• Title/Summary/Keyword: bovine.

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Purification of IgG1 Type Mouse Monoclonal Antibodies with DEAE-Trisacryl Chromatography (DEAE-Trisacryl 크로마토그래피법에 의한 IgG1 Type 쥐 단일클론 항체의 분리정제)

  • 최태부;정용근
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.335-342
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    • 1988
  • An anion exchange chromatography was employed for the purification of mouse monoclonal antibodies from ascitic fluid and in vitro cultivation media. After cultivation of hybridomas, Alps 25-3, HCGK, A4W, and KW, producing IgG1, the culture supernatants were harvested by centrifugation, precipitated with 50-60% ammonium sulfate, and dialyzed against 0.025 M Tris-HCI buffer (pH 8.2). Then the dialyzed samples were loaded into a DEAE-Trisacryl M anion exchange column. Monoclonal antibodies bound to the DEAE-Trisacryl M were eluted with 0.025 M Tris-HCI buffer (pH 8.2) containing 30-40 mM NaCl. In ammonium sulfate precipitation, the recovery of the monoclonal antibody was shown to be 90% and 84% from in vitro culture media containing 10% and 2% fetal bovine serum, respectively. On the other hand, the pretreatment by ultrafiltration enhanced the yield up to 91% whereas the purity was lower than that by ammonium sulfate treatment. Subsequently, in the DEAE-Trisacryl M chromatographic separation, the purities and recoveries of all the monoclonal antibodies from both the in vitro culture supernatants and ascitic fluids were 70-80% and 65% respectively. The monoclonal antibody, Alps 25-3 could be further purified with a purity of 95% through an immunoadsorbent chromatography.

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Generation of Newly Discovered Resistance Gene mcr-1 Knockout in Escherichia coli Using the CRISPR/Cas9 System

  • Sun, Lichang;He, Tao;Zhang, Lili;Pang, Maoda;Zhang, Qiaoyan;Zhou, Yan;Bao, Hongduo;Wang, Ran
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1276-1280
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    • 2017
  • The mcr-1 gene is a new "superbug" gene discoverd in China in 2016 that makes bacteria highly resistant to the last-resort class of antibiotics. The mcr-1 gene raised serious concern about its possible global dissemination and spread. Here, we report a potential anti-resistant strategy using the CRISPR/Cas9-mediated approach that can efficiently induce mcr-1 gene knockout in Escherichia coli. Our findings suggested that using the CRISPR/Cas9 system to knock out the resistance gene mcr-1 might be a potential anti-resistant strategy. Bovine myeloid antimicrobial peptide-27 could help deliver plasmid pCas::mcr targeting specific DNA sequences of the mcr-1 gene into microbial populations.

Effect of Carbonic Anhydrase on CO2 Absorption in Amine Solutions for CO2 Capture (CO2 포집용 아민 흡수제에서 탄산무수화 효소가 CO2 흡수에 미치는 영향)

  • Lee, In-Young;Kwak, No-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.39 no.11
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    • pp.607-612
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    • 2017
  • The effect of carbonic anhydrase on $CO_2$ absorption rates and the heat of reaction were evaluated in various amine solutions for post combustion $CO_2$ capture process. The $CO_2$ absorption rate was analyzed in 30 wt% MEA, AMP, DMEA, MDEA aqueous solutions with and without carbonic anhydrase (250 mg/L) from bovine erythrocyte. $CO_2$ absorption rates were increased in all solutions with carbonic anhydrase. The effect of carbonic anhydrase on absorption rates was more in tertiary amine (DMEA and MDEA) solutions than in primary amine (MEA) and hindered amine (AMP) solutions. The heat of reaction of MEA, DMEA, MDEA aqueous solutions with and without carbonic anhydrase were measured using reaction calorimeter. Carbonic anhydrase decreased the heat of absorption in all solutions. The results suggested that tertiary amines that have the excellent desorption ability were suitable for applying carbonic anhydrase to the post combustion $CO_2$ capture process and the effect of carbonic anhydrase was best in MDEA solution.

A STUDY ON THE MECHANICAL PROPERTIES OF EXPERIMENTAL, COMPOSITES CONTAINING ZIRCONIA FILLER (지르코니아 필러를 첨가한 복합레진의 기계적 성질에 관한 연구)

  • Rew, Kyung-Hee;Choi, Ho-Young;Choi, Kyoung-Kyu;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.25 no.3
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    • pp.421-434
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    • 2000
  • The purpose of this study was to evaluate the influences of incorporation of zirconium-silicate on diametral tensile strength, shear bond strength to the enamel, and depth of cure of 7 experimental composites. One group contained no filler(group 1 or control group), and the other 6 composites contain 75% filler in which zirconium-silicate(Zr-Si) were 0%, 2%, 4%, 6%, 8%, 10% with reduced contents of silica filler, respectively. Both of fillers were treated with 1% silane (${\gamma}$-methacryloxypropyltrimethoxy silane). Light curable monomers were prepared by mixing Bis-GMA and TEGDMA with 3:1 ratio and adding camphoroquinone(CQ) 0.6% with tertiary amine 0.3%. Diametral tensile strengths of specimens with $3mm{\times}6mm$ were measured with Instron (No.4467, USA) with 1mm/min crosshead speed. Shear bond strengths of composites which bonded to bovine enamel etched with 37% phosphoric acid were measured at Instron Testing Machine with as same speed as in diametral tensile strengths. Depth of cure were measured by a method that composite was filled in cylinder mold, illuminated at one side. and uncured composite was removed with acetone, and the residual thickness of composite was measured. Following results were obtained ; 1. Composites containing 0%, 2%, or 4% zirconium-silicate filler(group 2, 3 and 4) showed the statistically higher diametral tensile strength than the others. (p<0.05) 2. Increase of zirconium-silicate filler contents reduced the diametral tensile strength of experimental composites. ($r^2$=0.8721, p=0.0002) 3. Increase of zirconium-silicate filler contents did not affect the shear bond strength of experimental composites. ($r^2$=0.2815, p=0.4067) 4. Increase of zirconium-silicate filler contents reduced significantly the depth of cure of experimental composites. ($r^2$=0.9700, p<0.0001) These results mean that the mechanical properties of composites could not be improved by incorporation of small amount of zirconium-silicate filler. Also, the increased contents of zirconium-silicates fillers was found to reduce the diametral tensile strength and depth of cure.

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The Correlation of Antioxidative Effects of 5 Korean Common Edible Seaweeds and Total Polyphenol Content (한국산 5종 해조류의 항산화효과와 총 폴리페놀 함량과의 관련성)

  • Kwak, Chung-Shil;Kim, Sung-Ae;Lee, Mee-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1143-1150
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    • 2005
  • Antioxidative activities of 5 common edible seaweeds in Korea, three brown algae (seaweed fusiforme, sea mustard, sea tangle), one green algae (sea lettuce) and one red algae (laver), were examined. The antioxidative activities of ethanol extracts from these seaweeds were examined by measuring of inhibition rates against iron-induced linoleate peroxidation, DPPH (1,1-diphenyl -2-picrylhydrazyl) radical generation and MDA-BSA (malondialdehyde-bovine serum albumin) conjugation. Sea lettuce ethanol extract showed the strongest anti-oxidative activity among them, especially in inhibition against conjugation of lipid peroxide and protein. Second to sea lettuce, laver and sea tangle ethanol extracts showed high DPPH radical scavenging activity and inhibition against MDA-BSA conjugation. However, seaweed fusiforme and sea mustard ethanol extracts did not show antioxidative activities. Sea mustard contained the highest total flavonoids (11.33 mg/g dry wt) and sea lettuce contained the highest total polyphenol (8.97 mg/g dry wt) among these seaweeds. In addition, there was strong positive correlation between the antioxidative activity and total polyphenol content in these seaweeds, suggesting polyphenol compounds may contribute to antioxidative effect of seaweeds. From these data, it is suggested to consume much of seaweeds such as sea lettuce, laver and sea tangle to prevent age-related chronic diseases, and also develope neutraceutical products using polyphenol rich fraction from sea lettuce.

Hyaluronidase Inhibitory Activity of Extracts from Doenjang, Chungkookjang and Miso (된장, 청국장 및 미소추출물의 Hyaluronidase 저해활성)

  • Ahn, Sun-Kyung;Hong, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1119-1123
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    • 2005
  • The inhibitory effects of water and methanol extracts of Doenjang, Chungkookjang and Miso on bovine hyaluronidase were examined. The extracts were vacuum-dried and the resulting pellets were dissolved with 0.1 M acetate buffer with 1$\%$ DMSO to a final concentration of 50 mg/mL. The water extracts of Doenjang, Chungkookjang and Miso inhibited 62$\%$, 70$\% $, 56$\%$ of hyaluronidase activity, respectively, while the extract of crushed soybean inhibited 24$\%$ of the activity. Under the same condition, disodiumcromoglycate known as an anti-allergic drug inhibited 46$\%$ of hyaluronidase activity at the concentration of 0.35 mg/mL as a positive control. Also the methanol extracts of Doenjang, Chungkookjang and Miso inhibited 48$\%$, 52$\%$, 70$\%$ of hvaluronidase activity, respectively, while the extract of crushed soybean inhibited 46$\%$ of the activity. After heat treatment of the water extracts of Doenjang, Chungkookjang and Miso at 100"C for 10 min, hyaluronidase inhibitory effects of them were reduced approximately to half. However, hyaluronidase inhibitory effects of methanol extracts were maintained well even after heat treatment.

Effect of Lentinus edodes and Pleurotus eryngii Extracts on Proliferation and Apoptosis in Human Colon Cancer Cell Lines (표고와 새송이버섯이 대장암 세포 증식 및 세포사멸에 미치는 영향)

  • 황용주;남혜경;장문정;노건웅;김선희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.2
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    • pp.217-222
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    • 2003
  • We studied effects of hot water extract of Lentinus edodes (Berk.)sing. and Pleurotus eryngii (De Candolle ex Fries) Quel mushroom on proliferation and apoptosis of the human colon adenocarcinoma, HT-29 and Caco-2.. Cells were maintained with Dulbecco's modified Eagle medium/Ham's F-12 nutrient mixture supplemented with 10% fetal bovine serum at 37$^{\circ}C$ in a humidified $CO_2$. For cell proliferation experiments, cells were seeded in 35 mm dishes, treated with the various concentrations of the extract for the different time course. Apoptosis was measured by caspase-3 activity The more contents of the extract added in HT-29 and Caco-2 were, the more cell proliferation was suppressed. When we incubated HT-29 cells for 24, B\ulcorner72, and 96 hours after treatments, cell proliferation was markedly suppressed after 96 hours. Also, caspase-3 activity in HT-29 was increased by the treatment of Lentinus edodes and Pleurotus eryngii extracts. However, the treatment of the extract to SNU484, Korean stomach adenocarcinoma, did not show any influence on cell proliferation and caspase-3 activity Therefore, Lentinus edodes and Pleurotus eryngii are strongly recommended for the prevention and treatment of colon cancer.

Alkaline Pilot Processing for Recovery of Fish Muscle Protein and Properties of Recovered Protein (어육 단백질 회수를 위한 알칼리 Pilot 공정과 회수 단백질의 특성)

  • Jang, Young-Boo;Kim, Gun-Bae;Lee, Keun-Woo;Choi, Yeung-Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.8
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    • pp.1045-1050
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    • 2006
  • Optimum operation condition for pilot scale of alkaline processing for fish muscle was investigated by measuring protein solubility, yield, texture, and water-holding capacity. Recovered protein yield was 33.2% for whole fish and 61.8% for minced muscle. Optimum homogenized speed and time, using industrial scale homogenizer, were 3,000 rpm and 5 min, respectively. Limited centrifugal force of continuous cylinder type was 4,000 rpm for recovering soluble protein, and 2,000 rpm for recovering precipitated proteins. The pH control agents such as citric acid, sodium phosphate and calcium oxide decreased the breaking force and deformation of recovered protein gel. The breaking force and deformation of the recovered proteins were high compared to conventional surimi. The breaking force and deformation were decreased by addition of salt, starch and bovine plasma proteins. Whiteness of recovered protein gel was lower than that of conventional surimi. Alkaline processing greatly decreased nitrogen content and chemical oxygen demand in waste water. The results suggest that alkaline processing has a potential as industrial production for recovering the proteins from fish muscle.

Pharmacological Effects of ginseng Saponins on Receptor Stimulation-responses

  • Eiichi Tachikawa;Kenzo Kudo;Kazuho Harada;Takeshi Kashimoto;KatsuroFurumachi;Yoshikazu Miyate;Atsushi Kakizaki;Eiji Takahashi
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.40-46
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    • 1998
  • We investigated the influence of the root of Panax ginseng C. A. Meyer on the secretion of catecholamines from bovine adrenal chromaffin cells, which are used as a model of nervous systems. In two major parts extracted from the ginseng root, the crude saponin fraction, but not the non-saponin fraction, reduced the secretion from the cells, stimulated by acetylcholine (ACh). Ginseng saponins (ginsenosides) are classified into three groups, the panaxadiol, the panaxatriol and the oleanolic acid groups, on the basis of the chemical structures of their saponins. Both the panaxadiol and the panaxatriol saponins, excluding only one oleanolic acid saponin ginsenoside-Ro, generally reduced the ACh-evoked secretion. The inhibitory effects of the panaxatriol were much stronger than those of the panaxadiol. However, ginsenoside-Rg, and -Rh3 in the panaxadiol saponins were the potent inhibitors comparable to the panaxatriol saponins. Ginsenoside-Rg2 in the panaxatriol was the most effective. It is probable that the ginsenoside inhibition of the catecholamine secretion is due to the suppression of the function of the nicotinic ACh receptor-cation channels. On the other hand, ginsenoside-Rg2 did not affect the angiotensin II-, the bradykinin-, the histamine- and the neurotensin- induced catecholamine secretions from the chromaffin cells and the muscarine- and the histamine- induced contraction of the ileum in guinea-pigs. Ginsenoside-Rbl, a panaxadiol saponin, and ginsenoside-Ro had no or only a slight effect on them. On the contrary, ginsenoside-Rg3 not only competitively inhibited the muscarine-induced ileum contraction but also reduced the angiotensin R -, the bradykinin-, the histamine- and the neurotensin-induced catecholamine secretions. Thus, the ginseng root contains active ingredients, namely some ginsensides, which suppress the responses induced by receptor stimulation. The inhibitory effects of ginseng saponins may be one of the action mechanisms for the pharmacological effects of the Panax ginseng root.

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Fabrication, Estimation and Trypsin Digestion Experiment of the Thermally Isolated Micro Teactor for Bio-chemical Reaction

  • Sim, Tae-Seok;Kim, Dae-Weon;Kim, Eun-Mi;Joo, Hwang-Soo;Lee, Kook-Nyung;Kim, Byung-Gee;Kim, Yong-Hyup;Kim, Yong-Kweon
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.5 no.3
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    • pp.149-158
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    • 2005
  • This paper describes design, fabrication, and application of the silicon based temperature controllable micro reactor. In order to achieve fast temperature variation and low energy consumption, reaction chamber of the micro reactor was thermally isolated by etching the highly conductive silicon around the reaction chamber. Compared with the model not having thermally isolated structure, the thermally isolated micro reactor showed enhanced thermal performances such as fast temperature variation and low energy consumption. The performance enhancements of the micro reactor due to etched holes were verified by thermal experiment and numerical analysis. Regarding to 42 percents reduction of the thermal mass achieved by the etched holes, approximately 4 times faster thermal variation and 5 times smaller energy consumption were acquired. The total size of the fabricated micro reactor was $37{\times}30{\times}1mm^{3}$. Microchannel and reaction chamber were formed on the silicon substrate. The openings of channel and chamber were covered by the glass substrate. The Pt electrodes for heater and sensor are fabricated on the backside of silicon substrate below the reaction chamber. The dimension of channel cross section was $200{\times}100{\mu}m^{2}$. The volume of reaction chamber was $4{\mu}l$. The temperature of the micro reactor was controlled and measured simultaneously with NI DAQ PCI-MIO-16E-l board and LabVIEW program. Finally, the fabricated micro reactor and the temperature control system were applied to the thermal denaturation and the trypsin digestion of protein. BSA(bovine serum albumin) was chosen for the test sample. It was successfully shown that BSA was successfully denatured at $75^{\circ}C$ for 1 min and digested by trypsin at $37^{\circ}C$ for 10 min.