• Korean Journal of Agricultural Science
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• v.13 no.1
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• pp.82-89
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• 1986

This experiment was carried out to determine the optimum freezing and thawing rates of the hamster embryos. The female hamsters were induced to superovulate by intraperitoneal injections of 30 i.u. PMSG and mated with males of the same strain of 4 days the PMSG injection. They were killed and embryos were flushed from the oviduct and uterine horn on 3 days after mating. Embryos were flushed with a modified Dulbecco's phosphate-buffered saline and equilibrated with 1.5 M-dimethylsulphoxide by a 3-step procedure. The freezing rates of the samples were $1^{\circ}C/min$ from room temperature to $-6^{\circ}C$ and the samples were seeded at $-6^{\circ}C$. After being held for 3 min at the seeding temperature, the rates were $0.3^{\circ}C/min$ from $-6^{\circ}C$ to $-35^{\circ}C$. From $-35^{\circ}C$ to $-70^{\circ}C$, the rates were divided into $0.1^{\circ}C/min$, $1^{\circ}C/min$ and $10^{\circ}C/min$, respectively. At $-70^{\circ}C$ the samples were plunged directly into liquid nitrogen. The samples were thawed at $4^{\circ}C/min$ and $12^{\circ}C/min$ from $-196^{\circ}C$ to $37^{\circ}C$, and for 2 min in $37^{\circ}C$ water bath, respectively. The average numbers of ovulation points and embryos recovered were 35.1 and 27.0 appearing 77.0% recovery rates. Eight cell embryos in the embryos recovered were 24.8. The survival rates of embryos according to the freezing rates were 55.5~67.7% at $0.1^{\circ}C/min$, 58.8~64.9% at $1^{\circ}C/min$ and 40.5~44.7% at $10^{\circ}C/min$, respectively. The survival rates at $10^{\circ}C/min$ were significantly low. The survival rates of embryos according to the thawing rates were 53.5% at $4^{\circ}C/min$, 53.7% at $12^{\circ}C/min$ and 59.1% in $37^{\circ}C$ water bath. The survival rates, in $37^{\circ}C$ water bath were slightly higher, but we did not find any differences among them. In conclusion, the best freezing rates of hamster embryos were $1^{\circ}C/min$ from the room temperature to $-6^{\circ}C/min$, $0.3^{\circ}C/min$ from $-6^{\circ}C/min$ to $-35^{\circ}C$ and $-0.1^{\circ}C/min$ or $1^{\circ}C/min$ from $-35^{\circ}C$ to $-70^{\circ}C$. The hamster embryos thawed for 2 min in $37^{\circ}C$ water bath showed the best survival rates.

• Journal of Embryo Transfer
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• v.28 no.1
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• pp.19-24
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• 2013

This study assesses of efficiency of oocyte recovery and in vitro development for during the non breeding season in goat. Thirty-four matured goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injections of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotropin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes were activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I+II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytes. The clavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes. There was no significant difference in oocyte quality between seasonal treatments.

• Tuberculosis and Respiratory Diseases
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• v.48 no.3
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• pp.357-364
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• 2000

Background : Complicated exudative pleural fluid collections have traditionally been treated by either closed tube thoracostomy drainage or by open surgical drainage. Complete drainage is important in order to control pleural sepsis, restore pulmonary function, and entrapment. Recently intracavitary fibrinolytic therapy has been advocated as a method to facillitate drainage of complicated exudative pleural effusion and to allow enzymatic debridemant of the restrictive fibrinous sheets covering the pleural surface. The purpose of this study is to prospectively evaluate the effects of image-guided catheter drainage with high dose urokinase(UK) instillation in the treatment of complicated pleural effusions. Patients : Twenty complicated pleural effusion patients that poorly respond to image-guided drainage were allocated to receive UK. There were 8 pneumonia and 12 tuberculosis. Methods : Drugs were diluted in 250 mL normal saline and were infused intrapleurally through the chest tube or pig-tail catheter in a daily dose of 250,000 IU of UK. Response was assessed by clinical outcome, fluid drainage, chest radiography, pleural ultrasound and/or computed tomography. Results : The mean UK instillation time was $1.63{\pm}0.10$. The mean volume drained UK instillation was $381.3{\pm}314.4\;mL$, and post-UK was $321.6{\pm}489.5\;mL$. The follow up duration after UK therapy was mean $212.9{\pm}194.5$ days. We had successful results in 19 cases (95.0%). There were 12 pleural thickenings (60.0%), 2 markedly decreased effusions (10.0%) and 5 cases of no thickening or effusion. There was recurrence after treatment in only one patient(5%) with complicated pleural effusiondue to tuberculosis. Conclusions : Image-guided drainage with high dose UK instillation (250,000 U/day) in complicated pleural effusion is a safe and more effective method than closed thoracostomy drainage. And this management, in turn, can obviate surgery in most cases.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.61-68
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• 2003

Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to improve reproductive efficiency of artificial insemination with fresh- and frozen-semen following estrus induction in dog. Fifty infertilie dogs (age 2~3 years) were selected fur the study and divided into three different estrus induction treatment groups. Group 1 : dogs (n=15) were given clomifene (0.1 mg/kg) orally f3r five days at 12 hr intervals. Croup 2: dogs (n=15) were given bromocriptine (50 $\mu$g/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Croup 3, n=20) when pro-estrus occurred. After being treated, the dogs were evaluated fur the rates of estrus induction and time interval lapses from treatment to beginning of the pro-estrus. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The ejaculated semen was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM; TES, 209 mM; citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 co above the surface of liquid nitrogen (L$N_2$) for 23 min. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, there was no significant differences in the pregnancy rate of dogs between group treated with a combination of GnRH and bromocriptine and group treated clomifene or bromocriptine only. However, frozen-thawed semen can be used successfully fur artificial insemination in dog.

• Clinical and Experimental Pediatrics
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• v.46 no.1
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• pp.76-82
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• 2003

Purpose : The aim of this study was to determine whether differences exist in the presentation, clinical course, and outcome of Graves' disease between prepubertal children and adolescents. Methods : A retrospective chart review of 14 prepubertal(PREPUB, $7.2{\pm}0.9yr) and 38 pubertal (PUB, $12.4{\pm}1.5yr$) children with Graves' disease between January 1989 and November 1995 at St. Mary's Hospital and Kangnam St. Mary's Hospital was undertaken. Results : There were no significant differences in $T_3$, $T_4$, TSH between two the groups at diagnosis. The PUB group had significantly higher titers of antimicrosomal antibody(positive dilution factor $11,727.3{\pm}22,888.4$) than did the PREPUB group($2,111.5{\pm}2,285.0$, P<0.001). The PREPUB group had significantly higher titers of TSH-binding inhibitory immunoglobulin(TBII, $62.5{\pm}39.6$) than did the PUB group($44.9{\pm}10.4$, P<0.05) before treatment started. The duration(months) of medical therapy before thyroid function tests were normalized was longer in the PREPUB group than in the PUB group($T_3:6.8{\pm}5.0$ vs. $5.4{\pm}13.2$, $T_4:2.3{\pm}1.9$ vs. $2.1{\pm}2.2$, $TBII:26.7{\pm}24.0$ vs. $20.8{\pm}12.1$), especially that of TSH was significantly longer in the PREPUB group($14.6{\pm}11.0$ vs. $6.8{\pm}7.8$, P< 0.05). Total length of medical therapy was significantly longer in the PREPUB group than the PUB group($52.3{\pm}19.3$ vs. $37.9{\pm}16.3months$, P<0.01). During three years of antithyroid drug therapy, in the PREPUB group, the remission rate was lower and the relapse rate was higher than in the PUB group. Total length of treatment correlated negatively with chronological age(P=0.03). Conclusion : Prepubertal children require longer medical therapy to achieve a remission than do pubertal children. But there is an obvious need for more studies because of the small number of patients and the short duration of the follow-up.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.53-61
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• 2007

This study was investigated factors affecting the pregnancy rates after transfer of pronuclear microinjected embryos for the production of transgenic Korean black goats. Embryo transfer was carried out in 343 recipient Korean black goats from September 1999 to June 2000. Estrus was induced by the insertion of intravaginal progesterone devices $CIDR^(R)$ for 2 weeks. A single injection of 400 IU equine chorionic gonadotropin was administered at 48h before $CIDR^(R)$ removal to increase the proportion of does cycling and ovulation rate. Good quality embryos were prepared by microinjection of DNA into the pronuclei of fertilized goat oocyte and cultured in vitro. Pronuclear microinjected $1{\sim}8$ cell stage embryos were surgically transferred into the oviducts of the recipient at day 4 or 5 following $CIDR^(R)$ removal, and morula to blastocyst stage embryos were surgically transferred into uterus at day 9. Pregnancy was diagnosed by transrectal ultrasound scanning at $20{\sim}30d$ and 8 weeks following embryo transfer. The pregnancy rate was affected by several factors, such as estrus induction, the number of previous transfer, transfer site, stage of CL (corpus luteum), the number of recipient CL, stage of embryos and the number of transferred embryo. The pregnancy rate was significantly higher in recipients that came into estrus naturally than recipients that induced to come into estrus with $CIDR^(R)$(59.1% vs. 36.8%; P<0.05). The pregnancy rate was higher when the embryos were transferred into the left oviduct than transferred into the right oviduct (42.9% vs. 35.3%; P<0.05). The pregnancy rate of recipients with $CH_1$ (early) stage corpus hemorrhagicum in ovary was hi틴or than recipient with $CH_3$ (late) stage hemorrhagicum (47.5% vs. 17.9%; P<0.01). Higher pregnancy rates were obtained by transfer of 1-cell stage embryos into oviduct while late blastocysts (51.6% vs. 66.7%; P<0.01) into uterus. The pregnancy rates when 3 embryos were transferred to recipients were significantly higher than when 2 embryos we.e transferred (47.6% vs. 27.0%; P<0.05). Although there were no significant difference among the group, adhesion of reproductive organs, uterine size, ovulation rate of recipients, presence of large follicle and difficulty of transfer affected pregnancy rate of recipient. Higher pregnancy rates were obtained in the recipients with $8{\sim}15m$ diameter uterine horn as compared to the recipients with <5m diameter or >20mm diameter uterine hem (38.9%, 20% vs. 18.2%), in the recipients with large follicle in the ovulated ovary ipsilaterally (53.6% vs. 37.1%) and in the transfer which was carried out easily (39.2% vs. 27.8%, 0%). In conclusion, the high rate of pregnancy was achieved following transfer of pronuclear microinjected embryos when three or four 1-cell stage embryos were transferred into oviduct with $CH_1$ stage corpus hemorrhagicum in the ovary of recipient which came into estrus naturally.

• Journal of Embryo Transfer
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• v.26 no.3
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• pp.147-152
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• 2011

This study was conducted to examine whether efficiency of oocyte production from superovulated prepubertal goats. Fifteen prepubertal and twenty adult goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotrophin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes was activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at 38.5$^{\circ}C$ in an atmosphere of 5% CO$_2$, 5% O$_2$, 90% N$_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I + II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytcs. The cleavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.282-290
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• 1997

Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.7 no.1
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• pp.61-73
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• 2004

Purpose: Liver function test abnormalities have been reported frequently in patients receiving total parenteral nutrition (TPN). In adults, it is known that liver complications decrease with the use of cyclic parenteral nutrition (CPN), especially if the shift to cycling was not too late. However, there are few studies about the effects of cycling on liver injury in children beyond the neonatal period. The aim of this study is to evaluate the effect of the early use of CPN on total parenteral nutrition induced hepatic dysfunction. Methods: Twelve sets of CPN in 11 children (2 months to 17 years) were included in this study. Data on underlying diseases, age, length of time on TPN, macronutrient intake, complications, and biochemical parameters were collected from clinical records. All children had received CPN in the early period of persistent transaminase elevation or cholestasis complicated by previous continuous PN. The duration of infusion off-time in CPN was 2 hours in patients less than 3 months of age and 4 hours in the older children. Results: All 12 cases showed elevated aminotransferase and 5 of them also showed cholestasis. Serum total bilirubin concentration was normalized in all 5 cases with median periods of 8 days (p<0.05) after initiation of CPN. ALT either decreased significantly or was normalized in all cases with median periods of 30 days (p<0.05) on CPN. The CPN was well tolerated without significant complication except for one case of hyperglycemia. Conclusion: The early use of cyclic parenteral nutrition had a beneficial effect in improving hepatic dysfunction complicated by TPN in children.

• Journal of Yeungnam Medical Science
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• v.15 no.1
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• pp.29-35
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• 1998

Rats were studied during 45 minutes treadmill exercise to determine the effects of hyperglycemia and hyperlipidemia on the utilization of cardiac muscle glycogen, and the utilization of diaphragm muscle glycogen was also studied for comparing to cardiac muscle. The hyperglycemia was produced by ingestion of 25% glucose solution(lml/100gm, BW) and the hyperlipidemia by 10% intralipose ingestion(lml/l00gm, BW) with intraperitoneal injection of heparin(500 IU) 15 minutes before treadmill exercise. The mean blood glucose concentrations(mg/dL) in control and hyperglycemic rats were 110 and 145, respectively, and the mean plasma free fatty acid concentrations(${\mu}Eq/L$) in control, control exercise(control-E) and hyperlipidemia exercise(HL-E) rats were 247, 260 and 444, respectively. In the hyperglycemic trial, the cardiac muscle glycogen concentration was not significantly decreased by the exercise but the concentration in control rats was decreased to 73.9%(p<0.05). The glycogen concentration of diaphragm was significantly decreased in both groups by the exercise, but the hyperglycemia decreased the glycogen utilization by approximately 10% compared to the control. The cardiac muscle glycogen concentration was not decreased by the exercise in control and hyperlipidemic rats but the utilization of glycogen in hyperlipidemic rats is lower than that of the control. These data illustrate the sparing effect of hyperglycemia on cardiac muscle glycogen usage during exercise, but the effect of hyperlipidemia was not conclusive. In the skeletal muscle, the usage of glycogen by exercise was spared by both hyperglycemia and hyperlipidemia.