• Applied Biological Chemistry
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• v.6
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• pp.61-77
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• 1965

In this experiment, Akiochi was studied especially on plant growth on the degraded soils. Besides, such soils were carefully examined on its character and plant body was analysed to know the difference in various mineral contents. For this purpose, paddy cultivation was done with the variety Pal Dal at Suwon, Sosa and Pyungtak. Three plots were chosen at each location as the normal and 2 levels of akiochi, a-the stronger and b-the weaker. Harvests from these 9 plots were measured agronomically and also chemically analysised. As for soil, after an observation on vertical section of soil, samples from each layer were also studied both physically and chemically. The results are summarized as follows. 1. Outer changes in rice plant and changes in yield components. 1) Rice from Akiochi soil showed remarkably shortened culm length, head length, protrusoion length, blade length of boot leaf, and coleoptile length, compared with that from the normal paddy field. 2) There was a tendency for Akiochi rice to have more heads per plant. 3) Akiochi rice showed poorer intercalary growth of upper 3 internodes. The ratio of this upper internode length to total culm length was also smaller in this case. Consquently the ratio of lower internode length to total culm length became larger than that from normal peddy field. 4) Akiochi rice showed significantly fewer first spikelets and attached grains of head at main stem. 5) Maturing rate of both this main seem of whole plant body was remarkably lower than that of normal rice. 6) Akiochi rice showed lower head weight of main stem, total hulled rice weight, total grain yield, 1000-grain weight, straw weight and straw-hulled rice ratio. 2. Physical and chemical study on soil. 1) Akiochi soil showed thinner upper layer and total thickness of upper and lower parts was smaller than that of normal. 2) Akiochi soil of Suwon was mainly composed of sand, while that of Sosa and Pyungtak was composed of heavy clay. 3) Chemical analysis indicated that content of $SiO_2$ in upper layer is always lower than that of normal. But no other common tendencies were found. 4) This analysis further lillustrates lower content of Fe, & Mn at Suwon ; of Mn at Sosa and higher content of Fe at Sosa and organic matters at Pyungtak. 5) Some differences in the content of N in each plot could be marked though irregular. 3. Chemical Composition of plant body. 1) Chemical analysis on grain, boot leaf and straw did not suggest any remarkable differences between normal and Akiochi rice, except that the latter contains less Si in boot leaf and less Mn in straw. 2) Contents of each chemical element were measured in grain and straw to calculate the percentage of element content in grain to that of whole plant body including both grain and straw. Here, Akiochi rice always showed lower value in N, K and Mn. 4. Relationship between chemical composition of plant body and that of soil. Akiochi soil at Sosa marked lower content of Mn. This caused another lower content of this element in grain, boot leaf and straw. But except that, no remarkable relationship could be found in this study.

• Applied Biological Chemistry
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• v.20 no.1
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• pp.66-80
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• 1977

This research was carried as a part of the basic study, in which the aptitude of theKorean oak varieties as barrels for aging apple fine spirits was investigated, and thefollowing results were obtained. 1. Following was the result of the chemical analysis of the fruits which are now mass-produced and can be used as a substitute for raw materials for wine production. Apple (Malus pumila Miller var. domestica Schneider) : Total sugar. total acid, volatile acid and pectin of Jonathan (Hong-og) were 13.95%, 0.46%, 0.012%, 0.20% respectively. Total sugar, total acid, volatile acid and pectin of Ralls (Koog-kwang) were 13.35%, 0.43%, 0.011%, 0.45% respectively. 2. Because of low yield of apple juice due to cellulose, pectin, hemicellulose which are present besides sugars, acids in apples, the apple juice were treated with xylanase of Aspergillus niger SUAFM-430, cellulase and pectinase of Aspergillus niger SUAFM-6. This treatment increased the yield of apple juice. And the apple juice was sterilized by adding potassium metabisulfite $(K_2S_20_5)$ and Saccharomyces cerevisae var. ellipsoideus Rasse Johannisberg II (SUAFM-1018) as a cultivation yeast, which has a strong fermentation power was used to ferment. The yield of apple wine based on raw material was 86-87%. The amount of ethanol, extract and methanol obtained from Jonathan and Ralls were 13.5%, 5.4%, 0.04-0.05% respectively. 3. Wines were distilled for two times by the pot still method to make fine spirits. The yield of fine spirits from apple wine mash was 86.6%, and the pH of fine spirits from Jonathan and Ralls were 4.1, 4.2 respectively. 4. The oak chips made of inner part or outer part of 24 Korean oak varieties were used to select the barrel for aging fine spirits. Two oak chips (one oak chip: $1{\times}1{\times}5cm$) of the inner part or of the outer part of each oak variety were dipped into 300 ml of fine spirits, which was bottled in 640ml beer bottle, and followed aging. The colors, flavors and tastes of the fine spirits were checked during 6 months. A. As a criterion for the first screening of oak barrels for aging fine spirits, the rate five of color extraction was determined. The oak chips showed good results in their order as follows and the best 5 varieties were selected. Gal-cham: Quercus aliena Blume (Inner part), Gul-cham: Quercus variabilis Blume (Outer part), Gal-chain: Quercus aliena Blume (Outer part), Jol-cham: Quercus serrata Thumb (Inner and Outer part). Sin-gal-cham: Quercus mongolica Fisher (Outer and Inner part) Sang-su-ri: Quercus acutissima Carruthers (Outer and Inner part) B. To find out the influence of aging temperature on aging, apple fine spirits were aged by dipping each oak chip at room temperature $(24-25^{\circ}C)$) and $45^{\circ}C$. Aging at $45^{\circ}C$ gave the best result followed aging at $30^{\circ}C$ and then at room temperature. C. Apple fine spirits was aged for six months by dipping oak chips in Erlenmeyer flasks and was irradiated with U.V light. The U.V irradiation enhanced the aging effect by nearly two times, compared with the aging without U.V irradiation. D. In aging apple fine spirits by dipping two oak chips, it was observed that the extent of the extraction of most components of oak chips were strongly dependent upon the pH of fine spirits. E. Oak chips of five selected oak varieties and a Limousin white oak from France as a control were used. Each apple fine spitits was dipped by two oak chips, and was aged at room temperature $(24-25^{\circ}C)$, $30^{\circ}C$, $45^{\circ}C$, and with the U.V irradiation at room temperature shaking every week. After six months of aging, the panel test of these aged fine spirits (Young Brandy) showed the following result. Young brandy of apples aged at $45^{\circ}C$ by dipping oak chips of Gal-chain was almost as the fine spirits which were aged at room temperature by dipping Limousin white oak chips from France. Young brandy of with U.V. irradiation at room temperature which were aged by dipping oak chips of Gal-chain was a little worse than that from the fine spirits aged at room temperature by dipping Limousin white oak chips from France. And so, Korean oak varieties are thought to be able to be used for aging every apple fine spirit which was here investigated.

• Korean Journal of Agricultural Science
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• v.7 no.2
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• pp.87-102
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• 1980

In order to investigate the utilization probability of two years old laying hen for W.L. and R.I.R. breeds, carcass weight and percentage were examined and dried old chicken meat products were manufactured for experiments. The results obtained are as follows. 1. Average living body weight were 1,635.40g for the W.L. breeds and 2,289.29g for the R.I.R. breeds and percentage carcass and lean meat for the W.L. were 58.73% and 43.95%, for the R.I.R. 60.34%, 41.98%, respectively. 2. In constitution percentage of carcass on different parts for W.L. and R.I.R. breeds, head were 4.13% and 3.94%, wing 9.97% and 8.62%, breast 32.54% and 20.94%, back 11.35% and 9.75%, thigh 30.75% and 31.34%, hypordermic fat 11.37% and 17.34%, respectively. 3. In constitution percentage of lean meat on different parts for W.L. and R.I.R. breeds, head were 4.03% and 3.95%, wing 9.47% and 9.79%, breast 39.37% and 38.14%, back 11.24% and 9.40%, thigh 36.16% and 38.74%, respectively. 4. In chemical composition of old chicken meat for W.L. breed, moisture was 68.18%, crude protein 22.80%, crude fat 2.70%, extract 5.15% and crude ash 1.18% and for R.I.R. breed, moisture was 68.04%, crude protein 22.18%, crude fat 3.13%, extract 5.45% and crude ash 1.21%. 5. Weight loss in steaming for W.L. at $121^{\circ}C$ for 30min., 60min., and 90min. were 54.91, 56.43 and 58.42%, respectively, and for R.I.R. were 45.23, 47.68 and 49.68%, respectively. 6. The yield of old chicken meat product per a hen were 253.01g for W.L. and 368.64g for R.I.R., the ratio for fresh meat weight and for carcass weight were 35.47% and 26.34% for W.L. breed and 38.25 and 26.83% for R.I.R. breed. 7. In chemical composition of old chicken meat product for W.L., moisture was 16.69%, crude protein 66.16%, crude fat 12.81%, crude ash 4.35%, and R.I.R., moisture 16.11%, crude protein 65.95%, crude fat 13.78% and crude ash 4.57%. 8. To investigate the physical properties which was main factor affecting the product quality, tensile strength, tear strength and elongation rate were measured. The adhesive force of the product made under pressure of $70kg/cm^2$ was similar to those of chipo which was the control product. 9. When measured the color of each protein product, lightness of the product pressed at $70kg/cm^2$ was better than that at $35kg/cm^2$, and the lightness of breast muscle product at $70kg/cm^2$ and chipo was not significant as 16.7% and 16.4%, respectively. Dominant wavelength of product pressed at $70kg/cm^2$ was very similar to chipo which was yellowish orange. 10. In the results of sensory evaluation test containing taste, color, chewing texture and oder of the meat product, when index of chipo as control product was 100, index of breast meat product was higher than that as 118.4, but miscellaneous product was 99.7 and thigh product was 96.2. 11. Summing up the results written above, the meat product utilizing two years old laying hen was compared favorably with its similar food such as chipo on the point of nutrition and physical properties as high protein food, therefore, it was thought that industrialization must be highly appropriate.

• Applied Biological Chemistry
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• v.29 no.1
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• pp.62-72
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• 1986

At present, the definition and chemical analysis method of available soil phosphorus for plants have not been standardized because of the complexity of crop and soil characteristics in Korea and many analysis methods have been suggested with different extraction conditions. Suitable analytical method of available soil P should be established by the trial of various methods based on crop nutrition and soil conditions. To establish the most suitable analysis method of available soiIP, a pot experiment with young maize was conducted over 44 different upland soils collected over the land of Korea. The amount of uptaken P by the plant was determined by ten different chemical methods for the available soil P. The results obtained were as follows: 1. Total phosphorus content in the sample soils ranged ranged $533{\sim}4917\;ppm$, and showed significant positive correlation with the content of organic matter. 2. The P content was relatively low in the acid sulfate soil and very high in the volcanic ash soil although both types of soil contained high level of orgic matter. 3. The amount of extractable P determined by ten different methods were varied more or less, and the ratios of the extractable P to the total soil P were in the range of $1{\sim}48%$. 4. The relative values to the amount of extractable soil P by different methods were in the order of $H_2O(5\;min.)\;1.0\;<\;H_2O(60min.)\;2.27\;<\;NH_4HCO_3\;5.57\;<\;NaHCO_3\;7.42\;<\;Double\;lactate\;9.71\;<\;Bray\;No.1\;12.53\;<\;Lancaster\;17.63\;<\;Nelson\;25.96\;<\;AcOH\;27.6\;<\;CAL-method\;50.27$ 5. The amount of extractable P determined by all of applied methods was very low in acid sulfate soil, volcanic ash soil and coarse textured soil. 6. Soil pH and total soil P generally showed significant positive correlation with the chemically extracted P, and soil organic matter was negatively correlated with the determined by Nelson-and CAL-method. Olsen method which showed significant correlation with exchangeable calcium seemed to be recommendable for calcareous soils. 7. Total amount of uptaken P by Young maize through continuos twice cropping was 4.05% of total soil P in average, and the uptake in the second cropping was twice as much as that of the first cropping. 8. Three determination methods, i.e. Soltanpour-, Double lactate and Bray No. 1-method seemed to be more suitable than Lancaster method which is widely practiced at present in Korea. However, further study should be carried out with other crops and soils to most adequate chemical method for determination of available soil P.

• Journal of Korean Society of Forest Science
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• v.33 no.1
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• pp.33-58
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• 1977

A bark comprises about 10 to 20 percents of a typical log by volume, and is generally considered as an unwanted residue rather than a potentially valuable resourses. As the world has been confronted with decreasing forest resources, natural resources pressure dictate that a bark should be a raw material instead of a waste. The utilization of the largely wasted bark of genus Pinus, Quercus, and Populus grown in Korea can be enhanced by learning its physical and mechanical properties. However, the study of tree bark grown in Korea have never been undertaken. In the present paper, an investigative study is carried out on the bark of three genus, eleven species representing not only the major bark trees but major species currently grown in Korea. For each species 20 trees were selected, at Suweon and Kwang-neung areas, on the same basis of the diameter class at the proper harvesting age. One $200cm^2$ segment of bark was obtained from each tree at brest height. Physical properties of bark studied are: bark density, moisture content of green bark (inner-, outer-, and total-bark), fiber saturation point, hysteresis loop, shrinkage, water absorption, specific heat, heat of wetting, thermal conductivity, thermal diffusivity, heat of combustion, and differential thermal analysis. The mechanical properties are studied on bending and compression strength (radial, longitudinal, and tangential). The results may be summarized as follows: 1. The oven-dry specific gravities differ between wood and bark, further more even for a given bark sample, the difference is obersved between inner and outer bark. 2. The oven-dry specific gravity of bark is higher than that of wood. This fact is attributed to the anatomical structure whose characters are manifested by higher content of sieve fiber and sclereids. 3. Except Pinus koraiensis, the oven-dry specific gravity of inner bark is higher than that of outer bark, which results from higher shrinkage of inner bark. 4. The moisture content of bark increases with direct proportion to the composition ratio of sieve components and decreases with higher percent of sclerenchyma and periderm tissues. 5. The possibility of determining fiber saturation point is suggested by the measuring the heat of wetting. With the proposed method, the fiber saturation point of Pinus densiflora lies between 26 and 28%, that of Quercus accutissima ranges from 24 to 28%. These results need be further examined by other methods. 6. Contrary to the behavior of wood, the bark shrinkage is the highest in radial direction and the lowest in longitudinal direction. Quercus serrata and Q. variabilis do not fall in this category. 7. Bark shows the same specific heat as wood, but the heat of wetting of bark is higher than that of wood. In heat conductivity, bark is lower than wood. From the measures of oven-dry specific gravity (${\rho}d$) and moisture fraction specific gravity (${\rho}m$) is devised the following regression equation upon which heat conductivity can be calculated. The calculated heat conductivity of bark is between $0.8{\times}10^{-4}$ and $1.6{\times}10^{-4}cal/cm-sec-deg$. $$K=4.631+11.408{\rho}d+7.628{\rho}m$$ 8. The bark heat diffusivity varies from $8.03{\times}10^{-4}$ to $4.46{\times}10^{-4}cm^2/sec$. From differential thermal analysis, wood shows a higher thermogram than bark under ignition point, but the tendency is reversed above ignition point. 9. The modulus of rupture for static bending strength of bark is proportional to the density of bark which in turn gives the following regression equation. M=243.78X-12.02 The compressive strength of bark is the highest in radial direction, contrary to the behavior of wood, and the compressive strength of longitudinal direction follows the tangential one in decreasing order.

• Journal of Gastric Cancer
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• v.6 no.1
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• pp.36-42
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• 2006

Purpose: The $p21^{Waf1/Cip1}$ protein Inhibits the cell cycle by Inhibiting the phosphorylation at the $G1{\rightarrow}S$ check point, and the $p27^{kip1}$ protein similarly performs the suppressor function by controlling the p27-mediated G1 arrest. In this study, we analysed the clinical status and survival rates in correlations with p21 and p27 expression patterns in gastric cancer. Materials and Methods: Between 1993 and 1997, 192 patients who underwent surgeries in Catholic Medical Center were analysed retrospectively in this study. Immunohistochemical staining was performed and if the nuclei of the tumor cells were stained, we assumed those as positive results. Statistical analysis was based on clinicopathological findings and differences in survival rates. Results: The expression rate of p27 was 28.1% and 15.6% in p21 each. The ratio of T1-2(80.0%) was significantly high in p21 (+), but the ratio of T3-4 (50.6%) was slightly high in p21 (-). There was no statistical significance regarding other factors. The results in p27 was not much different from expression rate of p21 in T-stage. In addition, p27 expression in diffuse type (91.3%) was higher than in intestinal type (62.7%) by Lauren's classification (P<0.05). Also, there was no statistical significance in other factors. In the correlation of p21 and p27, p27 was positive when p21 was positive (53.5%). Conversely, p27 was negative when p21 was negative (76.5%, p<0.05). In the p21 and p27 combination test, there was higher rate of T1-2 (87.5%) in p21 (+)/p27 (+), and higher rate of T3-4 (58.1%) in p21 (-)/p27 (-) (P<0.05). Results showed higher rate of intestinal type (100%) in p21 (+)/p27 (+), and diffuse type (87.0%) was dominant in p21 (-)/p27 (-) (P<0.05) by Lauren's classification. Moreover, there was no statistical significance in the 5-year survival rate in the expression of p21 and p27, and the 5-year survival rate was highest in the case of p21 (+)/p27 (+) without statistical significance. Conclusion: In our study, $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ expressed similar patterns. The expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ affected the degree of invasiveness of the tumor, and. Combined examination result revealed the correlation of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ with Lauren's classification and depth of invasion of the tumor. However, we assumed that little difference between the survival rates depending on expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ has limited their value as predictable prognostic indicators.

• The Korean Journal of Air & Space Law and Policy
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• v.28 no.2
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• pp.223-258
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• 2013

1967 Outer Space Treaty(Treaty on Principles Governing the Activities of States in the Exploration and Use of Outer Space, including the Moon and Other Celestial Bodies; OST) is a treaty that forms the basis of international space law. OST is based on the 1963 Declaration of Legal Principles Governing the Activities of States in the Exploration and Use of Outer Space announced by UNGA resolution. As of May 2013, 102 countries are states parties to OST, while another 27 have signed the treaty but have not completed ratification. OST explicitly claimed that the Moon and Other Celestial Bodies are the province of all mankind. Art. II of OST states that "outer space, including the Moon and other celestial bodies, is not subject to national appropriation by claim of sovereignty, by means of use or occupation, or by any other means", thereby establishing res extra commercium in outer space like high seas. However 1979 Moon Agreement stipulates that "the moon and its natural resources are the Common Heritage of Mankind(CHM)." Because of the number of the parties to the Moon Agreement(13 parties) it does not affect OST. OST also established its specific treaties as a complementary means such as 1968 Rescue Agreement, 1972 Liability Convention, 1975 Registration Convention. OST bars states party to the treaty from placing nuclear weapons or any other weapons of mass destruction in orbit of Earth, installing them on the Moon or any other celestial body, or to otherwise station them in outer space. It exclusively limits the use of the Moon and other celestial bodies to peaceful purposes and expressly prohibits their use for testing weapons of any kind, conducting military maneuvers, or establishing military bases, installations, and fortifications. However OST does not prohibit the placement of conventional weapons in orbit. China and Russia submitted Draft Treaty on the Prevention of the Placement of Weapon in Outer Space and of the Threat or Use of Force against Outer Space Objects(PPWT) on the Conference on Disarmament in 2008. USA disregarded PPWT on the ground that there are no arms race in outer space. OST does not have some articles in relation to current problems such as space debris, mechanisms of the settlement of dispute arising from state activities in outer space in specific way. COPUOS established "UN Space Debris Mitigation Guidelines" based on "IADC Space Debris Mitigation Guidelines" and ILA proposed "International Instrument on the Protection of the Environment from Damage Caused by Space Debris" for space debris problems and Permanent Court of Arbitration(PCA) established "Optional Rules for Arbitration of Disputes Relating to Outer Space Activities" and ILA proposed "1998 Taipei Draft Convention on the Settlement of Space Law Dispute" for the settlement of dispute problems. Although OST has shortcomings in some articles, it is very meaningful in international law in considering the establishment of basic principles governing the activities of States in the exploration and use of outer space, including the Moon and Other Celestial Bodies. OST established the principles governing the activities of states in the exploration and use of outer space as customary law and jus cogens in international law as follows; the exploration and use of outer space shall be carried out for the benefit and in the interests of all countries and shall be the province of all mankind; outer space shall be free for exploration and use by all States; outer space is not subject to national appropriation by claim of sovereignty, by means of use or occupation, or by any other means. The principles of global public interest in outer space imposes international obligations erga omnes applicable to all States. This principles find significant support in legal norms dealing with following points: space activities as the "province of all mankind"; obligation to cooperate; astronauts as envoys of mankind; avoidance of harmful contamination; space activities by States, private entities and intergovernmental organisations; absolute liability for damage cauesd by certain space objects; prohibition of weapons in space and militarization of the celestial bodies; duty of openness and transparency; universal application of the international space regime.

• Sleep Medicine and Psychophysiology
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• v.17 no.2
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• pp.100-108
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• 2010

Objectives: "Internet-addiction" came into common use not only in clinical setting but also in everyday life. But, pathophysiology and diagnostic criteria of the internet addiction remain unknown. Because adolescents are in developing period, they might be vulnerable to the internet addiction, depression and sleep-related problem. The objectives of this study were to investigate the characteristics of internet addiction and its association with sleep pattern and depression in Korean adolescence. Methods: Subjects were 799 middle and high school students in Seoul, Korea. We administered a self-reported questionnaire including socio-demographic data, Korean versions of Young's Internet Addiction Scale (YIAS), Pittsburgh Sleep Quality Index (PS-QI), the Center for Epidemiologic Studies for Depression Scale (CES-D) and questions about internet using patterns. Data of 696 subjects were included in analysis. Chi-square tests were used to analyze proportional differences, and ANOVA with post-hoc tests were used to analyze differences among groups. Partial correlation analyses were performed to analyze the correlation of internet addiction with other variables (two-tailed, p<0.05). Results: Of the 696 participants (grade 2 of middle school; M2 135 vs. grade 1 of high school; H1 238 vs. grade 2 of high school; H2 323), 2.0% (n=14) were internet-addicted (IA), 27.7% (n=193) were over-using (OU) and 70.3% (n=489) were not-addicted (NA). The mean scores of YIAS, PSQI and CES-D scores were 35.24${\pm}$12.78, 5.53${\pm}$3.04 and 16.72${\pm}$8.69, respectively. In higher grade students, average total sleep time was shorter (M2 426.20${\pm}$67.68 min. vs. H1 380.47${\pm}$62.57 min. vs. H2 354.67${\pm}$73.37 min., F=51.909, p<0.001), and PSQI (4.69${\pm}$3.14 vs. 5.42${\pm}$3.15 vs. 5.97${\pm}$2.83, F=8.871, p<0.001) CES-D (13.53${\pm}$8.37 vs. 16.96${\pm}$8.24 vs. 17.87${\pm}$8.84, F=12.373, p<0.001) scores were higher than those of lower grade students. Comparing variables among IA, OU and NA groups, computer using time not for study (96.36${\pm}$63.31 min. vs. 134.92${\pm}$86.79 min. vs. 213.57${\pm}$136.87 min., F=34.287, p<0.001) and portable device using time not for study (84.22${\pm}$79.11 min. vs. 96.97${\pm}$91.89 min. vs. 152.31${\pm}$93.64 min., F= 5.400, p=0.005) were different among groups. PSQI (5.26${\pm}$2.97 vs. 6.08${\pm}$2.97 vs. 7.50${\pm}$4.41, F=8.218, p<0.001) and CES-D scores (15.40${\pm}$8.08 vs. 19.05${\pm}$8.42 vs. 30.43${\pm}$13.69, F=32.692, p<0.001) were also different among groups. YIAS score were correlated with computer using time not for study (r=0.356, p<0.001) and portable device using time not for study (r= 0.136, p<0.001). PSQI score (r=0.237, p<0.001) and CES-D score (r=0.332, p<0.001). YIAS score and PSQI score (r=0.131, p= 0.001), YIAS and CES-D score (r=0.265, p<0.001), PSQI score and CES-D score (r=0.357, p<0.001) were correlated each other. Conclusion: These results suggested that adolescents' internet-addiction was correlated with not only computer and portable device using time not for study but also depression and sleep-related problems. We should pay attention to depression and sleep-related problems, when evaluating internet-addiction in adolescents.

• Tuberculosis and Respiratory Diseases
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• v.45 no.4
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• pp.846-860
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• 1998

Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

• Tuberculosis and Respiratory Diseases
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• v.45 no.6
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• pp.1265-1276
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• 1998

Background : Nitric oxide(NO) is an endogenously produced free radical that plays an important role in regulating vascular tone, inhibition of platelet aggregation and white blood cell adhesion to endothelial cells, and host defense against infection. The highly reactive nature of NO with oxygen radicals suggests that it may either promote or reduce oxidant-induced cell injury in several biological pathways. Oxidant injury and interactions between pulmonary vascular endothelium and leukocytes are important in the pathogenesis of acute lung injury, including acute respiratory distress syndrome(ARDS). In ARDS, therapeutic administration of NO is a clinical condition providing exogenous NO in oxidant-induced endothelial injury. The role of exogenous NO from NO donor or the suppression of endogenous NO production was evaluated in oxidant-induced endothelial injury. Method : The oxidant injury in cultured rat lung microvascular endothelial cells(RLMVC) was induced by hydrogen peroxide generated from glucose oxidase(GO). Cell injury was evaluated by $^{51}$chromium($^{51}Cr$) release technique. NO donor, such as S-nitroso-N-acetylpenicillamine(SNAP) or sodium nitroprusside(SNP), was added to the endothelial cells as a source of exogenous NO. Endogenous production of NO was suppressed with N-monomethyl-L-arginine(L-NMMA) which is an NO synthase inhibitor. L-NMMA was also used in increased endogenous NO production induced by combined stimulation with interferon-$\gamma$(INF-$\gamma$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), and lipopolysaccharide(LPS). NO generation from NO donor or from the endothelial cells was evaluated by measuring nitrite concentration. Result : $^{51}Cr$ release was $8.7{\pm}0.5%$ in GO 5 mU/ml, $14.4{\pm}2.9%$ in GO 10 mU/ml, $32.3{\pm}2.9%$ in GO 15 mU/ml, $55.5{\pm}0.3%$ in GO 20 mU/ml and $67.8{\pm}0.9%$ in GO 30 mU/ml ; it was significantly increased in GO 15 mU/ml or higher concentrations when compared with $9.6{\pm}0.7%$ in control(p < 0.05; n=6). L-NMMA(0.5 mM) did not affect the $^{51}Cr$ release by GO. Nitrite concentration was increased to $3.9{\pm}0.3\;{\mu}M$ in culture media of RLMVC treated with INF-$\gamma$ (500 U/ml), TNF-$\alpha$(150 U/ml) and LPS($1\;{\mu}g/ml$) for 24 hours ; it was significantly suppressed by the addition of L-NMMA. The presence of L-NMMA did not affect $^{51}Cr$ release induced by GO in RLMVC pretreated with INF-$\gamma$, TNF-$\alpha$ and LPS. The increase of $^{51}Cr$ release with GO(20 mU/ml) was prevented completely by adding 100 ${\mu}M$ SNAP. But the add of SNP, potassium ferrocyanate or potassium ferricyanate did not protect the oxidant injury. Nitrite accumulation was $23{\pm}1.0\;{\mu}M$ from 100 ${\mu}M$ SNAP at 4 hours in phenol red free Hanks' balanced salt solution. But nitrite was not detectable from SNP upto 1 mM The presence of SNAP did not affect the time dependent generation of hydrogen peroxide by GO in phenol red free Hanks' balanced salt solution. Conclusion : Hydrogen peroxide generated by GO causes oxidant injury in RLMVC. Exogenous NO from NO donor prevents oxidant injury, and the protective effect may be related to the ability to release NO. These results suggest that the exogenous NO may be protective on oxidant injury to the endothelium.