• Asian-Australasian Journal of Animal Sciences
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• v.14 no.9
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• pp.1260-1266
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• 2001

The effect of protein supplementation, $O_2$ concentration and co-culture on the development of embryos produced by nuclear transfer using cultured cumulus cell was investigated. Recipient oocytes and cumulus cells were obtained from the ovaries of the slaughtered Hanwoo cows. Donor cumulus cells were cultured in Dulbecco's modified Eagle medium containing 10% fetal bovine serum at 5% $CO_2$ in air at $38.5^{\circ}C$. The 1 to 6 passages of cumulus cells were isolated and used as donor cells. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. One $15{\mu}s$ pulse of 180 volts was applied to induce the fusion between karyoplast and cytoplast. The fused embryos were activated with $10{\mu}M$ calcium ionophore for 5 min and 2 mM 6-dimethylaminopurine for 3 h. To examine the effect of protein supplementation, nuclear transfer (NT) embryos were cultured in one of the following 4 treatments : 1) CR1aa + 3 mg/ml BSA for 7 days ; 2) CR1aa + 10% FBS for 7 days ; 3) CR1aa + 1.5 mg/ml BSA + 5% FBS for 7 days ; and 4) CR1aa + 3 mg/ml BSA for first 3 days and then CR1aa + 1.5 mg/ml BSA + 5% FBS for 4 days. Culture took place at 5% $CO_2$, 5% $O_2$ and 90% $N_2$ at $38.5^{\circ}C$. Although there were no significant differences in cleavage rate among different protein supplements, the rates of blastocyst formation were significantly different. When NT embryos were cultured in the medium supplemented with only BSA, they could develop to only morula not to blastocyst. However, when FBS was supplemented, NT embryos developed to blastocyst stage. In order to investigate the effect of $O_2$ concentration and co-culture, NT embryos were cultured in CR1aa + 1.5 mg/ml BSA + 5% FBS with or without cumulus cell co-culture at an atmosphere of 5% $CO_2$ in air (20% $O_2$) or 5% $CO_2$, 5% $O_2$, 90% $N_2$ (5% $O_2$) at $38.5^{\circ}C$ for 7 days. The percentage of blastocyst development was significantly higher when the NT embryos were cultured at an atmosphere of 5% $O_2$ than that of 20% $O_2$ (p<0.05). However, there was no significant difference between with and without cumulus cell co-culture at an atmosphere of 5% $O_2$ or 20% $O_2$. Fifty embryos were transferred to 25 recipients and 5 recipients were pregnant at 100 days. From 5 pregnant cows, only one cow was delivered of female twin. In conclusion, the embryos reconstructed by enucleation of metaphase II oocytes and introduction of the cycling and quiescent cumulus donor cells in Hanwoo had developmental potential to term after embryo transfer to recipient cows.

• Clinical and Experimental Pediatrics
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• v.45 no.12
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• pp.1503-1511
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• 2002

Purpose : Although the short- and long-term outcomes of low birth weight(LBW) neonatal intensive care unit(NICU) survivors have been extensively studied, much less information is available for normal birth weight(NBW) infants(greater than 2,500 gm) who require NICU care. Methods : We retrospectively examined the neonatal hospitalizations and one year health status of 302 NBW and 131 LBW admissions to our NICU. Information on the neonatal hospitalization was obtained from a review of medical records. Postdischarge health status was collected by using telephone surveys and medical records. Results : After initial discharge, 21.2% of the NBW infants and 23% of the LBW infants required rehospitalization during the first year of life and there was no significant difference between the two groups. The reasons for rehospitalization of the NBW infants included respiratory disorders (32.1%), G-I problems(26.2%), genitourinary problems(11.9%), surgery(10.7%), cardiac problems(7.1%), and congenital/developmental problems(1.2%). For the LBW infants, the order of frequency was the same, with the percentages slightly different. Neonatal risk factors related to the rehospitalization of the NBW infants included mechanical ventilation, duration of mechanical ventilation, and congenital anomaly. But no positive significant correlation of neonatal risk factors with rehospitalization of LBW infants was found. Conclusion : Low and normal birthweight NICU survivors were rehospitalized at similar rates. The most common cause of rehospitalization was respiratory problems. Neonatal risk factors related to rehospitalization of NBW infants were mechanical ventilation, duration of mechanical ventilation, and congenital anomaly. However, no positive significant correlation of neonatal risk factors with rehospitalization of LBW infants was found. The data suggests that NBW infant survivors, as well as LBW infant NICU survivors, require close follow up.

• Parasites, Hosts and Diseases
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• v.31 no.1
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• pp.57-66
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• 1993

The potential of fumagillin dicyclohexylamine salt to treat and prevent intestinal giant-cystic disease in Israel carp, Cyprinus carpio nudus, was monitored in field experimental studies. In experiment 1 (therapeutic), most fish were already naturally infected with more advanced stage of Relohqnellu: kitnuet. Fumagillin was administered to ash (mean body weiht of 830 g for a Penod of one month at a dose of 10.62 mg in the first group and 5.3 mg in the second group per fi sh per day. In experiment 2 (fprophylactlcl), most flesh also were already naturally infected with an early developmental stage of the protozoa and fish (average body weight of 484 g) were administered fumagillln for 45 days at a dose of 3.95 mg per fish per day. In both experiments, the cumulative mortalities of fish and the extrusion rates of the polar filaments of the spores were significantly decreased in a dose-independent fashion. In experiment 2 no dead fish were observed. No adverse side effects of the drug were observed among fish from any dosage group. In experiment 2, an oval or dot-like concave lesion of most cysts developed at the 7th day and the vegetative form was never observed at the 17th day postmedication and the cysts were grossly reduced in size as compared with the control group, beginning at the 24th day until the end of the study. In contrast, it was scarcely effective to the cysts in experiment 1. Taking the seasonal development of the protozoa into consideration, the above results revealed that oral administration of fumagillin at 3.95 mg/500 g body weight/day for a month Is the optimal dose for the treatment and prevention of thelohanellosls caused by T kitnuei among C. carpio nudus.

• Korean Journal of Weed Science
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• v.10 no.4
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• pp.269-276
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• 1990

Weed species belonging to the broadleaf family prevailed in the experimental sites followed by Cyperucea and grass family. In terms of life cycle, the dominant weed species were the perennial weeds rather than annual weeds. Most of weeds were emerged between 30-60days after transplanting in Chuncheon and between 40-60 days after transplanting in Hongcheon and Hoengseong areas. Among agronomic characteristics of rice affected by weed emergence were decrease of plant height by 2-4㎝, panicle number by 1.3-2.9, spkelet by 3. 7-7.5, ripening rates by 3.3-6.5%, and milled yield by 12-17%.

• Korean Journal of Weed Science
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• v.10 no.2
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• pp.83-92
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• 1990

Experiment was conducted to establish the weed control system on the paddy fields by investigating species, period and amount of weeds in Gangwon province. Four locations were selected for these experiments : West plain area, Chunchon(74m above the sea level), mid-mountainous area, Hongchon(300 m), high alpine area, Hoengsong(450m), and east sea area, Myungju(14m). Weed emergence was investigated 6 times from 10 days after transplanting to 60 days with 10 days interval. From the experiments stated above, the results were summarized as follows ; The identity of weed species was small in grass family, but broad-leaf family was numerous. Also the dominant weed species was generally the perenial weeds. The weed emergence was initiated from 10 days after translanting. The amounts of weed counted the most in Chunchon. A large number of weeds were emerged between 30-60 days after transplanting in Chunchon and between 40-60 days after transplanting in Hongchon and Hoengsong. Among agronomic characteristics affected by weed emergence during rice development, plant height was decreased by 2~4cm, panicle number by 1.3~2.9, spikelet by 3.7~7.5, ripening rates 3.3~6.5%, and milled yield by 12~17%, respectively.

• Pediatric Infection and Vaccine
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• v.13 no.2
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• pp.137-146
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• 2006

Purpose : A regional lymphadenitis is the most frequent adverse reaction of BCG. In order to find out developmental factors and establish a strategy of management, we investigated the clinical courses of children with lymphadenitis following BCG on the aspect of BCG strains, suppurative rates according to the sizes of lymph node and the clinical difference with or without treatment. Methods : From January 1997 to June 2004, 52 children less than 24 month-age-old diagnosed as BCG lymphadenitis in Department of Pediatrics, St. Vincent's Hospital, The Catholic University of Korea were enrolled. The type of BCG strain, place of vaccination, location and size of lymphadenitis were assessed with medical records, retrospectively. Finally, we analysed the correlations between BCG strains or the sizes of lymph node and natural remission or suppuration. Results : The first detected mean age of BCG lymphadenitis was 5.5 month-age. The larger of the measurement was at the first visiting, the younger of age that was first presented. The most frequent location was the same sided axillary region of BCG injection. Among 52 subjects, 46 cases(88.5%) were vaccinated with intradermal Pastuer strain, and only 5 cases(9.6%) were done with percutaneous multipunctured Tokyo strain. Twenty eight cases(53.8%) were regressed naturally, otherwise 24 cases(46.2%) were suppurated. The larger those were sized, the higher freqeuncies those were suppurated on, significantly. Treatment with medications could not prevent the suppuration and could not shorten the healing periods. Conclusion : We predict that there are differences between the occurrent rate of BCG lymphadenitis and BCG strains or methods. Treatment with medication is not recommended owing to its ineffectiveness. Especially, in case of non-suppurative lymphadenitis should be onlyless influence on the tuberculin skin test, cause less adverse reactions, and is inexpensive. observed without treatment, because it could be regressed naturally. An ideal BCG makes a scar, We should make an effort to choose the best BCG strain that can fulfill such requirements.

• Development and Reproduction
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• v.5 no.1
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• pp.47-52
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• 2001

Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

• Reproductive and Developmental Biology
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• v.39 no.1
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• pp.13-21
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• 2015

The importance of genetic resource preservation has been highlighted in the literature as a means of maintaining genetic diversity. Investigations for hematologic values and the differential count of white blood cell count (WBC) for Korean indigenous cattle (KIC) and endangered indigenous cattle (EIC) are rarely performed. Therefore, the objective of this study was to investigate the hematologic values of total 40 EIC (White, Black, Mini cattle) and 35 KIC as control by analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of EIC were significantly decreased by age (p<0.05). The mean values of RBC, HCT, MCV and MCHC between EIC and KIC of the same age (2~3 years) showed the statistical significance (p<0.05). Also, in the WBC of EIC, the mean values were decreased according to the age from $13.9{\times}10^3/{\mu}L{\sim}12.7{\times}10^3/{\mu}L$ under 1 year to $9.1{\times}10^3/{\mu}L{\sim}11.5{\times}10^3/{\mu}L$ over 2 years respectively. In the differential count of WBC of EIC (White, Black, Mini cattle), it showed generally the rates of 40.2%, 52.2%, 49.0% lymphocyte and 27.2%, 33.9%, 32.0% segmented neutrophil from 2~3 years respectively. Result of this study will be used for establishing reference range for blood analysis in EIC such as white, black and mini cattle. This study reported hematological values which could serve as baseline information for comparison in conditions of nutrient deficiency, physiological and health status of endangered Korean native cattle. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2005.12a
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• pp.38-45
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• 2005

The egg bags of Korean salamander(Hynobius leechii) were collected from farmlands in Gyeongsangnam-do area. The assumed breeding time, numerical variation of embryos in each egg bag, mortality and the rates of abnormalities were investigated. The toxicity of benomyl, the metabolite carbendazim and BIC which were frequently spread in agricultural area and caused spontaneous embryonic malformation was investigated. The assumed breeding time between the end of February and the end of March has the difference about a month because of a habitat and it takes about 2 or 3 weeks from laying eggs to hatching. The length of each egg bag and the number of embryos were very varied in each area. It is due to geographical variation. Among egg bags in total study area, only 406 of egg bags(17.70% of total egg bags) developed all of embryos to normal larvae, and 78.49% of total embryos were normally developed. The patterns of spontaneous embryonic malformation were 26 species from A to Z and the abnormal patterns in individual were 8 species and above. the geographical differences about the abnormal pattern were identified and 11 habitats categorized 4 groups. The most frequent abnormality in Gyeongsangnam-do area is the dysplasia of external gill. The caudal dysplasia, abdominal blister and dysplasia of fin were also frequently observed. Individuals showing severe external defect were histologically studied and they showed retinal hypo-pigmentation, thyroid carcinoma, somatic muscular dysplasia, degeneration of cephalic neuron and various organ dysplasia. Benomyl and carbendazim were treated by 10pM$^{\sim}$10uM and BIC was treated by 1$^{\sim}$40ppm to know the effect of toxicity about toxic substance of salamander. After benomyl was treated, a survival rate was sharply dropped from 2 to 8 days. $LC_{100}$ identified in $1{\mu}M$, $LC_{50}$ identified between 100nM and $1{\mu}M$. $EC_{50}$ was assumed between 10nM and 100nM. The prevalent external malformation was abdomen swelled abnormally and histo-pathological effects were abdomen, neural tube and lens hernia. This suggests that benomyl is the toxicitic substance which inhibits the development of digestive system and nervous system. The result of treated carbendazim was similar to that of the treated benomyl. The survival rate is sharply dropped between 2 and 6 days. $LC_{100}$ was identified $1{\mu}M$ and $LC_{50}$ was identified between 10nM and 100nM. This shows that cabendazim has stronger lethal toxicity than benomyl. Ventral blister, eye dysplasia and cephalic dysplasia in the individual of external malformation mean that cabendazim affected nervous system much more than benomyl. Because the toxicity of BIC affected less in the beginning but affected more in the near hatching period, the period causing toxicity is somewhat different. $LC_{100}$ identified near 40ppm and $LC_{50}$ identified near 25ppm. The external defect shows mainly ventral blister and histo-pathological results show intestinal deformities. This result suggests the BIC inhibited strongly the development of digestive system. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of movement of neural crest cells of benomyl. These abnormal developments may be caused by the rupture of epithelium, the loss of microtubule, the reduction of spindle size, the inhibition of spindle assembly formation, the destruction of spindle poles of carbendazim. These abnormal developments may be caused cytotoxicity by inhibition of the synthesis of a number of macromolecules and similar reaction the inhibition of benomyl.

• Korean journal of applied entomology
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• v.39 no.3
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• pp.135-140
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• 2000

Development and reproduction of the cotton caterpillar, Palpita indica, were investigatedunder different temperatures (15 .O, 17.5, 20.0, 22.5, 25 .O, 27.5, 30.0, 32.5, and 35 .O$^{\circ}$C). Duration fromegg to pre-adult of the cotton caterpillar were ranged from 68.6 days at 175$^{\circ}$C to 19.7 days at 35.0% (3.5times shorter growth period compared with that at 17S$^{\circ}$C). At 15.0$^{\circ}$C, cotton caterpillar eggs developedto the last larval instar but were not able to go through the pupal stage. The lower developmentalthreshold temperatures and degree-days of egg, larva, pupa, and complete development were 13.4, 10.6,11.6, and 11.5"C and 55.3,251.5, 138.3, and 479.8 degree days, respectively. The hatching, pupation andemergence rates were higher at 25.0eC and 27.5"C compared with other temperatures. The survival ratefrom the hatched larva to adult was the highest at 27.5"C. The preoviposition and the adult longevity were11.5 and 30.6 days at 17.5"C and 1.5 and 9.2 days at 35.0$^{\circ}$C, respectively. The mean fecundity perfemales was greater at 25.0$^{\circ}$C and 27.5"C compared with other temperatures. Mean generation time indays (T) was shorter on higher temperature. Net reproductive rate per generation (R,) was the lowest atthe highest temperature as well as at the lowest, and it was 199.1 which was the highest at 27.5"C. Theintrinsic rate of natural increase (r,) was highest at 30.0$^{\circ}$C as 0.148. As a result, optimum ranges oftemperature for P. indica growth were between 25.0-32.5"C .emperature for P. indica growth were between 25.0-32.5"C .t;C .