• Ocean and Polar Research
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• v.37 no.1
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• pp.49-59
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• 2015

Spatial variation in the reproductive effort of Manila clam Ruditapes philippinarum is often closely associated with variation in the seawater temperature and food availability, which determines gonad maturity and the quantity of gamates produced during spawning. Previous studies also have reported that severe infection by the protozoan parasite Perkinsus olseni exerts a negative impact on clam reproduction, retarding gonad maturation or decreasing the reproductive effort. In the present study, we investigated impacts of P. olseni infection on the reproductive condition of Manila clam during a spawning season. Histology revealed that 54% of female clams in Wando off the south coast were in spawning, while only 10% of the female from Gomso and 0% of the female from Seonjaedo in Gyeonggi bay off the west coast were engaged in spawning at the end of May in 2004. Ray's fluid thioglycollate media (RFTM) assay was applied to assess P. olseni infection and indicated that the infection intensity in Wando ($3,608,000{\pm}258,000cells/g$ wet tissue) was significantly higher than the levels in Gomso ($1,305,000{\pm}106,000cells/g$ wet tissue) and Seonjaedo ($1,083,000{\pm}137,000cells/g$ wet tissue, p < 0.001). The size of the ripe female follicle determined from histology was significantly smaller in Wando ($0.032mm^2$) compared to the sizes in Gomso ($0.059mm^2$) and Seonjaedo ($0.052mm^2$, p < 0.05). Accordingly, the number of ripe eggs in the follicle was significantly fewer among clams in Wando (14) compared to the numbers determined in Gomso (23) and Seonjaedo (22). The absolute quantity of egg in ripe clams from Wando (31.01 mg) was also significantly smaller than Seonjaedo (61.79 mg) and Gomso (133.3 mg). Quantity of total protein, carbohydrate, and lipid in the tissue in the Wando samples was significantly smaller than the quantities determined in Gomso and Seonjaedo (p < 0.001). The observed poor reproductive condition and proximate tissue composition of the females in Wando were, in part, explained by the extremely high level of the parasites, sapping the ability to store energy in the host tissues, which is used in tissue growth and the egg production.

• Clinical and Experimental Reproductive Medicine
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• v.15 no.1
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• pp.40-52
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• 1988

Estradiol-17${\beta}$($E_2$) levels in the blood were estimated according to varying the time and amount of the administration of $Clomid^{(R)}$. $Clomid^{(R)}$ were administered on the 2nd, 3rd and 4th menstrual day corresponding to the recruitment period and on the 5th menstrual day corresponding to the selection period of the ovarian follicles, respectively. And $Clomid^{(R)}$ were administered 50 mg, 100 mg and 150 mg/day, repectively. The effects of the sequential HMG to $E_2$ levels in the blood were also estimated. The results were as following : 1. Blood $E_2$ levels according to the day and amount of administration of $Clomid^{(R)}$ were the highest in the group 3(D $2{\sim}6$, 150 mg/day, with HMG) and the lowest in the group 6(D $5{\sim]9$, 50 mg/day, without HMG). $E_2$ levels showed increasing tendency to 0 day. 2. In the cases of the administration of $Clomid^{(R)}$ during the $2nd{\sim}6th$ menstrual day, $E_2$ levels according to the amount were similar among groups and showed increasing tendency daily. 3. In the cases of administration of $Clomid^{(R)}$ during the $2nd{\sim}6th$ menstrual day, $E_2$ levels according to the sequential HMG independent of the amount of $Clomid^{(R)}$ were higher in the with HMG group than without HMG groups. 4. In the case of the administration of $Clomid^{(R)}$ during the $5th{\sim}9th$ menstrual day, $E_2$ levels according to the amount were the highest in the 100 mg/day group and the lowest in the 50mg/day group. 5. In the cases of administration of $Clomid^{(R)}$ independent of the amount during the 5th${\sim}$9th menstrual day, $E_2$ levels according to the sequential HMG were higher in the with HMG group than without HMG group. 6. $E_2$ levels according to the amount independent of the day of the administration of $Clomid^{(R)}$ were the highest in the 100 mg/day group and 150 mg/day, 50 mg/day group in low sequence. 7. $E_2$ levels according to the sequential HMG independent of the day and amount of the administration of $Clomid^{(R)}$ were higher in the with HMG group than the without HMG group. 8. $E_2$ levels according to the day of the administration of $Clomid^{(R)}$ independent of the amount of $Clomid^{(R)}$ and sequential HMG were the highest in the group D 2${\sim}$6 and the lowest in the group D 5${\sim}$9. According to the above results, there were higher $E_2$ levels in the group with sequential HMG than without HMG. Therefore, the hypothesis, postulated initially by the author, was not verified that sequential HMG would not affect the $E_2$ levels which were related to the process of the selection of the ovarian follicle in the connection with 'FSH window'. Because it may be the stimulation after the selection of later predominant follicle. And the highest level of $E_2$ was estimated in the $Clomid^{(R)}$ 150 mg/day group with sequential HMG on the 2nd${\sim}$6th day, and the higher levels were estimated in the 2nd${\sim}$6th day, 3rd${\sim}$7th day and 4th${\sim}$8th day groups than the 5th${\sim}$9th day group. The lower levels were estimated in the $Clomid^{(R)}$ 50 mg/day group without HMG than 100 mg/day and 150 mg/day on the 5th${\sim}$9th day. Therefore, further study will be needed that combines analyses of the E2 levels in the blood according to the various administration of $Clomid^{(R)}$ with or without sequential HMG and determination of the numbers and size of the ovarian follicles by ultrasonogram.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.63-70
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• 2009

Objective: This study was performed to investigate the effectiveness of minimal stimulation using rhFSH and GnRH antagonist compared with GnRH antagonist multidose protocol (MDP) in IVF treated patients with aged 40 and above. Methods: Seventy-five patients with aged 40 and above were equally randomized to minimal stimulation group (n=37) or GnRH antagonist MDP group (n=38). For minimal stimulation group, ultrasound monitoring was started on cycle day 7 or 8. Daily injections of 0.25 mg cetrorelix together with 150 IU rhFSH were started from the day at 13${\sim}$14 mm of a leading follicle diameter. For GnRH antagonist MDP group, daily injections of 225 IU rhFSH were initiated from cycle day 2 and GnRH antagonist was started at a dose of 0.25 mg/day on rhFSH stimulation day 6 or the day at 13${\sim}$14 mm of leading follicle diameter. In both groups, transvaginal ultrasound-guided oocyte retrieval was performed. According to cleavage and morphologic characteristics of embryos, embryos were transferred 3 to 5 days after oocyte retrieval. Results: There were no differences in patients' characteristics and cycle cancellation rate between the two groups. Total dose and duration of rhFSH used were significantly fewer and shorter in minimal stimulation group than those in GnRH antagonist MDP group. The numbers of oocytes retrieved, mature oocytes and transferred embryos were also lower in minimal stimulation group. However, there were no significant differences in the clinical pregnancy rate and miscarriage rate between the two groups. Conclusions: This study demonstrates that minimal stimulation protocol provides comparable pregnancy rates to GnRH antagonist MDP with fewer dose and days of rhFSH used, and thus can be a cost-effective alternative in women aged 40 and above.

• Korean Journal of Environmental Biology
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• v.20 no.3
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• pp.224-231
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• 2002

Photoperiod (length of light per day) is a major factor in regulating reproductive function in golden hamsters. The information of photoperiod is transmitted to the reproductive endocrine system by melatonin. Thus the effects of melatonin aye investigated in male golden hamsters exposed to photoperiods. Paired testicular weights were markedly reduced in the animals housed in short photoperiod $(SP,\le{12\;hours\;day^{-1})$ and injected with melatonin in the evening, but not in long photoperiod $(LP,\le{12.5}\;hours\;day^{-1})$ and injected with melatonin in the morning. The histological examination of regressed testes showed reduction of tubular lumen diameter including the numbers of cells and Leydig cell number. The mean values of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) were also lowered in the sexually inactive animals than in the sexually active animals. Melatonin receptor was identified by reverse-transcription polymerase chain reaction (RT-PCR) and its expression was examined in various tissues to scrutinize the action site of melatonin. It turned out 309 nucleotides and was definitely expressed in hypothalamus and pituitary including spleen, retina, and epididymis. And gonadotropin releasing hormone (GnRH) gene, which is a key element in regulating reproduction, was identified by RT-PCR but the expression of GnRH was not modified by the treatment of melatonin. Taken together, photoperiod via melatonin indirectly affects reproductive endocrine system, possibly through the release of GnRH, not the synthesis of GnRH.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.67-67
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• 2003

Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

• Development and Reproduction
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• v.18 no.3
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• pp.145-152
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• 2014

This study was performed to investigate the effect of of transdermal testosterone gel (TTG) on controlled ovarian stimulation (COS) and IVF outcomes and ovarian morphology according to pretreatment duration in poor responders. A total of 120 women were recruited for this pilot study. They were randomized into control, 2 weeks, 3 weeks or 4 weeks TTG treatment groups. For three TTG treatment groups, 12.5 mg TTG was applied daily for 2 weeks, 3 weeks or 4 weeks in preceding period of study stimulation cycle. After 3 weeks of TTG pretreatment, significant increase of antral follicle count (AFC) and significant decreases of mean follicular diameter (MFD) and resistance index (RI) value of ovarian stromal artery were observed (p=0.026, p<0.001, p<0.01, respectively). The total dose of rhFSH administered for COS significantly decreased after 3 and 4 weeks TTG treatment both compared with control group (p<0.001, p<0.001). The numbers of oocytes retrieved and mature oocytes were significanty higher in 3 and 4 weeks TTG treatment groups than control group (p<0.001, p<0.001 in the number of oocytes retrieved; p<0.001, p<0.001 in the number of mature oocytes). The clinical pregnancy rate and live birth rate were increased only in 4 weeks TTG treatment group compared with control group (p=0.030 and p=0.042, respectively). These data demonstrated that TTG pretreatment for 3 to 4 weeks increases AFC and ovarian stromal blood flow, thereby potentially improving the ovarian response to COS and IVF outcome in poor responders undergoing IVF/ICSI.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1534-1541
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• 2001

Forty-five Large White gilts were used to study the effect of energy intake from 28 to 176 d of age on body composition and reproductive development. From 28 to 60 d, the gilts were fed ad libitum a 16.6 MJ DE/kg, 24% crude protein and 1.3% total lysine diet. From 61 d of age three dietary treatments were used; 1) ad libitum access to feed (15.6 MJ DE/kg, 21% crude protein and 1.07% total lysine) (H), 2) feed offered at 75% (M) of the previous days intake of H, and 3) feed offered at 60% (L) of the previous days intake of H. ADG from 61 to 176 d of age was (p<0.05) affected by treatment. Although live weight at 176 d of age did not differ (p>0.1) the H gilts had higher (p<0.08) carcass weights than the M or L gilts. Back fat depths were similar (p>0.1) for all treatments at 115 d of age, however by 176 d of age M and H gilts were fatter (p<0.1) than L gilts. The mean lipid deposition (LD) from 115 to 176 d of age for L gilts (78.9 g/d) was less (p<0.05) than for M gilts (143.6 g/d) and H gilts (135.6 g/d). There were no differences between treatments for protein deposition (PD) over the same period. More (p<0.05) H gilts (n=8) attained puberty (first observed estrus) than either M gilts or L gilts (n=4 for both). Follicle numbers were similar (p>0.1) across treatments. For gilts that attained puberty, H gilts had fewer (p<0.05) follicles (13.5) than M gilts (19.7) and L gilts (21.3). For gilts with follicular development, H gilts had the heaviest (458.7 g) reproductive tract weight (RTW). However, for those that attained puberty, L gilts had the heaviest RTW. RTW were lowest for those with no follicular development. Energy restriction had a negative impact on puberty attainment, i.e. it took longer to reach puberty. However, for gilts that attained puberty, the number of follicles was greater for those on r feed intakes. It would appear that rate of fat deposition, but not necessarily the total amount of fat, plays an important role in puberty attainment.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.2
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• pp.133-139
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• 2004

Objective: To investigate the association of FSH receptor (FSHR) polymorphism at position 680 with outcomes of controlled ovarian hyper-stimulation for IVF-ET in Korean women. Design: Genetic polymorphism analysis. Materials and Methods: The FSHR polymorphism was analyzed by PCR-RFLP in 172 ovulatory women below the age of 40 year. Patients with polycystic ovary syndrome, endometriosis, or previous history of ovarian surgery were excluded. Results: Genotype distribution was 41.9% for the Asn/Asn, 47.7% for the Asn/Ser, and 10.5% for the Ser/Ser FSHR genotype group. There was no difference in age of subjects and infertility diagnosis between genotype groups. When the patients were grouped according to their FSHR genotype, the basal levels of FSH (day 3) were significantly different among the three groups ($6.0{\pm}0.3\;IU/L$ (mean $\pm$ SEM), $5.8{\pm}0.3\;IU/L$, and $8.6{\pm}1.2\;IU/L$ for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively, p=0.002). The Ser/Ser group showed a higher total doses of gonadotropins required to achieve ovulation induction, and a lower serum estradiol levels at the time of hCG administration compared with other two groups, but the differences were of no statistical significance. The numbers of oocytes retrieved were significantly different among the three groups ($8.6{\pm}0.8$, $9.9{\pm}0.6$, and $6.3{\pm}0.9$, for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively, p=0.049). Clinical pregnancy rates were 42.4%, 25.9%, and 29.4% for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively. Conclusion: Homozygous Ser/Ser genotype of FSHR polymorphism at position 680 was associated with decreased ovarian response to gonadotropin stimulation for IVF-ET.