• Title/Summary/Keyword: $^{15}N$ labeling

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$^{99m}Tc$ Labeling Kit Preparation and Characteristics of Anti-NCA-95 Monoclonal Antibody (항 NCA-95 단일클론항체의 $^{99m}Tc$표지 키트 제조 및 특성 연구)

  • Hong, Mee-Kyoung;Jeong, Jae-Min;Chung, June-Key;Choi, Seok-Rye;Kim, Chae-Kyun;Lee, Yong-Jin;Lee, Dong-Soo;Lee, Myung-Chul;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.4
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    • pp.541-547
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    • 1996
  • The previous monoclonal antibody labeling method for bone marrow immunoscintigrapy was complicated and laborious for clinical application. Also it showed a relatively low labeling efficiency. To improve this procedure, we compared several direct labeling methods of $^{99m}Tc$. 1) The labeling efficiency in the method using gluconate as a transchelator was low (40-70%), but immunoscintigraphy using this radiotracer produced a clear image. 2) To improve labeling efficiency, ${\beta}$-mercaptoethanol was removed after reduction. The labeling efficiency was improved up to 70-80%, but the radioactivity of the blood pool was high. 3) The higest labeling efficiency (>90%) and best quality images could be obtained by using MDP as a transchelating agent. It did not require additional procedures for separation of labeled antibodies. The immunoreactivity of this antibody was 60%. Residual MDP which can be taken up by the bone could be removed by PD-10 column. The reduced antibodies were stable with a high labeling efficiency (>90%) for up to 47 days by deep freezing. We concluded that the improved procedure for $^{99m}Tc$ labeling of anti-NCA-95 monoclonal antibody using MDP as a transchelating agent will be a simple and useful method for clinical application.

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Korean Semantic Role Labeling Using Domain Adaptation Technique (도메인 적응 기술을 이용한 한국어 의미역 인식)

  • Lim, Soojong;Bae, Yongjin;Kim, Hyunki;Ra, Dongyul
    • Journal of KIISE
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    • v.42 no.4
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    • pp.475-482
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    • 2015
  • Developing a high-performance Semantic Role Labeling (SRL) system for a domain requires manually annotated training data of large size in the same domain. However, such SRL training data of sufficient size is available only for a few domains. Performances of Korean SRL are degraded by almost 15% or more, when it is directly applied to another domain with relatively small training data. This paper proposes two techniques to minimize performance degradation in the domain transfer. First, a domain adaptation algorithm for Korean SRL is proposed which is based on the prior model that is one of domain adaptation paradigms. Secondly, we proposed to use simplified features related to morphological and syntactic tags, when using small-sized target domain data to suppress the problem of data sparseness. Other domain adaptation techniques were experimentally compared to our techniques in this paper, where news and Wikipedia were used as the sources and target domains, respectively. It was observed that the highest performance is achieved when our two techniques were applied together. In our system's performance, F1 score of 64.3% was considered to be 2.4~3.1% higher than the methods from other research.

NMR Studies on the Structure of Human Annexin I

  • Lee, Yeon-Hee;Han, Hee-yong;Oh, Jee-Young;Na, Doe-Sun;Lee, Bong-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.86-86
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    • 1997
  • Human annexin I is a member of annexin family of calcium dependent phospholipid binding proteins, which have been implicated in various physiological roles including phospholipase A$_2$ (PLA$_2$) inhibition, membrane fusion and calcium channel activity. In this work, the structure of N-terminally truncated human annexin I (Δ-annexin I) and its interactions with Ca$\^$2+/, ATP and cAMP were studied at atomic level by using $^1$H, $\^$15/N, $\^$l3/C NMR (nuclear magnetic resonance) spectroscopy. The effect of Ca$\^$2+/ binding on the structure of Δ-annexin I was investigated, and compared with that of Mg$\^$2+/ binding. The addition of Ca$\^$2+/ to Δ-annexin I caused some changes in the high field and low field regions of $^1$H NMR spectra. Whereas, upon addition of Mg$\^$2+/ to Δ-annexin I, almost no change could be observed. Also we found that the binding ratio of ATP to Δ-annexin I is 1. Because Δ-annexin I is a large protein with 35 kDa molecular weight, site-specific (carbonyl-$\^$l3/C, amide-$\^$15/N) labeling technique was used to determine the interaction sites of Δ-annexin I with Ca$\^$2+/ and ATP. Assignments of all the histidinyl carbonyl carbon resonances have been completed by using Δ-annexin I along with its specific 1,2-subdomain. The carbonyl carbon resonances originating from His52 and His246 of Δ-annexin I were significantly affected by Ca$\^$2+/ binding, and some Tyr and Phe resonances were also affected. The carbonyl carbon resonances originating from His52 is significantly affected by ATP binding, therefore His52 seems to be involved in the ATP binding site of Δ-annexin I.

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Neuronal Nitric Oxide Synthase-Immunoreactive Neurons In the Hamster Visual Cortex: Lack of Co-localization with Parvalbumin (햄스터 시각피질에서 Neuronal nitric oxide synthase-면역반응성 뉴런: parvalbumin과의 co-localization 부재)

  • Jin Mi-Joo;Lee Jee-Eun;Ye Eun-Ah;Jeon Chang-Jin
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.344-351
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    • 2005
  • Nitric oxide (NO) and calcium-binding proteins occur in various types of cells in the central nervous system. They are important signaling and calcium buffering molecules, respectively. In the present study, using immunocytochemistry we examined the distribution and the co-localization pattern of neurons containing neuronal nitric oxide synthase (nNOS) and parvalbumin in the visual cortex of hamster. The overall number of parvalbumin-immunoreactive (IR) neurons was 17 times higher than that of the nNOS-IR neurons in the hamster visual cortex. The highest differences were found in layer V, where parvalbumin-IR neurons were 54.7 times more abundant than nNOS-IR neurons. Many nNOS- and parvalbumin-IR neurons were similar in size, shape, and manner of distribution in the visual cortex. However, two-color immunofluorescence revealed that no neurons in the hamster visual cortex expressed both nNOS and parvalbumin. The present results indicate that there are subtle species differences in the co-localization pattern between nNOS and calcium-binding proteins. The present results also suggest not only the heterogeneity and functional diversity of nNOS-IRneurons in the visual cortex, but also the importance of understanding animal diversity

Effect of Dietary Fiber and Fat on Tumor Incidence and Cell Proliferation of Colonic Mucosa in DMH-Treated Rats (Dimethylhydrazine으로 처리한 쥐에서 식이섬유소와 지방종류가 대장의 종양발생율과 세포증식에 미치는 영향)

  • 최주선
    • Journal of Nutrition and Health
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    • v.31 no.4
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    • pp.697-707
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    • 1998
  • This study was designed to observe the effect of dietary fiber and fat on colon tumor incidence and cell proliferation. Male Sqraue Dawley rats(n=225) at 7 weeks of age, were divided into 3 groups depending on the type of fat b(beef tallow, corn oil and DHA-rich fish oil) and each group was again divided into 3 groups depending on type of fiber(fiber-free, perctin and cellulose) . The experimental diet containing dietary fat at 15%(w/w) and fiber at 6%(w/w) levels was fed for 25 weeks. At the same time, each rats was intramuscularly injected with DMH two times a week for 6 weeks to geive total dose of 180mg/kg body weight. Cell proliferation was measured by in vivo incroporation of bromodeoxyuridine (BrdU) into DNA. Fish oil decreased the tumor incidence (9.67%) compared with beef talow (33.39%) and corn oil (21.21%). Tumor incidence was decreased in all groups that fed cellulose (11.67%) compared with those of fiber-free(21.74%) and pectic(19.70%). Most of tumors was distributed at the site of the distal colon. The rats fed both fish oil and cellulose significantly decreased th enumber of tumors and tumor incidence compared to other groups. Fish oil was more effective in preventing cell prolofieration by decreasing crypt length and labeling index(LI) compared with beef tallow(p<0.05). Cell proliferation in distal colon was more developed to the upper part of the crypt compared to proximal colon. Overall tumor incidence and cell proliferation were more affected by dietary fat. But the effect of dietary fiber was different depending on type of fat in the experimental diet. These results suggest that a DHA -rich fish oil may has more decisive effect in inhibiting the cell proliferation in colon.

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Central energy metabolism remains robust in acute steatotic hepatocytes challenged by a high free fatty acid load

  • Niklas, Jens;Bonin, Anne;Mangin, Stefanie;Bucher, Joachim;Kopacz, Stephanie;Matz-Soja, Madlen;Thiel, Carlo;Gebhardt, Rolf;Hofmann, Ute;Mauch, Klaus
    • BMB Reports
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    • v.45 no.7
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    • pp.396-401
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    • 2012
  • Overnutrition is one of the major causes of non-alcoholic fatty liver disease (NAFLD). NAFLD is characterized by an accumulation of lipids (triglycerides) in hepatocytes and is often accompanied by high plasma levels of free fatty acids (FFA). In this study, we compared the energy metabolism in acute steatotic and non-steatotic primary mouse hepatocytes. Acute steatosis was induced by pre-incubation with high concentrations of oleate and palmitate. Labeling experiments were conducted using [$U-^{13}C_5$,$U-^{15}N_2$] glutamine. Metabolite concentrations and mass isotopomer distributions of intracellular metabolites were measured and applied for metabolic flux estimation using transient $^{13}C$ metabolic flux analysis. FFAs were efficiently taken up and almost completely incorporated into triglycerides (TAGs). In spite of high FFA uptake rates and the high synthesis rate of TAGs, central energy metabolism was not significantly changed in acute steatotic cells. Fatty acid ${\beta}$-oxidation does not significantly contribute to the detoxification of FFAs under the applied conditions.

Expression, Purification and NMR Studies on MC4R-TM2 Mutant

  • Oh, Dae-Seok;Yun, Ji-Hye;Lee, Weon-Tae
    • Journal of the Korean Magnetic Resonance Society
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    • v.16 no.1
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    • pp.34-45
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    • 2012
  • Melanocortin-4 receptor (MC4R) subtype is associated with obese humans. Especially, in a patient with severe early-onset obesity, novel heterozygous mutation in the MC4R gene was detected, resulting in an exchange of aspartic acid to asparagine in $90^{th}$ amino acid residue located in the predicted second trans-membrane domain (TM2). Mutations in the melanocortin-4 receptor (MC4R) gene are the most frequent monogenic causes of severe obesity which have been described as heterozygous with loss of function. In order to compare structure difference between MC4R wild type (MC4R-TM2-wt) and mutant (MC4R-TM2-D90N), we designed both MC4R-TM2-wt and MC4R-TM2-D90N construct in pET 21b vector. In this study, we optimized high-yield purification procedure for recombinant TM2-D90N. Eluted recombinant protein was resolubilized under urea condition for thrombin cleavage reaction and we conducted the high-performance liquid chromatography (HPLC) with reverse phase column under 1% acetonitrile, 0.01% TFA buffer solution. The molecular size of purified target peptide was confirmed by Tricine-SDS page analysis. To characterize MC4R-TM2-D90N, we have performed $^{15}N$-isotope labeling of peptide using M9 media and purified labeled target peptide for hetero-nuclear NMR spectroscopy.

Correlation of Preoperative Ki67 and Serum CA15.3 Levels with Outcome in Early Breast Cancers - a Multi Institutional Study

  • Rasmy, A;Abozeed, W;Elsamany, S;El Baiomy, M;Nashwa, A;Amrallah, A;Hasaan, E;Alzahrani, A;Faris, M;Alsaleh, K;AlFaraj, A
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3595-3600
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    • 2016
  • Background: To investigate the association between preoperative pathological Ki-67 labeling index and serum tumor marker cancer antigen 15-3 (CA 15-3) with clinic-pathological parameters and treatment outcomes in early breast cancer. Materials and Methods: A retrospective study at 4 cancer centers in Saudi Arabia and Egypt was performed. Data were collected for female patients diagnosed with unilateral early breast cancer between March 2010 and October 2013. Cases treated with neoadjuvant chemotherapy (NACT) followed by surgery and radiotherapy were included. NACT included 6-8 cycles of anthracycline and taxane based regimens. Trastuzumab and hormonal treatments were added according to HER2 and hormone receptor status. Baseline serum CA15.3 and pathological Ki67 levels were evaluated and correlated with disease free survival (DFS) and overall survival (OS). Results: A total of 280 pts was included. The median age was 49 years (38-66 y) and median overall survival was 35 (20-38) months (mo). Estrogen receptors (ER), progesterone receptors (PR) and HER 2 receptors were positive in 233 (83.2%), 198 (70%) and 65 cases (23.2%), respectively. High preoperative Ki67 and CA15.3 were noted in 177 (63.2%) and 131 (46.8%). A total of 45 (16%) patients had distal or local recurrence and 24 (8.6%) died of their disease. Most of the relapsed cases had high preoperative Ki-67 (n=41, 91%) and CA15.3 (n=28, 62%) values. All of the patients who died had a high Ki-67 but CA15.3 was high in 9 (37%) only. Mean DFS/OS in patients with high preoperative Ki-67 was 32 months /32 months as compared to 37 months/35 months in those with normal Ki-67 (p<0.001). Correlation of preoperative CA15.3 and survival was statistically not significant. Conclusions:Preoperative Ki-67 can be a predictive and prognostic marker. Higher levels are associated with poor DFS and OS in patients with early BC.

Status of serving labeling of home meal replacement-soups and stews, and evaluation of their energy and nutrient content per serving (가정간편식-국·탕·찌개류의 인분표시 및 영양표시 실태와 1인분 제공량 당 열량 및 영양성분 함량 평가)

  • Kim, Mi-Hyun;Choi, In-Young;Yeon, Jee-Young
    • Journal of Nutrition and Health
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    • v.54 no.5
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    • pp.560-572
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    • 2021
  • Purpose: In this study, the serving size of home meal replacement (HMR)-soups (Guk, Tang) and stews (Jjigae) available in the Korean market was investigated, and an evaluation of the nutrition per serving was conducted based on the nutrition labeling. Methods: The market research was conducted from March to August 2021 on products sold on the internet, convenience stores, supermarkets, and hypermarkets. A total of 370 products were investigated and classified into 3 types: Guk (n = 129), Tang (n = 132), and Jjigae (n = 109). Results: An analysis of the survey revealed that 72.9% of Guk, 71.2% of Tang, and 79.8% of Jjigae had labels with servings per container, and 89.2% of Guk, 91.7% of Tang, and 99.1% of Jjigae had labels with nutrition facts. The nutritional evaluation per serving of Guk, Tang, and Jjigae was conducted for 259 products (87 Guk, 86 Tang, and 86 Jjigae) having labels containing both the servings per container and nutrition facts. The average serving size of Tang was 367.6 g, which was significantly higher than Guk (325.3 g) and Jjigae (305.1 g) (p < 0.001). The calorie content of Jjigae (171.4 kcal) and Tang (162.3 kcal) was significantly higher than Guk (90.8 kcal) (p < 0.001), and the protein content was the highest in Tang (16.3 g) (p < 0.001). The sodium content per serving of Jjigae (1,479.0 mg) was significantly higher than Guk (1,073.3 mg) and Tang (959.8 mg) (p < 0.001). The percent daily value per serving of all three types was less than 10% on average for calories and 15-30% for protein, whereas for sodium showed an average of around 50% (48-74%). Conclusion: The serving size and nutritional value per serving of the HMR-soups and stews found in this study can be used as basic data to establish the reference serving size.

The Effect of Jininwhachul-tang-ga-wasong on the Hepatic Cancer in Mouse by N-Nitrosodiethylamine (진인화철탕가와송(眞人化鐵湯加瓦松)이 N-Nitrosodiethylamine으로 유발된 흰쥐의 간암에 미치는 영향)

  • Lim, Dong-Seok;Choi, Chang-Won
    • The Journal of Internal Korean Medicine
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    • v.29 no.3
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    • pp.582-594
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    • 2008
  • Objective : The purpose of our study was to investigate the effect of JinlnWhaChul-tang-ga-wasong (JIN) on NDEA-induced liver tumorigenesis. Materials and Methods : We investigated the possible protective effects of Jininwhachul-tang-ga-wasong (JIN) as an anticancer against NDEA-induced liver injury in mice. Experimental mice were classified into 3 groups; normal, saline administered group (control group), and JIN extract (0.15g/kg/every other day) administered group (JIN group) after being injected with NDEA over 12 weeks. We examined the state of differentiation of these tumors and the effects of JIN after 6 weeks. To confirm the induction of apoptosis, the cells were analyzed by terminal deorynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, acridine orange staining and flow cytometric analysis. To investigate inhibitory effect on the expression of COX-2 by JIN, we performed COX-2 immunohistochemistry and reverse transcription polymerase chain reaction analysis. Results : Body weights significantly decreased in the control and JIN groups compared with the normal group. The levels of cholesterol, hemoglobin and testosterone decreased in the control compared with the normal group. The level of estradiol significantly increased in the control compared with the normal group. The control group reacted with TUNEL assay more than the normal and JIN groups. Upon naked eye, light and electron microscopic examination, JIN improved the morphological and histopathological changes of the liver caused by NDEA-induced hepatic neoplasm. COX-2 immunoreactivity decreased in the JIN group compared with the control group, mRNA expression of the control group was greater than the normal and JIN groups. Conclusion : these results suggest the possibility that JIN may exert an anti-tumor effect on NDEA-induced liver tumorigenesis.

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