• Title/Summary/Keyword: C5 Protein

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Expression of TIMP1, TIMP2 Genes by Ionizing Radiation (이온화 방사선에 의한 TIMP1, TIMP2 유전자 발현 측정)

  • Park Kun-Koo;Jin Jung Sun;Park Ki Yong;Lee Yun Hee;Kim Sang Yoon;Noh Young Ju;Ahn Seung Do;Kim Jong Hoon;Choi Eun Kyung;Chang Hyesook
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.171-180
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    • 2001
  • Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.

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Functional Screening for Cell Death Suppressors and Development of Multiple Stress-Tolerant Plants

  • Moon Hae-Jeong;Baek Dong-Won;Lee Ji-Young;Nam Jae-Sung;Yun Dae-Jin
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.143-148
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    • 2003
  • Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

Quality Characteristics of Yakju with Nuruk Extracts (누룩추출물로 제조한 약주의 품질특성)

  • Kang, Ji-Eun;Choi, Han-Seok;Kim, Jae-Woon;Kim, Chan-Woo;Yeo, Soo-Whan;Jung, Seok-Tae
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.223-230
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    • 2016
  • We investigated the fermentation characteristics of yakju made with nuruk extracts. Alcoholic fermentation was performed in an incubator set at $25^{\circ}C$ for 14 days. The study results are as follows: the alcohol content of yakju made with modified nuruk (18.67%) was higher than that made with other nuruk extracts (14.80-17.50%). The pH value of this modified nuruk (4.13) was higher than that of traditional nuruk (3.70-4.00). The yakju containing modified nuruk showed the highest acidity (3.64 mL/10 mL), amino acidity (4.72 mL/10 mL), soluble solid ($1.20^{\circ}Bx$), and color difference (39.78). The concentration of organic acids in traditional nuruks was 212.25-312.97 mg/100 mL, whereas that in modified nuruk was 295.92 mg/100 mL. Nitrogen compound concentrations in modified nuruk (2285.89 mg/L) were 2-14 times higher than that in traditional nuruk. Nitrogen compound concentration and amino acidity are the physiochemical characteristics with the highest impact on off-flavor and browning of yakju. In order to reduce these unfavorable characteristics, material with a lower protein content should be used.

Application of mix LPS-ELISA for monitoring of antibodies to major serogroups of Salmonella in animal (Mix LPS-ELISA법을 이용한 살모넬라균 복합감염항체 동시 모니터링)

  • Lee, Hee-soo;Im, Suk-kyoung;Woo, Seng-ryong;Yi, You-young;Kim, Jong-yeom;Joo, Yi-seok;Kim, Jong-man
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.793-802
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    • 1998
  • Salmonellosis caused by a number of serotypes of Salmonella is an infectious, acute or chronic, zoonotic disease and characterized by enteritis and diarrhea, septicemia in animal. In these studies we investigated the prevalent serotypes of Salmonella causing animal salmonellosis in Korea and the 71 strains of Salmonella spp. were isolated from materials such as mesenteric lymph nodes, fecal samples from slaughtered animal. With the identification test results, the most prevalent serotypes were, in order, S stanley 31 strains (43.7%), S typhimurium 19 strains (26.8%) and S montevideo 11 strains (15.5%), respectively. And we could establish the method for detection of antibodies to broad variety of Salmonella serotypes. Lipopolysaccharide(LPS) antigen extracted from Salmonella was more sensitive and specific than outer membrane protein antigen from that for detection of Salmonella antibody by using an indirect ELISA. The optimal concentration of antigen was 100ng/ml of LPS, the dilutions of conjugate and serum were 1 : 1,000~2,000 and 1 : 200~400, respectively. The mix LPS-ELISA which was used by mixing LPS from S typhimurium (group B), S choleraesuis (group C) and S enteritidis (group D) were more rapid and effective than that used LPS from individual strain for detection of Salmonella serogroup O4, O7 and O9 antibody at the same time. We could obtain the high values of optical density ($0.73{\pm}0.32$) by mix LPS-ELISA on the farm which had occurred salmonellosis, but very low values of $0.17{\pm}0.06$ on the negative farm of salmonellosis. So, the mix LPS-ELISA may be used to monitor the serological surveillance for the presence of infection with a number of serotypes of Salmonella and would be useful for prevention and control of salmonellosis in animal.

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Evaluation of different milking practices for optimum production performance in Sahiwal cows

  • Aslam, Naveed;Abdullah, Muhammad;Fiaz, Muhammad;Bhatti, Jalees Ahmad;Iqbal, Zeeshan Muhammad;Bangulzai, Nasrullah;Choi, Chang Weon;Jo, Ik Hwan
    • Journal of Animal Science and Technology
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    • v.56 no.4
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    • pp.13.1-13.5
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    • 2014
  • The production performance of multiparous lactating Sahiwal cows (n = 24) was evaluated according to both milking frequency and method. Selected animals were randomly divided into four groups containing six animals each under a completely randomized design. Cows in groups A & B were milked by the hand milking method three times per day, respectively. Similarly, cows in groups C & D were milked by the machine milking method two and three times per day, respectively. All animals were maintained under uniform feeding and management conditions. Dry matter intake was high in animal groups milked three times per day, and it remained unchanged between the hand and machine milking methods. Milk yield was higher (P < 0.05) in cows milked three times compared to those milked twice per day, and it did not differ between hand and machine milking methods. Milk fat percentage was higher (P < 0.05) in cows milked twice per day compared to those milked three times using both machine and hand milking methods. The percentage of total solids showed a similar pattern as the fat percentage. However, percentages of protein, lactose, and non-fat solids in milk were not significantly different (P > 0.05) among the treatment groups. Collectively, the results show that milking three times per day instead of twice at 8-hour intervals can enhance milk yield in Sahiwal cows using both hand and machine milking methods.

Effects of Post-harvest Storage Duration and Variety on Nutrient Digestibility and Energy Content Wheat in Finishing Pigs

  • Guo, P.P.;Li, P.L.;Li, Z.C.;Stein, H.H.;Liu, L.;Xia, T.;Yang, Y.Y.;Ma, Y.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1488-1495
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    • 2015
  • This study was conducted to investigate the effects of post-harvest storage duration and wheat variety on the digestibility and energy content of new season wheat fed to finishing pigs. Two wheat varieties (Shi and Zhong) were harvested in 2013 and stored in the warehouse of the Fengning Pig Experimental Base at China Agricultural University for 3, 6, 9, or 12 mo. For each storage period, 12 barrows were placed in metabolism crates and allotted to diets containing 1 of the 2 wheat varieties in a randomized complete block design. The experimental diets contained 97.34% wheat and 2.66% of a vitamin and trace mineral premix. With an extension of storage duration from 3 mo to 12 mo, the gross energy (GE) and crude protein (CP) of the wheat decreased by 2.0% and 12.01%, respectively, while the concentration of neutral detergent fiber (NDF), acid detergent fiber (ADF) and starch content increased by 30.26%, 19.08%, and 2.46%, respectively. Total non-starch polysaccharide, total arabinose, total xylose and total mannose contents decreased by 46.27%, 45.80%, 41.71%, and 75.66%, respectively. However, there were no significant differences in the chemical composition between the two wheat varieties with the exception of ADF which was approximately 13.37% lower in Shi. With an extension of storage duration from 3 mo to 12 mo, the digestible energy (DE), metabolizable energy (ME) content and the apparent total tract digestibility of GE, CP, dry matter, organic matter, ether extract, ADF and metabolizability of energy in wheat decreased linearly (p<0.01) by 5.74%, 7.60%, 3.75%, 3.88%, 3.50%, 2.47%, 26.22%, 27.62%, and 3.94%, respectively. But the digestibility of NDF changed quadratically (p<0.01). There was an interaction between wheat variety and storage time for CP digestibility (p<0.05), such that the CP digestibility of variety Zhong was stable during 9 mo of storage, while the CP digestibility of variety Shi decreased (p<0.05). In conclusion, the GE, DE, and ME of wheat was stable during the first 3 to 6 mo of post-harvest storage, and decreased during the following 6 to 12 mo of storage under the conditions of this study.

Nutritional Status and Hair Mineral Content of Elementary School Children with Behavioral Problems (문제 행동을 가진 초등학생의 영양 상태와 모발 미네랄 함유량)

  • Kang, Seung-Wan;Kim, Jin-Young;Cho, Sang-Woon;Park, Yoo Kyoung
    • Journal of the Korean Dietetic Association
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    • v.18 no.2
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    • pp.97-114
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    • 2012
  • This study was performed to assess the nutritional status and hair mineral content of children with behavioral problems and compare the values with a gender, age-matched control group. The subjects were recruited from S elementary school children in Seoul, Korea. Students scored higher than 60 points were diagnosed with behavioral problems according to the Korean-Child Behavior Checklist. Nutritional assessment of the two groups (behavior problem group: male n=15, female n=24; $9.6{\pm}1.6$ years, control group: male n=16, female n=18; $9.5{\pm}1.9$ years) was performed using a nutritional survey and by measuring hair mineral contents. In the results, food frequency questionnaire analysis showed that the intakes of anchovy (P<0.05), soybean curb (P<0.01), radish (P<0.05), bean sprouts (P<0.05), spinach (P<0.05), carrot (P<0.05), pumpkin (P<0.05), lettuce (P<0.05), cabbage (P<0.01), apple (P<0.05), and milk (P<0.01) were higher in the control group than the behavior problem group, whereas intakes of ramyeon (P<0.05), cookies (P<0.05), and coke (P<0.01) were higher in the behavior problem group than the control group. Intakes of most nutrients such as plant-derived protein (P<0.05), fiber (P<0.05), plant-derived calcium (P<0.05), phosphorus (P<0.05), plant-derived iron (P<0.05), vitamin B2 (P<0.05), vitamin B6 (P<0.05), vitamin C (P<0.01), vitamin E (P<0.05), and folate (P<0.05) were significantly higher in the control group than the behavior problem group. Hair analysis showed that the levels of arsenic (P<0.05), mercury (P<0.001), uranium (P<0.05), iron (P<0.001), boron (P<0.01), and germanium (P<0.001) were lower, but the levels of phosphate (P<0.05), chromium (P<0.001), sodium (P<0.05), and sulfur (P<0.001) were higher in the behavior problem group than the control group. Conclusively, behavioral problems constitute a complicated condition in which nutritional factors may play major roles. However, it is still under investigation as to whether or not modification of dietary habits or nutritional supplementation can improve children's behavior, since symptoms require a broad understanding of the environmental and genetic interactions.

Influence of proton pump inhibitor therapy on intestinal inflammation assessed by fecal calprotectin in pediatric patients

  • Kim, Su Yeong;Lee, Na Mi;Yun, Sin Weon;Chae, Soo Ahn;Lim, In Seok;Choi, Eung Sang;Yi, Dae Yong
    • Clinical and Experimental Pediatrics
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    • v.62 no.10
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    • pp.400-404
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    • 2019
  • Background: An increase in the numbers of patients with gastrointestinal symptoms has recently been observed. Purpose: To investigate the effects of proton pump inhibitor (PPI) therapy on intestinal inflammation in children and adolescents as confirmed by clinical manifestations and objectively assessed by fecal calprotectin (FC) level measurement. Methods: Consecutive children (aged 3-18 years) who presented with gastrointestinal symptoms and were treated with or without PPI for at least 1 month were enrolled. Patients were divided into PPI and non-PPI groups. The PPI group was further subdivided by treatment duration and type of PPI used. Stool samples were collected for FC evaluation at baseline and after treatment and clinical data and FC levels were compared between the groups. Results: Fifty-one patients (15 boys, 36 girls) were enrolled in the study. The PPI group included 37 patients, while the non-PPI group included 14 patients. Clinical symptoms were not significantly different. FC levels and laboratory results, including C-reactive protein levels, white blood cell count, and absolute neutrophil count, were not statistically different before versus after PPI treatment. After treatment, FC levels decreased to 8.1 mg/kg (-575.4 to 340.3 mg/kg) in the PPI group and increased to 5.6 mg/kg (-460.0 to 186.9 mg/kg) in the non-PPI group compared to those before treatment (P=0.841). The number of patients with increased FC levels was not significantly different between the 2 groups (48.6% vs. 64.3%, P=0.363), similar to that observed in patients with an FC level > 50 mg/kg (24.3% and 7.1%, P=0.250). PPI therapy type and duration did not affect the FC levels (P=0.811 and P=0.502, respectively). Conclusion: Although we aimed to confirm the evidence of intestinal inflammation due to PPI use in children and adolescents through clinical symptoms and FC measurement, no significant changes were observed.

Strategies of Improve Elderly Nutrition through Nutrition Education : Evaluation of the Effectivenss of the Program (영양교육에 의한 노인 영양증진전략연구 : 효과분석)

  • 임경숙;민영희;이태영;김영주
    • Korean Journal of Community Nutrition
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    • v.4 no.2
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    • pp.207-218
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    • 1999
  • The increased number of elderly in Korea presents great challenges for community nutrition programs. Strategies to enhance elderly nutrition are needed, as well as dietary advice that are reality-based and tailored to the need of the elderly. The purpose of this study was to evaluate the overall effectiveness of an elderly nutrition improvement program(ENIP), by assessing changes in nutrition knowledge, attitudes, and behavior after program completion. ENIP was conducted in Suwon for 4 months in 1998 at 5 centers for the elderly. The program provided individual nutrition counseling and brief biweekly group education sessions. To stimulate participation and motivation, the elderly in two centers were served free milk packs(200ml) three time a week during the program(milk+education group), and the elderly in 3 other centers were served yoghurt on the program day(education group). Altogether, 102 subjects finished the program(milk+education group 67 ; education group 35). Data about nutrition knowledge, dietary attitudes, diet records(24 hour recall) and dietary habit was collected before(baseline) and after the program(follow-up). The Drop-out rate for the milk+education group(2.9%) was significantly lower than that for the education group(30.0%)(p<.001). The Nutrition Knowledge fo females milk+education group increased significantly after the program, but no significant differences were observed in other groups. Over 60% of all the elderly showed positive responses to the healthy eating attitudes. Energy, protein, vitamin A, riboflavin, Calcium and Potassium intakes improved in the male milk+education group. The Vitamin C intake improved in the male education group. Energy and vitamin A intakes improved in the female milk+education group and energy, vitamin A, thiamin, riboflavin, niacin, Ca nad P in the female education group. A stepwise multiple regression analysis was performed to examine the effects of food intake changes on overall nutrient quality. For all elderly, the fish intake explained 12.6% of the variance, followed by candy intake and vegetable intake(model $R^2$=19.6%). Dietary habits such as meal time regularity and salty food reduction were markedly improved in both groups. Sixty-one percent of the milk+education subjects stated their participation was voluntary and active, while 51% of education subjects did. Sixty four percent of the subjects stated their interest on diet increased remarkably through program participation. Considering the results, it is conjectured that ENIP had a great impact on nutrient intake, dietary attitudes and habits. Milk supplement showed no direct effects on elderly nutrition improvement, but it encouraged the program by reducing drop-out rates and inducing voluntary participation. Therefore milk supplement could enhance the community nutrition education program. Moreover it is strongly suggested that the ENIP be expanded to other communities.

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A PCR Method for Rapid Detection of Buckwheat Ingredients in Food (식품에서 메밀 성분의 검출을 위한 PCR 방법)

  • Jeon, Young-Jun;Kang, Eun-Sil;Hong, Kwang-Won
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.276-280
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    • 2007
  • Buckwheat often causes severe allergic reactions in sensitive people. One of the major allergenic proteins in common buckwheat (Fagopyrum esculentum) has been found to be a BW10KD protein. In this study, we developed a PCR method to detect buckwheat ingredients in food using primers corresponding to the allergenic BW10KD gene. Five pairs of oligonucleotide primers successfully enabled PCR amplification of the specific regions of the genomic BW10KD DNA from buckwheat, but no amplification from seven other cereals and beans (barley, wheat, German millet, African millet, soybean, red bean, and black bean). The proposed PCR method was applied to analyze 12 processed foods (buckwheat flour, buckwheat noodle, buckwheat jelly, wheat noodle, instant noodle, black sesame gruels, sunsik, cookie, misutkaru, and three kinds of cereal); among them, only three samples including buckwheat flour, buckwheat noodle and buckwheat jelly showed a positive reaction to the detection. This PCR method was able to detect as little as 1 ng of common buckwheat DNA. This rapid and specific PCR method would be applicable to detect allergenic buckwheat ingredients in food.