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Biomedical subjects

B S Minhas

Publications and source records attributed to B S Minhas.

26 records · Page 2Linked to original sources

Survival and cell acquisition rates after preimplantation embryo biopsy: use of two mechanical techniques and two mouse strains.

Two strains of mouse embryos at the four- and eight-cell stages had biopsy specimens obtained by means of two different mechanical techniques: aspiration and displacement. Embryos and biopsy specimen cells were evaluated for survival and development. Blastomere acquisition rates were significantly higher with the displacement biopsy technique; however, no difference in survival or developmental rates was found in blastomere biopsy specimens removed from either four-cell or eight-cell embryos. A maximum of one blastomere can be removed from a four-cell embryo, whereas three blastomeres can be taken at biopsy from an eight-cell mouse embryo without significantly affecting embryo development, although mouse strain differences were noted. Intact, viable, biopsied blastomeres will develop in vitro when cocultured with morphologically intact embryos. Births of live offspring after embryo biopsy are reported.

Animals↗

Spontaneous zona reaction in the mouse as a limiting factor for the time in which an oocyte may be fertilized.

This study evaluated the effect of ovum aging on the in vitro fertilizability of mouse ova. Over 1347 ova were evaluated. Serial trypsin digestion of in vitro and in vivo aged ova revealed an increase in zona digestion time (0.25% trypsin) beginning at 40 hr, which increased over a 40-hr period and resulted in the unfertilized zona becoming as "hard" as the fertilized embryo zona. In vitro fertilizability showed a rapid decrease as zona hardening occurred with loss of cortical granules as assessed by electron microscopy. These data suggest that the window of fertilizability is "closed" by a spontaneous zona reaction occurring at about 55 hr post-human chorionic gonadotropin with loss of cortical granules and zona hardening as manifested by increasing zona digestion time with 0.25% trypsin.

Animals↗

Effects of platelet activating factor on mouse oocyte fertilization in vitro.

Platelet activating factor is rapidly gaining acceptance as a potent mediator in many reproductive processes. This study presents data that indicate a direct role of platelet activating factor in fertilization. Platelet activating factor was shown to significantly increase (p less than 0.001) the fertilization rate of mouse oocytes in vitro. Furthermore, CV3988, an inhibitor of platelet activating factor, was noted to significantly decrease in vitro fertilization rates at 10(-5) and 10(-4) mol/L concentrations.

Animals↗

The effects of platelet-activating factor on the motility of human spermatozoa.

Our laboratory has recently detected the presence of platelet-activating factor (PAF)-like activity in human spermatozoa. To gain further insight into the role of PAF on the male reproductive system, this study, using videomicroscopy, evaluated the effects of synthetic PAF on the motility of human spermatozoa. Treatment of 20 human semen samples with 3.69 x 10(-7) to 3.69 x 10(-13) M PAF resulted in statistically significant increases in motility. Treatment of spermatozoa with lyso-PAF (the biologically inactive form of PAF) showed no change in motility. Treatment of spermatozoa with PAF in severely asthenozoospermic males may be of therapeutic value.

Cell Survival↗

Superovulation and recovery of zygotes from Nubian and Alpine dairy goats.

Thirty-five purebred dairy goats (18 Alpines and 17 Nubians) were subjected to a superovulating hormone program consisting of an 11-d 6alpha-methyl-17alpha-acetoxy-progesterone; (MAP; 60 mg) intravaginal sponge treatment; 125 ug i.m. injections of the prostaglandin F(2alpha) analogue cloprostenol on d 1 and 9 of vaginal sponge treatment; and a 3-d, twice-a-day injection of 2.5 mg of pituitary follicle stimulating hormone (FSH-P) i.m. starting at day 9. Vaginal sponges were pulled the morning of day 11 at the time of the fifth FSH-P injection. Of 40 initiated superovulatory cycles, 33 does (10 Alpines and 23 Nubians) responded with an average of 17.7 (range 1 to 29) ovulations. There was no significant difference between the breeds with respect to corpora lutea (CLs) plus follicles ovarian response. A significantly greater (P< 0.05) number of Nubian does were in estrus and mated by 36 h after MAP sponge removal. All does that responded to treatment had done so within 72 h of sponge removal. Of the seven (17.5%) does that showed no estrous response to hormone treatment, six were Alpines (P < 0.01). Six goats (two Alpines and four Nubians) were subjected to a second hormone treatment cycle after a 45-d rest. Five of six does responded to a second hormone treatment cycle with four of five responding with a lower total ovarian response. The interval from sponge removal to mating did not affect the stage or quality of eggs harvested. Rather, the interval from mating to surgical flushing determined the stage of egg development. All animals examined from 24 to 32 h after initial mating had not ovulated. By 50 h, 20 of 22 does had ovulated. A total of 242 ovulated eggs (63%) was harvested, of which 199 (82%) were fertilized. Day 7 flushings yielded 36 eggs (67%), of which 28 (78%) were fertilized. This rate of superovulation, fertilization, and embryo recovery lends credibility to this technique in its ultimate objective of rapidly increasing the number of offspring from superior animals.

Journal Article↗

Visualization of pronuclei in living bovine zygotes.

Bovine embryos were surgically collected from the oviducts of superovulated crossbred heifers 48 h postonset of estrus. The one-celled ova were treated with 4'-6'-diamidino-2-phenylindole (DAPI) and observed under ultraviolet light by fluorescence microscopy. Both male and female pronuclei were visualized, identified and subjected to micromanipulation.

Animals↗

Platelet-activating factor production from in vitro and in vivo fertilized murine embryos is similar.

PROBLEM: This study measured platelet-activating factor (PAF) production of in vitro and in vivo fertilized murine embryos to ascertain whether site of fertilization impacted subsequent embryonic PAF production. METHOD: Oocytes and embryo were collected from stimulated B6D2F1 mice. PAF production throughout the preimplantation phase from the two-celled stage through expanded blastocysts was measured and compared among normally developing embryos with the only difference being site of fertilization. RESULTS: A striking increase in PAF production was noted for both in vivo and in vitro fertilized embryos over the four days of culture. Significantly higher (P < .001) levels of PAF production were noted with development from the two-celled stage (47.22 +/- 4.13; 44.30 +/- 2.43) to expanded blastocysts (254.31 +/- 24.01; 255.11 +/- 5.35 ng/embryo/24 h) for both in vitro and in vivo fertilized embryos, respectively. There was no significant difference (P > .05) in PAF production between the two groups of embryos. CONCLUSIONS: Embryonic PAF production in the mouse increases in a stage-specific manner, and specifically the site of fertilization in vitro versus in vivo does not affect PAF production in normally developing embryos.

Animals↗