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1
Kilani, Suha School of Medicine UNSW. "The use of polarised light microscopy as a non-invasive tool for early assessment of human oocytes and embryos." Awarded by:University of New South Wales. School of Medicine, 2007. http://handle.unsw.edu.au/1959.4/35247.
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The overall aim was to evaluate a non-invasive technique for the assessment of oocytes and embryos using polarized light microscopy (PolScope-LC) with the goal of improving success rates in IVF. A literature review revealed little validation of the PolScope techniques in published work. A reproducible and accurate method for measuring the zona pellucida (ZP) thickness and density involving the PolScope computer software was validated by achieving low coefficient of variance and small inter/intra observer errors. Utilizing this method, 1477 oocytes from 211 stimulated cycles were analysed in this thesis. Results showed that increasing age has an adverse effect on the ZP thickness and density. Study of extended culturing of embryos showed that the ZP starts thinning as early as day 3 and embryos tend to have denser zonas over time. Standardisation of timing of PolScope observations in relation to the meiotic spindle was studied. Metaphase II oocytes were examined sequentially in culture from aspiration until microinjection using the PolScope The spindle is a highly dynamic structure that can appear and disappear over time in culture. A visible spindle was detected in 58% of the oocytes immediately after aspiration. This percentage increased until it stabilised at 39-40hrs post hCG and then declined significantly. Average spindle signal intensity increased over time reaching its peak at 39-40hrs post hCG, then declined significantly by 40.5hrs post hCG. The importance of spindle presence and morphology was investigated by following up embryos created after sperm injection at 39-40hrs post hCG. There was a significant relationship between normal meiotic spindle shape and density and embryo quality. A higher percentage of ???usable??? embryos, and all of pregnancies, arose from oocytes with a normal barrel shaped spindle. Finally, the impact of two issues related to spindle formation - the type of hCG used to trigger oocyte maturation and the site of microinjection during ICSI were assessed using the PolScope. The results showed a biological difference in spindle formation and embryo quality between rhCG and uhCG. In a separate randomised trial embryo quality was better when injecting the sperm in the vegetal pole away from the spindle during ICSI. The results from this thesis suggest that PolScope, if appropriately applied, may assist in improving IVF outcome.
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Pfender, Sybille Helen. "Studies of asymmetric oocyte division and new genes controlling oocyte maturation." Thesis, University of Cambridge, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.648232.
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James, Candace Renee Berry Wallace D. "Method for isolating immature chicken oocytes." Auburn, Ala., 2009. http://hdl.handle.net/10415/2025.
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Li, Tin-Chiu. "A study of human endometrial function in the peri-implantation period." Thesis, Click to view the E-thesis via HKUTO, 2002. http://sunzi.lib.hku.hk/hkuto/record/B31981823.
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Mullen, Steven Francis. "Advances in the fundamental cryobiology of mammalian oocytes." Diss., Columbia, Mo. : University of Missouri-Columbia, 2007. http://hdl.handle.net/10355/4804.
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Thesis (Ph. D.)--University of Missouri-Columbia, 2007.<br>"May 2007" The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. Includes bibliographical references.
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Lingenfelter, Brandon Michael. "The effect of follicular aging on gene expression in oocytes and granulosal cells." Morgantown, W. Va. : [West Virginia University Libraries], 2008. https://eidr.wvu.edu/etd/documentdata.eTD?documentid=5783.
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Thesis (Ph. D.)--West Virginia University, 2008.<br>Title from document title page. Document formatted into pages; contains xii, 169 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.
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Fazio, Cynthia Marie. "The influence of meiotic onset on and the role of apoptosis in oocyte death during the meiotic prophase /." Thesis, McGill University, 2005. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=97951.
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Loss of germ cells that entered meiosis at different developmental stages was compared. Mice were injected with BrdU at 13.3, 14.3 or 15.3 days post coitum (dpc) and sacrificed either 3 days after BrdU injection or 4 days post partum (dpp). BrdU-labeled germ cells were detected in ovarian sections through double immunofluorescent staining for BrdU and either GCNA-1 or MVH as a germ cell marker. The results show that the loss of germ cells that entered meiosis at 13.3 or 15.3 dpc was excessive compared to the loss of total germ cells. Such preferential elimination was not found for germ cells that entered meiosis at 14.3 dpc. We conclude that oocyte loss during meiotic prophase is influenced by the timing of meiotic onset.<br>The mechanism of germ cell loss during ovarian development was tested by the presence of markers for apoptosis. Mouse ovaries were isolated at 12.5 dpc, 18.5 dpc and 2 dpp and cultured with doxorubicin (DXR) to induce cell death. Ovarian histological sections were double immunofluorescent stained for GCNA-1 and cleaved caspase-3 or PARP-1. The results suggest that caspase-3 is not activated in germ cells throughout ovarian development whereas PARP-1 is activated in germ cells at 12.5 dpc and 2 dpp but not at 18.5 dpc. Thus, no evidence has yet been provided to support the hypothesis that oocyte death during the meiotic prophase is mediated by apooptosis.
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Nieuwburg, Ross Willem. "Analysis of the role of dynactin in the polarisation of the cytoskeleton of the Drosophila oocyte." Thesis, University of Cambridge, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.610258.
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Raposo, Alexandre Augusto da Silva Figueiredo. "Nuclear migration and the regulation of microtubules in the Drosophila oocyte." Thesis, University of Cambridge, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.611219.
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Antelman, Jennifer L. "Involvement of mitochondrial transcription factor A (TFAM) in porcine gametogenesis and preimplantation embryo development." Diss., Columbia, Mo. : University of Missouri-Columbia, 2007. http://hdl.handle.net/10355/5022.
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Thesis (M.S.)--University of Missouri-Columbia, 2007.<br>The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on April 3, 2009) Vita. Includes bibliographical references.
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Chen, Ying. "The role of steroids in the regulation of oocyte cyst breakdown and primordial follicle assembly in the neonatal mouse ovary." Related electronic resource: Current Research at SU : database of SU dissertations, recent titles available, full text:, 2008. http://wwwlib.umi.com/cr/syr/main.
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Gibbons, John R. "Ultrasound-guided transvaginal follicular aspiration to provide a source of bovine oocytes for gene microinjection." Thesis, This resource online, 1994. http://scholar.lib.vt.edu/theses/available/etd-12162009-020346/.
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Egonmwan, Rosemary Iriowen. "Reproductive biology and growth of the land snails Archachatina marginata ovum and Limicolaria flammea." Thesis, University of Oxford, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.443146.
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Ramsoondar, Jagdeece J. (Jagdeece Jagdeo). "The regulation of sperm-egg interaction in vitro by a porcine follicular fluid protein /." Thesis, McGill University, 1986. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=65510.
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Alton, Michelle. "Control of the oocyte population in mouse ovaries." Thesis, McGill University, 2005. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=81585.
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Oocyte loss and meiotic prophase progression was studied in XY sex-reversed and XO female mice, two mouse models that lack pairing between their sex chromosomes. An arrest at the pachytene stage of meiosis was not observed, nor was a significant loss of oocytes at this stage compared to normal XX control mice. Thus, it was concluded that a pairing checkpoint either does not exist in oocytes or is not as stringent as the one observed in males.<br>The effect of mutating the pro-apoptotic Bax molecule was studied at three distinct ages corresponding to the time when female germ cells are premeiotic, in meiotic prophase, and arrested in dictyotene. Although it appeared that more germ cells were retained in the Bax homozygous mutant compared to the wild-type and heterozygous mice at 18.5 dpc, by 24.5 dpc all of the mice possessed similar numbers of germ cells. These results indicate a role for Bax in germ cell death but also support the idea that an alternative pathway can compensate for the elimination of this molecule.
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趙志昂 and Chi-ngong Philip Chiu. "Investigation on the spermatozoa-zona binding inhibitory factors from human follicular fluid." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1999. http://hub.hku.hk/bib/B31222407.
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Chiu, Chi-ngong Philip, and 趙志昂. "An investigation into the biochemical and biological properties of zona-binding inhibitory factor 1 from human follicular fluid." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2003. http://hub.hku.hk/bib/B31245249.
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Fung, Chun-kit. "An investigation on the effects of glutamine in culture meida on the preimplantation mouse embryo /." Hong Kong : University of Hong Kong, 1999. http://sunzi.lib.hku.hk/hkuto/record.jsp?B21629821.
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Chiu, Chi-ngong Philip. "Investigation on the spermatozoa-zona binding inhibitory factors from human follicular fluid /." Hong Kong : University of Hong Kong, 1999. http://sunzi.lib.hku.hk/hkuto/record.jsp?B21415249.
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Ying, Ying. "Male accessory sex glands and oocyte activation at fertilization in the golden hamster /." Hong Kong : University of Hong Kong, 1998. http://sunzi.lib.hku.hk/hkuto/record.jsp?B20792712.
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Cebrián, Serrano Alberto. "Factors affecting the in vitro embryo production in cattle associated to ovum pick up sistem." Doctoral thesis, Universitat Politècnica de València, 2013. http://hdl.handle.net/10251/27646.
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La producción de embriones mediante la recuperación de ovocitos inmaduros por ovum pick up (OPU), y su posterior maduración, fecundación y cultivo en el laboratorio in vitro, presenta numerosos beneficios para optimizar el potencial reproductivo, tanto de hembras como de machos. Además, frente a la superovulación convencional mediante tratamiento hormonal y la recogida de embriones in vivo, la producción in vitro de embriones (PIVE) con ovocitos de OPU ofrece considerables ventajas. Sin embargo, actualmente la PIVE continua siendo ineficiente e incapaz de producir embriones de calidad similar a los in vivo, lo cual ha limitado una aplicación más amplia de esta tecnología. Así pues, el objetivo de esta tesis fue la optimización de la PIVE en ganado vacuno, condicionado por las peculiaridades y deficiencias de la PIVE cuando los ovocitos son recuperados por la técnica de OPU. Con este fin, cinco experimento se llevaron a cabo en esta tesis. En el primero de ellos se estudió el efecto del fluido oviductal bovino (FOb) sobre el desarrollo y la calidad embrionaria (Experimento 1). Las fases del proceso de PIVE en las cuales el cultivo de ovocitos/embriones, bien individualmente o bien en número reducido, pudiera perjudicar el posterior desarrollo hasta el estadio de blastocisto y/o a su calidad, se estudiaron en el Experimento 2. En el Experimento 3 se testó si el desarrollo y la calidad de embriones cultivados in vitro en número reducido podría ser mejorada con la adición conjunta de factor de crecimiento epidérmico, insulina, transferrina y selenio (FCE-ITS) o por el sistema de cultivo de embriones llamado well of well (WOW). Las propiedades protectoras de la melatonina frente a los daños causados por el estrés oxidativo, subsecuentes de las condiciones de PIVE o de un estrés térmico durante la maduración ovocitaria, fueron evaluadas en el Experimento 4. Por último, en el Experimento 5 usamos ovocitos recolectados por OPU para evaluar el efecto del semen sexado sobre<br>Cebrián Serrano, A. (2013). Factors affecting the in vitro embryo production in cattle associated to ovum pick up sistem [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/27646<br>Palancia
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Rothstein, Dianne M. Marcolongo Michele S. "Protein mediated attachment mechanisms associated with blastocyst implantation /." Philadelphia, Pa. : Drexel University, 2008. http://hdl.handle.net/1860/2814.
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Fung, Chun-kit, and 馮俊傑. "An investigation on the effects of glutamine in culture meida on the preimplantation mouse embryo." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1999. http://hub.hku.hk/bib/B31222560.
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Ying, Ying, and 應嬴. "Male accessory sex glands and oocyte activation at fertilization in the golden hamster." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1998. http://hub.hku.hk/bib/B31239663.
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Chung, Man-kin, and 鍾文健. "Biological characterization of cumulus glycodelin on humanspermatozoa-zona pellucida interaction." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B42182633.
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Cheung, Tsz-yan, and 張芷恩. "A study of endocrine disrupting chemicals (TCDD and Bisphenol A) on endometrial receptivity and implantation." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hdl.handle.net/10722/207984.
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The endocrine disrupting chemicals (EDCs) are exogenous compounds that mimic natural hormones and disrupt endocrine functions in humans and animals. Accumulating evidence suggests that EDCs may have detrimental impact on human reproduction including infertility, distortion of sex ratios and menstrual problems. TCDD, one of the most toxic man-made chemicals, was found to affect embryo maturity, implantation and reproduction. Bisphenol A (BPA), another EDC commonly used nowadays in plastic products, exhibits weak estrogenic activity in human bodies. However, TCDD and BPA have distinct chemical structures, chemical properties and receptor binding, and their mechanistic actions on human body remain largely unknown.
The present study is to delineate the effects of EDCs on embryo implantation and development under in vitro exposure. It is hypothesized that EDCs (TCDD and BPA) may modulate fertility of animals by affecting early pre-implantation embryo development and attachment onto uterus. The effect of EDCs on the expression of receptors (AhR, ERE, ERR,,and ERa) and downstream reporter genes (CYP1A1 and C3) were studied by real-time PCR and Western blotting. An in vitro spheroid (JEG-3)-endometrial cells (Ishikawa) co-culture assay was established to study the effect of EDCs on spheroid attachment. Since microRNA, Wnt-signaling and adhesion molecules play important roles in implantation process, the expression of selected miRNA, Wnt-signaling and adhesion molecules was studied after EDCs treatment. Specific activities of the EDCs receptors were confirmed by inhibitor treatment or siRNA knockdown studies. Furthermore, EDC-treated embryos were transferred to pseudo-pregnant surrogate mice to determine the effect of EDCs on implantation outcome on day 8.
It was found that Ishikawa and JEG-3 cells expressed AhR, ERIIand ERRE. TCDD treatment induced CYP1A1 expression in both cell lines (P<0.05). TCDD at 10nM significantly suppressed (P<0.005) spheroid attachment (74% compared to control 97%). When both cell lines were treated with AhR antagonists DMF (10 aaa/ANF (1 /////with TCDD (10nM), the suppressive effect of TCDD on spheroid attachment in co-culture assay was nullified, suggesting that TCDD acts through AhR receptor. In BPA exposure, BPA (0.001 to 10 tt) induced the expressions of ER))and C3ain Ishikawa but reduced the expressions of EReeand C3ain JEG-3 cells. BPA (10 iiiisuppressed spheroids attachment (74% vs Control 94%, P<0.005); while co-treatment with ER antagonists (ICI 182,780), ERaaantagonistaaMPP), ERMMantagonist (PTHPP) or ERE///siRNA, the suppressive effect of BPA (10sM) on spheroid attachment was nullified, suggesting that BPA acts through ER receptors. Moreover, TCDD and BPA suppressed the expressions of B-catenin and E-cadherin for cell-cell adhesion. Activation of Wnt-signaling pathway by Wnt3a conditioned medium or LiCl, rescued the low spheroid attachment rate induced by EDCs. Both EDCs reduced embryo implantation rate in pseudo-pregnant mice, suggesting the adverse effect of EDCs on embryo implantation and/or endometrial receptivity.
Taken together, TCDD and BPA affected the JEG-3 spheroid attachment onto the Ishikawa endometrial cells and mouse embryo implantation. These effects might be mediated through the action of receptors and Wnt/β-catenin signaling pathway.<br>published_or_final_version<br>Obstetrics and Gynaecology<br>Doctoral<br>Doctor of Philosophy
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Halley-Stott, Richard Paul. "Quantitative transcriptional reprogramming of somatic cell nuclei with Xenopus leavis oocytes." Thesis, University of Cambridge, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.610108.
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Swanson, Willie J. "The molecular evolution of abalone fertilization proteins /." Diss., Connect to a 24 p. preview or request complete full text in PDF format. Access restricted to UC campuses, 1998. http://wwwlib.umi.com/cr/ucsd/fullcit?p9907825.
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Chung, Man-kin. "Biological characterization of cumulus glycodelin on human spermatozoa-zona pellucida interaction." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B42182633.
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Wang, Lei. "Cloning and characterization of a novel oocyte-specific gene Fbos encoding an F-Box protein in rainbow trout (Oncorhynchus mykiss)." Morgantown, W. Va. : [West Virginia University Libraries], 2009. http://hdl.handle.net/10450/10786.
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Thesis (M.S.)--West Virginia University, 2009.<br>Title from document title page. Document formatted into pages; contains vii, 51 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 46-51).
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Black, David H. "Development of ovum pickup and in vitro embryo production to assess fertility responses for mineral intervention studies." Thesis, University of Nottingham, 2018. http://eprints.nottingham.ac.uk/52598/.
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As nutrition is of central importance to cattle fertility, this study sought to assess how veterinarians and nutritional advisers manage trace element imbalance in the UK; diagnosis and treatment. The study also sought to develop a robust system for oocyte recovery (ovum pick-up (OPU)) and in vitro embryo production (IVP) for commercial use, and to identify key factors influencing success, including oestrus synchrony and ovarian stimulation prior to OPU. The intention originally was to use OPU/IVP to investigate the impact of mineral imbalances on bovine oocyte quality, early embryo development and pregnancy establishment following embryo transfer (ET). In the first survey of its kind in the UK, the understanding and approach of advisers to mineral nutrition on farms was investigated. Of the 173 respondents, 78% were vets in practice. The overall importance of minerals was recorded by vets as low 33%, medium 37%, and high 30%, while non-vets scored importance as low 17%, medium 48%, and high 35%. There was little consensus amongst the advisers, or within the vet and non-vet subgroups about mechanisms and interactions associated with deficiency, and particularly of copper responsive conditions. The most frequently identified deficiencies were selenium, copper and iodine, while the most commonly identified toxicity was molybdenum. For copper responsive conditions, all of the listed treatments were used at least "occasionally"; the most frequently being glass boluses, in-feed supplementation, matrix boluses, and then copper injections. While there was a diverse choice of treatments, altering the ration was relatively rarely selected. This thesis also provides the first large-scale retrospective analysis of factors influencing the establishment of a commercially robust ovum pick up (OPU) and in vitro embryo production (IVP) platform in the UK. Over a 5-year period, a system was developed and validated for use in the UK with 2,138 cycles of OPU. These cycles were analysed as four sets of data and included two IVP laboratories and 6 OPU teams. Factors in these analyses included OPU team, IVP laboratory, ovarian stimulation protocol and semen type (unsorted vs sex-sorted). The mean number of follicles aspirated by the OPU teams ranged from 6.5 to 14.9 (P < 0.001), while the number of oocytes collected was between 4.0 and 12.4 (P < 0.001). There was an indication (P=0.055) that the blastocyst per oocyte rate varied between teams. The proportion of blastocysts from oocytes that cleaved was higher (P=0.01) for unsorted than sexed semen. Two commercial products containing different ratios of follicle stimulating hormone (FSH) to luteinising hormone (LH) (Folltropin® and Pluset®) were compared in ovarian stimulation programs. The addition of 'coasting' (short-term (typically 48h) hormonal withdrawal after FSH stimulation), prior to OPU was also investigated. Pluset® resulted in a greater (P < 0.001) mean number of follicles aspirated, more (P=0.003) blastocysts per oocyte matured and more (P < 0.001) embryos per cycle (2.45), compared with Folltropin® (1.17) or with no stimulation (1.24). Throughout the study there was a steady improvement in blastocyst production per OPU cycle. In a separate analysis, Grade 1 cumulus oocyte complexes (COCs) as a proportion of COCs recovered, oocytes that cleaved as a proportion of total COCs, and blastocysts as a proportion of total COCs, were all greater (P < 0.05) for stimulated than non-stimulated cycles, irrespective of FSH/LH product. A composite score of oocyte quality and quantity was proposed (sCOC); Log Total Mean sCOC was correlated (P < 0.001) with both the proportion of blastocysts per oocyte collected, and the total number of embryos produced per cycle. Finally, twelve peri-pubertal heifers (approximately 10 months old) participated in a crossover trial which compared PRID® (Delta®) vs CIDR® progesterone releasing intravaginal devices for use in OPU/IVP cycles. Vaginoscopic examination found higher vaginal inflammation grades for PRID® than CIDR® (P < 0.001). There was evidence of vaginal inflammation continuing for at least 2 weeks after device withdrawal. The proportion cleaved of oocytes inseminated was higher for PRID® than CIDR® (P < 0.05). Numerically but not significantly there was a higher proportion of blastocysts per cycle and a higher Log Total Mean sCOC score per cycle with PRID® than CIDR® treatments, but blastocyst yield was low throughout, suggesting a need to repeat the trial. Data collection and analyses are ongoing, to identify other key performance indicators within the OPU/IVP embryo transfer (ET) system, with a view to refining the sCOC composite score model. A robust OPU/IVP/ET system has been developed and this could be used to investigate further how mineral imbalances impact oocyte competence and blastocyst yield.
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Denadai, Renan [UNESP]. "Características qualitativas de oócitos obtidos por Laparoscopic Ovum Pick-up (LOPU) e da maturação nuclear em ovinos." Universidade Estadual Paulista (UNESP), 2013. http://hdl.handle.net/11449/128043.
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000847150.pdf: 860477 bytes, checksum: 76e841df35fa2f2e934e0f4bedb91cbc (MD5)<br>O presente estudo visou avaliar a resposta ovariana a superestimulação utilizando 80UI de FSH e 300UI de eCG após ablação por laparoscopic ovum pic-up (LOPU1) de todos os folículos visíveis presentes em ambos os ovários. Determinar o melhor momento para realização de uma nova LOPU (LOPU2), mediante a avaliação ultrassonográfica dos ovários a cada 12 horas após LOPU1. E comparar a respostas ovariana do tratamento Duplo LOPU (DLOPU) com o tratamento One Shot adaptado de Baldassarre (1996), quanto a quantidades dos Complexos Cumulos-Oócito (COCs), e qualidade das estruturas obtidas e após processo de maturação in vitro (MIV) por 24 horas. Foi determinado o momento para realização da LOPU2 em 36 horas após a superestimulação. A média de folículos presentes na avaliação ultrassonográfica imediatamente antes a LOPU em ambos os tratamentos foi cerca de 9 estruturas, não diferindo da media de folículos aspirados, a taxa de recuperação comum aos dois tratamentos foi de 62,5%. A avaliação imediata das estruturas obtidas bem como a após 24 de MIV demonstrou que não houve diferença qualitativa COCs entre os tratamentos<br>The present study aimed to evaluate the ovarian response to superstimulation using 80UI FSH and eCG 300UI after ablation LOPU (LOPU1) of all follicles present in both ovaries. Determine mlehor time to make a new LOPU (LOPU2) by ultrasound evaluation of the ovaries every 12 hours LOPU1. And comparing the responses ovarian DLOPU treatment with the treatment Ono Shot adapted Baldassarre (1996) with the DLOPU described here, the amounts of COC and quality of the structures obtained after process and IVM for 24 hours. It was determined the time for completion of LOPU2 in 36 hours after the overstimulation. The mean number of follicles present in the ultrasonographic evaluation immediately before the LOPU in both treatments was about 9 structures did not differ from follicles aspirated media, the recovery rate common to the two treatments was 62.5%. Immediate assessment of the structures obtained as well as after 24 IVM demonstrated that COCs do not hear qualitative difference between treatments
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Denadai, Renan. "Características qualitativas de oócitos obtidos por Laparoscopic Ovum Pick-up (LOPU) e da maturação nuclear em ovinos /." Botucatu, 2013. http://hdl.handle.net/11449/128043.
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Orientador: Sony Dimas Bicudo<br>Banca: Eunice Oba<br>Resumo: O presente estudo visou avaliar a resposta ovariana a superestimulação utilizando 80UI de FSH e 300UI de eCG após ablação por laparoscopic ovum pic-up (LOPU1) de todos os folículos visíveis presentes em ambos os ovários. Determinar o melhor momento para realização de uma nova LOPU (LOPU2), mediante a avaliação ultrassonográfica dos ovários a cada 12 horas após LOPU1. E comparar a respostas ovariana do tratamento Duplo LOPU (DLOPU) com o tratamento One Shot adaptado de Baldassarre (1996), quanto a quantidades dos Complexos Cumulos-Oócito (COCs), e qualidade das estruturas obtidas e após processo de maturação in vitro (MIV) por 24 horas. Foi determinado o momento para realização da LOPU2 em 36 horas após a superestimulação. A média de folículos presentes na avaliação ultrassonográfica imediatamente antes a LOPU em ambos os tratamentos foi cerca de 9 estruturas, não diferindo da media de folículos aspirados, a taxa de recuperação comum aos dois tratamentos foi de 62,5%. A avaliação imediata das estruturas obtidas bem como a após 24 de MIV demonstrou que não houve diferença qualitativa COCs entre os tratamentos<br>Abstract: The present study aimed to evaluate the ovarian response to superstimulation using 80UI FSH and eCG 300UI after ablation LOPU (LOPU1) of all follicles present in both ovaries. Determine mlehor time to make a new LOPU (LOPU2) by ultrasound evaluation of the ovaries every 12 hours LOPU1. And comparing the responses ovarian DLOPU treatment with the treatment Ono Shot adapted Baldassarre (1996) with the DLOPU described here, the amounts of COC and quality of the structures obtained after process and IVM for 24 hours. It was determined the time for completion of LOPU2 in 36 hours after the overstimulation. The mean number of follicles present in the ultrasonographic evaluation immediately before the LOPU in both treatments was about 9 structures did not differ from follicles aspirated media, the recovery rate common to the two treatments was 62.5%. Immediate assessment of the structures obtained as well as after 24 IVM demonstrated that COCs do not hear qualitative difference between treatments<br>Mestre
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姚元慶 and Yuanqing Yao. "The effects of human oviductal cells and follicular fluid on sperm functions." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1998. http://hub.hku.hk/bib/B31239614.
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Artus, Jérôme François Nicolas. "Analyse fonctionnelle du gène Ovum mutant candidate gene 1 chez la souris : du développement précoce à la régulation du cycle cellulaire." Paris 7, 2006. http://www.theses.fr/2006PA077223.
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Bien que très conservé au cours de l'évolution, le cycle cellulaire a été modifié pour répondre à de nouveaux programmes de développement. Récemment, les analyse de souris mutantes ont démontré que la plupart des régulateurs importants du cycle cellulaire ne sont pas nécessaires durant le développement ou bien ont fonction restreinte à un tissu, soulignant le fait qu'/n vivo de nombreux aspects du cycle restent encore à élucider. Durant la recherche de gènes candidats pour une mutation létale conditionnelle portée par la lignée murine DDK, un nouveau gène a été identifié : il s'agit du gène Ovum mutant candidate gène 1. L'inactivation d'Omcgl entraîne la mort des embryons aux alentours de l'implantation. De manière intéressante, cette létalité est précédée par défauts de cycle cellulaire notamment un allongement de la phase M accompagné de désorganisation du fuseau mitotique et de la plaque métaphasique. Le gène Omcgl apparaît donc comme un régulateur important dans le contrôle des premières divisions embryonnaires. Cette létalité très précoce ainsi que l'impossibilité de générer des cellules ES déficientes pour Omcgl ne permettent pas de définir plus précisément sa fonction. C'est la raison pour laquelle nous avons généré des lignées de MEFs et de cellules ES dans lesquelles l'expression de ce gène peut être contrôlée finement. Ces approches perte et gain de fonction ont été réalisées à partir du système inductible à la tétracycline et du système CRE/LoxP. Les conséquences de la modulation d'expression d'Omcgl ont été examinées en terme de prolifération/survie, capacité de différenciation et analyse globale du transcriptome<br>While highly conserved through evolution, the cell cycle has been extensively modified to adapt to new developmental programs. Recently, analyses of mouse mutants revealed that several important cell cycle regulators are either dispensable for development or have a tissue- or cell-type-specific function, indicating that many aspects of cell cycle regulation during mammalian embryo development remain to be elucidated. During the search for candidate genes implicated in an early conditional letal mutation carried by the DDK strain of mice, we discovered a new gene, Ovum mutant candidate gene 1. Disruption of Omcg1 by homologous recombination results in an early embryonic lethality at the end of the preimplantation development. Interestingly, mutant embryos exhibit alterations of the cell cycle including lengthening of the M phase, abnormal mitotic spindles and misalignment of metaphase chromosomes indicating that Omcgl plays a crucial role in the control of mouse early embryonic cell cycle. Early embryonic death of Omcg1 embryos and the failure to establish a homozygous null ES cell line have prevented us from defining more precisely its role. Thus, we generated genetically modified ES and MEFs cells in which the level of expression of Omcg1 can be modulated at will. We adopted both gain- and loss-of-function strategies based on the use of the tetracycline inducible System and Cre/LoxP system. The consequences of the modulation of Omcg1 expression in ES cells and MEFS were examined at the level of cell parameters (proliferation, cell cycle progression, apoptosis,. . . ), cell differentiation both in vitro and in vivo and global gene expression
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Makkar, Guneet. "The Role of conventional sperm parameters, quantitative motile characteristics and acrosome reaction of spermatozoa in predicting successful outcome following artificial insemination." Hong Kong : University of Hong Kong, 2000. http://sunzi.lib.hku.hk/hkuto/record.jsp?B22505507.
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Yao, Yuanqing. "The effects of human oviductal cells and follicular fluid on sperm functions /." Hong Kong : University of Hong Kong, 1998. http://sunzi.lib.hku.hk/hkuto/record.jsp?B20868212.
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Tejomurtula, Jyothsna. "Identification of a novel importin [alpha] predominantly expressed in bovine oocytes and early embryos." Morgantown, W. Va. : [West Virginia University Libraries], 2007. https://eidr.wvu.edu/etd/documentdata.eTD?documentid=5488.
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Thesis (M.S.)--West Virginia University, 2007.<br>Title from document title page. Document formatted into pages; contains vi, 45 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 40-45).
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Zhang, Lin. "Microbial pathogen contamination in mouse gametes and embryos." Diss., Columbia, Mo. : University of Missouri-Columbia, 2008. http://hdl.handle.net/10355/5671.
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Thesis (M.S.)--University of Missouri-Columbia, 2008.<br>"May 2008" The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Includes bibliographical references.
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Makkar, Guneet. "The Role of conventional sperm parameters, quantitative motile characteristics and acrosome reaction of spermatozoa in predictingsuccessful outcome following artificial insemination." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2000. http://hub.hku.hk/bib/B31224933.
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Mingoti, Rodolfo Daniel. "Qualidade oocitária e embrionária e perfil hormonal e metabólico de vacas repetidoras de serviço submetidas à secagem e indução de lactação." Universidade de São Paulo, 2018. http://www.teses.usp.br/teses/disponiveis/10/10131/tde-29112018-114206/.
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A hipótese do presente estudo sugere que a baixa fertilidade de vacas Holandesas (Bos taurus) repetidoras de serviço (RS) está relacionada à baixa qualidade oocitária que, por sua vez, é associada ao quadro de resistência periférica a insulina (RPI). Ainda, a indução de lactação (IL) em vacas Holandesas (Bos taurus) RS pode reverter o quadro de RPI e, consequentemente, melhorar a qualidade oocitária e a produção in vitro de embriões (PIVE). Para testar a hipótese proposta, este estudo objetivou avaliar o efeito da fase da lactação e da gestação [Exp. 1], o efeito da secagem de RS [Exp. 2] e o efeito da IL [Exp. 3] sobre a RPI, a qualidade oocitária e a PIVE em vacas da raça Holandesa (Bos taurus). Nos três estudos foram realizados testes de tolerância à glicose (TTG) para avaliar a RPI através do perfil hormonal sérico de insulina e glicose. Além disso, avaliou-se o perfil bioquímico sérico e folicular e a qualidade oocitária através da PIVE. Verificou-se que, em resposta à infusão de 0,3mg glicose/kg PV, as vacas RS no final da lactação secretaram 53% mais insulina e captaram 40% menos de glicose quando comparadas as vacas no terço inicial de lactação (Exp 1). Esses achados são indicativos do estabelecimento do quadro de RPI nas vacas RS em lactação. Durante o período seco, as vacas RS secretaram 96% mais insulina e captaram 56% menos glicose que as vacas no terço inicial da lactação e as vacas RS em lactação, respectivamente (Exp. 2). Ainda, as vacas com lactação induzida secretaram 11% menos insulina e captaram a mesma quantidade de glicose que vacas paridas em fases semelhantes de lactação (Exp 3), demonstrando que o protocolo de IL foi eficiente em alterar o perfil metabólico, revertendo o quadro de RPI presente nas vacas RS. Nos Exp. 1, 2 e 3 foram verificadas maiores concentrações plasmática de triglicérides (TG; P < 0,05), colesterol total (COL; P < 0,05) e LDL (P < 0,05) no soro sanguíneo em vacas RS quando comparadas com vacas no terço inicial de lactação. Durante o período seco (Exp. 2 e 3), observou-se incremento desses metabólitos, destacando aumento na concentração de TG (P < 0,05), COL (P < 0,05) e LDL (P < 0,05) plasmático em vacas secas quando comparadas as vacas em lactação [início e final (RS) da lactação]. No liquido folicular foram observadas variações no perfil bioquímico para COL e TG. Nos Exp. 1, 2 e 3, verificou-se que vacas RS possuem maior concentração de TG (P < 0,05) e COL (P < 0,05) no fluido folicular do que vacas no terço inicial de lactação. Contrariando a hipótese inicialmente proposta, as vacas RS em lactação e as vacas secas apresentaram maior taxa de blastocisto (P < 0,05) e número de blastocistos por OPU (P < 0,05) que as vacas no terço inicial de lactação (Exp. 1, 2 e 3). Através do perfil de insulina circulante em resposta ao TTG foi possível demonstrar o estabelecimento do quadro de RPI em vacas RS (P < 0,05). Além disso, constatou-se agravamento da RPI em vacas secas (P < 0,05). Esse quadro foi associado ao aumento do escore de condição corporal (P < 0,05) e do peso vivo (kg; P < 0,05) nas vacas RS. Em conclusão, não foi verificada associação negativa entre RPI, qualidade oocitária e PIVE em vacas Holandesas (Bos taurus) RS. Apesar da indução de lactação em vacas Holandesas (Bos taurus) RS alterar o metabolismo e diminuir o quadro de RPI, não foi verificado efetivo positivo na qualidade oocitária e na PIVE.<br>The hypothesis of the present study suggests that low fertility of repeat breeders (RB) Holstein (Bos taurus) cows is related to low oocyte quality, which is associated with peripheral insulin resistance (PIR). Also, induction of lactation (IL) in RB Holstein (Bos taurus) cows can revert PIR and, consequently, improve oocyte quality and in vitro embryo production (IVEP). In order to test the proposed hypothesis, the objective of this study was to evaluate the effect of phase of lactation and gestation [Exp. 1], effect of drying RB [Exp. 2] and effect of IL [Exp. 3] on PIR, oocyte quality and IVEP of Holstein (Bos taurus) cows. In all three studies, glucose tolerance tests (GTT) were performed to evaluate PIR through the serum hormonal insulin and glucose profile. In addition, we evaluated the serum and follicular biochemical profile and oocyte quality through IVEP. It was verified that, in response to a 0.3mg glucose/kg of body weight (BW), RB cows at the end of lactation secreted 53% more insulin and captured 40% less glucose when compared to cows in the initial third of their lactation (Exp. 1). These findings are indicative of the establishment of PIR in RB lactating cows. During the dry period, RB cows secreted 96% more insulin and captured 56% less glucose than cows in the initial third of their lactation and RB lactating cows, respectively (Exp. 2). Also, cows with induced lactation secreted 11% less insulin and captured the same amount of glucose than calved cows in similar lactation phase (Exp. 3), demonstrating that the IL protocol was efficient to alter the metabolic profile, reverting PIR present in RB cows. In Exp. 1, 2 and 3 higher plasmatic concentrations of triglycerides (TG; P<0.05), total cholesterol (COL; P<0.05) and LDL (P<0.05) were verified in the blood serum in RB cows when compared to cows in the initial third of their lactation. During the dry period (Exp. 2 and 3), we observed the increment of these metabolites, and a notable elevation of the plasmatic TG (P < 0.05), COL (P < 0.05) and LDL (P < 0.05) in dry cows when compared to lactating cows [beginning and end (RB) of lactation]. In the follicular fluid, it was possible to observe variations in the biochemical profile for COL and TG. In Exp. 1, 2 and 3, it was verified that RB cows have higher concentration of TG (P < 0.05) and COL (P < 0.05) in the follicular fluid than cows in the initial third of their lactation. Contrary to the initially proposed hypothesis, RB lactating cows and dry cows presented higher blastocyst rate (P<0.05) than cows in the initial third of lactation (Exp. 1, 2 and 3). Through the circulating insulin profile in response to the GTT it was possible to demonstrate the establishment of PIR in RB cows (P<0.05). Also, it was observed worsening of the PIR in dry cows (P<0.05). This condition was associated with an increase in body condition score (P<0.05) and BW (kg; P<0.05) in RB cows. In conclusion, no negative association between PIR, oocyte quality and IVEP was observed in RB Holstein (Bos taurus) cows. Although induction of lactation in RB Holstein (Bos taurus) cows altered the metabolism and reduced PIR, no positive effect was observed in oocyte quality and IVEP.
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Salinas-Flores, Liliana, and n/a. "Understanding and improving the cryopreservation of pacific oyster (Crassostrea gigas) oocytes via the use of two approaches : modification of an existing cryopreservation protocol and manipulation of the lipis fraction of the oocytes." University of Otago. Department of Food Science, 2008. http://adt.otago.ac.nz./public/adt-NZDU20080305.143446.
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Cryopreservation of gametes is a valuable tool for the fast-growing aquaculture industry in New Zealand. In the present study, research was aimed to improve the cryopreservation of Pacific oyster (Crassostrea gigas) oocytes. For this, two main approaches were used: the modification of an existing published (standard) cryopreservation protocol for oyster oocytes and the modification of the oocytes themselves prior to cryopreservation. The objectives in the chapters of this thesis were: (a) determination of the cryobiological characteristics of oyster oocytes; (b) assessment and reduction of intracellular ice formation (IIF) in oocytes; and (c) modification of the lipid fraction (cholesterol and fatty acids) of oocytes prior to cryopreservation.
Knowledge of the membrane permeability parameters in response to concentrations of water and ethylene glycol (EG), the influence of temperature upon these parameters, and the osmotic tolerance limits of oyster oocytes were used to develop computer models that simulated the cellular volume changes that oocytes underwent during EG addition and removal. The models predicted that when one part of EG was added in one step to one part of oocyte suspension and equilibrated for 20 min at 20 �C, similar volume changes in oocytes would be obtained, compared to a more complicated multi-step addition method. This method of addition resulted in similar post-thaw fertilization rates to those obtained by using the multi-step addition method, thus reducing oocyte handling.
Cryomicroscopy was used to assess the effect of cooling rates and EG concentration on the temperature at which oocytes underwent IIF. It was found that IIF occurred at higher subzero temperatures when fast cooling rates were used (30 and 5 �C min⁻�) and at EG concentrations ranged between 0 and 15%. At a relatively slower cooling rate of 0.3 �C min⁻� and with 10% EG, which are the conditions employed in the standard cryopreservation protocol, no IIF occurred.
The steps of the standard protocol that were more likely to cause oocyte damage were identified by evaluating the fertilization rate of oocytes at each step. Results showed that oocytes were most damaged by cooling them to -35 �C and followed by plunging them in liquid nitrogen. Contrary to what had been observed under the cryomicroscope, transmission electron microscopy (TEM) analysis revealed that all oocytes cryopreserved by the standard protocol contained cytoplasmic ice. In addition, it was also observed that oocytes were at two developmental stages when frozen (prophase and metaphase I). These observations prompted the development of alternative cooling programmes aimed to reduce intracellular ice. The effect of cooling rate, plunge temperature and time held at the plunge temperature were thus evaluated, based on post-thaw fertilization rate of oocytes. Overall, neither the cooling rate nor the holding time had an effect on oocyte fertility. However, the plunge temperature had an effect, where oocytes plunged at -60 �C had lower post-thaw fertilization rates than oocytes plunged at -35 �C. Through the slowing of the cooling rate, lengthening of the holding time and lowering of the plunge temperature, it was possible to reduce the amount of ice in the cytoplasm. However, the reduction of intracellular ice did not improve the post-thaw fertilization rate of the oocytes; on the contrary, post-thaw fertilization decreased notoriously. From these results, it can be suggested that oyster oocytes are more likely to be damaged by exposure to high intra and extracellular solute concentration than IIF during cryopreservation.
In an effort to modify the lipid content of oyster oocytes prior to cryopreservation and thus, making them more resistant during cryopreservation, oocytes were incubated in solutions that would add or remove cholesterol or in solutions rich in long chain fatty acids (EPA or DHA). Oocytes incubated in cholesterol-rich solutions showed a positive uptake of fluorescently labelled cholesterol and this effect was dose dependent. Nevertheless, this uptake did not improve the post-thaw fertilization rate nor did it increase the total cholesterol content of the oocytes. When oocytes were incubated in non-conjugated or conjugated EPA or DHA, no increase in the proportion of these fatty acids was identified in the fatty acid profiles of whole oocytes and no improvement of the post-thaw fertilization rate was recorded.
Given that there was no uptake of fatty acids from the incubation media by the oocytes, a different approach was taken. This involved the supplementation of lipid-rich diets to the oyster broodstock during gametogenesis (cold-conditioning) and vitellogenesis (warm-conditioning). Despite results showing that lipid content and, indeed, fatty acid profile was altered through the diet, the results also showed that fresh oocytes from broodstock fed during cold-conditioning did not show any improvement in their fertilization rates, nor did they benefit from a lipid-rich diet during warm-conditioning. On the other hand, cryopreserved oocytes did have higher post-thaw fertilisation rates when broodstock were fed during cold-conditioning and, although no effect was found from feeding broodstock with either of the lipid-rich diets during warm-conditioning, trends indicated that a diet consisting of fresh microalgae or the commercial supplement Algamac would yield the highest post-thaw fertilization rates.
This thesis has furthered the understanding of some of the factors that determine cryosurvival in oyster oocytes and has demonstrated that both physical and biological issues must be taken into consideration for cryopreservation. Specifically, the results in this thesis helped to modify an empirically developed cryopreservation protocol for Pacific oyster oocytes. In addition, the results also showed strong evidence of the survival of oyster oocytes to intracellular ice and highlighted the importance of supplying the broodstock with lipid-rich food during the periods of gamete formation and maturation in order to obtain oocytes that are more amenable to cryopreservation. These benefits could be of significant practical importance and may be extended for the development or refinement of cryopreservation protocols for other shellfish species of commercial importance to the aquaculture industry of New Zealand.
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Stavréus-Evers, Anneli. "Implantation : morphological and biochemical characterization of the receptive human endometrium /." Stockholm, 2002. http://diss.kib.ki.se/2002/91-7349-313-9.
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Kodithuwakku, Kankanamge Suranga Pradeep Kodithuwakku. "Olfactomedin-1 (OLFM-1) in human endometrium and fallopian tube: its roles on endometrial receptivity andtubal ectopic pregnancy." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46541603.
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張韻怡 and Wan-yee Ana Cheong. "MicroRNA let-7a regulates integrin beta-3, vav3, and dicer to modulate trophoblast activities and hence embryo implantation." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hdl.handle.net/10722/193411.
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MicroRNAs are small regulatory RNAs that bind to the seeding regions within the 3’-untranslated region (3’-UTR) of their target transcripts to modulate transcript stability and/or inhibit protein translation. MicroRNA Let-7a belonging to the Lethal-7 (Let-7) family is down-regulated at the blastocyst stage, suggesting its suppression is crucial for embryo implantation. Yet, the underlying mechanism on how Let-7a modulates blastocyst implantation remains largely unknown. In silico analysis identified attachment related integrin beta-3, outgrowth related vav3, and the microRNA processing dicer, as Let-7a targets. Therefore, it is hypothesized that down-regulation of Let-7a promotes embryo implantation by stimulating these target proteins.
Let-7a is down-regulated during blastulation and at 3-hour post-estradiol activation of the dormant blastocysts. Force-expression of Let-7a in mouse blastocysts suppressed blastocyst attachment, outgrowth on fibronectin-coated plates and compromised pregnancy in vivo. Dual luciferase assay using the 3’-UTR reporter constructs of the integrin beta-3, vav3, and dicer confirmed the interaction between Let-7a and the three targets. Force-expressing or inhibiting Let-7a expression in mouse blastocysts by electroporating the Let-7a precursor or inhibitor respectively illustrated post-transcriptional regulation of Let-7a on integrin beta-3 and vav3, and transcriptional regulation on dicer. Dormant blastocysts retrieved from the delayed implanting mice expressed high Let-7a levels, which was suppressed in the first 12-hours of estradiol activation. Concomitantly, dormant blastocysts expressed low levels of integrin beta-3, vav3, and dicer, yet, their protein expression was up-regulated from 3 hours-post estradiol activation. Furthermore, addition of integrin beta-3 antibody suppressed attachment of trophoblast spheroids (blastocyst surrogate) onto endometrial epithelial cells in a co-culture model and the outgrowth of the spheroids on fibronectin-coated plates.
Knockdown of Vav3 with siRNA decreased blastulation, hatching, and outgrowth rates of the embryos in vitro. Although the loss of vav3 activities did not affect embryo implantation, it suppressed trophoblast migration on fibronectin-coated plates and invasion into collagen matrix. In contrast, force-expression of vav3 enhanced blastocyst outgrowth, and promoted cell proliferation. Blocking integrin beta-3 activities in the vav3 knock-down embryos further suppressed blastocyst outgrowth, suggesting the intertwining effect of the integrins and vav3.
Dicer knock-down in mouse blastocysts decreased mature Let-7a expression and suppressed blastulation and hatching in vitro and implantation in vivo. Dicer knock-down in estradiol activated mouse blastocysts decreased the epidermal growth factor receptor expression and lowered the affinity of the embryos to EGF, and suppressed the implantation potential to a level similar to that of dormant blastocysts.
Taken together, the suppression of Let-7a by estradiol up-regulates integrin beta-3, vav3, and dicer. The increased Itgb3 expression promotes blastocyst attachment and intertwined with the up-regulated vav3 expression to enhance blastocyst outgrowth. The increased vav3 expression further stimulates cell proliferation and confers blastocyst invasion into the collagen matrix. Dicer, by altering microRNA processing, facilitates blastulation and thereby embryo implantation. Thus, the loss of Let-7a biological activities during blastulation is crucial to enhance blastulation and stimulate trophoblast activities for successful embryo implantation.<br>published_or_final_version<br>Obstetrics and Gynaecology<br>Doctoral<br>Doctor of Philosophy
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Gan, Yong Chen Zhen. "Model-based simulations of the piercing process in piezo-ICSI." Diss., Columbia, Mo. : University of Missouri--Columbia, 2008. http://hdl.handle.net/10355/7111.
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Title from PDF of title page (University of Missouri--Columbia, viewed on Feb. 23, 2010 ). The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. Dr. Zhen Chen, Dissertation Supervisor. Vita. Includes bibliographical references.
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Oliveira, Louise Helen de. "Associação da insulina circulante com a função ovariana e qualidade oocitária em vacas holandesas." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/11/11139/tde-03022016-153026/.
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O objetivo do primeiro estudo foi avaliar a produção in vitro de embriões (PIVE) em vacas holandesas não lactantes submetidas a aspiração oocitária (OPU) posteriormente ao protocolo de superestimulação folicular similar ao descrito por Nivet et al. (2012) em comparação à realização da OPU em dia aleatório do ciclo estral. Para tal, vacas holandesas não lactantes e não gestantes foram distribuídas aleatoriamente em delineamento tipo crossover em Controle (n = 35), em que as vacas não foram tratadas com FSH, mas submetidas a uma sessão de aspiração em dia aleatório do ciclo estral; ou p-FSH (n = 35), em que, 36 horas após a OPU para sincronização da onda folicular, as vacas foram tratadas com p-FSH por 3 dias e 44 horas após, submetidas a sessões de OPU. O número total de complexos cumulusoócito (CCO) recuperados e o número de oócitos viáveis foram semelhantes entre os grupos controle e p-FSH. Além disso, não houve aumento na proporção de CCO viáveis (CCO viáveis / CCO total recuperado). Da mesma forma, não se detectaram diferenças no número de embriões / sessão de OPU e taxa de blastocistos. O protocolo de superestimulação folicular não melhorou a PIVE em vacas holandesas não lactantes. O experimento 2 testou a hipótese de que vacas leiteiras de alta produção se tornam cada vez mais resistentes à insulina com o avançar da lactação, e consequentemente, a qualidade do oócito é comprometida. Foram utilizadas vacas holandesas em 50 (51,5 ± 3,7; n = 30), 100 (102,3 ± 9,4; n = 30) e 150 (154,5 ± 18,9; n = 30) dias em lactação (DEL). Durante o teste de tolerância à glicose (TTG), não houve diferença entre grupos para qualquer variável relacionada à glicose circulante. No entanto, medidas de insulina circulante foram diferentes em vacas aos 150 DEL em comparação com 50 ou 100 DEL, tais como: maior insulina basal, pico, Δ máx de insulina e AUC 5-60. Porém, não houve diferença entre os grupos para o número ou percentagem de oócitos viáveis. Assim, as vacas desenvolveram resistência à insulina com o aumento do DEL. No entanto, o aumento da resistência à insulina não foi associado com alteração detectável na qualidade dos oócitos aspirados de folículos pequenos e médios. O experimento 3 foi para avaliar se o aumento de insulina circulante durante os períodos de pré e pós desvio folicular aumenta o desenvolvimento inicial e final, do folículo, bem como do corpo lúteo (CL). Além disso, por induzir a ovulação de um folículo maior, o CL resultante de vacas com alta insulina circulante também é maior e mais esteroidogênico, refletindo em maiores concentrações circulantes de progesterona (P4). O delineamento experimental utilizado foi o quadrado latino em arranjo fatorial 2x2, em quatro grupos experimentais: 1) CC = água pré e pós desvio folicular (n = 16); 2) CP = água e propilenoglicol (PPG) pré e pós desvio folicular, respectivamente (n = 16); 3) PC = PPG e água pré e pós desvio folicular, respectivamente (n = 16) e 4) PP = PPG pré e pós desvio folicular (n = 16). O aumento agudo e transitório, durante os períodos de pré e pós desvio não aumentou o desenvolvimento folicular, luteal e concentrações plasmáticas de P4.<br>The aim of the first study was to evaluate the in vitro embryo production (IVEP) in nonlactating Holstein cows subjected to ovum pick-up (OPU) after ovarian superstimulation with a protocol similar to that described by Nivet et al. (2012) in comparison with OPU at a random day of the estrous cycle. Nonlactating Holstein cows were randomly assigned in a crossover design to: Control (n = 35) in which cows were not treated with p-FSH, but subjected to OPU at a random day of the estrous cycle; or p-FSH (n = 35), in which, 36 hours after OPU to synchronize follicle wave, the cows were treated with p-FSH for 3 days and 44 hours later, subjected to OPU sessions. The total number of cumulus-oocyte complex (COC) recovered and the number of viable oocytes were similar between control and p-FSH groups. In addition, there was no increase in the proportion of viable COC (viable COC / overall COC recovered). Likewise, we detected no differences in the number of embryos / OPU session and blastocyst rate. Follicle superstimulation protocol with p-FSH did not improve IVEP in nonlactating Holstein cows. Experiment 2 tested the hypothesis that high-producing dairy cows become increasingly resistant to insulin with advancing lactation, and consequently oocyte quality is compromised. We used Holstein cows at 50 (51.5 ± 3.7; n = 30), 100 (102.3 ± 9.4; n = 30) and 150 (n = 30 154.5 ± 18.9) days in milk (DIM). During the glucose tolerance test (GTT), there was no difference between groups for any variable related to circulating glucose. However, circulating insulin measurements such as basal insulin, peak insulin, Δ max and AUC 5-60 were higher for cows at 150 DIM. Nevertheless, there was no difference between groups for the number or percentage of viable oocytes. Therefore, although cows developed insulin resistance with increasing DIM, this has not been associated with detectable change in the quality of oocytes aspirated from small and medium follicles. The third experiment assessed whether the increase in circulating insulin during periods of pre- and post-follicle deviation increases the initial and final follicle size and corpus luteum (CL) volume. Moreover, by inducing ovulation of greater follicles, resulting in greater CL, cows with high circulating insulin also have higher circulating progesterone (P4). The experimental design was a Latin square in a 2x2 factorial arrangement in four groups: 1) CC = water pre and post follicle deviation (n = 16); 2) CP = water pre and propylene glycol (PPG) post follicle deviation (n = 16); 3) PC = PPG and water pre and post follicle deviation, respectively (n = 16), 4) PP = PPG pre and post follicle deviation (n = 16). Acute and transient circulating insulin increase during periods of pre and post follicle deviation has not affected follicle development, luteal volume or plasma concentrations of P4.
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Cheung, Hoi-yan, and 張凱恩. "The study of Chinese herbal medicinal compound on implantation : in vitro spheroid-endometrium co-culture." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hdl.handle.net/10722/196547.
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Traditional Chinese medicine (TCM) plays an important role in the Chinese healthcare system for over five thousand years. It includes the use of herbal medicine, acupuncture, Tui Na (推拿), and diet therapy. TCM helps to maintain a balance of Yin-Yang (阴阳), Five Phases (五行), Meridians (经络) and Qi (气) inside the body. In practise, pregnant women take tocolytic drugs to tonify the blood and qi to provide a continuous supply of nutrients for baby.
Traditional Chinese herbal medicines usually prescribed as a complex formula to produce synergistic or agonistic effect to maintain a well balance of the above components in human bodies. Moreover, TCM usually cannot produce immediate effect on patients, therefore, the efficacy of individual component remains largely unknown. This study aims to investigate whether Chinese tocolytic drug components could modulate fertility by affecting the in vitro spheroid (blastocyte surrogate) attachment process by using trophoblastic (JEG-3) and endometrial epithelial (Ishikawa) cells to mimic the embryo-endometrial implantation process.
Nine Chinese herbal medicinal compounds (Atractylenolide I(白术内酯), Atractylenolide II(白术内酯II), Atractylenolide III(白术内酯III), Paeoniflorin(芍药苷), Albiflorin(芍药内酯苷), Nuzhenide(女贞子甙), Ecliptasaponin A(旱莲甙A), Wedelolactone(蟛蜞菊内酯) and Columbianadin(二氢欧山芹醇当归酸酯)) which are commonly found in traditional Chinese tocolytic drug formula were selected to study (1) the toxicity of the drugs on trophoblastic (JEG-3) and endometrial epithelial (Ishikawa) cells growth, (2) the effect of three tocolytic drugs (Atractylenolide I, Atractylenolide II and Atractylenolide III) on spheroid attachment, and (3) their effect of the expression of Wingless (Wnt) signaling molecules (Active-β-Catenin, Axin-2, β-catenin, E-cadherin, GSK-3β, and Mucin-1).
It was found that the nine compounds, Atractylenolide I, Atractylenolide II, Atractylenolide III, Paeoniflorin, Albiflorin, Nuzhenide, Ecliptasaponin A, Wedelolactone and Columbianadin did not affect cell viability at 25μM, 25μM, 5μM, 0.2μM, 125μM, 125μM, 125μM, 5μM and 25μM, respectively, by cell proliferation assay. However, at these concentrations, the spheroid attachment was not significantly increased by Atractylenolide I, Atractylenolide II and Atractylenolide III. Interestingly, the protein expression of GSK-3β and Active-β-catenin were up-regulated by the three compounds in both cells and JEG-3 cells respectively. The expressions of Axin-2 and E-cadherin were up-regulated by Atractylenolide III in Ishikawa cells and Atractylenolide II in JEG-3 cells. Atractylenolide I and Atractylenolide III increase the Ishikawa cells expression of Active-β-catenin and β-catenin respectively and together suppress the JEG-3 cells Mucin-1 and β-catenin expression.
In conclusion, the nine tocolytic compounds have different effect on cell proliferation. Atractylenolide I, Atractylenolide II and Atractylenolide III did not enhance the attachment rate of JEG-3 spheroid onto Ishikawa monolayer. However, they affected Wnt-signaling molecules expression, suggesting that they may modulate endometrial receptivity. Further experiments are needed to study their combined effect on co-culture and expression of Wnt-signaling molecules.<br>published_or_final_version<br>Obstetrics and Gynaecology<br>Master<br>Master of Medical Sciences
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Moon, Yeoncheol. "A study of piezo-drill vibration for intracytoplasmic sperm injection /." free to MU campus, to others for purchase, 2004. http://wwwlib.umi.com/cr/mo/fullcit?p1420943.
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Dórea, Marcus Dantas. "Folículos residuais subsequentes à aspiração folicular em bovinos." Universidade Federal do Espírito Santo, 2012. http://repositorio.ufes.br/handle/10/5107.
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Previous issue date: 2012-03-30<br>Objetivou-se com este estudo avaliar a taxa de folículos residuais pós-aspiração de folículos ovarianos de 8 e 12 mm de diâmetro, por meio da avaliação dos fatores endócrinos, analisando a interferência destas estruturas no desenvolvimento normal da dinâmica de crescimento. Foram selecionadas vacas da raça Holandesa (N=13) não lactantes e com dois ciclos estrais normais consecutivos; estas foram tratadas com 500mg de cloprostenol sódico (Sincrocio®) e 3 mg de norgestomet (Crestar®) (D0), 2 mg de benzoato de estradiol (Sincrodiol®) (D1), após a emergência da onda folicular foram distribuídas aleatoriamente em dois tratamentos caracterizados pela ausência ou presença de implantes de norgestomet, cada tratamento dividido em dois grupos: presença do implante e folículo de 8 mm (G1), ausência do implante e folículo de 8 mm (G2), presença do implante e folículo de 12 mm (G3) e ausência do implante e folículo de 12 mm (G4). O monitoramento ultrassonográfico (Esaote Pie Medical®) foi realizado duas vezes ao dia com intervalo de 12 horas. Os folículos ovarianos foram aspirados com 8 e 12mm e avaliados quanto à presença do resíduo ao processo de aspiração, quando residual, eram aspirados novamente. Os dados de dinâmica de crescimento folicular, perfil hormonal do plasma sanguíneo e do fluído folicular foram verificados quanto à distribuição normal, pelo teste de Shapiro-Wilk e comparadas nos diferentes tratamentos pelo teste t de student. Os dados de perfil de esteroidogênico dos folículos aspirados em diferentes diâmetros foram analisados pelo SAS GLM procedure para o principal efeito de tratamento e as comparações foram feitas pelo teste F. O nível de significância adotado foi de 5%. O tratamento com progestágeno não afetou o crescimento folicular. A taxa de folículo residual foi de 74.6%. O tratamento com progestágeno não afetou (P>0.05) o percentual de folículos residuais, independente do diâmetro do folículo, no entanto, o efeito do diâmetro folicular aproximou a significância estatística (P=0.07). Os folículos residuais cresceram cerca de 5 mm ao dia. Dos folículos residuais aproximadamente 65% se tornaram folículos ativos, 12% folículos luteinizados e 23% folículos inativos. Conclui-se que a presença de folículos residuais ocorre na maioria dos folículos aspirados com maior frequência em 12 mm e grande parte ativos e com elevada concentração de estradiol.