• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.215-223
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• 2003

This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.27 no.4
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• pp.371-384
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• 1982

Highly cold tolerant varieties are requested not only at high latitute cool area but also tropical high elevated areas, and the required tolerance is different from location to location. IRRI identified 6 different types of cold tolerance required in the world for breeding purpose; a) Hokkaido type, b) Suweon type, c) Taipei 1st season type, d) Taipei 2nd season type, e) Tropical alpine type and, f) Bangladesh type. The cold tolerance requested in Korea is more eargent in Tongil group cultivars and their required tolerance is the one such as the physiological activities at low temperature are as active as in Japonica group cultivars at least during young seedling stage and reproduction stage. With conventional Japonica cultivars, such cold tolerant characters are requested as short growth duration but stable basic vegetative growth, less sensitive to high temperature and less prolonged growth duration at low temperature. The methods screening for cold tolerance were developed rapidly after the Tongil cultivar was reliesed. The facilities of screening for cold tolerance, such as, low temperature incubator, cold water tank, growth cabinet, phytotron, cold water nursery in Chuncheon, breeding nursery located in Jinbu, Unbong and Youngduk, are well established. Foreign facilities such as, cold water tank with the rapid generation advancement facilities, cold nurseries located in Banaue, Kathmandu and Kashimir may be available for the screening of some limitted breeding materials. For the reference, screening methods applied at different growth stages in Japan are introduced. The component characters of cold tolerance are not well identified, but the varietal differences in a) germinability, b) young seedling growth, c) rooting, d) tillering, e) discolation, f) nutrition uptake, g) photosynthesis rate, h) delay in heading, i) pollen sterility, and j) grain fertility at low temperature are reported to be distinguishable. Relationships among those traits are not consistent. Reported studies on the inheritance of cold tolerance are summarized. Four or more genes are controlling low temperature germinability, one or several genes are controlling seedling tolerance, and four or more genes are responsible for the pollen fertility of the rice treated with cold air or grown in the cold water nursery. But most of those data indicate that the results may come out in different way if those were tested at different temperature. Many cold tolerant parents among Japonicas, Indicas and Javanicas were identified as the results of the improvement of cold tolerance screening techniques and IRTP efforts and they are ready to be utilized. Considering a) diversification of germ plasm, b) integration of resistances to diseases and insects, c) identification of adaptability of recommending cultivars and, d) systematic control of recommending cultivars, breeding strategies for short term and long term are suggested. For short term, efforts will be concentrated mainly to the conventional cultivar group. Domestic cultivars will be used as foundation stock and ecologically different foreign introductions such as from Hokkaido, China or from Taiwan, will be used as cross parents for the adjustment of growth durations and synthsize the prototype of tolerances. While at the other side, extreme early waxy Japonicas will be crossed with the Indica parents which are identified for their resistances to the diseases and insects. Through the back corsses to waxy Japonicas, those Indica resistances will be transfered to the Japonicas and these will be utilized to the crosses for the improvement of resistances of prototype. For the long term, efforts will be payed to synthsize all the available tolerances identified any from Japonicas, Indicas and Javanicas to diversify the germ plasm. The tolerant cultivars newly synthsized, should be stable and affected minimum. to the low temperature at all the growing stages. The resistances to the diseases and insects should be integrated also. The rapid generation advancement, pollen culture and international cooperations were emphasized to maximize the breeding efficiency.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.425-435
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• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.166-179
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• 1999

Apical sealing is essential for the success of surgical endodontic treatment. Root-end cavity is apt to be contaminated with moisture or blood, and is not always easy to be dried completely. The purpose of this study was to evaluate the influence of dry methods of retrocavity on the apical seal in endodontic surgery. Apical seal was investigated through the evaluation of apical leakage and adaptation of filling material over the cavity wall. To investigate the influence of various dry methods on the apical leakage, 125 palatal roots of extracted human maxillary molar teeth were used. The clinical crown of each tooth was removed at 10 mm from the root apex using a slow-speed diamond saw and water spray. Root canals of the all the specimens were prepared with step-back technique and filled with gutta-percha by lateral condensation method. After removing of the coronal 2 mm of filling material, the access cavities were closed with Cavit$^{(R)}$. Two coats of nail polish were applied to the external surface of each root. Apical three millimeters of each root was resected perpendicular to the long axis of the root with a diamond saw. Class I retrograde cavities were prepared with ultrasonic instruments. Retrocavities were washed with physiologic saline solution and dried with various methods or contaminated with human blood. Retrocavities were filled either with IRM, Super EBA or composite resin. All the specimens were immersed in 2% methylene blue solution for 7 days in an incubator at $37^{\circ}C$. The teeth were dissolved in 14 ml of 35% nitric acid solution and the dye present within the root canal system was returned to solution. The leakage of dye was quantitatively measured via spectrophotometric method. The obtained data were analysed statistically using one-way ANOVA and Duncan's Multiple Range Test. To evaluate the influence of various dry methods on the adaptation of filling material over the cavity wall, 12 palatal roots of extracted human maxillary molar teeth were used. After all the roots were prepared and filled, and retrograde cavities were made and filled as above, roots were sectioned longitudinally. Filling-dentin interface of cut surfaces were examined by scanning electron microscope. The results were as follows: 1. Cavities dried with paper point or compressed air showed less leakage than those dried with cotton pellet in Super EBA filled cavity (p<0.05). However, there was no difference between paper point- and compressed air-dried cavities. 2. When cavities were dried with compressed air, dentin-bonded composite resin-filled cavities showed less apical leakage than IRM- or Super EBA-filled ones (p<0.05). 3. Regardless of the filling material, cavities contaminated with human blood showed significantly more apical leakage than those dried with compressed air after saline irrigation (p<0.05). 4. Outer half of the cavity showed larger dentin-filling interface gap than inner half did when cavities were filled with IRM or Super EBA. 5. In all the filling material groups, cavities contaminated with blood or dried with cotton pellets only showed larger defects at the base of the cavity than ones dried with paper points or compressed air.

• Tuberculosis and Respiratory Diseases
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• v.53 no.5
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• pp.497-509
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• 2002

Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.

• Korean journal of applied entomology
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• v.9 no.2
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• pp.57-74
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• 1970

These experiments were conducted to investigate the :actors influencing the systemic action of Dimethoate (O,O-dimethyl-S-(N-methylcarhamoylmethyl) photphorodithioate) to rice seeds and the phytotoxic effects on the seed germination. Dimethoate $(Roxion^{(R)})$ $40\%$ emulsion was used. The varieties tested were Jinheung. Nongkwang,Suwon #82, Norm #6, Paltal, Shirogane, Suseong, Pungkwang, Shin #2, Fujisaka #5, Kwanok, and Jaekeun. The permeated Dimethoate was extracted from the treated seeds by chloroform and quantities were determined by Spectrophotometer. The phytotoxicity was evaluated from the effects on the germination of the treated seeds which were kept in an incubator. The oxygen consumption was measured by Warburg Manometer at $30^{\circ}C$ for 60 minutes. Indices of KOH disintegration of seeds and chemical composition of the seeds were also determined. The results obtained were as followings; 1) The amount of permeated Dimethoate in the seeds showed remarkable differences with varieties. The amount of Dimethoate per 100 grains was greater as in the ascending order of Suseong, Kwanok, Nongkwang, Jinheung, Paltal, Fujisaka #5, Suwon #82, Norm #6, Shirogane, Shin #2, Pungkwang and Jaekeun. 2) It was observed that the total amount of Dimethoate in the seeds(mg./100 grains) were greater among the varieties with large grain than those with small grains, while reverse cases were true in the amount of Dimethoate in a gramme of seeds, probably because of the greater surface areas In a small grains for a gramme weight. 3) There was no significant correlation between the permeated amount of Dimethoate and amount of absorbed water by the seeds when the seeds were treated with $0.1\%$ Dimethoate for 24 and 48 hours. 4) The permeability of Dimethoate to seeds significantly increased in the prolonged soaking periods, higher concentration, and higher temperature. 5) When the seeds were treated with $0.1\%$ Dimethoate for 24 and 48 hours at $15^{\circ},\;20^{\circ},\; 20^{\circ},\; and \;30^{\circ}C$, the permeated amount of Dimethoate were increased at higher temperature. It seems to be that the more active penetration of Dimethoate was involved at the higher temperature. 6) The phytotoxic effects of Dinethoate on the seed germination varied with the varieties. An descending order of varietal tolerance of seeds was as followings: Jinheung, Fujisaka #5, Suwon #82, Paltal, Nongkwang, Jaekeun, Shin #2, Kwanok, Shirogane, Pungkwang, Suseong, and Norm #6. 7) There was a positive correlation between the amount of Dimethoate permeated into the seeds (mg./gram. of seeds) and phytotoxicity of seeds. 8) The Phytotoxic effects of Dimethoate showed close correlation with the degree of KOH disintegration of seeds, average germination periods, and oxygen respiration of seeds. 9) It was observed that higher protein contents of the seeds decreased the phytotoxic effects of Dimethoate. 10) Relatively high negative correlation between the degree of KOH disintegration of seeds and crude protein content of the seeds was observed. 11) The average germination period was delayed for about 2 days when the seeds were treated with $0.2\%$ Dimethoate for 24 hours at $30^{\circ}C$. 12) The oxygen consumption of the seeds treated with $0.2\%$ Dimethoate for 24 hours at $30^{\circ}C$ was greatly decreased when compared with that of the normal seeds. 13) The amount of oxygen consumption of the seeds (in 24 hours after 24 hours water soaking) was negatively correlated with the average germination periods of the seeds.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.457-473
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• 2002

The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

• Journal of Wetlands Research
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• v.20 no.2
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• pp.155-160
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• 2018

We investigated to know the growth patterns of Aedes albopictus larva at the four different temperature conditions. Each of 120 individuals was placed into a $20m{\ell}$ vial and 12 sets (a set of 10 vials) were separated into 12 water tanks ($17{\times}24{\times}18cm^3$). Each water tank was composed of 3 the $1^{st}$ instar, 3 the $2^{nd}$ instar, 2 the $3^{rd}$ instar, and 2 the $4^{th}$ instar. Three sets of water tanks were placed under the four different incubator temperatures ($17^{\circ}C$, $21^{\circ}C$, $24^{\circ}C$, $28^{\circ}C$). We found that the eclosion rates were $20.00{\pm}5.77%$ at $21^{\circ}C$ and $3.33{\pm}3.33%$ at other temperatures. For the mosquito larva mortality rate, $1^{st}$ instar was $19.24{\pm}3.65%$, $2^{nd}$ instar was $16.48{\pm}3.25%$, $3^{rd}$ instar was $23.54{\pm}5.06%$, and $4^{th}$ instar was $40.74{\pm}7.08%$. The lowest mortality rate in growth stages according to temperature was $13.33{\pm}6.67%$ at $17^{\circ}C$ in $1^{st}$ instar larva, $7.41{\pm}7.41%$ at $21^{\circ}C$ at $2^{nd}$ instar larva, $10.74{\pm}6.43%$ at $24^{\circ}C$ in $3^{rd}$ instar larva, and $20.37{\pm}5.46%$ at $28^{\circ}C$ in $4^{th}$ instar larva. The survival period of mosquitoes in underwater were $26.33{\pm}0.67days$ at $17^{\circ}C$, $23.33{\pm}1.33days$ at $21^{\circ}C$, $20.00{\pm}2.52days$ at $24^{\circ}C$, and $11.67{\pm}1.20days$ at $28^{\circ}C$. From our results the most effective temperature to the normal growth of mosquito larva was $21^{\circ}C$, and the highest mortality rate was shown at the $4^{th}$ instar stage of larva growth. Our results would provide the basic data for the mosquito larva's growth pattern.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 1998.05a
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• pp.37-54
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• 1998

This study was conducted to investigate the effects of mugwort extracts on the blood ethanol concentration, liver function and low level of cadmuim(Cd) in rats. The effects of mugwort extracts on the blood ethanol concentration was studied in Sprague-Dawley rats (10 weeks old) administered p.o. with 25% ethanol (5g/1kg body weight) and then injected with mugwort extracts (at the 2% levels of daily feed consumption compared with the concentration of catechins level in mugwort extracts) in caudal vein. SD rats were divided into five groups : control group (CON-E, only ethanol and 0.85% saline sol'n treated instead of each extracts), water extracts of mugwort treated to the control (MDW-E), ethanol extracts of mugwort treated to the control (POH-E). And then rat plasma of each time (0hr, 1hr, 2hr, 3hr) was investigated ethanol concentration by gas chromatography. Another rats were measured at the time of 0 and 5hr for the test of GOD(Glutamic Oxaloacetic Transaminase) and GPT(Glutamic Pyruvic Transaminase). Components of each extracts were analyzed by using high performance liquid chromatography. The effects of mugwort extracts on the liver function were studied in culture of rat hepatocyte composed of three groups : Control group and two groups treated with each extracts (1% & 2% MDW, 1% & 2% MOH). Condition of rat hepatocytes cultured for 36hr at $37^{\circ}C$(5% $CO_2$ incubator), number of cells, GOT and GPT activity were investigated. The results obtained were summarized as follows ; 1. Catechins level of mugwort extracts was $8{\sim}10mg/100g(MDW)$, $3{\sim}4mg/100g(MOH)$ 2. The contents of (-)-Epigallocatechin was high in MDW 3. The effects of mugwort extracts on the blood ethanol concentration were as follows; 1) The order in ethanol degradation efficiency was MDW-E > MOH-E > CON-E. 2) Ethanol concentration significantly decreased (p<0.05) in MDW-E and MOH-E. 4. The effects of mugwort extracts on the liver function were as follows; (rat hepatocytes cultured for 36hr at $37^{\circ}C$) 1) Cells condition of MDW-L was better than other groups. 2) The order in number of cells (rat hepatocytes) was 2% MDW-L >1% MDW-L >1% MOH-L > Con-L > 2% MOH-L 5. Cd treatment increased concentrations of hepatic GSH level, and decreased GOT activity in plasma. Therefore, this results suggest that the effects of mugwort extracts may an important rols in degradation ethanol and recovery liver function in body. Also, Mugwort extracts may modify the toxicities of Cd in Cd-treated rats and play an important roles in preventing the liver from various toxicants including Cd in Cd treated rats.

• Korean Journal of Soil Science and Fertilizer
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• v.9 no.1
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• pp.17-23
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• 1976

In order to investigate the fate of nitrogen in the paddy soil, Suchang, Hwasoon and Susan soil which have different properties, were treated with several nitrogen fertilizers such as ammonium chloride, ammonium sulfate, urea and SCU (sulfur-coated urea), and incubated under water-logged condition in $30^{\circ}C$ incubator. $NH_4-N$, $NO_3-N$, $Fe^{++}$ and pH in soil and stagnant water, were determined at 10, 20, 30, 40 and 50 days after incubation. The obtained results were summarized as follows: 1. The effect of rising temperature was increased in order of Hwasoon>Suchang>Susan and the effect of air drying soil was risen in order of Susan>Hwasoon>Suchang, while the rate of ammonication was in order of Susan>Suchang>Hwasoon. 2. The changes of $NH_4-N$ in stagnant water was dependent upon the nitrogen concentration of $NH_4Cl$ and $(NH_4)SO_4$ plat was high and decreased after 30 days incubation, but increased after 40 days and then decreased again. In contrast with the above, $NH_4-N$ concentration of urea and SCU plot was low but the change showed slightly through the incubation period. 3. Accumulation of $NH_4-N$ in the oxidative layer of the $NH_4Cl$ and $(NH_4)_2SO_4$ plot was higher than that of urea and SCU plot and $NH_4-N$ content was decreased with the incubation period. The change of $NH_4-N$ in the reductive layer showed the same pattern. 4. The changes of $NO_3-N$ in the stagnant water were different according to soil properties and nitrogen fertilizer. $NO_3-N$ concentration in stagnant water of urea and SCU plot was higher than in the $NH_4-Cl$ $(NH_4)_2SO_4$ plot and nearly disappeared after 30 to 40 days incubation. 5. The $NO_3-N$ concentration in the oxidative layer of soil was higher than reductive layer. The pattern of change was different in accordance with soil properties and nitrogen fertilizers. In general, nitrification in urea and SCU plot was more increased than $(NH_4)_2SO_4$ plot. In reductive layer, the concentration of $NO_3-N$ was very low until 30 days incubation and thereafter increased slightly. 6. Upon the concentration of $NH_4-N$ and $NO_3-N$ in stagnant water and soil, it was assumed that denitification of urea and SCU plot was higher than $NH_4Cl$ and $(NH_4)_2SO_4$ plot and denitrified nitrogen in incubation period was above 50%.