• Title/Summary/Keyword: Protein fractions

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Effect of Experimental Electrization Stress on the Pathogenesis of Escherichia coli in Mice (실험적 감전 Stress 가 Escherichia coli 감염마우스의 기병에 미치는 영향)

  • Lee, Bang Whan
    • Korean Journal of Veterinary Research
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    • v.13 no.2
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    • pp.119-129
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    • 1973
  • A hypothesis that is a stress condition in animal may cause either enhancement or reduction of the host resistance against microbial infection was experimentally studied. Among of many processes for stress formation an experimental electrization in mice was devised, on the bases of blood picture analysis, and studied the effect of experimental electrization of mice on E. coll infection. The results obtained were as follows. 1. Electrization with ordinary current, A. C. 60 cps., on the path of symmetrical line of both posterior limbs at 20 to 100 volts (less than 10 mA) for 15 to 30 seconds was able to induce a stress reaction in blood pattern without showing any dangers of electrocution, electric burns and other residual signs, and no correlation between blood pattern of the reaction and an amount of current between 20 to 100 volts was observed. As the electrodes, two of 21 gauge hypodermic needles were used, when the electrization each of them were inserted into the center of toe tissue of the both legs. 2. Serum protein fractions following the experimental electrization showed a tendency of a low A/G ratio and a high value of ${\alpha}$-globulin. 3. In the studies on the effect of electrical stress on the pathogenesis of E. coli in mice, a group in which a simultaneous electrization and infection, and a group infected two hours after electrization showed 80 per cent mortality. On the other hand, infection after 20 hours electrization and control groups showed their mortality of 40 and 60 per cent respectively.

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Studies on the Adenosinetriphosphatase in the Mushroom (I) -Purification and Properties of $F_{1}-ATPase$ in Lentinus edodes (Berk.) Sing- (버섯의 Adenosinetriphosphatase(ATPase)에 관한 연구(I) -표고버섯(Lentinus edodes) 중 $F_{1}-ATPase$의 정제 및 그 성질-)

  • Min, Tae-Jin;Park, Hye-Lyoun
    • The Korean Journal of Mycology
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    • v.19 no.3
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    • pp.214-219
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    • 1991
  • Adenosine-5'-triphosphatase$(F_1-ATPase)$ in the Lentinus edodes was fractionated by ammonium aulfate 30% saturation and purified by Sephadex G-200 gel filtration in three times. Three kinds of protein fractions of $F_1-ATPase$ were isolated from this mushroom, and fraction I and ll showed its activity for the substrate, adenosine-5'-triphosphate. Its optimum pH and temperature were found to be pH 7.6 and $58^{\circ}C$, and its thermal stability was stabled for 30 min. at $20-30^{\circ}C$. The Km value of this enzyme was 1.81 mM.

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Effects of heavy metals on contents of various phosphate compounds and organic compounds in chlorella cells (Chlorella의 인산화합물 및 유기물함량에 미치는 중금속의 영향)

  • 이종삼;임영복
    • Korean Journal of Microbiology
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    • v.20 no.1
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    • pp.27-40
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    • 1982
  • The effects of heavy metals on the growth rate and phosphate metabolism of Chlorella elliposidea cells were investigated. Chlorella cells were cultured in the media treated with Hg(0.3, 0.7, 0.9 ppm), Cd(1, 5, 15ppm), and Zn(1, 5, 50ppm) for 6days. Aliquots cells were taken out at the inoculation and at intervals during the culture, and measured packed cell vlolume and optical density. The inhibitions of heavy metals on the growth rate and chlorophyll contents were traced. Also after 6 days culture, the amounts of inorganic phosphate and organic compounds of various fractions in Chlorella cells were observed. The turbid effects of heavy metals on the growth rate and chlorphyll contents of Chlorella cells were in order of Hg>Cd>Zn. Because heavy metals depressed the biosynthesis of inorganic polyphosphates and nucleic acids and turn over of inorganic phosphates, the amounts of various phosphate compounds were decreased. The inhibitory effect of photosynthesis by heavy metals resulted in lower contents of carbohydrate. Due to the turbidity of biosynthesis of amino acids by heavy metals, contents of protein were reduced in comparison with those of control. It is suggested conciusively that the minimum concentrations affected by heavy metals on the growth rate and phosphate metabolism of Chlorella cells were 0.7 ppm Hg, 15ppm Cd, 50ppm Zn.

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Inhibitory Effect of the Leaves of Rumex crispus L. on LPS-induced Nitric Oxide Production and the Expression of iNOS and COX-2 in Macrophages

  • Im, Nam-Kyung;Jung, Yeon-Seop;Choi, Jun-Hyeok;Yu, Mi-Hee;Jeong, Gil-Saeng
    • Natural Product Sciences
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    • v.20 no.1
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    • pp.51-57
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    • 2014
  • In this study, the anti-inflammatory effects of ethanol extract of Rumex crispus L. and its fractions were investigated in RAW 264.7 macrophages. To evaluate the anti-inflammatory effects of extract, we studied nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), and tumor necrosis factor-alpha ($TNF-{\alpha}$) levels in RAW 264.7 cells. The ethanol extract of R. crispus L. significantly decreased NO production and the levels of other inflammatory factors, such as PGE2 and $TNF-{\alpha}$, in lipopolysaccharide (LPS)-stimulated macrophages in a dose-dependent manner. We also assessed the inducible nitric oxide synthase (iNOS) and the cyclooxygenase 2 (COX-2) protein expression by western blot. Ethyl acetate fraction of R. crispus L. had the strongest anti-inflammatory effect. These results suggest that ethyl acetate extract of R. crispus L. might be beneficial in the treatment of chronic inflammatory diseases.

Enzymes of Carbohydrate Metabolism in Cowpea (Vigna unguiculata L. Walp. cv. Caloona) Nodules

  • Lee, Hoi-Seon;Ahn, Young-Joon
    • Applied Biological Chemistry
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    • v.41 no.4
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    • pp.222-227
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    • 1998
  • The activities of enzymes of carbohydrate metabolism have been determinated in the host cytosolic and bacteroid fractions of cowpea (Vigna unguiculata) nodules formed with B. japonicum I 16 and in roots of nodulated cowpeas. The host cytosolic fraction of the nodules contained the enzymes of glycolytic pathway and the pentose phosphate pathway, whereas the bacteroids had only limited capacity for carbohydrate metabolism and appeared to be insufficient for the complete glycolytic pathway as well as starch synthesis and degradation. In a time-course study, using plants grown in a glasshouse, the acetylene-reducing activity (ARA) of the nodules increased in parallel with the total N content of plants and protein of the nodules until approximately 8 weeks after planting. Subsequently, the weight and size of the nodules and the weight of the plants continued to increase, but there was a sharp decrease in the ARA and the total N content of the plants.

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Immunochemical Studies on Expression of Quinoproteins in Escherichia coli

  • Ryou, Chong-Suk;Kim, Jae-Beom;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.95-98
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    • 2000
  • An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

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Leaves of Raphanus sativus L. Shows Anti-Inflammatory Activity in LPS-Stimulated Macrophages via Suppression of COX-2 and iNOS Expression.

  • Park, Hye-Jin;Song, Minjung
    • Preventive Nutrition and Food Science
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    • v.22 no.1
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    • pp.50-55
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    • 2017
  • Raphanus sativus L. (RS) is a cruciferous vegetable that is widely consumed in Korea. The anticancer activity of leaves of RS (RSL) extract has been investigated; however, no studies focused on its anti-inflammatory effects. Therefore, the aim of the current study was to evaluate the anti-inflammatory effects of RSL extract. In brief, RSL powder was fractionated into n-hexane, chloroform, ethyl acetate, n-butanol, and water-soluble fractions. Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were treated with each fraction for initial screening. It was found that the chloroform fraction significantly inhibited nitric oxide release in LPS-stimulated RAW264.7 cells with a half maximal inhibitory concentration value of $196{\mu}g/mL$. In addition, the mRNA and protein expression levels of inducible nitric oxide synthase, measured using reverse transcriptase-polymerase chain reaction and western blotting, respectively, were reduced in a concentration-dependent manner. Moreover, the inflammatory cyclooxygenase-2 enzyme expression decreased. Furthermore, the expression of nuclear factor-kappa B ($NF-{\kappa}B$), the key regulator of the transcriptional activation of the inflammatory cytokine genes, was reduced by the RSL chloroform fraction. Therefore, the results of our study suggest that RSL exhibits anti-inflammatory effects in LPS-stimulated macrophages via $NF-{\kappa}B$ inactivation.

Polysaccharide Characteristics from Hot Water Extract of Aloe saponaria Callus (Aloe saponaria 캘러스의 열수 추출물 유래 다당의 특성)

  • Baek, Jin-Hong;Kim, Myung-Uk;Kang, Tae-Su;Hur, Won;Lee, Shin-Young
    • KSBB Journal
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    • v.24 no.1
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    • pp.59-64
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    • 2009
  • The callus formation from inferior leaf of Aloe saponaria was induced in M & S medium supplemented with 10-30 ${\mu}M$ NAA (${\alpha}$-naphthalene acetic acid) and 3-7 ${\mu}M$ kinetin under incubation in the dark at $25^{\circ}C$ for 6 weeks. The hot water extract ($100^{\circ}C$, 24 hrs) from cultured callus was obtained and the components analysis for the extract were examined to determine the callus can synthesized the bioactive component such as Aloe polysaccharide. The freeze dried extract contained the sugar of 53.2%, protein of 7.3%, ash of 18.5% and water of 21% (w/w). Two fractions (Fr-I and Fr-II) were obtained by Sepharose CL-4B gel permeation chromatography and Fr-I, major fraction was further purified with dialysis. From sugar analysis by TLC and GC, the purified Fr-I fraction consisted of glucose (77.6%), galactose (17.7%), mannose (4.7%, w/w) and uronic acid (trace). The molecular weight of purified Fr-I fraction determined by GPC was about 110 kDa.

Inhibitory Effects of Extracts of Houttuynia cordata Thumb (Saururaceae) on Cytokine Release of Human Monocytic THP-1 Cells and Human Eosinophilic EoL-1 Cells

  • Lee, Ji-Sook
    • Biomedical Science Letters
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    • v.16 no.4
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    • pp.341-347
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    • 2010
  • In the present study, we investigated whether Houttuynia cordata Thumb (Saururaceae; HC) extracts have an anti-inflammatory effect in human monocytic THP-1 cells and human eosinophilic EoL-1 cells. The dried and powdered whole plants of HC were extracted with 80% EtOH. The combined extract (HC-1) was concentrated under reduced pressure. The residue was diluted with water, and then successively partitioned with n-hexane, EtOAc, and BuOH to produce the n-hexane (HC-2), EtOAc (HC-3), BuOH (HC-4), and the water-soluble fractions (HC-5), respectively. HC extracts have no cytotoxicity on THP-1 cells and EoL-1 cells at a high concentration of $10\;{\mu}g/ml$ for 24 h, except HC-2 extract ($10\;{\mu}g/ml$). Interleukin-6, Interleukin-8 and Monocyte chemoattractant protein-1 in THP-1 cells were increased after the treatment with the extract from house dust mite or LPS. The increase of cytokine production was strongly suppressed by HC-3 extract, in comparision with other extracts. HC-3 also had inhibitory effect on Interleukin-6 production increased by mite extract and LPS in EoL-1 cells. However, HC-3 extract increased Interleukin-8 production induced by mite extract and LPS in EoL-1 cells. These results suggest that HC extracts may be used as useful agents for treating allergic disorders such as asthma and atopic dermatitis.

Subcellular Distribution of Heavy Metals in Organs of Bivalve Modiolus Modiolus Living Along a Metal Contamination Gradient

  • Podgurskaya, Olga V.;Kavun, Victor Ya.
    • Ocean Science Journal
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    • v.41 no.1
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    • pp.43-51
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    • 2006
  • Concentration and distribution of Fe, Zn, Cu, Cd, Mn, Pb, Ni among subcellular fractions (cellular membrane structures and cytosol) and Zn, Cu, Cd among cytoplasmic proteins in the kidney and digestive gland of mussel Modiolus modiolus living along a polymetallic concentration gradient were studied. It was found in the kidney of M. modiolus from contaminated sites that the Fe percent increased in the "membrane" fraction, whereas Zn, Pb, Ni and Mn percent increased in the cytosol compared to the kidney of the control mussel. Note kidney cytosol of M. modiolus from clean and contaminated sites sequestered major parts of Cu and Cd. In the digestive gland of M. modiolus from contaminated sites Fe, Zn, Cd, Mn, Ni percent increased in the "membrane" fraction, whereas Cu, Pb percent increased in the cytosol compared to digestive gland of control mussel. Gel-filtration chromatography shows kidney of M. modiolus contains increased metallothionein-like protein levels irrespective of ambient dissolved metal concentrations. It was shown that the metal detoxification system in the kidney and digestive gland of M. modiolus was efficient under extremely high ambient metal levels. However, under complex environmental contamination in the kidney of M. modiolus, the metal detoxification capacity of metallothionein-like proteins was damaged.