• Title/Summary/Keyword: MOLECULAR WEIGHT

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Ultrastructure and Metallothionein Expression in Rat Liver Treated with Di-(2-ethylhexyl)phthalate (Di-(2-ethylhexyl)phthalate에 의한 흰쥐 간세포 미세구조와 metallothionein 발현에 미치는 영향)

  • Kim, Da-Ham;Moon, Seung-Hoon;Lee, Mi-Young;Lee, Jong-Hwa;Park, Young-Hyun;Shin, Kil-Sang;Kim, Wan-Jong
    • Korean Journal of Environmental Biology
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    • v.25 no.4
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    • pp.289-296
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    • 2007
  • Di-(ethyhexyl) phthalate (DEHP), commonly used as a plasticizer for manufacturing flexible vinyl products, has been the topic of extensive research, especially concerning endocrine disrupting properties. Metallothionein (MT) is a low molecular weight (6,000$\sim$7,000 Da), cysteine-rich (22$\sim$23%), metal-binding protein and is known to be induced by extrinsic factors such as chemical agents and stresses. Some of the known function of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Nonetheless, the definitive physiological function of MT are still unknown. This study was carried out to investigate the effects of DEHP on the ultrastructural changes and the expression of MT of the rat liver. The rats were orally intubated with either corn oil (experimental control) or 0.5 mg, 1.5 mg and 4.5 mg DEHP kg$^{-1}$ day$^{-1}$ in 0.5 mL of corn oil for 15 days before sacrificing and sampling. DEHP induced mild ultrastrctural changes of some cell organelles such as rough endoplasmic reticulum, mitochondria, lysosomes and peroxisomes in the rat liver treated with DEHP. In the respect of immunogold labelling and Western blotting, MT expression of the liver tissue was up-regulated by DEHP. In conclusion, DEHP has effects on the ultrastructures and hepatic function for MT expression in rat.

The Effects of Needle Electrode Electrical Stimulation on Cellular Necrosis Blocking the Hippocampus after Induction of Ischemia (침전극저주파 자극이 해마의 세포사 차단에 미치는 영향)

  • Lee, Jung-Sook;Kim, Byung-Ki;Kim, Sung-Won;Kim, Ji-Sung;Kim, Dong-Il;Song, Chi-Won
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.2
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    • pp.111-120
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    • 2009
  • Objectives : This study was performed to investigate the effects of Needle Electrode Electrical Stimulation (NEES) on ischemia-induced cerebrovascular accidents. After obstruction and reperfusion of ${\ast}{\ast}$ arteries in white mice, the amounts of necrosis and inflammation related substances IL-6, Caspase-3, and PARP, C-fos were measured in neurons of the hippocampus. The following results were obtained. Methods : This study used 21 male specific pathogen free (SPF) SD (Sprague Dawley) rats, 8 weeks of age and approximately 300 g in weight, that were given at least 1 week to adapt to the lab environment Each exposed artery was completely occluded with non-absorbent suture thread and kept in that state for 5 minutes. The sutures were then removed to allow reperfusion of blood. Test group is control group for comparison with the common carotid artery occlusion models, a GI group that underwent common carotid artery occlusion, and a needle electrode electrical stimulation (NEES) group that underwent NEES after artery occlusion. The GI and NEES groups were given 12, 24, or 48 hours of reperfusion before NEES. NEES device (PG6, ITO, Japan, 9V, current, 2Hz) was used to stimulate the right and left acupoint ST36 of the SD rats for 30 minutes while they were sedated with 3% isoflurane. An immunohistochemistry test was done on the forebrains of the GI induced rats. All the data collected from this study was symbolized and analyzed using a statistics processing program (SPSS 12.0K/PC). The level of significance was set at ${\alpha}$=0.05 and a T-TEST analysis was used to find out the effects of treatment on each of the groups: the normal group, the CVA induced group, and the treatment after CVA induction group. Results : Both PARP and C-fos immuno-reactive cells, related to apoptosis, were greater in the GI groups than the NEES group. Caspase and IL-6 immuno-reactive cells, related to inflammation, were greater in the GI and NEES groups than the control group. Conclusions : This research was conducted to study the effects of NEES on CVA due to ischemia. Occlusion and reperfusion was performed on the common carotid arteries of white rats, after which amounts of substances related to neuron necrosis and inflammation - PARP, IL-6, Caspase-3, and C-fos - were measured in the Hippocampus

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Molecular Mechanism of Tetrabromobisphenol A (TBBPA)-induced Target Organ Toxicity in Sprague-Dawley Male Rats

  • Choi, Jae-Seok;Lee, Young-Jun;Kim, Tae-Hyung;Lim, Hyun-Jung;Ahn, Mee-Young;Kwack, Seung-Jun;Kang, Tae-Seok;Park, Kui-Lea;Lee, Jae-Won;Kim, Nam-Deuk;Jeong, Tae-Cheon;Kim, Sang-Geum;Jeong, Hye-Gwang;Lee, Byung-Mu;Kim, Hyung-Sik
    • Toxicological Research
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    • v.27 no.2
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    • pp.61-70
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    • 2011
  • Brominated flame retardants (BFRs) are present in many consumer products ranging from fabrics to plastics and electronics. Wide use of flame retardants can pose an environmental hazard, which makes it important to determine the mechanism of their toxicity. In the present study, dose-dependent toxicity of tetrabromobisphenol A (TBBPA), a flame retardant, was examined in male prepubertal rats (postnatal day 18) treated orally with TBBPA at 0, 125, 250 or 500 mg/kg for 30 days. There were no differences in body weight gain between the control and TBBPA-treated groups. However, absolute and relative liver weights were significantly increased in high dose of TBBPA-treated groups. TBBPA treatment led to significant induction of CYP2B1 and constitutive androstane receptor (CAR) expression in the liver. In addition, serum thyroxin (T4) concentration was significantly reduced in the TBBPA treated group. These results indicate that repeated exposure to TBBPA induces drug-metabolising enzymes in rats through the CAR signaling pathway. In particular, TBBPA efficiently produced reactive oxygen species (ROS) through CYP2B1 induction in rats. We measured 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of DNA oxidative damage, in the kidney, liver and testes of rats following TBBPA treatment. As expected, TBBPA strongly induced the production of 8-OHdG in the testis and kidney. These observations suggest that TBBPA-induced target organ toxicity may be due to ROS produced by metabolism of TBBPA in Sprague-Dawley rats.

Antigenic protein fractions reacting with sera of sparganosis patients (스파르가눔 항원단백질에 대한 스파르가눔증 환자 혈청의 반응 양상)

  • Choi, Sung-Ho;Kang, Shin-Yong;Kong, Yoon;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.26 no.3
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    • pp.163-168
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    • 1988
  • To observe the antigenic protein fractions in saline extract of Spirometra mansoni plerocercoid (sparganum), the crude extract was separated in reducing conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE). The proteins, transferred by celctrophoresis to introcillulose paper, were reacted with sera from 15 surgically confirmed sparganosis and 24 cysticercosis patients for immunoblotting. Out of 30 identified protein bands in the extract, bands of 29 and 36 kilodaltons (kDa) were the strongest and the most frequently reacting with specific antibody (IgG) in sparganosis sera. Bands of highter molecular weight also reacted with the sera but their frequency of reactions was lower. Sera of cysticercosis reacted with different protein bands in saline extract of sparganum, but the cross reactions were observed in strong antigenic bands of 29 and 36 kDa.

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Identification of Immunodominant B-cell Epitope Regions of Reticulocyte Binding Proteins in Plasmodium vivax by Protein Microarray Based Immunoscreening

  • Han, Jin-Hee;Li, Jian;Wang, Bo;Lee, Seong-Kyun;Nyunt, Myat Htut;Na, Sunghun;Park, Jeong-Hyun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.53 no.4
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    • pp.403-411
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    • 2015
  • Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes. Although many P. vivax proteins have been discovered, their functions are largely unknown. Among them, P. vivax reticulocyte binding proteins (PvRBP1 and PvRBP2) recognize and bind to reticulocytes. Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes. PvRBP1 and PvRBP2 are large (>326 kDa), which hinders identification of the functional domains. In this study, the complete genome information of the P. vivax RBP family was thoroughly analyzed using a prediction server with bioinformatics data to predict B-cell epitope domains. Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray. The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa. Evaluation of the humoral immune response to each recombinant PvRBP protein indicated a high antigenicity, with 38-88% sensitivity and 100% specificity. Of them, N-terminal parts of PvRBP2c (PVX_090325-1) and PvRBP2 like partial A (PVX_090330-1) elicited high antigenicity. In addition, the PvRBP2-like homologue B (PVX_116930) fragment was newly identified as high antigenicity and may be exploited as a potential antigenic candidate among the PvRBP family. The functional activity of the PvRBP family on merozoite invasion remains unknown.

Transgenic Plants with Enhanced Tolerance to Environmental Stress by Metabolic Engineering of Antioxidative Mechanism in Chloroplasts (엽록체 항산화기구 대사조절에 의한 환경스트레스 내성 식물)

  • Kwon Suk-Yoon;Lee Young-Pyo;Lim Soon;Lee Haeng-Soon;Kwak Sang-Soo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.151-159
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    • 2005
  • Injury caused by reactive oxygen species (ROS), known as oxidative stress, is one of the major damaging factors in plants exposed to environmental stress. Chloroplasts are specially sensitive to damage by ROS because electrons that escape from the photosynthetic electron transfer system are able to react with relatively high concentration of $O_2$ in chloroplasts. To cope with oxidative stress, plants have evolved an efficient ROS-scavenging enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX), and low molecular weight antioxidants including ascorbate, glutathione and phenolic compounds. To maintain the productivity of plants under the stress condition, it is possible to fortify the antioxidative mechanisms in the chloroplasts by manipulating the antioxidation genes. A powerful gene expression system with an appropriate promoter is key requisite for excellent stress-tolerant plants. We developed a strong oxidative stress-inducible peroxidase (SWPA2) promoter from cultured cells of sweetpotato (Ipomoea batatas) as an industrial platform technology to develop transgenic plants with enhanced tolerance to environmental stress. Recently, in order to develop transgenic sweetpotato (tv. Yulmi) and potato (Solanum tuberosum L. cv. Atlantic and Superior) plants with enhanced tolerance to multiple stress, the genes of both CuZnSOD and APX were expressed in chloroplasts under the control of an SWPA2 promoter (referred to SSA plants). As expected, SSA sweetpotato and potato plants showed enhanced tolerance to methyl viologen-mediated oxidative stress. In addition, SSA plants showed enhanced tolerance to multiple stresses such as temperature stress, drought and sulphur dioxide. Our results strongly suggested that the rational manipulation of antioxidative mechanism in chloroplasts will be applicable to the development of all plant species with enhanced tolerance to multiple environmental stresses to contribute in solving the global food and environmental problems in the 21st century.

Purification and Characterization of β-Lactamase Secreted from Bacillus sp. J105 Strain having β-Lectam Antibiotics Resistance. ((β-lactam계 항생물질 저항성을 지닌 Bacillus sp. J105 균주로부터 분비되는 베타 락탐 분해효소의 정제 및 특성)

  • Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Min-Jeong;Lee, Young-Choon;Lee, Jai-Heon;Joo, Woo-Hong;Choi, Yung-Hyun;Lim, Hak-Seob;Kim, Jeong-In;Seo, Kwon-Il;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.845-851
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    • 2008
  • ${\beta}-Lactamase$, secreted from Bacillus sp. J105 strain was purified to a single band on SDS-PAGE by ammonium sulfate precipitation, ion exchange column chromatography and gel-filtration. The molecular weight of the purified enzyme was 31 kDa on SDS-PAGE and its isoelectric point was 7.35. Optimal pH and temperature for enzymatic reaction were 5 and $40^{\circ}C$, respectively. As a result of total amino acid composition analysis of the purified enzyme, Gly and Ala were occupied 14.1 and 13.3 mole %, respectively. Km and Vmax value of purified enzyme were 1.33 mM and 0.36 mM/ml using ampicillin as a substrate, respectively.

Isolation of Anticarcinogenic Isoflavone-conjugated Glycoproteins from a Submerged Liquid Culture of Agaricus blazei Mycelia by the Autolysis Process (신령버섯균사체 액체배양물의 자가분해에 의한 항암성 isoflavone-conjugated glycoprotein 분리)

  • Kim, So Young;Kim, Young Suk;Jang, Joung Soon;Kim, Boh Hyun;Rakib, Abdur Md.;Kim, Gon Sup;Kim, Jeong Ok;Ha, Yeong Lae
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1316-1324
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    • 2014
  • Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone-conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at $53^{\circ}C$ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with $^1H$-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.

Synthesis of Poly(alkyl methacrylate)s Containing Various Side Chains for Pour Point Depressants (서로 다른 측쇄 구조를 가진 폴리(알킬 메타크릴레이트)계의 저온유동성 향상제 합성)

  • Hong, Jin-Sook;Kim, Young-Wun;Chung, Keun-Wo;Jeong, Soo-Hwan
    • Applied Chemistry for Engineering
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    • v.21 no.5
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    • pp.542-547
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    • 2010
  • n-Paraffin and saturated fatty acid methyl esters in the diesel and bio-diesel fuel crystallize at low temperature. Many articles have addressed various solutions for the low temperature crystallization problem and one of them is the use of methacrylate copolymers. In this work, we synthesized a series of copolymers in the reaction condition of 70 : 30 molar ratio of lauryl methacrylate (LMA) (or stearyl methacrylate (SMA)) and alkyl methacrylates. The structures of the copolymers were characterized by $^1H$-NMR and FT-IR spectroscopy, and the molecular weight of copolymers were obtained from Gel Permeation Chromatography (GPC) method. The concentrations of additives were 500~1000 ppm and 1000~10000 ppm in diesel fuels and bio-diesel fuel (BD5 and BD20), respectively. The addition of copolymers changes the many properties of fuel such as the pour point (PP), cloud point (CP) and cold filtering plugging point (CFPP). For example, the low temperature properties of the copolymers containing SMA ($PSMAmR_2n$) were excellently improved about 15, 7, and $10^{\circ}C$ for PP, CP and CFPP, respectively.

Isolation and Purification of Fibrinolytic Enzyme of Edible Mushroom, Sarcodon aspratus(Berk.)S. Ito (능이버섯으로부터 Fibrin 분해활성이 있는 단백질의 분리 및 정제)

  • 이종호;양정례;정청송;김희숙;조재선
    • Journal of Life Science
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    • v.11 no.6
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    • pp.561-567
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    • 2001
  • To isolate and purify fibrinolytic active substance from Sarcodon aspratus(N $H_4$)$_2$S $O_4$ precipitation, DE52 anion exchange column chromatography, Sephacryl-S 200gel filtration chromatography and Mono S cation FPLC were carried out and the characterizations of the purified enzyme were investigated. The bound active fraction on DE52 anion exchange column chromatography were eluted with 0.2 M NaCI and the fibrionlytic enzyme was purified after following Sephacryl-S200 gel fitration chromatography and Mono S cation EPLC. The specific activity of purified enzyme was 55.2 U/mg protein and increased 11.3 fold comparing crude extract and the yield was 49.5%. 12% SDS-PAGE electrophoresis and gel filtration chromatography revealed that Sarcodon aspratus fibrionloytic enzyme was highly purified and had 29.300 Da molecular weight. Enzyme activity of the purified fibrinolytic enzyme from Sarcodon aspratus was increased on higher pH and was stable until pH 10.5. On temperature dependent stability, the enzyme activity was decrease sharply but remained 25% relative activity on 8$0^{\circ}C$. This enzyme activity was inhibited by heavy metal ion, C $U^{2+}$ and $Co^{3+}$ with 68% and 38%, respectively. And also, the enzyme activity was inhibited with $Ca^{2+}$ chelator EDTA and serine protease inhibitor PMSF. These results from this study suggested that the fibrinolycit enzyme from Sarcodon aspratus is a serine protease and the enzyme activity was increased by $Ca^{2+}$ or $Mg^{2+}$ ion.n.ion.n.

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