This study was carried out to assess the effect of superovulation response and quality of embryos recovered from donor cows after a single subcutaneous injection of FSH dissolved in polyethylene glycol (PEG). Cows were allocated into control and 3 experimental treatment groups. In control, cows were injected intramuscularly 50 mg FSH twice daily for 4 days. Group 1 were injected subcutaneously with a single dose of 400 mg FSH dissolved in 30% PEG solution. Group 2 were injected subcutaneously with a single dose of 200 mg FSH dissolved in 30% PEG solution. Finally in group 3, cows were injected twice 200 mg FSH dissolved in 30% PEG solution by subcutaneous. Superovulation was initiated by injection of FSH between Day 8 and 14 of the estrus cycle (Day 0, the day of estrus), and followed by injection of 25 mg PGF$_2$$\alpha$ at 48 h after first FSH injection. Cows were then artificially inseminated (AI) with semen twice at 48 and 60 h after PGF$_2$$\alpha$ injection. At 7 days after the second AI, embryos collected non-surgically by flushing the uterine horns and were counted and compared morphologically as being transferable and degenerated among different superovulation treatments. Furthermore, progesterone and estradiol -17 $\beta$ in plasma were measured by radioimmunoassay following different treatments at given days All cows of treated groups were observed heat. but control group was showed 77.8%. Superovulation response was observed as 77.8, 87.5, 88.9, and 100% in control, Groups 1, 2 and 3 The mean number of corpus lutea (CL) detected in Group 1 were 19.6, which was, respectively significantly (P<0.05) higher than those of other groups (11.1, 13.4 and 7.6, respectively). However, there did not differ on the mean number of total embryos recovered and of transferable embryos between control and treated groups.
Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
Reproductive and Developmental Biology
/
v.28
no.3
/
pp.181-185
/
2004
This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.
Lung transplantation is the established treatment for the end stage lung disedse find preservation of the organ is a major obstacle In performing lung transplantation. For solving this problem, we evaluated the histopathologic changes for various preservation solutions. Male mongrel dogs of similar size and weight (15∼20 kg) were used. The dog lungs were flushed with 4fl normal saline(group 1 'n:5): Modified Euro-Collins solution(group 2 n:5) and University of Wisconsin solution (group 3 : n=6), 60m11kg through a catheter placed in the main pulmonary artery aft r flushing of PGE 1 (20ng1kg). The lungs were preserved for 60 hours and measured dry and wet weights. Histologic specimens were taken every 6 hours and %toed for light microscopic evaluation. The edema ratio of the lungs peaked in 12 hours although there was no difference between the groups. Histologically, alveolar septal changes developed in one case (20%) after 1 hour preservation with normal saline. In case of the University of Wisconsin solution, the alveolar septal distortions and swellings were seen in 1 cases (20%) after 6 hours preservation compared with 3 cases (60%) after 6 hours preservation with Modified Euro-Collins solution. Changes of the pneumocytes were observed after 24 hours preser- vation in group 1, after 48 hours preservation in group 2 and after 60 hours preservation in group 3. We conclude that University of Wisconsin solution might have a superior preservation effect compare to normal saline and Modified Euro-Collins solutions.
Park, H.-S.;Jung, S.-Y.;Kim, T.-S.;Lee, M.-Y.;Jin, J.-I.;Hong, S.-P.;Lee, J.-S.;Kim, C.-H.
Journal of Embryo Transfer
/
v.19
no.2
/
pp.113-119
/
2004
The purpose of the present study was to examine whether collection time affects results of oocyte recovery from superovulated goats. Fiftyty-one mature Korean native goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen impregnated CIDR for 10 days and then the goats were divided into two groups. One group of the goats received a single intramuscular injection of 1,000 IU PMSG on Day 8 of CIDR insertion. The other group 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 IU hCG in the afternoon on Day 10. For oocyte recovery, donor goats were fasted 24 h before operation. Anesthesia was induced by intravenous injection of 2% xylazine(0.2 mg/kg body weight) and ketamin(11 mg/kg body weight). In vivo oocytes were recovered by follicle aspiration or oviduct flushing at 29 to 34, 35 to 40 and 41 to 50 h after hCG injection through mid-ventral incision. There was no significant difference in the mean number of CL and oocytes recovered. Oocyte collection at 29 to 40 h after hCG increased(P<0.05) the recovery rate of ovulated oocytes in oviducts compared to 41 to 50 h. The same results were also observed in the recovery of follicular oocytes. Oocyte grade was not affected by collection time. When oocytes were collected from follicular oocytes at 41 to 50 h after hCG, the recovery rate of Grade II oocytes was the lowest(P<0.05). From these results, it is suggested that oocyte recovery at 35 to 40 h after hCG will be successful for further use.
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.
Protection against ischemia-reperfusion injury is crucial for successful transplantation of the lung. It has been known that nitric oxide has many favorable effects on the donor lungs but at the same time, has some potential side effects of cytotoxicity. In this regards, we investigated whether the administration of nitroglycerin could decrease ischemia-reperfusion injury in isolated rat lung reperfusion model for the confirmation of the effect of nitroglycerin, a donor of nitric oxide, on lung transplantation. Material and Method: 35 Sprague-Dawley species male white rats were used for this experiment. For nitroglycerin group (n=18), nitroglycerin was administered intravenously followed by mixed in flushing solution for preservation. As a control group (n=17), we used the same amount of normal saline. To evaluate the effect of nitroglycerin on the lung, heart-lung block was obtained, weighed and stored in University of Wisconsin Solution at 1$0^{\circ}C$ for 24 hours. In each group of the isolated lungs, reperfusion was carried out with Krebs-Hensleit-diluted human blood for 60 minutes. As parameters of the state of the isolated lung, peak inspiratory and pulmonary arterial pressures were continuously recorded. Oxygen and carbon dioxide tension of reperfusing blood were measured before and after 30, 60 minutes of reperfusion. After sixty minutes of reperfusion, protein content in bronchoalveolar lavage fluid was measured also for the evaluation of the degree of alveolar flooding. Lung myeloperoxidase activity was determined to verify the accumulation of neutrophils. Results: Although statistically significant differences were not noted in peak inspiratory and pulmonary arterial pressure between control and nitroglycerin group, latter group showed lowering tendency of pulmonary arterial pressure during the entire reperfusion period. Oxygen tension was higher (p<0.05) in nitroglycerin group compared with that of the control group, in contrast, there were no differences in carbon dioxide tension, protein content in bronchoalveolar lavage fluid and myeloperoxidase activity between the groups. In the examination of ultrastructural changes, nitroglycerin denoted the protective effect on the pulmonary architecture compared with that of control group. Conclusion: Collectively, on the bases of these experimental results, prior treatment of donor lung with nitroglycerin could result in better preservation of the lung. Consequently, these nitroglycerin preserved lungs are thought to be more suitable for successful transplantation of the lung.
Purpose : The aims of this study were to verify the incidence of hypohidrosis and to determine the predictive value of noninvasive indicator test ($Neurocheck^{TM}$) for sweating after administration of topiramate in newly diagnosed pediatric epileptic patients. Methods : A total of 46 epileptic patients (22 boys; 24 girls) on topiramate treatment were evaluated in this study at the Department of Pediatrics, Chonbuk National University Hospital, from October 2004 to July 2005. We measured sweating functions using a noninvasive sweating test ($Neurocheck^{TM}$) before topiramate medication, and after 3 months when topiramate reached its target dosage. We performed a direct questionnaire survey for the hypohidrosis related symptoms during topiramate treatment. Results : The mean age was $7.8{\pm}3.2year$. The mean dosage of topiramate was $4.5{\pm}0.8mg/kg/day$. Among the patients, there were 40 complex partial seizures, one simple partial seizure, two partial seizures with secondarily generalization, two generalized seizures, and one Lennox-Gastaut syndrome case. Of the 46 epileptic patients, 17 patients (37.0 percent) experienced hypohidrosis and hypohidrosis related symptoms, 12 (26.1 percent) had facial flushing, four (8.7 percent) had heat intolerance, one (2.2 percent) had lethargy, but no one had anhidrosis. Among the 17 patients, the mild group numbered 12 and the severe group totalled five. Hypohidrosis by $Neurocheck^{TM}$ was diagnosed in 16 patients. The overall measures of agreement between $Neurocheck^{TM}$ and the survey was 76.5 percent. The specificity of this test was 89.7 percent. Patients who showed a time delay after medication, especially over 3 minutes, were seen only in the severe group. Conclusion : $Neurocheck^{TM}$ could be clinically useful to detect and predict topiramate induced hypohidrosis in pediatric epileptic patients. We recommend that patients who show a delay over 3 minutes in $Neurocheck^{TM}$ test after topiramate initiation should be monitored for hypohydrosis.
For Improvement of lung preservation, many tripes of preservation solution were developed and tested. The aim of this study was to compare the effect of the most frequently used extracellular type pieservation solution (Low Potassium Dextran, LPD) with a newly developed trehalose containing extracellular type preservation solution(ET-Kyoto, ETK) on postischemic lung function. Twelve New-Zealand white rabbit lungs were harvested and studied on an isolated, blood-perfused model of lung function after 4 hours of cold ischemia at $10^{\circ}C$ In group I (n=6), lungs were preserved with 100 mL/kg of LPD solution; in group II(n=6), lungs were preserved with 100 mL/kg of ETK solution. A few minutes before flushing with preservation solutions, 20$\mu\textrm{g}$ of PGEI were injected into main pulmonary artery. Functions of the preserved lung were compared with PO2, PA pressure, t acheal air pressure, and drylwet ratio. The pulmonary efferent blood oxygen tension at the end of the 60-minute reperfusion period was higher in group II compacted with group I(486.5 $\pm$ 80.3 mmHg versus $432.5\pm82.9$ mmHg at FiO2 1.0, p-value = NS). The mean pulmonary arterial pressure was similar in both $groups.(33.7\pm2.2$ mmHg versus $35.5\pm2.0$ mmHg, p-value : NS). The peak inspiratory airway pressure was significantly lower in group 11(8.010.6 mmHg versus 11.8 $\pm$ 1.4 mmHg, p-value=0.02) The water content of the lung was lower in group II $(70.2\pm6.9%$ versus 78.5 $\pm6.1%),$ but not significant. These data demonstrate that a newly-developed trehalose-containing ET-Kyoto solution yield equal or slightly superior lung function after reperfusion compared with LPD solution.
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.
This study explored all domestic and international theories for maintenance and health enhancement of an old and big tree, and carried out the anatomical survey of the operation part of the tree toward he current status of domestic surgery and the perception survey of an expert group, and drew out following conclusion through the process of suggesting its reform plan. First, as a result of analyzing the correlation of the 67 subject trees with their ages, growth status. surroundings, it revealed that they were closely related to positional characteristic, damage size, whereas were little related to materials by fillers. Second, the size of the affected part was the most frequent at the bough sheared part under $0.09m^2$, and the hollow size by position(part) was the biggest at 'root + stem' starting from the behind of the main root and stem As a result of analyzing the correlation, the same result was elicited at the group with low correlation. Third, the problem was serious in charging the fillers (especially urethane) in the big hollow or exposed root produced at the behind of the root and stem part, or surface-processing it. The benefit by charging the hollow part was analyzed as not so much. Fourth, the surface-processing of fillers currently used (artificial bark) is mainly 'epoxy+woven fabric+cork', but it is not flexible, so it has brought forth problems of frequent cracks and cracked surface at the joint part with the treetextured part. Fifth, the correlation with the external status of the operated part was very high with the closeness, surface condition, formation of adhesive tissue and internal survey result. Sixth, the most influential thing on flushing by the wrong management of an old and big tree was banking, and a wrong pruning was the source of the ground part damage. In pruning a small bough can easily recover itself from its damage as its formation of adhesive tissue when it is cut by a standard method. Seventh, the parameters affecting the times of related business handling of an old and big tree are 'the need of the conscious reform of the manager and related business'. Eighth, a reform plan in an institutional aspect can include the arrangement of the law and organization of the old and big tree management and preservation at an institutional aspect. This study for preparing a reform plan through the status survey of the designated old and big tree, has a limit inducing a reform plan based on the status survey through individual research, and a weak point suggesting grounds by any statistical data. This can be complemented by subsequent studies.
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