• Journal of Veterinary Clinics
• /
• v.13 no.2
• /
• pp.144-148
• /
• 1996

The effectiveness of sodium carboxymethylcellulose (SCMC) and deztran in the prevention of adhesion formation on the uterus and embryo collection in rabbits was elucidated. Following induction of adhesion on uterus and uterine horns by abrasion and retrograde flushing of embryos in gonadotropins primed rabbits, the solutions of saline (for control), 1% SCMC, 10% dextran and a synthetic solution of 1% SCNC and 10% dextran in saline were infused in the abdominal cavity at the dose of 5 ml/kg of body weight. The average percentage of adhesion was 11.1, 28.6, 41.7 and 73.3% in the rabbits infused with the synthetic, 1% SCMC, 10% dextran and saline solutions, respectively. The synthetic solution was more effective than other solutions in the rabbits. The average number and recovery rate of embryos were significantly (P<0.01) higher in the synthetic solution group than 1% SCMC, 10% dextran or saline solution groups. Among the collected embryos in the groups, the distribution of the normal embryos was higher in the synthetic solution group (99%) and the 10% dextran solution group (95.7%) than the 1% SCMC solution group (78.4%) and saline (66.2%). Theretore, a synthetic solution which is combined with 1% SCMC and 10% deztran in saline can be effectively used for the prevention of adhesion formation after uterine surgery and embryo collection.

• Journal of Chest Surgery
• /
• v.27 no.9
• /
• pp.723-731
• /
• 1994

We have modified the isolated perfused working rabbit lung model [IPWL] by perfusing the isolated lung with a hollow fiber membrane deoxygenator.For assessment the stored lung was ventilated with FIO2 0.4 and perfused with 37$^{\circ}$C deoxygenated circulating blood at a rate 5ml/kg/min for several hours until lung failure.We chose to compare our developing solution which contained low potassium and pentastarch with the modified Euro-Collins solution .Experiments were divided into four groups[n=6] based on the type of flushing preservation solution and preservation time.The flushed lungs were then preserved into same solution at 8~10$^{\circ}$C with 100% O2 inflated condition for 1 or 20 hours.These following results were obtained.The IPWL model requires only one animal per experiment and allows for the continuous assessment of aerodynamic performance. This should therefore be used as screening test in lung preservation.One hour preservation groups, there were no significant difference in recovery rates of PaO2, PAP and Paw. Survival time in the one hour preservation groups were very significant long in the Group II[LPPS, p<0.01]. Twenty hours preservation groups, there were no significant difference in the recovery rates of PAP and Paw between Group III[m-ECS] and Group IV[NS], but PaO2 was significantly worse at onset of reperfusion in Group III when compared with Group IV [p<0.05]. And also survival time in the 20 hours preservation groups were significant long in the Group IV [p<0.05].

• Journal of Soil and Groundwater Environment
• /
• v.6 no.1
• /
• pp.45-52
• /
• 2001

This study investigated the effectiveness of ultrasonic waves on the relative permeability under a range of soil type, flushing rate, and sonication power. This study was conducted in the laboratory using a specially designed and fabricated equipment, and the laboratory study was simulated by ECLPISE 100 which is a commercial black oil simulator. The test results indicated the sonication increased contaminant extraction significantly. From analytical standpoint, sonication caused a change in the relative permeability of the test samples, a reduction in residual oil saturation and an increase in both irreducible water saturation and wettability. These three parameters are highly related with $(C_{10})^2$. The computer software ECLIPSE 100 can be used to analyze the change of the relative permeability due to sonication in two phase immiscible flow.

• Korean Journal of Ecology and Environment
• /
• v.38 no.spc
• /
• pp.39-43
• /
• 2005

The dynamics of water quality with the storm events were analyzed in a small reservoir for irrigation, Lake Wangkung. Water quality of the inflowing stream fluctuated seasonally with the variation of flow rate. Thermal stratification was consistent from April to October below 2 m depths and anoxic layer was developed below 2 m depth in summer. The unique feature of temperature showed that thermal stratification was disrupted by a heavy rain event during monsoon, but hypolimnetic hypoxia were reestablished after a few days. Phosphorus and nitrogen increased immediately following storm events. The marked increase may be due to the input of P-rich storm runoff from the watershed. Internal phosphorus loading can be one of the explanations for TP increases in summer. When there was a storm, total populations of phytoplankton and zooplankton was reduced immediately following the storm, indicating possible flushing of algae and zooplankton. After a lag period of low-density the plankton population bloomed to a peak again within five days after the storm. Turbid water in lake became clear again which coincided with the time of the phytoplankton buildup. The results demonstrate that water quality is regulated greatly by rainfall intensity in Lake Wangkung.

• Korean Journal of Animal Reproduction
• /
• v.4 no.1
• /
• pp.75-82
• /
• 1980

This experiment was carried out to improve a simple and effective procedure of egg transfer which is considered to be the most useful technique for the improvementand proliferation of domestic animals. Several experiment procedures such as superovulation, surgical recovery of ovulated egg, in vitro capacitation of ejaculated spermatozoa, in vitro fertilization and culture of embryo were conducted. The results obtained in this experiment were summarized as follows: 1. In rabbits treated with PMS in combination with estradiol and HCG, 10 to 43 eggs were obtained in one rabbit and the average ovulation number of 20 rabbits was 21 eggs. 2. Six to 24 eggs (average 13 eggs) were recovered from the removed oviducts by the methods of flushing technique and the average recovery rate of 20 rabbits was 61.9 percent. 3. Most rabbit spermatozoa ejaculated by artificial vagina were capacitated in vitro by the culture of the spermatozoa with PBI medium at 37$^{\circ}C$ for 4 hours after removal of seminal plasma. 4. Normal fetilization following in vitro fetilization was observed in 88 (42.7%) of 206 eggs. 5. The number of eggs developed to 2-, 4-, and 8-cell stage following in vitro culture with PBI medium at 37$^{\circ}C$ was 26 (70.3%), 20 (54.1%) and 17 (45.9%) of 37 eggs used for in vitro culture.

• Korean Journal of Veterinary Research
• /
• v.34 no.2
• /
• pp.395-406
• /
• 1994

To study the conditions to enhance success of embryo transfer in the dog, 20 mixed-breed bitches were used for the experiment along with 4 male dogs for mating. The bitches were paired according to synchronism of natural estrus, or the counterpart as donor or recipient was treated with gonadotropin as FSH (follicular stimulating hormone) or PMSG (pregnant mare serum gonadotropin) for induction of estrus to be synchronized with estrus of the other bitch in natural estrus. Embryo recovery was performed in two ways for comparison, either by flushing each uterine horn after ovariohysterectomy or by flushing each horn in the state of non-ovariohysterectomy. In addition, the result of pregnancy according to the embryo stage and the repeatability of the experimental animals as donor or recipient were also investigated. FSH or PMSG was administered to the bitches which had passed over 4 months from last estrus, resulting in estrus-positive in 3 dogs of 6 FSH-treated dogs (50.0%), and in 5 dogs of 9 PMSG-treated dogs (55.6%), determined by proestrus signs and vaginal smear test. Estrus-positive bitches induced with gonadotropin were used as donor or recipient resulting in one embryo-recovered bitch as donor and one offspring-delivered bitch as recipient in 5 PMSG-treated dogs, whereas no result was obtained from 3 FSH-treated dogs. The rate of embryo recovery to be compared with number of corpus luteum was 68.2% in ovariohysterectomized dogs and 55.2% in non-ovariohysterectomized dogs, respectively. The number of dogs from which embryo was collected were 4 dogs of 6 ovariohysterectomized dogs (66.7%) and 6 dogs of 7 non-ovariohysterectomized dogs (85.7%), respectively. The result of parturition was obtained from one dog of 5 estrus-induced recipients, whereas no result was obtained from 3 natural-estrus recipients. The only dog which delivered a male puppy had been transferred 3 morulae and 2 blastocysts. Of 6 repeat-used bitches in canine embryo transfer, 3 dogs showed repeatability either as donor or recipient. These results indicated that inducing estrus of a dog with gonadotropin is feasible in canine embryo transfer to be synchronized with that of a natural-estrus dog, that embryo recovery is also possible in non-hysterectomized dogs, that the estrus-induced dog is also usable as recipient to result in parturition, and that repeat-use of a bitch as donor or(and) recipient is possible in canine embryo transfer.

• Journal of Embryo Transfer
• /
• v.13 no.2
• /
• pp.97-106
• /
• 1998

The pronuclear injection of metallothionein-human growth hormone (MT-hGH) gene into rabbit zygotes was performed to establish in vitro developmental system and to detect the presence of the injected gene by nested PCR. Mature female New Zealand White rabbits were superovulated by eGG and hCG treatments. The rabbits were mated and the zygotes were collected from the oviducts 18-22 h after hCG injection by flushing with D-PBS. Two to three picoliters of MT-hGH gene was microinjected into male pronuclei. The foreign gene-injected zygotes were cultured in TCM-199 or RD mediurn containing 10% FCS with a monolayer of rabbit oviductal epithelial cefls in a 5% $CO_2$ incubator. The presence of injected DNA in rabbit embryos or blastomeres at different developmental stages .vas detected by a nested PCR analysis. The results are summarized as follows ; 1.The developmental rate of the MT-hGH gene-injected zygotes to blastocyst was significantly higher in TCM-199 medium (68.1%) than in RD medium (42.9%). 2.The gene injection into pronuclei at 18 or 22 hours post hCG treatment during pronuclear stage did not much affect on the in vitro development of the rabbit embryos. 3.The rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased when they developed to blastocysts. The results indicate that the screening of transgene in rabbit embryos by nested PCR analysis could be a prornisible method for the preselection of transgenic embryos. Furthermore, the preselection of transgenic embryos would greatly reduce hoth the cost and effort of production of transgenic animals.

• Clinical and Experimental Reproductive Medicine
• /
• v.42 no.3
• /
• pp.94-100
• /
• 2015

Objective: The goal of this study was to ascertain optimal assisted hatching (AH) method in frozen embryo transfer. We compared the effect of depending on whether mechanical or laser-AH was performed before or after the vitrification of embryo development rate and blastocyst cell numbers. Methods: In order to induce superovulation, pregnant mare's serum gonadotropin followed by human chorionic gonadotropin were injected into 4- to 5-week-old female mice. 2-cell embryos were then collected by flushing out the oviducts. The Expanded blastocysts were recovered after the collected embryos were incubated for 48 hours, and were then subjected to artificial shrinkage (AS) and cross-mechanical AH (cMAH) or quarter-laser zona thinning-AH (qLZT-AH) were carried out using the expanded blastocysts before or after vitrification. After 48 hours of incubation, followed by vitrification and thawing (V-T), and blastocysts were fluorescence stained and observed. Results: The rate of formation of hatched blastocysts after 24 and 72 hours of incubation was significantly higher in the AS/qLZT-AH/V-T group than in the other groups (p<0.05). The cell number of the inner cell mass was higher in AS/V-T/non-AH and AS/V-T/cMAH groups than those of others (p<0.05). In the control group, the number of trophectoderm and the total cell number were higher than in the AS-AH group (p<0.05). Conclusion: The above results suggest that AS and AH in vitrification of expanded blastocysts lead to the more efficient formation of hatched blastocysts in mice.

• Journal of Embryo Transfer
• /
• v.9 no.2
• /
• pp.153-160
• /
• 1994

This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 $\mu$sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.

• Clinical and Experimental Reproductive Medicine
• /
• v.27 no.3
• /
• pp.275-282
• /
• 2000

Objective: This study was to determine the effect of different concentration of calcium III medium on the preimplantational development of zygotes and early 2 cell embryos. Methods: Female mice of ICR strain ($5{\sim}8$ weeks old) were superovulated and mated with fertile males. Zygotes or early 2-cell embryos were collected by flushing the oviducts $31{\sim}32$ hours after hCG injection. The embryos were cultured in various concentrations of $Ca^{2+}$ in medium or with EDTA, EGTA and $Ni^{2+}$. Result and Conclusion: Treatment of high concentraion of $Ca^{2+}$ (3.42 mM $(2X){\sim}17.l$ mM (10X)) in medium didn't develop well compared to the control. Low concentrations of $Ca^{2+}$ (0.214mM $(1/8X){\sim}0.855$ mM (1/2X) were deterimental to development beyond 2-cell stage. EDTA, $Ca^{2+}$ chelating agent was treated with ranged concentrations of EDTA (0.014 $mM{\sim}0.107$ mM) to medium contaning 1.71 mM $Ca^{2+}$ showed beneficial effect to development to blastocyst compared to the control. EGTA, extracellular $Ca^{2+}$ chelator, was treated with ranged concentrations of EGTA ($0.014{\sim}0.107$ mM) to the medium contaning 1.71 mM $Ca^{2+}$. There is no significant difference with the control. $Ni^{2+}$ (50 ${\mu}M$), T-type $Ca^{2+}$-channel blocker was treated to medium contaning low concentration of $Ca^{2+}$. It overcame 2-cell block significantly. Rate of degenerated embryos decreased and developmental rate to morula and blastocyst increased more than low $Ca^{2+}$ concentration alone. Further studies are needed for the overcoming effect of 2-cell block by $Ni^{2+}$.