• Clinical and Experimental Reproductive Medicine
• /
• v.31 no.2
• /
• pp.95-103
• /
• 2004

Objective: To compare COH characteristics and IVF outcomes among IVF-ET patients who were treated with various therapeutic modalities for ovarian endometriomas and to propose effective pre-cyclic therapeutic modalities to improve IVF-ET outcomes in the patients with ovarian endometriomas. Methods: All cases that had undergone IVF-ET after laparoscopy between January 1997 to August 2003 were reviewed. Forty-eight patients with tubal factor were assigned to Group I. Twenty seven, 22 and 38 patients diagnosed as severe pelvic adhesion with ovarian endometriomas by laparoscopy received only medical therapy (Group II), cyst aspiration (Group III), and sclerotherapy (Group IV), respectively. Laparoscopic cystectomy was performed in 20 patients (Group V). Resistance index was measured on day administering hCG. Results: As compared with Group I, in Group II resistance index increased (p<0.05) but number of oocytes, good-quality oocyte ratio (mature and intermediate oocytes/total retrieval oocytes), fertilization rate, and embryo development rate decreased (p<0.05). In Group III fertilization rate and embryo development rate decreased (p<0.05). There was no difference between Group IV and Group I in all parameters except basal FSH which increased (p<0.05). In Group V basal FSH, and resistance increased (p<0.05) and number of oocytes and good-quality oocytes ratio decreased (p<0.05). Conclusion: Sclerotherapy is an effective therapeutic option which can be done prior to IVF-ET cycles in the patients with ovarian endometriomas. Further studies on a large scale are necessary to confirm these data.

• Journal of Nutrition and Health
• /
• v.36 no.2
• /
• pp.154-166
• /
• 2003

To investigate the effects of isoflavone supplementation on both bone mineral density and hormone variation in premenopausal women who had decreased bone mass, the 24 subjects were divided into two groups: one was the underweight group, consisting of 13 subjects, and the other was the normal weight group consisting of 11 subjects. For each group, we investigated the effects of isoflavone supplementation of 90 mg/day on both bone mineral density and hormone variation during 3 menstrual cycles. Anthropometric measurements, dietary recall, and analyses of blood and urine were assessed from baseline to post-treatment. The results were as follows: The average age of the underweight group was 21.8 years old and that of the normal weight group was 23.2 years old. The comparative results for the two groups at baseline were as follows: Onset of menarche, menstrual cycle, and menstrual length were not significantly different between the groups. Serum protein, total, HDL-, LDL-cholesterol, triglyceride, Ca, P, Mg, Cu, and Zn level were not significantly different between the groups. Serum estradiol, SHBG, LH, and FSH level were also not significantly different between the groups. Lumbar spine BMD by T scores of the underweight group was significantly lower than that of the normal weight group. Serum osteocalcin, urinary DPD, and urinary pH were not significantly different between the groups. The comparative results for the two groups at post-treatment were as follows: From baseline to post-treatment, the intake of energy, nutrients and isoflavone in food did not significantly change in either group. Serum protein, total cholesterol, HDL-, LDL-cholesterol, and triglyceride levels did not significantly change in either group. Serum Ca, Cu, and Zn levels were significantly lower in both groups and serum Mg level significantly decreased only in the underweight group. Serum estradiol levels were significantly lower in both groups, but serum SHBG, LH, and FSH levels did not significantly change in either group. Lumbar spine BMD by T score of the underweight group significantly increased to 15%, but that of the normal weight group did not significantly change. Serum osteocalcin of the underweight group significantly increased to 28%, while that of the normal weight group significantly increased to 40%. Urinary DPD of the normal weight group significantly increased to 12%. The results show that the BMD of the underweight group was lower than that of the normal weight group. Therefore, the underweight group had a disadvantage in obtaining maximum bone mineral density. The results also show that isoflavone supplementation during 3 menstrual cycles was effective in increasing the bone mineral density of the lumbar spine and affected bone metabolism markers in premenopausal underweight women. Therefore, it can be concluded that sufficient intake of isoflavone could be helpful in preventing decreases in bone mass among premenopausal women, especially underweight women.

• Clinical and Experimental Reproductive Medicine
• /
• v.17 no.2
• /
• pp.173-184
• /
• 1990

At infertility clinic, department of Obstetrics and Gynegology, Kyung Hee Medical Hospital, 80 patients who underwent IVF-ET from January to July, 1989 were evaluated for the prediction of endogenous LH Surge and its effects on outcome of controlled ovarian hyperstimulation (COH) were compared among LH Surge group without hCG given (N=18), with hCG given (N=5), and no-LH Surge group with hCG given (N=57). LH Surge were occurred in 23(28.7%) out of 80 patients studied. Serum E2 levels on Day-1, Day 0, Day+1, were no significant different among three groups. When basal serum LH/FSH ratio is above 1.0, the possibility of endogenous LH Surge is much higher (56.3% in LH Surge group without hCG given). Serum P4 levels on Day 0 were significantly increased in LH Surge group without hCG given. Cycles which serum P4 level is higher than l.0ng/ml were 70.6% of LH Surge group without hCG given. But there was no significant interrelationship between endogenous LH Surge and serum P4 rising rate as an efficient predictor of the occurrence of endogenous LH Surge in COH for IVF. There was no significant differences in number of follicles, follicular size on Day-1, Day 0, Day+1, and number of oocyte collected per cycle. The oocyte fertilization rate of No-LH surge group with hCG given was significantly higher than LH Surge group without hCG given. There was no significant difference in oocyte cleavage rate among three groups.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.22-22
• /
• 2001

To produce reconstituted rabbit embryos with fetal fibroblasts, the present study was evaluated the efficiencies of the different fusion and activation conditions as assessments of subsequent development and chromosome in the embryos. New Zealand White rabbits were used throughout the study. Fetal fibroblasts collected from 22-d of fetuses were cultured in DMEM ＋ 10% FBS in 5% $CO_2$ in air. The culture was maintained for 10 passages. In every passage half of cell suspension were kept In frozen. From rabbits treated with FSH in 30% PVP solution and hCG, oocytes were surgically collected from oviducts at 14 h post-hCG injection and stripped off their cumulus cells by re-pipetting in a 300 IU hyaluronidase solution. Oocytes with an extruded first polar body and dense cytoplasm were enucleated by micromanipulation in Ham's F-10 medium＋7.5 g/$m\ell$ cytochalasin B. Euncleation was confirmed under a fluorescence microscope after staining with 5 g/$m\ell$ bisbenzimide for 2 min. Each enucleated oocyte was injected with a fetal fibroblast into a perivitelline space. Reconstructed eggs were compared fusion rates either at 2.0 ㎸/cm or 1.6 ㎸/cm(60 sec, double pulses). After fusion, all eggs were activated with the combination of 5 M ionomycin (5 min) and 10 g/$m\ell$ cycloheximide (CHX, 3h), and cultured in CRlaa medium and transferred into TCM199＋10% FBS on day 3. Although there was not significantly differ in fusion rate between treatments (60%, 2.0 ㎸/cm vs. 79.4%, 1.6 ㎸/cm), none of them in the eggs fused with 2.0 ㎸/cm developed to blastocyst. In comparison of development and chromosome status between different activation treatments (Group 1; 5 M ionomycin/10 g/$m\ell$ CHX, Group 2; 5 M ionomycin/5 g/$m\ell$ CHX ＋ 2 mM DMAP after fusion with 1.6 ㎸/cm), there were not differ in cleavage and development rates (67.3% and 28.9% in Group 1; 67% and 33% in Group 2). All out of 8 embryos evaluated in Group 1 appeared a normal diploid chromosome sets and mean number of cells (Mean SEM) on day 4.5 of culture was 141.5 23.15 (n=8). It can be concluded that the use of cycloheximide has not happened in chromosome abnormalities, and fetal fibroblasts can be used for cloning in rabbit.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.10
• /
• pp.1403-1406
• /
• 2005

The present study was undertaken to study the effect of thermal stress on sexual behaviour of superovulated ewes. Fourteen adult Bharat Merino ewes with an average body weight of 29.4${\pm}$2.34 kg were randomly allocated into two groups of 7 each. All the animals were grazed on natural pasture in the morning and evening hours and housed in shed during night. Animals of Group-1 were housed in shed from 10:00 to 16:00 h while the animals of Group-2 were exposed to thermal stress in a hot chamber ($40^{\circ}C$ /6 h/day). All the animals were offered drinking water once a day at 16:30 h. Meteorological observations i.e. dry bulb, wet bulb, minimum and maximum temperature were recorded daily inside the shed as well as in hot chamber throughout the experimental period. For superovulation of animals, standard protocol developed at the Institute, using FSH (Ovagen 5.4 mg in eight injections) and PMSG (200 IU) was followed. Various sexual behaviour parameters (circling, tail fanning, head turning, standing and approaching to ram) and estrus incidence (onset of estrus and estrus duration) were observed in both the groups. The different estrus symptoms were graded subjectively on arbitrary scale of 0-5 where 0 representing no sexual behaviour (0%) and 5 representing maximum intensity in sexual behaviour (100%). Estrus was detected with the help of a marked aproned ram of proven vigor at six hourly intervals. The average percent values for sexual behaviour parameters recorded in Group-1 and Group-2 animals were 53.7${\pm}$3.76 vs. 41.1${\pm}$2.18 for circling, 71.8${\pm}$5.42 vs. 49.0${\pm}$4.41 for tail fanning, 64.7${\pm}$3.30 vs. 44.5${\pm}$4.34 for head turning, 90.1${\pm}$3.16 vs. 75.8${\pm}$4.02 for standing and 63.8${\pm}$4.8 vs. 41.9${\pm}$4.58 for approaching to ram. Animals exposed to thermal stress had significantly lower values of these sexual behaviour parameters. The animals kept in shed exhibited estrus earlier (25.4${\pm}$1.12 h) and duration was higher (37.7${\pm}$1.59 h) as compared to animals exposed to thermal stress i.e. 30.6${\pm}$1.16 h and 31.7${\pm}$3.57 h, respectively. The findings of the study indicate that thermal stress reduces the intensity of sexual behaviour in ewes and may result in failure of the animal to mate and conceive.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2002.11a
• /
• pp.80-80
• /
• 2002

This study was carried out that to investigate the effects of amino acids supplemented with culture medium on development of porcine embryos cultured in vitro. Cumulus oocyte complexes (COCs) were cultured in the maturation medium containing hormones (0.5$\mu\textrm{g}$/$m\ell$ LH, 0.5$\mu\textrm{g}$/$m\ell$ FSH and 1$\mu\textrm{g}$/$m\ell$ estradiol-17${\beta}$) for 20-22 h at 39$^{\circ}C$ in an atmosphere of 5% CO$_2$in air. Subsequently, COCs were cultured in hormone-free maturation medium for 20-22 h. After maturation for 40-44h, oocytes were removed cumulus cells by pipetting and cultured with epididymal sperm for 5 h in the mTBM. Embryos obtained were divided in 4 groups (1) cultured in NCSU 23 containing 0.4% BSA to blastocyst stage(Control), (2) essential amino acids (EA), (3) non-essential amino acids (NA), (4) mixture of essential and non essential amino acid (EA+NA). All treated groups(2-4) were used a glucose free NCSU 23 medium supplemented with pyruvate (0.33 mM), lactate (4.5 mM) to morula stage. From morula to blastocyst stage embryos of all treated groups were cultured in NCSU 23 containing 0.4% BSA. The rates of cleaved oocytes at 48 h after IVF were from 82% to 88% in the groups of control, EA, NA and EA+NA, respectively. The in vitro developmental rates into blastocysts in the groups of EA and EA+NA were significantly (P<0.05) higher than those of group of control (35.1, 35.4 vs. 19.4%, respectively), however, no significant (P<0.05) between control and NA. In conclusion, supplemented with essential amino acid or mixture of essential and non essential amino acid in the culture medium at morula stage increased the rate of development to blastocyst on in vitro produced porcine embryos.

• Journal of Embryo Transfer
• /
• v.7 no.2
• /
• pp.125-131
• /
• 1992

The nudear changes of bovine oocytes during 24 hrs. of culture for mejotic maturation were examined. Bovine oocytes were collected from small(<2 mm), medium(2~6 mm) and large(>6mm) follicles and classified into three grades by their morphological characteristics. A total of 242 oocytes collected were obtained:from 184 small, 157 medium and 1 large follicles, respectively and were classified into 95 grade I, 155 grade H and 92 grade III oocytes. All the bovine oocytes collected and graded were washed with a basal medium and incubated in groups of 10 for 24 hrs in 5% $CO_2$ and 39$^{\circ}C$. The basal medium used was composed of TCM-199 supplemented with sodium bicarbonate, sodium pyruvate, streptomycin, penicillin G and 10% FCS. The oocytes were cultured in drops of 50,$\mu$l basal medium supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH and 1$\mu$g /ml estradiol-17$\beta$. The oocytes were fixed and examined on their chromosomal status by 1% acetorcein staining in the interval of 3 hrs. Most of the grade I oocytes developed to germinal vesicule stage at 0 to 3 hrs., germinal vesicle breakdown at 6 hrs., metaphase I at 9 to 15 hrs., anaphase I and telophase I at 18 hrs., and metaphase II and the first polar body at 24 hrs. after culture for meiotic maturation. However, it was found that compared to grade I oocytes, grade H and W oocytes reached earlier to germinal vesicle breakdown and most of them developed earlier to M II stage at 21 hrs. after culture.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.6
• /
• pp.842-849
• /
• 2007

The purpose of the present study was to produce live offspring from in vitro fertilized goat embryos. Oocytes were collected from abattoir ovaries and kept in oocyte collection medium. Oocytes were washed 4-5 times with maturation medium containing medium-199 with 5 ${\mu}g/ml$ FSH, 100 ${\mu}g/ml$ LH, 1 ${\mu}g/ml$ estradiol-$17{\beta}$ 50 ${\mu}g/ml$ gentamycin, 10% inactivated estrus goat serum, and 3% BSA (fatty acid free). Oocytes were placed in 100 ${\mu}l$ drops of maturation medium containing granulosa cell monolayer and incubated in a 5% $CO_2$ incubator at $38.5^{\circ}C$ for 27 h. For capacitation of spermatozoa fresh semen was processed and mixed in 3 ml fertilization TALP medium containing 50 ${\mu}g/ml$ heparin and kept in the above incubator for 2 h. The capacitated spermatozoa were coincubated with matured oocytes for fertilization. Cleaved embryos were separated and cultured in embryo development medium with oviductal cells and 494 embryos were produced. Recipient goats were synchronized with two injections of 15 mg $PGF_{{2}{\alpha}}$/goat 10 days apart. Eighty early stage embryos were transferred into the uterotubal junction of 14 surrogate mothers using laparoscopy techniques. One recipient delivered twin kids, whereas another two recipients each.delivered a single kid The parentage of these kids was evaluated using highly polymorphic co-dominant microsatellites markers. From the present study, it was concluded that live goat kids can be produced from in vitro matured and fertilized goat embryos, to the best of our knowledge for the first time in India.

• Clinical and Experimental Reproductive Medicine
• /
• v.15 no.1
• /
• pp.25-34
• /
• 1988

The intrafo1licular echoes of cumulus oophoruses within ovarian follicles were assessed with the use of ultrasound in 86 women taking part in an in vitro fertilization(IVF) or gamete intrafallopian transfer(GIFT) program, stimulated with pure follicle-stimulating hormone(FSH)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin (hCG). When intrafo1licular echoes were clearly separated from the follicular wall or relatively dispersed within the follicle, they were considered to be a dissociated cumulus, and when they were only slightly prominent from the follicular wall, they were suspected to be a nondissociated cumulus. A cumulus was seen in 62.1% of the follicles larger than 10 mm diameter and 75.1% of them were dissociated. The larger the follicles in size, the more the cumuluses in number and dissociation. The number of follicles and intrafollicular echoes per woman was not different whether or not she would be pregnant, but the number of dissociated cumuluses was significantly more in pregnant women. The number of observed dissociated cumuluses correlated significantly with the number of recovered mature oocytes. When an intrafollicular echo is seen, it can be taken as evidence of a sign of maturity of that particular follicle and oocyte. Ultrasonographic monitoring of intrafollicular echoes and follicular size is very helpful to predict follicular maturation in ovulation stimulation cycles.

• Clinical and Experimental Reproductive Medicine
• /
• v.23 no.3
• /
• pp.277-282
• /
• 1996

Serum inhibin concentrations, determimed by radioimmunoassay, were measured in women undergoing pituitary suppression with Decapeptyl and subsequently ovarian stimulation with Highly Purified-Metrodin(HP-FSH) to appraise follicular development. Early follicular basal serum inhibin level correlated with the number of oocytes retrieved(r=0.89, n=8, p<0.05). The number of oocytes retrieved showed a significant correlation with serum inhibin level on the day of hCG administration(r=0.73, n=8, p<0.05). The number of mature oocytes showed a significant correlation with serum inhibin level on the day of hCG administration(r=0.73, n=8, p<0.05). These data suggest that: (1) In the early follicular phase, basal serum inhibin may be a valid index to predict ensuing follicular growth : (2) In the preovulatory phase, maximum serum inhibin may be one of the indexes of follicular development during hyperstimulation cycles.