28 which had not yet been investigated. First, we show that lin-28(lf) mutants exhibit reduced fertility associated with abnormal somatic gonadal morphology. In particular, the abnormal spermatheca-uterine valve morphology of lin-28(lf) hermaphrodites traps embryos in the spermatheca, which disrupts ovulation and causes embryonic lethality. The same genes downstream of lin-28 in the regulation of hypodermal developmental timing also act downstream of lin-28 in somatic gonadal morphogenesis and fertility. Importantly, we find that hypodermal expression, but not somatic gonadal expression, of lin-28 is sufficient for restoring normal somatic gonadal morphology in lin-28(lf) mutants. We propose that the abnormal somatic gonadal morphogenesis of lin-28(lf) hermaphrodites results from temporal discoordination between the accelerated hypodermal development and normally timed somatic gonadal development. Thus, our findings exemplify how a cell-intrinsic developmental timing program can also control proper development of other interacting tissues, cell non-autonomously. We also investigated the expression patterns and functions of two lin-28 isoforms in C. elegans. Our analysis of spatial expression patterns suggests that lin-28a and lin-28b are co-expressed in diverse tissues. Consistently, neither of isoform specific knock-out mutant, lin-28a(lf) or lin-28b(lf), exhibits defects in hypodermal development, somatic gonad, or fertility, indicating functional redundancy of two isoforms. Our study will contribute to further investigation of lin-28 isoforms by providing the mutants of each isoform as well as the primary analysis of their phenotypes.

Orientador: Irene Bastos Franceschini Vicentini / Banca: Elizabeth Romagosa / Banca: Maíra Aparecida Stefanini / Banca: Bruno Cezar Stefanini / Banca: Silvana Martinez Baraldi Artoni / Resumo: A carcinicultura de água doce tem sido reconhecida como uma forma para a produção de crustáceos com baixo impacto ambiental. O camarão Macrobrachium amazonicum apresenta grande potencial para a aqüicultura. Nos últimos anos, vários trabalhos sobre a biologia dessa espécie têm sido realizados com o intuito de desenvolver um pacote tecnológico sustentável para o cultivo de Macrobrachium amazonicum. Esse trabalho teve como objetivo descrever a estrutura microscópica do sistema reprodutor masculino (testículos, ductos deferentes e glândulas androgênicas) e do hepatopâncreas dos diferentes morfotipos de Macrobrachium amazonicum. Para tanto 40 machos adultos foram divididos em quatro grupos morfotípicos, sendo denominados de TC (Translucent Claw), CC (Cinnamon Claw), GC1 (Green Claw 1) e GC2 (Green Claw 2). Os animais foram pesados e foram realizadas medições do comprimento corpóreo total e do comprimento do segundo quelípodo direito. Após, os animais foram mortos e tiveram seus testículos, ductos deferentes, glândulas androgênicas e hepatopâncreas retirados e submetidos à rotina histológica de historesina. Foi observada a diferenciação de três morfotipos, segundo as análises realizadas, sendo eles, TC (Translucent Claw), CC (Cinnamon Claw) e GC(Green Claw). Entretanto estudos enfatizando o comportamento reprodutivo, assim como, a histoquímica e a ultraestrutura do sistema reprodutivo e do hepatopâncreas, durante a diferenciação morfotípica devem ser realizados para que se possa conhecer a dinâmica morfotípica dos machos adultos de Macrobrachium amazonicum. / Abstract: The culture of the freshwater prawns has been recognized as a way to produce crustaceans with a low enviromental impact. Macrobrachium amazonicum is the South American prawn with the greatest potencial for aquaculture. Since 2001 a multidisciplinary and multi-institutional research program has been developed technology for Macrobrachium amazonicum culture in Brazil. Thus, this study attemped to characterize the light microscope structures of the reproductive system (testes, vas deferens and androgenic gland) and the hepatopancreas of the differents morphotypes of the Macrobrachium amazonium. Specimens were collected and classified in four morphotypical groups: TC, CC, GC1 and GC2. The specimens were weight, and the total length and the cheliped length were measured. Then the testes, vas deferens, androgenic gland and hepatopancreas were fixed for histological routine of historesin. Three distinct morphological morphotypes were identified, TC (Translucent Claw), CC (Cinnamon Claw) and GC (Green Claw) morphotypes. Therefore more studies concerning to the reproductive behavior and the histochemical and ultrastrucutral studies need to be accomplished to elucitade the morphotypic differentiation of the male adults of Macrobrachium amazonicum. / Doutor