• Title/Summary/Keyword: Analysis on Labeling

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Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

Dietary total sugar intake of Koreans: Based on the Korea National Health and Nutrition Examination Survey (KNHANES), 2008-2011 (한국인의 총 당류 섭취실태 평가: 2008~2011년 국민건강영양조사 자료를 이용하여)

  • Lee, Haeng-Shin;Kwon, Sung-Ok;Yon, Miyong;Kim, Dohee;Lee, Jee-Yeon;Nam, Jiwoon;Park, Seung-Joo;Yeon, Jee-Young;Lee, Soon-Kyu;Lee, Hye-Young;Kwon, Oh-Sang;Kim, Cho-Il
    • Journal of Nutrition and Health
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    • v.47 no.4
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    • pp.268-276
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    • 2014
  • Purpose: The aim of this study is to estimate total sugar intake and identify major food sources of total sugar intake in the diet of the Korean population. Methods: Dietary intake data of 33,745 subjects aged one year and over from the KNHANES 2008-2011 were used in the analysis. Information on dietary intake was obtained by one day 24-hour recall method in KNHANES. A database for total sugar content of foods reported in the KNHANES was established using Release 25 of the U.S. Department of Agriculture National Nutrient Database for Standard Reference, a total sugar database from the Ministry of Food and Drug Safety, and information from nutrition labeling of processed foods. With this database, total sugar intake of each subject was estimated from dietary intake data using SAS. Results: Mean total sugar intake of Koreans was 61.4 g/person/day, corresponding to 12.8% of total daily energy intake. More than half of this amount (35.0 g/day, 7.1% of daily energy intake) was from processed foods. The top five processed food sources of total sugar intake for Koreans were granulated sugar, carbonated beverages, coffee, breads, and fruit and vegetable drinks. Compared to other age groups, total sugar intake of adolescents and young adults was much higher (12 to 18 yrs, 69.6 g/day and 19 to 29 yrs, 68.4 g/day) with higher beverage intake that beverage-driven sugar amounted up to 25% of total sugar intake. Conclusion: This study revealed that more elaborated and customized measures are needed for control of sugar intake of different subpopulation groups, even though current total sugar intake of Koreans was within the range (10-20% of daily energy intake) recommended by Dietary Reference Intakes for Koreans. In addition, development of a more reliable database on total sugar and added sugar content of foods commonly consumed by Koreans is warranted.

Expression of Hypoxia-inducible Factor-1 $\alpha$ in Esophageal Squamous Cell Carcinoma: Relationship to Prognosis and Tumor Biomarkers (식도 편평세포암에시 Hypoxia-inducible Factor-1 $\alpha$의 발현: 예후와 종양표지자와의 상관성)

  • 양일종;김종인;이해영;천봉권;조성래
    • Journal of Chest Surgery
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    • v.37 no.8
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    • pp.691-701
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    • 2004
  • Background: Tissue hypoxia is a characteristic of many human malignant neoplasms, and hypoxia inducible factor-1 (HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increased oxygen delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-1 a has been reported in many human malignancies, but in esophageal squamous cell carcinoma, the influence of HIF-1 a on tumor biology, including neovascularization, is not still defined. Material and Method: The influence of HIF-1 a expression on angiogenic factors, correlation between the tumor proliferation and HIF-1 a expression, interaction of HIF-1 a expression and p53, and correlation between HIF-1 a expression and clinicopathological prognostic parameters were investigated, using immunohistochemical stains for HIF-1 a, VEGF, CD34, p53, and Ki-67 on 77 cases of resected esophageal squamous cell carcinoma. Result: HIF-1 a expression in cancer cells was found in 33 of 77 esophageal squamous cell carcinoma cases. The 33 cases (42.9%) showed positive stain for HIF-1 a. High HIF-1 a expression was significantly associated with several pathological parameters, such as histologic grade (p=0.032), pathological TMN stage (p=0.002), the depth of tumor invasion (p=0.022), regional lymph node metastasis (p=0.002), distant metastasis (p=0.049), and lymphatic invasion (p=0.004). High HIF-1 a expression had significant VEGF immunoreactivity (p=0.008) and Ki-67 labeling index (p<0.001), but was not correlated with microvascular density within tumors (p=0.088). The high HIF-1 a expression was correlated with aberrant p53 accumulation with a marginal significance (p=0.056). The overall 5-year survival rate was 34.9%. The survival rate of patients with a high HIF-1 a expression was worse than that of patients with low-expression tumors (log-rank test, p=0.0001). High HIF-1 a expression was independent unfavorable factors although statistical significance is marginal in multivariate analysis. Conclusion: It is suggested that (1) high HIF-1 a expression in esophageal squamous cell carcinoma is associated with tumor hypoxia, or with genetic alteration in early carcinogenesis and progressive stages, (2) high HIF-1 a expression may be associated with intratumoral neovascularization through HIF-VEGF pathway, and (3) high HIF-1 a expression is associated with poor prognosis in patients with esophageal squamous cell carcinoma and may playa role as biomarker for regional lymph node metastasis.

Current status, perception and practicability of restaurant staffs related to reducing sodium use in Seongnam, Korea (성남 지역 외식업소의 나트륨 저감화 현황과 종사자의 저감화 실행에 대한 인식 및 실천용이도)

  • Ahn, So-Hyun;Kwon, Jong Sook;Kim, Kyungmin;Lee, Yoonna;Kim, Hye-Kyeong
    • Journal of Nutrition and Health
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    • v.52 no.5
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    • pp.475-487
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    • 2019
  • Purpose: With the increase of going out to eat, reducing the sodium in restaurant foods has a crucial impact on reducing sodium intake. This study aimed to assess the current status and perceptions of restaurant staffs related to reducing sodium use in restaurants. Methods: Restaurant managers and chefs (n = 312) in Seongnam area completed a questionnaire on the current status related to sodium use, the barriers to practice for reducing sodium use, support needs, and the practicability of methods for reducing sodium use in restaurants. Results: The percentage of restaurants in the preaction stage (including the precontemplation, contemplation, and preparation stages) for reducing sodium use was 79.7%. Logistic regression analysis showed that measuring salinity while cooking was associated with measuring seasoning (OR, 4.761; 95% CI, 2.325 ~ 9.751), action/maintenance stages of behavior change (OR, 2.829; 95% CI, 1.449 ~ 5.525) and providing salinity information of restaurant foods (OR, 6.314; 95% CI, 2.964 ~ 13.45). Maintaining taste and hindering the cooking process were the main barriers to reduce sodium use. The total practicability of actions for reducing sodium was higher in staffs who worked in restaurants that measured seasoning and salinity while cooking (p < 0.05 and p < 0.01, respectively). The hardest item to practice was 'purchase foods after comparing sodium content in the nutrition labeling'. 'Avoid serving salt-fermented foods as side dishes', 'serve small portions of kimchi and less salty kimchi', and 'put up promotional materials for reducing sodium intake' were selected as easy items to perform. The majority (82%) was willing to reduce sodium in restaurant foods under the support of local government and they desired the promotion of participating restaurants and education on cooking skills to reduce sodium. Conclusion: Measuring seasoning and salinity while cooking is a meaningful practice that is associated with stages of behavior change and the practicability of actions for reducing sodium. It is necessary to provide support and education with a gradual approach to staffs for reducing sodium in restaurant foods.

Hsp90 Inhibitor Induces Cell Cycle Arrest and Apoptosis of Early Embryos and Primary Cells in Pigs

  • Son, Myeong-Ju;Park, Jin-Mo;Min, Sung-Hun;Hong, Joo-Hee;Park, Hum-Dai;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.33-45
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    • 2011
  • Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

Genotype $\times$ Environment Interaction of Rice Yield in Multi-location Trials (벼 재배 품종과 환경의 상호작용)

  • 양창인;양세준;정영평;최해춘;신영범
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.6
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    • pp.453-458
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    • 2001
  • The Rural Development Administration (RDA) of Korea now operates a system called Rice Variety Selection Tests (RVST), which are now being implemented in eight Agricultural Research and Extension Services located in eight province RVST's objective is to provide accurate yield estimates and to select well-adapted varieties to each province. Systematic evaluation of entries included in RVST is a highly important task to select the best-adapted varieties to specific location and to observe the performance of entries across a wide range of test sites within a region. The rice yield data in RVST for ordinary transplanting in Kangwon province during 1997-2000 were analyzed. The experiments were carried out in three replications of a random complete block design with eleven entries across five locations. Additive Main effects and Multiplicative Interaction (AMMI) model was employed to examine the interaction between genotype and environment (G$\times$E) in the biplot form. It was found that genotype variability was as high as 66%, followed by G$\times$E interaction variability, 21%, and variability by environment, 13%. G$\times$E interaction was partitioned into two significant (P<0.05) principal components. Pattern analysis was used for interpretation on G$\times$E interaction and adaptibility. Major determinants among the meteorological factors on G$\times$E matrix were canopy minimum temperature, minimum relative humidity, sunshine hours, precipitation and mean cloud amount. Odaebyeo, Obongbyeo and Jinbubyeo were relatively stable varieties in all the regions. Furthermore, the most adapted varieties in each region, in terms of productivity, were evaluated.

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Risk Factor Analysis for Preventing Foodborne Illness in Restaurants and the Development of Food Safety Training Materials (레스토랑 식중독 예방을 위한 위해 요소 규명 및 위생교육 매체 개발)

  • Park, Sung-Hee;Noh, Jae-Min;Chang, Hye-Ja;Kang, Young-Jae;Kwak, Tong-Kyung
    • Korean journal of food and cookery science
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    • v.23 no.5
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    • pp.589-600
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    • 2007
  • Recently, with the rapid expansion of the franchise restaurants, ensuring food safety has become essential for restaurant growth. Consequently, the need for food safety training and related material is in increasing demand. In this study, we identified potentially hazardous risk factors for ensuring food safety in restaurants through a food safety monitoring tool, and developed training materials for restaurant employees based on the results. The surveyed restaurants, consisting of 6 Korean restaurants and 1 Japanese restaurant were located in Seoul. Their average check was 15,500 won, ranging from 9,000 to 23,000 won. The range of their total space was 297.5 to $1322.4m^2$, and the amount of kitchen space per total area ranged from 4.4 to 30 percent. The mean score for food safety management performance was 57 out of 100 points, with a range of 51 to 73 points. For risk factor analysis, the most frequently cited sanitary violations involved the handwashing methods/handwashing facilities supplies (7.5%), receiving activities (7.5%), checking and recording of frozen/refrigerated foods temperature (0%), holding foods off the floor (0%), washing of fruits and vegetables (42%), planning and supervising facility cleaning and maintaining programs of facilities (50%), pest control (13%), and toilet equipped/cleaned (13%). Base on these results, the main points that were addressed in the hygiene training of restaurant employees included 4 principles and 8 concepts. The four principles consisted of personal hygiene, prevention of food contamination, time/temperature control, and refrigerator storage. The eight concepts included: (1) personal hygiene and cleanliness with proper handwashing, (2) approved food source and receiving management (3) refrigerator and freezer control, (4) storage management, (5) labeling, (6) prevention of food contamination, (7) cooking and reheating control, and (8) cleaning, sanitation, and plumbing control. Finally, a hygiene training manual and poster leaflets were developed as a food safety training materials for restaurants employees.