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Preimplantation embryo morphology following early luteal phase anti-nidatory treatment with mifepristone (RU486) in the rhesus monkey 总被引:6,自引:0,他引:6
Ghosh D Lalitkumar PG Wong VJ Hendrickx AG Sengupta J 《Human reproduction (Oxford, England)》2000,15(1):180-188
The ultrastructural characteristics of peri-implantation stage embryos recovered on day 6 after ovulation from rhesus monkeys with or without mifepristone (RU486) treatment during the early luteal phase were examined in the present study. Monkeys were randomly allocated to two groups; group 1 animals were injected s.c. with 2 ml vehicle (1:4, benzyl benzoate: olive oil, v/v, n = 21) and group 2 animals received a single dose of mifepristone (2 mg/kg body weight, w/v, n = 30) in the same volume of vehicle on day 2 after ovulation in mated cycles. On day 6 after ovulation, female monkeys of both groups were laparotomized and their reproductive tracts were flushed to retrieve preimplantation stage embryos. Embryos that showed frank degeneration or desynchrony on gross microscopical examination were not included in the present study. Preimplantation embryo growth on day 6 after ovulation was significantly (P < 0.05) affected in the morula-blastocyst transition stage in mifepristone-treated monkeys compared with that in the control group of monkeys. Ultrastructurally, administration of mifepristone on day 2 after ovulation depressed preimplantation stage embryo development, characterized by loss of cell polarity, lack of mitochondrial maturity, and lack of differentiation in trophoblast cells. Furthermore, preimplantation embryos from mifepristone-treated animals displayed a higher occurrence of inter-blastomere space, intra-cytoplasmic vacuoles, myelinoid bodies, accumulation of lipid droplets, lysosomes, lipofuscins, autophagosomes and multivesicular bodies. Collectively, it appears that the developmental potential of preimplantation embryos was significantly compromised in mifepristone-treated cycles. 相似文献
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Ghosh D Lalitkumar PG Wong V Dhawan L Rosario JF Hendrickx AG Lasley BL Overstreet JW Sengupta J 《Contraception》2001,63(2):95-102
Intravaginal administration of an anti-angiogenic agent, fumagillin, during blastocyst implantation, inhibits pregnancy establishment in a dose-related manner in the rhesus monkey. In the present study, mated female rhesus monkeys were vaginally inserted with tampons containing vehicle (group 1; n = 5) and test agent (fumagillin, 4 mg/animal; group 2; n = 6) on cycle day 20, and endometrial tissue samples were collected on cycle day 24 from all monkeys and processed for morphometric and ultrastructural analysis. Concentrations of estradiol-17beta, progesterone and chorionic gonadotrophin in peripheral circulation were determined. From serum profiles of hormones, two monkeys in group 1, and one animal in group 2 appeared pregnant. Endometrial morphology revealed histologic evidence of pregnancy in three of six fumagillin-treated animals, while other three fumagillin-treated animals showed degenerative changes in glands and venules along with marked extravasation. It is possible that the function of corpus luteum was affected by fumagillin treatment resulting in inadequate progesterone production (p <0.05), and consequent inadequate endometrial secretory preparation and receptivity, as revealed from decline in apical movement of vacuoles (p <0.05) and increase (p <0.05) in extravasation of red cells and leukocytes. 相似文献
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D. Ghosh Latika Dhawan P. G. L. Lalitkumar Viviana Wong J. F. Rosario A. G. Hendrickx B. L. Lasley J. W. Overstreet Jayasree Sengupta 《Contraception》2001,63(6):197
Intravaginal administration of an anti-microbial agent, (Ala8,13,18)-magainin II amide, during blastocyst implantation inhibits pregnancy establishment in a dose-related manner in the rhesus monkey (Macaca mulatta). In the present study, mated female rhesus monkeys were vaginally inserted with tampons containing vehicle (Group 1; N = 5) and test agent (magainin, 0.5 mg/animal; Group 2; N = 6) on cycle day 20. Endometrial tissue samples were collected on Cycle Day 24 from all monkeys and processed for morphometric and ultrastructural analysis. Concentrations of estradiol-17β, progesterone, and chorionic gonadotrophin in peripheral circulation were determined, which revealed that two monkeys in Group 1 were pregnant while no animals were pregnant in Group 2. Endometrial morphology, however, revealed histologic evidence of pregnancy in three out of the six magainin-treated animals. It appears that intra-vaginal administration of magainin II amide had a marginal effect on the implantation stage endometrium and the initiation of the implantation process in the rhesus monkey. 相似文献
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Successful blastocyst implantation depends upon the synchronous dialogue between age- and stage-matched embryo and adequately primed maternal endometrium. Endometrial signals present in the uterine lumen influence the growth and the viability of preimplantation stage embryo. Thus, uterine secretion of embryotoxic cytokines may affect the preimplantation stage embryo. Our previous study in the rhesus monkey has indicated that tumor necrosis factor-alpha (TNF-alpha) is one such candidate present in the uterine lumen, which may act as an embryotoxic agent. In the present study, the effect of TNF-alpha on de novo protein synthesis by mouse morulae (n = 100) and blastocysts (n = 100) in vitro was investigated by 2D-polyacrylamide gel electrophoresis. A total of 35 and 40 protein spots were detected in lysates of control morulae and blastocysts, respectively. Exposure of embryos to TNF-alpha (50 ng/ml) reduced the number of protein spots to 15 and 17 compared to that of control morulae and blastocysts. Seven spots in morula and 13 protein spots in blastocyst flourograms showed quantitative changes in their expressions with exposure to TNF-alpha. Morulae and blastocysts exposed to TNF-alpha expressed 8 and 17 protein spots, respectively, that were not seen in control embryos. It appears from the present study that exposure of preimplantation stage embryos to TNF-alpha affects their protein synthesis both quantitatively and qualitatively. 相似文献
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Vayeda Chintan Manranjan Devendra Singh Yadav Hitesh Amrutlal Jogia Praful Lalitkumar Chauhan 《Scientia pharmaceutica》2013,81(4):1043-1056
A simple, fast, and sensitive reversed-phase HPLC method with UV detection was developed for the quantitation of omeprazole and its eleven related compounds (impurities) in pharmaceutical formulation using the Thermo Accucore C–18 (50 mm × 4.6 mm, 2.6 μm) column. The separation among all the compounds was achieved with a flow rate of 0.8 mL min−1 employing a gradient program of mobile phase A [0.08 M glycine buffer pH 9.0: acetonitrile; 95:05 (v/v)] and mobile phase B [acetonitrile: methanol; 65:35 (v/v)]. The chromatographic detection was carried out at a wavelength of 305 nm. The method was validated for specificity, linearity, and recovery. The huskiness of the method was determined prior to validation using the Design of Experiments (DOE). The ANOVA analysis of DOE with a 95% confidence interval (CI) confirmed the buffer pH of mobile phase A (p <0.0001) and column temperature (p<0.0001) as significant Critical Method Parameters (CMPs). 相似文献
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Lalitkumar PG Sengupta J Dhawan L Sharma DN Lasley BL Overstreet JW Ghosh D 《Contraception》2000,62(3):155-159
In the present study, fumagillin, which is an antibiotic with potent angiostatic activity secreted from Aspergillus fumigatus was administered intravaginally during peri-implantation stage in the rhesus monkey and its effects on ovarian function, blastocyst implantation and pregnancy outcome in the rhesus monkey were investigated. Female monkeys (n = 18) showing normal menstrual cycles were vaginally inserted with tampons containing fumagillin (0 mg/animal in group 1; 1 mg/animal in group 2; 2 mg/animal in group 3; 4 mg/animal in group 4) on cycle day 20 of the mated treatment cycle, and these were removed on day 26 of the treatment cycle. Pregnancy was found to occur in animals treated with 1 mg and 2 mg fumagillin. However, animals treated with 4 mg fumagillin remained non-pregnant along with decreased (p <0.001) concentration of progesterone in circulation during the luteal period compared with that in normal, non-mated, ovulatory cycle. 相似文献
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There are various cellular mediators which can affect the process of blastocyst implantation by regulating the proliferation and differentiation of conceptus and maternal endometrial cells. Insulin-like growth factors I (IGF-I) and II (IGF-II) are potent mitogenic and differentiation-promoting growth factors. However, the role of IGF peptides at implantation in primate species is not well understood. The objective of the present study was to immunohistochemically localize IGF-I and IGF-II peptides in trophoblast cells and maternal endometrial cells during lacunar and villous stages of placentation in the Rhesus monkey. Female animals (n = 10) were laparotomized on estimated days 13-16 after fertilization to collect primary implantation sites which were subjected to immunohistochemical staining for IGF-I and IGF-II peptides. Cell-type specificity for IGF-I and IGF-II was evident with a very low level of IGF-I peptide immunolocalized in trophoblast cells lining lacunae, and primary and secondary villi, while moderate to high amounts of IGF-II peptide were detected in lamellar syncytiotrophoblast cells lining lacunae, early villi and cell columns, as well as in migrating trophoblast cells in the extravillous compartment and in endovascular trophoblast cells. The observed presence of IGF-II peptide in differentiated lamellar syncytiotrophoblast cells during the very early stages of implantation and placentation in the Rhesus monkey may be important in their transition to this differentiated cell population. Maternal endometrial cells showed similar distribution profiles for IGF-I and IGF-II. In conclusion, we report differential distribution of IGF-I and IGF-II peptides in trophoblast cell populations at the feto-maternal interface during lacunar and villous stages of gestation in the Rhesus monkey. 相似文献
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