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Prenatal Deaths and External Malformations Caused by X-Irradiation during the Preimplantation Period of ddy Mice (임신 ddy Mice에서 착상전기 방사선 조사에 따른 산전 사망 및 외부 기형 발현)

  • Ro, Hee-Jeong;Choi, Ihl-Bhong;Gu, Yeun-Wh
    • Radiation Oncology Journal
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    • v.16 no.3
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    • pp.233-243
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    • 1998
  • Purpose : To evaluate the effects of X-irradiation on prenatal deaths, i.e., preimplantation deaths, embryonic deaths and fetal deaths, and on external malformations in precompacted preimplantation ddy mice Materials and Methods : Pregnant mice (n=85) obtained by limiting the mating time to from 6 to 9 A.M., were segregated into 11 groups. The first five groups (n=26) were irradiated with X-ray doses of 0.1 0.5, 0.75, 1.5, and 3 Gy, respectively, at 24 h post conception (p.c.) of the preimplantation Period. The second five (n=27) groups were irradiated at the same X-ray doses, respectively, but at 48 h p.c. of the preimplantation period. The last group (n=32) was the control group. The uterine contents were examined on the 18th day of gestation for prenatal deaths and external malformations. Results : 1) A statistically significant increase in preimplantation deaths with increasing dose was observed in the experimental groups irradiated at 24 h p.c. and in the groups irradiated at 48 h p.c., as compared to the control group. The threshold dose was close to 0.05 Gy and 0.075 Gy for the irradiations at 24 h p.c. and 48 h p.c. respectively. 2) A statistically significant increase in embryonic deaths with increasing dose was observed in all irradiation groups, except the group irradiated with a dose of 0.1 Gy at 48 h p.c.. 3) No fetal deaths were found in any experimental group. 4) In the experimental groups irradiated at 24 h p.c. anomalies increased with statistical significance, as compared with the control group : 2 exencephalies, 2 open eyelids, 3 anophthalmias, 2 cleft Palates, 2 gastroschisis, 1 abdominal wall defect. 1 leg defect, and 2 short tail anomalies: the threshold dose for external malformations was close to 0.2 Gy at 24 h p.c.. In the groups irradiated at 48 h p.c., 1 open eyelid and 2 short tail anomalies were observed, but there was no statistical significance in those malformations. Conclusion : The results of this study reveal that X-irradiation of precompacted preimplantation ddy mice causes not only preimplantation deaths and embryonic deaths but also external malformations. In addition, external malformations were observed in our experiments at diagnostic doses, including 0.1 and 0.5 Gy. For this reason, we recommend that irradiation should be avoided during the preimplantation period by applying Rugh's 10-day rule.

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Physicochemical, Microbiological and Sensory Properties of Food Additive-Free Grilled Fork Products during Cold Storage (식품 첨가제 미함유 그릴 돈육햄의 냉장저장 중 물리화학, 미생물학 및 관능적 품질 특성)

  • Kim, Il-Suk;Jin, Sang-Keun;Park, Ki-Hoon;Jung, Gi-Jong;Kim, Dong-Hun;Yang, Mi-Ra;Hah, Kyung-Hee;Lee, M.
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.269-275
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    • 2006
  • The objective of this study was to investigate the physicochemical, microbiological and sensory properties of food additive-free grilled pork products manufactured using loin (T1), tender loin (T2) and ham (T3). The samples were heated for 30 min at $60^{\circ}C$, and then 50 min for $150^{\circ}C$. After cooling, vacuum packaged grilled pork samples was stored at $4{\pm}1^{\circ}C$ for 40 days. The pH values of grilled pork samples ranged from 5.92 (T1) to 6.10 (T3) at the initial storage time, and from 6.28 (T1) to 6.60 (T3) after 40 days. The water holding capacities(%) was $85.99{\sim}93.24%$ for T1, $95.26{\sim}93.89%$ for T2 and $89.11{\sim}94.67%$ for T3, all of which were slightly higher than those of other pork products. The shear force values of T2 were significantly higher(p<0.05) than those of the other pork products throughout the storage period. The TBARS and VBN values of T2 were significantly higher(p<0.05) than those of T1 and T3. With regard to microorganisms, all grilled pork samples was in good condition, showing $1.93{\sim}3.48\;log_{10}$ CFU/g via total plate counts, and $1.74{\sim}3.48\;log_{10}$ CFU/g far lactic acid bacteria throughout the storage period. Regarding sensory evaluation, the scores of overall acceptability in all products were above 5.0 points through 40 days of storage.

The Effect of Seasoning on the Intestinal Absorption -Absorption by Passive Transport and the Effect of Red Pepper- (조미료가 창자 운동과 흡수기능에 미치는 영향 -소장의 피동적 흡수에 대한 고추의 영향-)

  • Shin, Dong-Hoon;Kim, Joong-Soo;Koh, Jae-Pyong;Ahn, Seung-Woon
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.23-31
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    • 1973
  • Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

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Improvement in Antagonistic Ablility of Antagonistic Bacterium Bacillus sp. SH14 by Transfer of the Urease Gene. (Urease gene의 전이에 의한 길항세균 Bacillus sp. SH14의 길항능력 증가)

  • 최종규;김상달
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.122-129
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    • 1998
  • It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.

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Optimization of Culture Conditions for D-Tagatose Production from D-Galactose by Enterobacter agglomerans. (Entrobacter agglomerans에 의한 D-Galactose로부터 D-Tagatose 생산조건의 최적화)

  • 오덕근;노회진;김상용;노봉수
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.250-256
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    • 1998
  • D-Tagatose production from D-galactose was investigated using 35 type strains of American Culture Type Collection (ATCC) and Korean Collection for Type Cultures (KCTC) which have potential to produce D-tagatose. Enterobacter agglomerans ATCC 27987 was selected as a D-tagatose producing strain due to its short fermentation time and high production of D-tagatose. Optimization of the culture conditions for D-tagatose production by E. agglomerans ATCC 27987 was performed. Among various carbon sources, D-galactose was the most effective carbon source for D-tagatose production. As the D-galactose concentration was increased, cell growth and D-tagatose production increased. Effect of nitrogen sources on D-tagatose production was studied. Of inorganic nitrogen sources, ammonium sulfate was effective one for D-tagatose production and yeast extract was the most suitable organic nitrogen nutrient. The concentrations of inorganic compounds such as KH$_2$PO$_4$, K$_2$HPO$_4$, and MgSO$_4$$.$7H$_2$O were also optimized for D-tagatose production. The optimal medium was determined to contain D-galactose of 20 g/l, yeast extract of 5.0 g/l, (NH$_4$)$_2$SO$_4$ of 2.0 g/l, KH$_2$PO$_4$ of 5.0 g/l, K$_2$HPO of 5.0 g/l, and MgSO$_4$$.$7H$_2$O of 5 mg/l. The optimal environmental conditions in a 250-$m\ell$ flask were found to be pH of 6.0, temperature of 30$^{\circ}C$, and agitation speed of 150 rpm. D-tagatose of 0.41 g/l could be obtained in 24 h from 20 g/l D-galactose at the optimal culture condition without induction and cell concentration.

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A Study on the Actual Condition of the Adult-smoking in a Region (일 지역 성인의 흡연실태)

  • Chong Young-sook
    • Journal of Korean Public Health Nursing
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    • v.13 no.1
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    • pp.97-113
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    • 1999
  • To investigate the actual condition of the adult-smoking in Chinan County. I used self-reporting questionnaires among 923 residents living in nine districts selected at random among the sites of eleven eup-myons from December 28. 1998 to January 6. 1999. collected data and analyized using SPSS. The smoking rate of adults in Chinan County was $36.9\%$. There was a significant difference in smoking rate according to the age. gender, education and job among general characteristics. The smoking rate in group of above 40s was about $40\%$ and that of the male took $55.5\%$ which was higher than that of female and the smoking rate of the people having high-school education was the highest$(46.2\%)$. With regard to the career. the smoking rate of the farmers ranked first$(46.1\%)$. Therefore anti-smoking business for smokers should be focused on above 40 years old, males. people having high school education and farmers. As the result of the survey of smoking habits of 340 people who smoke currently, the average smoking begining age was 21.8 and $81.5\%$ among them was between 16-25 years old. $4.4\%$ was under 15. during around the elementary school. Most motives to smoke were as followed; curiosity or taste$(33.6\%)$, relief of stress $(31.2\%)$, peer presure$(26.5\%)$ And considering the amount of the cigarette which is smoked per day, the case which is less than a pack of cigarettes was highest as $75.5\%$ and the people who smoke over one pack of cigarettes took $24.5\%$. As for the kind of tobacco which is smoked, how to smoke and the desire for the smoking, most case was toxin was moderate$(47.8\%)$ or mild$(46.9\%)$. shallowly$(49.0\%)$ or deeply$(46.3\%)$ and under stress$(33.4\%)$. after meals$(27.8\%)$, during drink$(15.7\%)$ and so on. The highest point marked among the factors of smoking motives was 'the reduce of negative emotion' $(3.27\pm1.00)$. followed by 'uncomfortable habits' $(2.87\pm1.02)$, 'addiction' $(2.84\pm1.06)$. 'habit' $(2.74\pm1.12)$. 'pleasure' $(2.70\pm1.04)$. 'stimulus' $(2.59\pm.90)$, 'sensation-exercise satisfaction' $(2.42\pm.97)$. Smokers smoke to reduce the negative emotions when angry in most common case. depressed. anxious. uncomfortable. lone. ashamed or embarrased. and intend to solve the certain problem. etc. Other motives are uncomfortable habit. addiction. habit. pleasure and the pursuit of stimulus. The level of nicotine dependence of adults m Chinan County was 10.57 which amount to 'high' wholly. As the resulf of the level of nicotine dependence score. the people who are low in the level of nicotine was $33.5\%$. the people who are high was $48.2\%$. very high was $18.4\%$. The approach for anti-smoking for smokers should be conducted differently according to the level of the nicotine. For the people who are in low level of nicotin dependence the prohibition of the smoking should be guided through the approach to foster strong will. for those who are in 'high' by acquiring proper method for the prohibition of smoking. and for those who are 'very high' the anti-smoking should be induced by providing proper program because of the possibility of the suffer from abstinence syndrome. The difference of the level of nicotine with the general characteristics of the objects had not statistically significant difference. The difference of the level of nicotine dependence accompanied by smoking habit had statistically significant difference according to the amount of smoke, the kind of tobacos. smoke inhale habit. In other words, the group of heavy smokers had higher level of the nicotine dependence than that of the light smokers relatively and the group which smoke strong taboaco has higher level of nicotine than that of which smoke mild or moderate. And the group of smokers who smoke deeply has higher level of nicotine than that who smoke shallowly or nonswallow. Aa a result of the analysis of the correlation between smoking motive factors and the level of nicotine, there was the indication that people who smoke for the decrease of the negative emotion. habit, pleasure. stimulus. sensation-exercise satisfaction had high level of the nicotine dependence. As the result of the anti-smoking will of smokers. $65.0\%$ of them had prohibition of smoking will. $29.3\%$ had no will to quit smoke. The most important reason for anti-smoking was health. $67.9\%$ had experience to try to quit smoke and the biggest reason to fail to quit smoking was the lack of the will power to keep anti-smoking. $52.8\%$ of them were advised to stop smoking from their spouses or children. only $2.8\%$ were by medical. The people who have the opinion to need anti-smoking education were $69.6\%$. Therefore when the business for the hygine of the mouth for adult is set. it should be centered on the people who have intention of prohibition of the smoking and help to quit smoking by way of other affirmative counter-program not smoking under stress.

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Study on the Processing and Compositions of Salted and Dried Mullet Roe (영암산 염건 숭어알의 가공과 조성에 관한 연구)

  • Joe, Sang-June;Rhee, Chong-Ouk;Kim, Dong-Youn
    • Korean Journal of Food Science and Technology
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    • v.21 no.2
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    • pp.242-251
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    • 1989
  • The salted-dried mullet(Mugil japonicus) roe is a kind of traditional food particulary in the area of Young-am gun, Chunnam province. This study was conducted to conform the scientific processing conditions and to evaluate the nutritional quality and changes of major components during storage times. The manufacturing method was that the fresh roe was salted for about 20 hours for the preparation of salted-dried roe, washed by clean waters, drained, shaped a flat piece with 1.2cm thickness by pressing, and spreaded sesame oils on the surface of the salted roe periodically during wind drying for 20 days. The dried roe was blanched in heated water$(80^{\circ}C/3min)$ and packaged the dried product for storages. The fractional compositions of free lipid of wind dried roe were 40% of neutral lipids, 12% of glycolipids and 9% of phospholipids and those of bound lipids were 13% of neutral lipids. 10% of glycolipids and 13% of phospholipids respectively. The major fatty acids of the roe were $C_{16:0}$, $C_{18:0}$, $C_{18:1}$, $C_{18:2}$ and $C_{20:0}$ which was consisted of free and bound lipids in wind drying method during processing and storages. Total amino acids were 99.87g/100g and major amino acids were Glu, Pro, Leu, Lys and CySH and the protein score was average 155% and the chemical score was average 109%. Free amino acids was 1,376mg% that had 50.61% of Pro and the major kinds of those were Tyr and CySH.

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Studies on Stability for the Quality of Ginseng Products -3. Determination of Sorption Properties and Optimum Moisture Contents in Extract, Extract Powder and Extract Tea of Red Ginseng- (인삼제품(人蔘製品)의 품질안정성(品質安定性)에 관한 연구(硏究) -3. 홍삼정(紅蔘精), 정분(精粉) 및 정차(精茶)의 흡습특성(吸濕特性)과 적정수분함량(適正水分含量) 기준설정(基準設定)-)

  • Choi, Jin-Ho;Byun, Dae-Seok;Park, Kil-Dang;Kim, Mu-Nam
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.2
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    • pp.215-221
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    • 1984
  • To investigate stability for the quality of selected ginseng products, their sorption properties were clarified in red ginseng extract(RGEP), and red ginseng extract powder(RGEP), and red ginseng extract tea(RGET). Simultaneously, the BET monolayer value of each product was determined in order to inquire out the possibility of establishment as a criterion for the optimum moisture content of the ginseng products. Based on the BET monolayer moisture level of spray dried RGEP which ranged from 4.08 to 4.65%, it would be desirable to establish the optimum moisture content of the products at 4.4${\pm}$0.3%. This is 1.3 to 1.9% lower than the criter on, "less than 6.0%". The optimum moisture level for RGET of which monolayer value ranged 0.93 to 1.37% would be 1.2${\pm}$0.17%. In this case, the maximum permissible limit of moisture content could presumably be raised up to 1.37% in place of current criterion, "less than l.2%". From the results of a study on the growth of molds, the optimum moisture content for RGE assumed to be extended up to 40.0${\pm}$1.07 despite 36.0${\pm}$1.0% of the present criterion. On the other hand, a storage study under the maltreated condition, $48{\pm}2^{\circ}C$ 75%RH, was also carried out in order to make it clear whether the BET monolayer values were able to be used as indices for optimum moisture level of the products. In all samples tested adsorption occurred at even higher levels of moisture than the monolayer values. However, since there are many other possible factors affecting the quality of products the optimum moisture content is preferable to be reduced to the monolayer value. As a result, it was proved that the optimum level of moisture for both RGEP and RGET could be established by the monolayer values.

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Effects of Dropwort Powder on the Quality of Castella (미나리가루의 첨가가 Castella의 품질에 미치는 영향)

  • Park, Sang-Jun;Lee, Kwang-Suck;An, Bye-Lyung
    • Journal of the East Asian Society of Dietary Life
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    • v.17 no.6
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    • pp.834-839
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    • 2007
  • This study was designed to determine the optimal ratio of dropwort powder in castella by adding the powder at levels of 0, 3, 6, 9, and 12% respectively. The properties of the castella were analyzed by specific gravity, specific volume, color determinations, texture properties and sensory evaluation. The Specific gravity increased with increasing amount of dropwort powder. However, the specific volume decreased with increasing dropwort powder. For the color values, as more dropwort powder was added, the L-value decreased. The castella with 9% dropwort powder had a higher hardness, gumminess, and chewiness. A sensory panel perceived that the external and internal color of the castella become darker with the dropwort powder substitution and the grain size decreased with increasing amount dropwort powder, while sweet taste showed no significant difference. The order of overall preference was DP 9>DP 6>DP 12>CON>DP 3. Therefore, the substitution of 9% of wheat flour with dropwort powder was recommended in the production of castella.

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Protective Effects on A2Kb Transgenic Mice That Were Immunized with Hepatitis B Virus X Antigen Peptides by the Activation of CD8+ T Cells; XEP-3 Specific CTL Responses in the in vitro Culture (B형 간염 바이러스 X 항원을 면역한 A2Kb Transgenic Mice에서 CD8+ T Cell의 활성화에 의한 X 항원 표현 재조합 Vaccinia Virus에 대한 방어 효과; in vitro 배양을 통한 XEP-3 특이적인 CTL의 반응)

  • Hwang, Yu Kyeong;Kim, Hyung-Il;Kim, Nam Kyung;Park, Jung Min;Cheong, Hong Seok
    • IMMUNE NETWORK
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    • v.2 no.1
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    • pp.41-48
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    • 2002
  • Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.