• Title/Summary/Keyword: L(2, 1)-labeling

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Conjugation Process in Spirogyra varians Monitored with FITC-lectins(Zygnemataceae, Chlorophyta)

  • Yoou, Min-Chul;Kim, Man-Kyu;Kim, Gwang-Hoon
    • ALGAE
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    • v.24 no.1
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    • pp.39-45
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    • 2009
  • The conjugation processes of a filamentous freshwater green alga Spirogyra varians were examined using FITC-lectins. Conjugation comprised five steps: 1) aligning with adjacent filaments, 2) formation of conjugation protru-sion (papilla), 3) fusion of the protrusions, 4) formation of conjugation tube,and 5) formation of zygotes. Three lectins, ConA, RCA and UEA, showed considerable labeling during the progression of conjuation. FITC-ConA labeled the surfaces of filaments throughout the whole conjugation processes. FITC-RCA labeling was observed at the conjugation protrusions only after the papilla formation. Strong labeling continued until formationg of zygotes at the contacting area where the conjugation tube developed, but no labeling was detected on the surface of vegetative filaments. The labeling decreased gradually over time and disappeared when zygotes were formed. FITC-UEA showed similar labeling pattern with FITC-RCA except that weak labeling remained after zygote formation. Inhibition experiments using RCA, UEA which are complementary to sugars L-fucose and D-galactose, showed considerable decrease of conjugation (<32% vs. 70% in control). These results suggested that the lectin-carbohydrate recognition system might be involved in the conjugation of spirogyra varians.

A Study on Nutrition Composition Labeling and Nutrition Claim Practices for the Processed Foods (시판 가공식품의 영양성분 및 영양강조 표시 실태에 대한 연구)

  • 오세인;장영애
    • Korean journal of food and cookery science
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    • v.20 no.1
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    • pp.100-111
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    • 2004
  • The purpose of this study was to investigate the status of current nutrition labeling on the packaging of the processed foods that provide consumers with a reliable and consistent source of information, which has been considered as a useful aid for food selection and a potent educational tool for nutrition in daily life. The 2,160 processed foods purchased at the supermarket on September, 2002, were divided by food category issued from the 2002 food codes and assessed in the terms of the nutrition composition labeling and nutrition claims. Nutrition composition labeling was found on 356 of the 2160 processed foods items. Milk and dairy products had 49.7% of nutrition composition labeling, which was the largest number among the food category. Tables were most frequently used as the type of nutrition composition labeling (79.8%). Nutrition composition including many different ways of expression, such as a table of nutrition composition, indication of nutrition composition, analysis table of nutrition composition and comparative table of nutrition composition, made frequent use of nutrition composition labeling titles (78.7%). The various unit of measures were use in the nutrition labeling of the processed foods, per l00g or 100$m\ell$ was the highest (44.6%) under the currently practiced nutrition labeling. The correct labeling standard with nutrient content and % RDA except energy, was used on 47.8% of labels, and those with only liability indication nutrient and liability indication nutrients plus discretion indication nutrients were 25.3 and 22.5% respectively. The processed foods with nutrition claims were 8.0% (172 items). Nutrition claims were divided in two ways: nutrient content claims and nutrient comparative claims. The most frequently used claims were contained in the former (44.4%) and more or plus in the latter case (16.3%). Ca was the most popular item as a nutrition claim nutrient (50.6%).

The Evaluation of Factors Which Influence Binding Efficiency of Modified in Vivo Erythrocyte Labeling Technique (변형 체내 표지법에 의한 적혈구 표지시 결합효율에 영향을 미치는 인자 평가)

  • Seo, Han-Kyung;Kim, Min-Woo;Lim, Seok-Tae;Sohn, Myung-Hee
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.4
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    • pp.300-305
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    • 2004
  • Purpose: We underwent this study to evaluate the factors which influence labeling efficiency when modified in vivo erythrocyte labeling technique was used. Materials and methods: Thirty healthy volunteers (M:F=19:11, age:$25{\pm}2$ yrs) were enrolled in this study. Totally, two hundred ten samples were obtained from them. The 1 mg of stannous pyrophosphate was injected intravenously at the beginning of labeling. After suitable tinning time (5 min, 20 min, 35 min) passed by, blood (5 mL, 3 mL or 1 mL) was withdrawn into 10 mL syringe previously containing Tc-99m (740 MBq) and anticoagulant (heparin, ACD or CPDA) through 19-gauged scalp needle. The generator ingrowth time of Tc-99m was within 24 hrs in each case. The blood samples were placed on rotating invertor during incubation (10 min, 25 min, 40 min) but some of them were not. Immediately after the conclusion of incubation, the labeled blood specimens to analyze were centrifuged. and then %Unbound Tc-99m was calculated. Statical analysis was used paired T-test and one way ANOVA with SPSS 10.0. Results: The binding efficiency at 1 mL of blood volume was $73{\pm}32%,\;91{\pm}10%$ at 3 mL and $96{\pm}7%$ at 5 mL (p<0.01). The binding efficiency at 5 min of tinning time was $45{\pm}23%,\;98{\pm}6%$, at 20 min and $97{\pm}8%$ at 35 min (p<0.001). The binding efficiency at 10 min of incubation time was $96{\pm}7%,\;95{\pm}12%$ at 25 min and $98{\pm}3%$ at 40 min (p>0.05). The binding efficiency in case of using rotating invertor was $96{\pm}7%$ and the binding efficiency in case of not using it was $87{\pm}18%$ (p>0.05). There was no significant difference between them. In binding efficiency according to kinds of anticoagulants, ACD was $98{\pm}4%$, CPDA was $97{\pm}6%$ and heparin was $89{\pm}20%$ (p<0.001). Conclusion: When modified in vivo erythrocyte labeling technique is used with Tc-99m, the methods to obtain the highest labeling efficiency are as follow. The withdrawing blood volume should be over 3 mL, tinning time should be kept between 20 min and 35 min, and incubation time should be kept between 10 min and 40 min. ACD or CPDA have to be used as a anticoagulant except heparin and the blood samples should be placed on rotating invertor during incubation.

Radiolabeling of NOTA and DOTA with Positron Emitting $^{68}$Ga and Investigation of In Vitro Properties (양전자 방출핵종 $^{68}$Ga을 이용한 NOTA와 DOTA의 표지 및 시험관내 특성 연구)

  • Jeong, Jae-Min;Kim, Young-Ju;Lee, Yun-Sang;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.330-336
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    • 2009
  • Purpose: We established radiolabeling conditions of NOTA and DOTA with a generator-produced PET radionuclide $^{68}$Ga and studied in vitro characteristics such as stability, serum protein binding, octanol/water distribution, and interference with other metal ions. Materials and Methods: Various concentrations of NOTA 3HCl and DOTA 4HCl were labeled with 1 mL $^{68}$GaCl$_3$ (0.18$\sim$5.75 mCi in 0.1 M HCl in various pH. NOTA 3HCl (0.373 mM) was labeled with $^{68}$GaCl$_3$(0.183$\sim$0.232 mCi/0.1 M HCl 1.0 mL) in the presence of CuCl$_2$, FeCl$_2$, InCl$_3$, FeCl$_3$, GaCl$_3$, MgCl$_2$ or CaCl$_2$ (0$\sim$6.07 mM) at room temperature. The labeling efficiencies of $^{68}$Ga-NOTA and $^{68}$Ga-DOTA were checked by ITLC-SG using acetone or saline as mobile phase. Stabilities, protein bindings, and octanol distribution coefficients of the labeled compounds also were investigated. Results: $^{68}$Ga-NOTA and $^{68}$Ga-DOTA were labeled optimally at pH 6.5 and pH 3.5, respectively, and the chelates were stable for 4 hr either in the reaction mixture at room temperature or in the human serum at 37$^{\circ}C$. NOTA was labeled at room temperature while DOTA required heating for labeling. $^{68}$Ga-NOTA labeling efficiency was reduced by CuCl$_2$, FeCl$_2$, InCl$_2$, FeCl$_3$ or CaCl$_3$, however, was not influenced by MgCl$_2$ or CaCl$_2$. The protein binding was low (2.04$\sim$3.32%). Log P value of $^{68}$Ga-NOTA was -3.07 indicating high hydrophilicity. Conclusion: We found that NOTA is a better bifunctional chelating agent than DOTA for $^{68}$Ga labeling. Although, $^{68}$Ga-NOTA labeling is interfered by various metal ions, it shows high stability and low serum protein binding.

Colocalization of ${\alpha}$of Gq Protein with Actin Filaments in L8E63 Cells

  • Chae, Sungsuk;Park, Dongeun
    • Animal cells and systems
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    • v.1 no.1
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    • pp.93-98
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    • 1997
  • The present study investigated the cellular localization of a-subunit of Gq (Gaq) protein in developing L8E63, rat skeletal muscle cell line. The colocalization of Gaq with actin cytoskeleton was demonstrated by double-labeling experiments. In mononucleated myoblasts, the immuno-fluorescence staining pattern of Gaq was almost identical with that of F-actin visualized with rhodamine-conjugated phalloidin. However, this colocalization of Gaq with cytoskeleton was not maintained in multinucleated myotubes. The staining pattern of Gaq in myotubes did not match with any specific subcellular structure, but appeared as a uniformly distributed diffuse staining throughout the whole cell surface. Interestingly, change in the expression level of Gaq was not detected during myoblast differentiation, suggesting that actin-associated Gaq protein might dissociate from the cytoskeleton as cells differentiate. Immunocytochemical experiments using specific antibodies directed against several G proteins indicated that the subcellular localizations of Gai1, Gai2, Gai3, and Gao were different from those obtained with Gaq.

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Characteristics of the Inhibitory Action of Protease Inhibitors on the Glucose-6-phosphate Transporter

  • Choi, Joon-Sig;Shin, Jeong-Sook;Choi, Hong-Sug;Park, Jong-Sang
    • BMB Reports
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    • v.30 no.2
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    • pp.157-161
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    • 1997
  • The present paper reports characteristics and specificity of the inhibitory action of $N^{\alpha}-tosyl-L-lysine-chloromethyl\;ketone$ (TLCK) and $N^{\alpha}-tosyl-L-phenylalanine-chloromethyl\;ketone$ (TPCK) on the glucose6-phosphate transporter of rat liver microsomes. The TLCK-induced inhibition was pH dependent. The inhibition constants for TPCK were determined by following pseudo-Lst order reaction mechanism. The inhibition was protected by preincubation with excess amount of glucose-6-phosphate. The results proved that (a) TLCK inactivates the microsomal glucose-6-phosphate transporter, (b) the inhibition results from the modification of sulfhydryl groups of the transporter.

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Paratope Mapping of Anti-Ex-A IgG as Studied by NMR (NMR에 의한 anti-Ex-A IgG의 항원결합부위 해석)

  • Kim, Ha-Hyeong;Lee, Gwang-Pyo
    • YAKHAK HOEJI
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    • v.40 no.4
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    • pp.422-427
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    • 1996
  • The anti-Ex-A IgG was specifically labeled with stable isotopes, DL-His-2,4-$d_2$, L-Phe-$d_5$, L-Trp-$d_5$, L-Tyr-2,6-$d_2$ and L-[1-$^{13}C$]Trp, by growing hybridoma cell in serum-free medium. By use of NMR spectroscopy with selectively labeled Fab fragment, we applied a paratope mapping on antigen-antibody complex. Assignments of the observed carbonyl carbon resonances have been determined by using $^{13}C$-$^{15}N$ double labeling method in order to assign the Trp resonances. Photo CIDNP was also applied to investigate the antigen-binding site(s) on the surface residues of antibody. We found that Trp 36, which is located at the $V_H$ domain, is an important residue to bind to Ex-A, however, two Tyr on the surface of anti-Ex-A IgG plays no crucial role to bind to antigen. On the basis of these results, we demonstrate that stable isotope-aided NMR strategy can be extended to molecular structural analyses of the complex of an Fab fragment and a protein antigen.

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Evaluation of 99mTc-MAG3-2-nitroimidazole for hypoxic tumor imaging

  • Lee, Yun-Sang;Kim, Young Joo;Jeong, Jae Min
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.5 no.1
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    • pp.18-25
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    • 2019
  • 2-Nitroimidazole derivatives have been reported to accumulate in hypoxic tissue. We prepared a novel $^{99m}Tc-MAG_3$-2-nitroimidazole and evaluated the feasibility for hypoxia imaging agent. $Bz-MAG_3$-2-nitroimidazole was synthesized by direct coupling of $Bz-MAG_3$ and 2-nitroimidazole using dicyclohexylcarbodiimide. $Bz-MAG_3$-2-nitroimidazole was labeled with $^{99m}Tc$ in the presence of tartaric acid and $SnCl_2-2H_2O$ at $100^{\circ}C$ for 30 min. And the reaction mixture was purified by $C_{18}$ Sep-pak cartridge. The labeling efficiency and the radiochemical purity were checked by ITLC-SG/acetonitrile. The tumor was grown in balb/c mice for 8~13 days after the subcutaneous injection of tumor cells, CT-26 (murine colon adenocarcinoma cell). Biodistribution study and tumor autoradiography were performed in the xenografted mice after i.v injection of 74 kBq/0.1 mL and 19 MBq/0.1 mL of $^{99m}Tc-MAG_3$-2-nitroimidazole, respectively. In vivo images of $^{99m}Tc-MAG_3$-2-nitroimidazole in tumor bearing mice were obtained 1.5 hr post injection. The labeling efficiency was $45{\pm}20%$ and the radiochemical purity after purification was over 95%. Paper electrophoresis confirmed negative charge of $^{99m}Tc-MAG_3$-2-nitroimidazole. $^{99m}Tc-MAG_3$-2-nitroimidazole was very stable at room temperature and its protein binding was 53%. The $^{99m}Tc-MAG_3$-2-nitroimidazole exhibited high uptake in the liver, stomach and intestine. In biodistribution study using tumor bearing mice, the uptakes (% ID/g) of the tumor were $0.5{\pm}0.1$, $0.4{\pm}0.0$, $0.2{\pm}0.1$ and $0.1{\pm}0.1$ at 5, 15, 30 min and 4 hrs. Tumor/muscle ratio were $1.4{\pm}0.1$, $2.2{\pm}0.83$, $3.0{\pm}0.9$, and 3.7 (n=2) for 5, 15, 30 min and 4 hrs. The uptake in hypoxic area was found higher than in non-hypoxic area of tumor tissue by autoradiography. In vivo images showed the relatively faint uptake to the hypoxic tumor region. $^{99m}Tc-MAG_3$-2-nitroimidazole was successfully synthesized and found feasible for imaging hypoxia.

Conformance of Caffeine Content and Labeling of Ready-to-drink Coffee (Ready-to-drink (RTD) 커피 음료의 카페인 함량과 표시사항의 적합성)

  • Ye Eun Seo;Yoojeong Lee;Jung Hoan Kim
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.537-543
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    • 2023
  • This study monitored the caffeine content of ready-to-drink coffee and verified the appropriateness of the labeling. The caffeine content was analyzed using HPLC. The average caffeine content of cold brew coffee was 0.31-1.04 mg/mL, with an average of 0.55 mg/mL. The average content of product was 147.27 mg/bottle, and taking into account the recommended daily intake, an adult can consume 2.7 bottles. Americano coffee was 0.15-0.38 mg/mL, with an average of 0.28 mg/mL. The average content of product was 110.42 mg/bottle, and considering the recommended daily intake, an adult can consume 3.6 bottles. The caffeine content of decaffeinated cold brew coffee was 5.14 mg/bottle and compared to Americano coffee, more than 95% of the caffeine was removed. In addition, we verified the tolerance level of the total caffeine content in ready-to-drink coffee, and none of them exceeded 120%, signifying that all commercial products were effectively managed.

Acute and Subchronic Inhalation Toxicity Evaluation of Methyl Formate in Rats (Methyl formate의 랫드를 이용한 급성 및 아만성 흡입독성 평가)

  • Kim, Hyeon-Yeong;Lee, Sung-Bae;Han, Jeong-Hee;Kang, Min-Gu;Yang, Jeong-Sun
    • Environmental Analysis Health and Toxicology
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    • v.25 no.2
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    • pp.131-143
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    • 2010
  • We performed the tests of acute and subchronic inhalation toxicity of methyl formate, which has limited toxicological data in spite of its widespread use and enhanced hazard consequent on its high volatility. The median lethal concentration ($LC_{50}$) was evaluated to be above 5,000ppm(12.27 mg/L). In the test with subchronic inhalation, there are no deaths, but with reduction of body weight, food intake, organ weight by exposure to 400 (0.98 mg/L) and 1,600 (3.92 mg/L) ppm, dose-dependently. There were statistical differences in some hematological and blood biochemical parameters as compared to control (e.g. neutrophile and lymphocyte in the 1,600 ppm group, calcium and A/G in 1,600 ppm group). Methyl formate under the exposure of 1,600 ppm showed the respiratory findings with nasal, it was confirmed that the chemical has respiratory hazard with 1,600 ppm inhalation exposure, induces nasal epithelial atrophy, olfactory cell degeneration/regeneration and the contraction of olfactory cells, etc. According to the notification with Ministry of Labor (No. 2009-68) for classification, labeling and MSDS of chemicals, it is suggested for methyl formate to be classified as category 4 in acute (10.0$4\leq20.0$ mg/L), category 2 (0.2$\leq$1.0 mg/L/6h, 90 days) in specific target organ-repeated exposure.