• Microbiology and Biotechnology Letters
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• v.36 no.2
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• pp.128-134
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• 2008

In order to examine efficient L-threo-2,3-Dihydroxyphenylserine (L-threo-DOPS) synthesis process using whole cell biocatalyst, a thermostable L-threonine aldolase (L-TA), which cloned from Streptomyces coelicolor A3(2) and improved for stability, was expressed in a Corynebacterium glutamicum R strain. The constructed Corynebacterium expression vector, pCG-H44(1) successfully expressed L-TA in C. glutamicum R strain, but showed very low expression level. In order to improve the expression level, the expression vector named pCG-H44(2) was reconstructed by eliminating 1 nucleotide between SD sequence and start codon of L-TA. The pCG-H44(2) vector plasmid was able to overexpress L-TA approximately 3.2 times higher than pCG-H44(1) in C. glutamicum R strain (CGH-2). When the whole cell of CGH-2 was examined in a repeated batch system, L-threo-DOPS was successfully synthesized with a yield of 4.0 mg/ml and maintain synthesis rate constantly after 30 repeated batch reactions for 130 h.

• Animal Bioscience
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• v.37 no.6
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• pp.1007-1020
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• 2024

Objective: We increased the nuclear maturation rate of antral follicle derived oocytes by using a pre-in vitro maturation (IVM) culture system and improved the developmental potential of these porcine pathenotes by supplementing with melatonin. Furthermore, we investigated the expression patterns of genes involved in cumulus expansion (HAS2, PTGS2, TNFAIP6, and PTX3) derived from small and medium antral follicles before and after oocyte maturation. Methods: Only the cumulus oocyte-complexes (COCs) derived from small antral follicles were induced with [Pre-SF(+)hCG] or without [Pre-SF(-)hCG] the addition of human chorionic gonadotropin (hCG) during the last 7 h of the pre-IVM period before undergoing the regular culture system. The mature oocytes were investigated on embryonic development after parthenogenetic activation (PA). Melatonin (10^-7 M) was supplemented during in vitro culture (IVC) to improve the developmental potential of these porcine pathenotes. Results: A pre-IVM culture system with hCG added during the last 7 h of the pre-IVM period [Pre-SF(+)hCG] effectively supported small antral follicle-derived oocytes and increased their nuclear maturation rate. The oocytes derived from medium antral follicles exhibited the highest nuclear maturation rate in a regular culture system. Compared with oocytes cultured in a regular culture system, those cultured in the pre-IVM culture system exhibited considerable overexpression of HAS2, PTGS2, and TNFAIP6. Porcine embryos treated with melatonin during IVC exhibited markedly improved quality and developmental competence after PA. Notably, melatonin supplementation during the IVM period can reduce and increase the levels of intracellular reactive oxygen species (ROS) and glutathione (GSH), respectively. Conclusion: Our findings indicate that the Pre-SF(+)hCG culture system increases the nuclear maturation rate of small antral follicle-derived oocytes and the expression of genes involved in cumulus expansion. Melatonin supplementation during IVC may improve the quality and increase the blastocyst formation rate of porcine embryos. In addition, it can reduce and increase the levels of ROS and GSH, respectively, in mature oocytes, thus affecting subsequent embryos.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.53-59
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• 1997

This study was conducted to investigate the effect of hormones on in vitro maturation of porcine follicular oocytes. The results obtained were as follows : 1. The maturation rates of oocytes cultured in medium containing FSH 1, 10, 50 and 100$\mu\textrm{g}$/ml were 44.6, 58.5, 42.6 and 37.9%, respectively. And those were no difference to maturation rates among the medium containing FSH. However, the maturation rate(13.7%) of oocytes cultured without FSH was significantly(P<0.05) lower than those of oocytes cultured with FSH. 2. The maturation rates of oocytes cultured in medium containing hCG 0, 1, 10, 50 and 100IU/ml were 12.2, 11.9, 17.9, 21.9 and 45.6%, respectively. The maturation rate of oocytes cultured with 100IU/ml hCG was significantly(P<0.05) higher than those of oocytes cultured with 0~50IU/ml hCG concentrations. 3. The maturation rates of oocytes cultured in medium containing E2 0, 1, 10, 50 and 100$\mu\textrm{g}$/ml were 13.7, 10.5, 14.9, 11.4 and 5.9%, respectively. There were no differences to maturation rates among the E2 concentrations. 4. The FSH＋hCG treatment was the highest maturation rate in medium containing different combination of FSH, hCG and E2.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.39-45
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• 2007

This study was performed to induce the estrus in 9 anestrus Shih-tzu bitches by intramuscular injection of pregnant mare serum gonadotrophin (PMSG) 50IU/kg for 10 consecutive days and by intravenous injection of human chorionic gonadotrophin (hCG) 1,000IU/Head on Day 10. The day when the first injection of PMSG was counted as Day 0 of experiment. All of the bitches were monitored by vaginal discharges, displays the perineal region, vaginal swelling and male acceptances. The 9 bitches (100%) showed vaginal discharges and vaginal swelling, and were mated. The 5 bitches out of 9 bitches were pregnant (55.6%) and 4 bitches were non-pregnant (44.4%). The 3 bitches out of 5 pregnant bitches were spontaneously delivered (33.3%) and litter size were $1.66{\pm}1.15\;(1{\sim}3\;pups)$ pups. The 2 bitches were diagnosed as early embryonic death on days 38 and 41 after first injection of PMSG. These results indicated that rates of estrus induction, pregnancy and delivery were 100%, 55.6% and 33.3%, respectively, using PMSG and hCG.

• Korean Journal of Exercise Nutrition
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• v.23 no.1
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• pp.48-54
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• 2019

[Purpose] The combined effect of different types of post-exercise treatment has not been fully explored. We investigated the effect of combined cold water immersion (CWI) and compression garment (CG) use after maximal eccentric exercise on maximal muscle strength, indirect muscle damage markers in the blood, muscle thickness, and muscle soreness score 24 h after exercise. [Methods] Ten men performed two trials (CWI + CG and CON) in random order. In the CWI + CG trial, the subjects performed 15 min of CWI (15℃), followed by wearing of a lower-body CG for 24 h after exercise. In the CON trial, there was no post-exercise treatment. The exercise consisted of 6 × 10 maximal isokinetic (60°·s^-1) eccentric knee extensions using one lower limb. The maximal voluntary contraction (MVC) and maximal isokinetic (60°·s^-1) strength during knee extension, as well as the indirect muscle damage markers, were evaluated before exercise and 24 h after exercise. [Results] The maximal muscle strength decreased in both trials (p < 0.001), with no difference between them. The exercise-induced elevation in the myoglobin concentration tended to be lower in the CWI + CG trial than in the CON trial (p = 0.060). The difference in the MVC, maximal isokinetic strength, muscle thickness, and muscle soreness score between the trials was not significant. [Conclusion] CWI followed by wearing of a CG after maximal eccentric exercise tended to attenuate the exercise-induced elevation of indirect muscle damage markers in the blood.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.235-241
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• 1994

The study was conducted to determine the optimal hormone and glucose levels during the in vitro culture of bovine oocytes matured and fertilized in vitro for blastocyst development. Oocytes matured in TCM 199 + 10% FCS + hormones and glucose were fertilized in vitro in a TALP medium with swim up separated and heparin-treated epididymal cauda spermatozoa. Oocytes were cultured for 2~5 days in synthetic oviduct fluid medium (SOFM) supplemented with 10% FGS and with different hormone and glucose levels, and further cultured 5 days same medium in SOFM. The results are summarized as follows : The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + $\beta$ estradiol, PMSG + $\beta$ estradiol 0 to20 hours after insemination were 88.0% and 81.8%, 82.6% and 68.4%, 80.0% and 75.0%, 80.0% and 65.0%, 77.3% and 64.7%, respectively. The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + $\beta$ estradiol, PMSG + $\beta$ estradiol 20 to 40 hours after insemination were 92.0% and 87.0%, 92.0% and 82.6%, 91.3% and 81.0%, 85.2% and 73.9%, 87.5% and 81.0%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose lelvels 0~3 days after insemination were 31.5~48.1% and 10.0~16.7%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose levels 4~8 days after insemination were 30.0~53.8% and 8.7~19.2%, respectively. The cleavage and in vitro developmental rates to blastocyst were higher in TCM 199 media containing various glucose levels 0~3 days after insemination than 4~8 days.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.177-182
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• 2014

Feline ovulation time after LH surge have not been defined because its LH surge is occurred by several times of coital vaginal induction and cat has relatively longer time between LH surge and ovulation compared with other mammalian species. This study was performed to investigate the feline ovulation time after LH surge that was induced by hCG injection for superovulation with PMSG. For superovulation, all cats were received an initial injection of PMSG (200 IU, i.m.) followed 80 hrs later with an injection of hCG (200 IU, i.m.). And then, sampling of both ovaries was surgically performed at each 6 different times (10, 18, 22, 26, 29, and 32 hrs) after hCG injection. Cumulus-oocyte-complexes (COCs) were collected from 2 sides of oviducts and ovaries were fixed for ovarian histology. Total 38 COCs were collected only at hCG 32 hrs and no COCs were shown at earlier 5 times. However, in the ovarian histology, corpus haemorrhagicum or corpus luteum was not shown in all groups including ovary at hCG 32 hrs that COCs were collected. In conclusion, it was suggested that feline ovulation was occurred at 29~32 hrs after LH surge and taken relatively long time for CL formation after ovulation.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.4
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• pp.300-305
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• 2020

Objective: The feasibility of a gonadotropin-releasing hormone agonist (GnRHa) trigger in normal responders is still a matter of debate. The aim of this study was to compare the number of mature oocytes, the number of good-quality embryos, and the live birth rate in normal responders triggered by GnRHa alone, GnRHa and human chorionic gonadotropin (hCG; a dual trigger), and hCG alone. Methods: A retrospective cohort study was conducted at the infertility clinic of a university hospital. Data from 200 normal responders who underwent controlled ovarian hyperstimulation and intracytoplasmic sperm injection with a GnRH antagonist protocol between January 2016 and January 2017 were reviewed. The first study group consisted of patients with cycles triggered by GnRHa alone. The second study group consisted of patients with cycles triggered by both GnRHa and low-dose hCG (a dual trigger). The control group consisted of patients with cycles triggered by hCG alone. Results: The groups were comparable in terms of demographics and cycle characteristics. The numbers of total oocytes retrieved and metaphase II oocytes were similar between the groups. The total numbers of top-quality embryos were 3.2±2.9 in the GnRHa group, 4.4±3.2 in the dual-trigger group, and 2.9±2.1 in the hCG group (p=0.014). The live birth rates were 21.4%, 30.5%, and 28.2% in those groups, respectively (p=0.126). Conclusion: In normal responders, a dual-trigger approach appears superior to an hCG trigger alone with regard to the number of top-quality embryos produced. However, no clinical benefit was apparent in terms of live birth rates.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.2
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• pp.153-162
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• 1991

The present study was undertaken to clarify the role of calcium ion as a factor for the maturation of follicle-enclosed mouse oocytes. Follicles were isolated with two sharp needles under a stereomicroscope from mouse(ICR) ovaries which were treated PMSG 5 IU 45 hours previously. Isolated follicles were cultured for 14-16 hours in an organ culture system at $37^{\circ}C$, 5% $CO_2$ in air and in a 100% humidified incubator by treatment of hCG, EDTA and $^{45}Ca^{++}$. Culture medium was Modified Hank's Balanced Salt Sol. (MHBS) and addition of hCG (human chorionic gonadotropin) was made into two doses level 0.4 IU and 0.8IU from the stock sol. and also $^{45}Ca^{++}$ was treated in the culture medium. To explain the role of calcium, calcium chelating agent EDTA was treated to the culture of the mouse follicle-enclosed oocytes. Two observations were made in the present study; nucleus phase and $^{45}Ca^{++}$ uptake into the oocyte. HCG induced oocyte maturation in the follicle about two folds as much as the control group, whereas there is no difference in oocyte maturation between 0.4 IU and 0.8 IU of hCG. Optimum level of hCG seems to be 0.4 IU/ml in the mouse follicle culture. HCG stimulated $^{45}Ca^{++}$ uptake into the oocyte of the follicles by two folds. $^{45}Ca^{++}$ uptake in the control group is about 2.5 folds in comparison of the EDTA(1.71mM) treated group. However, calcium uptake in the EDTA treated groups tends to increase depending on the decrease of EDTA concentration. These observations suggest that firstly, hCG stimulates maturation of the oocyte of the follicle, secondly, $Ca^{++}$ influx is induced by hCG and thirdly, $Ca^{++}$ influx by the treatment of EDTA decreases as a dosage-dependent process. This $Ca^{++}$ uptake may take place by the changes of permeability which was induced by hCG treatment. That is, $Ca^{++}$ influx may trigger the resumption of oocyte maturation. It is further necessary in the future study how this $Ca^{++}$ uptake is induced by hCG and increases permeability of the follicle and oocyte.

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