Kim, Kwang-Dae;Kim, Ki-Hyung;Na, Yong-Jin;Lee, Kyu-Sup
Clinical and Experimental Reproductive Medicine
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v.26
no.3
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pp.419-432
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1999
Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.
Objective: This study was designed to evaluate the effects of endogenous LH surge, GnRH agonist (GnRH-a) or human chorionic gonadotropin (hCG) as ovulation trigger on pregnancy rate by intrauterine insemination (IUI). Method: Patients received daily 100 mg of clomiphene citrate (CC) for 5 days starting on the third day of the menstrual cycle followed by human menopausal gonadotropin (hMG) for ovulation induction. Follicles larger than >16 mm in diameter were present in the ovary, frequent LH tests in urine were introduced to detect an endogenous LH surge. Final follicular maturation and ovulation were induced by GnRH-a 0.1 mg (s.c.) or hCG $5,000{\sim}10,000$ IU (i.m.) administration except natural ovulation. Pregnancy was classified as clinical if a gestational sac or fetal cardiac activity was seen on ultrasound. Results: There were no differences in age, duration of infertility and follicle size, but more ampules of hMG were used in GnRH-a group compared to hCG 10,000 IU treated group (p<0.05). Lower level of estradiol ($E_2$) on the day of hCG or GnRH-a injection was observed in hCG 10,000 IU group than other treatment groups (p<0.01). The overall clinical pregnancy rate was 19.8% per cycle (32/162) and 22.2% per patient (32/144). Pregnancy rate was higher in natural-endogenous LH surge group (37.5%, 9/24) than GnRH-a (18.8%) or hCG treated group (20.9% & 13.9%), but this difference was not statistically significant. No patient developed ovarian hyperstimulation. Abortion rate was 22.2% (2/9) in hCG 5,000 IU group. Delivery or ongoing pregnancy rate was 37.5% (9/24), 18.8% (3/16), 16.3% (7/43) and 13.9% (11/79) in endogenous LH surge, GnRH-a, hCG 5,000 IU and hCG 10,000 IU treatment groups, respectively. Conclusion: These results support the concept that use of natural-endogenous LH surge in stimulated cycles may be more effective to obtain pregnancies by IUI than GnRH-a or hCG administration.
To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.
We developed a novel water-soluble camptothecin analobue, CKD602, and evaluated the inhibition of topoisomerase I and the antitumor activities against mammalian tumor cells and human tumor xenografts. CKD602 was a nanomolar inhibitor of the topoisomerase I enzyme in the cleavable complex assay. CKD602 was found to be 3 times and slightly more potent than topotecan and camptothecin as inhibitors of topoisomerase, respecitively. In tumor cell cytotoxicity, CKD602 was more potent than topotecan in 14 out of 26 human cancer cell lines tested, while it was comparable to camptothecin. CKD602 was tested for the in vivo antitumor activity against the human tumor xenograft models. CKD602 was able to imduce regression of established HT-29, WIDR and CX-1 colon tumors, LX-1 lung tumor, MX-1 breast tumor and SKOV-3 ovarian tumor as much as 80, 94, 76, 67, 87% and 88%, respectively, with comparable body weight changes to those of topotecan. Also the therapeutic margin (R/Emax: maximum tolerance dose/$ED-{58}$) of CKD602 was significantly higher than that of topotecan by 4 times. Efficacy was determined at the maximal tolerated dose levels using schedule dependent i.p. administration in mice bearing L1210 leukemia. On a Q4dx4 (every 4 day for 4 doses) schedule, the maximum tolerated dose (MTD) was 25 mg/kg per administration, which caused great weight loss and lethality in <5% tumor bearing mouse. this schedule brought significant increase in life span (ILS), 212%, with 33% of long-term survivals. The ex vivo antitumor activity of CKD602 was compared with that of topotecan and the mean antitumor index (ATI) values recorded for CKD602 were significantly higher than that noted for topotecan. From these results, CKD602 warrants further clinical investigations as a potent inhibitor of topoisomerase I.
Kim, Sung-Min;Kwak, Dong-Hoon;Kim, Sun-Mi;Jung, Ji-Ung;Lee, Dae-Hoon;Lee, Seoul;Jung, Kyu-Yong;Do, Su-Il;Choo, Young-Kug
Archives of Pharmacal Research
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v.29
no.8
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pp.666-676
/
2006
Gangliosides are widely distributed in mammalian cells and play important roles in various functions such as cell differentiation and growth control. In addition, diabetes and obesity cause abnormal development of reproductive processes in a variety of species. However, the mechanisms underlying these effects, and how they are related, are not fully understood. This study examined whether the differential expression of gangliosides is implicated in the abnormal follicular development and uterine architecture of streptozotocin (STZ)-induced and db/db diabetic mice. Based upon the mobility on high-performance thin-layer chromatography, mouse ovary consisted of at least five different ganglioside components, mainly gangliosides GM3, GM1, GD1a and GT1b, and diabetic ovary exhibited a significant reduction in ganglioside expression with apparent changes in the major gangliosides. A prominent immunofluorescence microscopy showed a dramatic loss of ganglioside GD1a expression in the primary, secondary and Graafian follicles of STZ-induced and db/db diabetic mice. A significant decrease in ganglioside GD3 expression was also observed in the ovary of db/db mice. In the uterus of STZ-induced diabetic mice, expression of gangliosides GD1a and GT1b was obviously reduced, but gangliosides GM1, GM2 and GD3 expression was increased. In contrast, the uterus of db/db mice showed a significant increase in gangliosides GM1, GD1a and GD3 expression. Taken together, a complex pattern of ganglioside expression was seen in the ovary and uterus of normoglycemic ICR and $db/^+$ mice, and the correspoding tissues in diabetic mice are characterized by appreciable changes of the major ganglioside expression. These results suggest that alterations in ganglioside expression caused by diabetes mellitus may be implicated in abnormal ovarian development and uterine structure.
This paper examines the American Civil Liberties Union(ACLU)'s recent legal challenge on patents held by Myriad Genetics on two genes (BRCA1 and BRCA2) associated with a high risk of breast and ovarian cancer. Instead of analyzing the ACLU's objections to the BRCA patents in terms of its legal technicalities and normative ethical principles, this paper seeks to situate this legal case in the broader historical context of the shifting understanding of the relationship between private ownership, economic development, and the public interest in academic sciences. This paper first briefly chronicles a series of scientific developments and key legal decisions involving patenting of life forms, including genetically engineered micro-organisms animals and biological materials of human origins like cell cultures and genes, that led to the US Patent and Trademark Office(USPTO)'s official guidelines on human gene patenting in 2001. At another level, this paper analyzes the expansion of the scope of intellectual property rights in the life sciences in terms of shifting economic and legal assumptions about public knowledge and its role for economic development in the 1970s. I then show how these economic, legal, and ethical ideas that linked private ownership and the public interest have been challenged from the 1990s, calling for revisions in intellectual property laws regarding a wide array of life forms. The tragedy of the anticommons in human gene patenting, according to ACLU, has severely undermined creative scientific activities, medical innovations, access to health care and rights to life among cancer patient groups. ACLU's objection to human gene patenting on several US-constitutional grounds in turn suggests issues regarding intellectual property are critically linked to vital issues pertinent to the creative communities in arts and sciences, such as free exchange of ideas, censorship and monopoly, and free expression and piracy etc.
Objective: This study was performed to examine the maturation and the development to the blastocyst stage of immature oocytes collected from patients with high risk of ovarian hyperstimulation syndrome (OHSS). Materials and Methods: Cumulus-oocyte complexes (COCs) were collected following only HCGpriming for non stimulated IVF-ET cycles of the patients. At the time of oocyte collection, COCs were classified into three groups in accordance with their appearance (Group I: oocytes with dispersed cumulus cells; Group II: oocytes with compacted cumulus cells; Group III: oocytes with sparse cumulus cells). The in vitro maturation and blastocyst development rates of the COCs were compared among these groups. From August 2001 to June 2002, 48 IVM/IVF-ET cycles from 42 patients (mean age: $32.4{\pm}3.8$ years) were performed. To prevent the occurrence of OHSS, the patients were primed with 10, 000 IU HCG alone 36 h before oocyte collection without gonadotropin stimulation. Oocytes were aspirated on cycle days from 7 to 13. The normal COCs were classified into three groups according to their appearance. The aspirated immature oocytes were cultured in YS maturation medium containing 30% (v/v) human follicular fluid (HFF), 1 IU/ml FSH, 10 IU/ml HCG and 10 ng/ml rhEGF. Fertilization was induced by intracytoplasmic sperm injection (ICSI). All zygotes were co-cultured with cumulus cells in $10{\mu}l$ YS medium containing 10% HFF until day 7 after oocyte collection. Blastocyst transfer was performed on day 5 after ICSI. Results: Th e mean number of oocytes cultured in the IVM/IVF cycles was $24.7{\pm}10.6$. Of 1185 COCs, those assigned to Group I, II and III were 470 (39.7%), 414 (35.0%) and 301 (25.4%), respectively. The maturation rate (94.5%, 444/470, p<0.05) in Group I was significantly higher than those of Group II (62.8%, 260/414) and Group III (73.1%, 220/301). Especially, 30.9% of COCs in Group I (145/470) was matured on the day of oocyte aspiration. There were no differences in the rates of fertilization and cleavage among the three groups. The development rate to the blastocyst stage in Group I (54.6%, 206/377, p<0.05) was also significantly higher than those in Group II (33.0%, 68/206) and Group III (30.1%, 52/173). Twenty-four clinical pregnancies (50.0%) was obtained and 22 pregnancies (45.8%) are ongoing. Implantation rate in the present study was 24.6%. Conclusion: These results suggest that there is a positive correlation between the appearance of COCs and the developmental competence of the immature oocytes in non stimulated IVM/IVF cycles.
35 patients of normal reproductive age group (22-37 yrs) were selected for ovulation induction using $Clomid^{(R)}$ and HMG. Correlation between canalization of cervical mucus and estradiol level in plasma. morphological ovarian follicular change by ultrasonogram, Ferning and Spinnbarkeit were studied in the induced ovulatory menstrual cycles. By retrospective analysis, the results were following; 1. Canalization appeared on the eighth day of the menstrual cycle in $Clomid^{(R)}$ group, and seventh day of HMG group. And the number of the channels increased gradually until the ovulation time. 2. The mean number of channels increased from 12${\pm}$12 in $Clomid^{(R)}$ group, 19${\pm}$14 in HMG group (Day-6) to a maximum of 46${\pm}$22 in $Clomid^{(R)}$ group (Day-1). 3. The maximum plasma estradiol(E2) level was 812${\pm}$722pg/ml in $Clomid^{(R)}$ group (Day-1), 1792${\pm}$504 pg/ml in HMG group (Day-1), and there was a good statistical correlation between the number of channels and the plasma E2 level. 4. The maximal follicular diameter was 19.4${\pm}$6.7 mm in $Clomid^{(R)}$ Group (Day-1) 19.3${\pm}$4.6mm in HMG group (Day-1) and after than decreased. There was a good statistical correlation between the number of channels and the follicular diameter. 5. When grade of ferning was I, the number of channels were 19.4${\pm}$6.7 in $Clomid^{(R)}$ group, 38${\pm}$26 in HMG group. When grade of ferning was III, the number of channels were 54${\pm}$30 in $Clomid^{(R)}$ group, 71${\pm}$36 in HMG group. 6. The maximum score of Spinnbarkeit was 2.6${\pm}$0.5 in Clomid group (Day-0), 2.6${\pm}$0.5 in HMG group (Day-2). In conclusion, the phenomenon of cervical mucus canalization showed the value of the clinical use as an ovulatin marker.
Many oocytes fail to fertilize and cleave in vitro and many embryos transferred back to uterus fail to implant or maintain implantation. Chromosomal abnormalities in the male and female gametes may contribute to this loss. The higher incidence of meiotic chromosomal abnormalities bas been found in oocytes than in sperm. The wide range of incidence of chromosomal abnormalities in unfertilized oocytes has been reported in human IVF program (26-63%). However, factors affecting chromosomal abnormalities are not well understood. The present study has been conducted to investigate effects of the method for ovarian hyperstimulation, women's age, and the number of oocytes retrieved per patients on the incidence of numerical chromosomal abnormalities. Five hundred eighty four unfertilized metaphase II oocytes were subjected to chromosomal analysis. Included unfertilized oocytes were from 220 patients (mean $age=32.7{\pm}3.0$) and three hundred thirty oocytes were legible for analysis. Two hundred fourty five oocytes out of 330 (73.3%) were normal, while 38 (11.5%) were hyperploidy, 35 (10.6%) were hypoploidy, and 12 (3.6%) were diploidy. Significant difference in chromosomal abnormalities was not found between two patient groups stimulated by follicular stimulating hormone/human menopausal gonadotrophin (FSH/HMG) (25.9%) and gonadotrophin-releasing hormone agonist/follicular stimulating hormone/human menopausal gonadotrophin (GnRHa/FSH/HMG) (28%). There was a tendency of increasing chromosomal abnormalities in unfertilized oocytes from older patients (<30 yrs: 20.3%, 30-34yrs: 26.9%, >34 yrs: 35.3%). The number of oocytes retrieved per patient had no effect the incidence of chromosomal abnormalities (1-5: 31. 4%, 6-10: 29.8%, 11-15: 28.6%, > 15: 16.5%). These results from the present study suggest that the chromosomal abnormalities observed in the unfertilized oocytes has not affected by the stimulation methods, patient's age, and the number of oocytes retrieved per patients.
Objective: The objective of this retrospective study was to compare the in vitro fertilization (IVF) outcomes of gonadotropinreleasing hormone (GnRH) agonist and GnRH antagonist protocols in poor responders. Methods: A total of 172 cycles in subjects with less than 5 oocytes retrieved treated with either GnRH agonist long protocols or antagonist protocols were included. The outcome variables such as numbers of growing follicles and retrieved oocytes, and the fertilization rate were evaluated as the main outcome measures. Results: There was no difference in regard to the numbers of growing follicles and oocytes, and fertilization rate between the two groups. $E_2$ level on Day 7/8, mean gonadotropin dose, and the days of stimulation were shown to be statistically different (p<0.01, respectively). Conclusion: Considering that similar results were observed with less time and gonadotropin dose, GnRH antagonist protocol may be considered as a preferable choice over GnRH agonist protocols in poor responders.
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