• Title/Summary/Keyword: $AgNO_3$ addition

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Production of Leptin in E. coli and Its Effect on Glucose and Acetate Transport and Expression of Uncoupling Protein-2 Gene in Adipose Tissues of Korean Cattle (Hanwoo)

  • Kim, K.S.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1062-1068
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    • 2004
  • Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

Antimicrobial Activity of Nano Materials against Acidovorax citrulli and Other Plant Pathogens (나노 화합물을 이용한 Acidovorax citrulli 및 식물병원성 미생물의 항균활성 효과 검정)

  • Kim, Sang Woo;Adhikari, Mahesh;Yadav, Dil Raj;Lee, Hyun Goo;Um, Young Hyun;Kim, Hyun Seung;Lee, Youn Su
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.12-19
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    • 2015
  • Antimicrobial activities of nano-materials were tested against several plant pathogens. Twelve different nano-materials were used to observe the antagonistic activity against three kinds of mold and sixteen different kinds of watermelon fruit rot pathogens (Acidovorax citrulli). According to the results, no antagonism have been found against the pathogen, Cylindrocarpon destructans. However in the case of Pythium ultimum, combination of Brass/Glucose 1,000 ppm confirmed the mycelial growth reduction by 94%. In addition, little effect was found against Rhizoctonia solani by Ag/Glucose 3,000 ppm. The remaining other nano-materials have different antimicrobial effect depending on the strains of A. citrulli. But in the case of lime (Cu/Salt 1,000 ppm) highest antimicrobial activity was observed with 97%. Moreover growth of five different strains of A. citrulli was checked by 99% with the combination of Ag/Glucose 1,000 ppm. 92% reduction of A. citrulli growth was observed with $Brass/CaCO_3$ 3,000 ppm. Tested nano-materials against different plant pathogens in this study showed the antimicrobial activity at the range of 24-70%.

Enzymatic Properties of Intracellular Adenosine Deaminase from Nocardioides sp. J-326TK

  • Hong-Ki Jun;Tae-Sook Kim
    • Journal of Life Science
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    • v.9 no.1
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    • pp.64-68
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    • 1999
  • The properties of purified intracellular adenosine deaminase (adenosine aminohydrolase, EC 3.5.4.4) of Nocardioides sp. J-326TK isolated from soil have been studied. The enzyme deaminated adenosine and 2`-deoxyadenosine and the respective {TEX}$K_{M}${/TEX} values were 4.0×{TEX}$10^{-4}${/TEX} M and 5.0× {TEX}$10^{-4}${/TEX} M, but the enzyme was not active on 8-bromoadenosine, 6-methylaminopurine riboside, ATP, ADP, 2`-AMP, 3`-AMP, 5`-AMP, dAMP, cAMP, NAD, FAD, NADP and adenine. The enzyme activity was strongly inhibited by the addition of {TEX}$Hg^{2+}${/TEX} and {TEX}$Ag^{+}${/TEX}, {TEX}$Cu^{2+}${/TEX}, {TEX}$Co^{2+}${/TEX} and {TEX}$Mn^{2+}${/TEX} also inhibited the activity but much less extent. The effect of alkyl reagents, metal chelating reagents and certain other compounds on the enzyme activity were also examined. No reagent activated the enzyme. On the contrary, the enzyme reaction was slightly inhibited by o-phenanthroline and 6-benzyladenosine.

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Production of Hydroxymethylfurfrual by Sesamum indicum L. Root Cultures (참깨 뿌리배양에 의한 hydroxymethylfurfrual 생산)

  • Chun, Jae-An;Lee, Jin-Woo;Yi, Young-Byung;Hong, Seong-Sig;Cho, Kang-Jin;Chung, Chung-Han
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1836-1840
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    • 2009
  • Recently, hydroxymethylfurfrual (HMF) has been highlighted as a key intermediate for the production of liquid biofuels and other valuable compounds. We used sesame roots as a biocatalyst to synthesize HMF using flask cultures. The synthesis of HMF was identified by GC-mass analysis. The highest root growth was observed in cultures with 1.0 mg/l NAA at $30^{\circ}C$, while root growth was not found in those without NAA treatment. When silver nitrate ($AgNO_3$) was added, the root growth was greatest in those treated with 0.5 mg/l $AgNO_3$ and cultured at $30^{\circ}C$. In the case of HMF synthesis, its highest yield was obtained in those treated with 0.5 mg/l NAA at $25^{\circ}C$, but low HMF was detected in those treated without naphthaleneacetic acid (NAA). The addition of $AgNO_$ to the culture medium showed a 8-10% reduction in HMF yield compared to that of the control, indicating its inhibitory effect on the synthesis of HMF. On the whole, an optimal culture temperature for HMF synthesis seemed to be between $25-30^{\circ}C$.

Effect of Growth Regulators, Carbon Sources and Silver Nitrate on Callus Formation and Plant Regeneration of Turf Grass (잔디의 캘러스 형성 및 재분화에 끼치는 식물생장조절제, 탄소원 및 AgNO3의 영향)

  • Han, S.S.;Rim, Y.S.;Jeong, J.H.
    • Korean Journal of Weed Science
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    • v.16 no.3
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    • pp.221-229
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    • 1996
  • This study was carried out to determine the effects of growth regulators, carbon sources and silver nitrate on callus formation and plant regeneration of turfgrass. The results were summarized as fallows : Callus from Korean lawngrass (Zoysia japonica Steud.) and pencross creeping bentgrass (Agrostis palustrir Huds.) induced better in MS medium than in N6 medium and by addition of 2,4-D than by that of NAA. Callus formation from Korean lawngrass and penncross creeping bentgrass was very effective at MS medium adding 1mg/L 2,4-D and at the medium adding 2mg/L 2,4-D, repectively. Growth of callus was good at MS medium containing 2mg/L 2,4-D+0.2mg/L NAA. Callus growth of Korean lawngrass and penncross creeping bentgrass was good when kinetin was added 0.2mg/L and 0.3mg/L, individually, to MS medium containg 2mg/L 2,4-D+0.2mg/L NAA. Regeneration rate from leaf and stock callus of Korean lawngrass was 44% at MS medium adding 2,4-D 2mg/L+NAA 0.2mg/L+kinetin 0.3mg/L and 32% at the medium containing 2,4-D 2mg/L+NAA 0.2mg/L+kinetin 0.3mg/L, each and that from leaf and stock callus of penncross creeping bentgrass was 80% and 67%, each, at the medium adding 2,4-D 2mg/l+NAA 0.2mg/L+kinetin 0.3mg/L. Regeneration rate of Korean lawngrass and penneross creeping bentgrass increased by 3 to 4% and by 10 to 16%, respectively, when added $AgNO_3$ 1~2mg/L to the above-mentioned regeneration medium.

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Cloning and Characterization of a Cellulase Gene from a Plant Growth Promoting Rhizobacterium, Bacillus subtilis AH18 against Phytophthora Blight Disease in Red-Pepper (고추역병을 방제하는 PGPR균주 Bacillus subtilis AH18의 항진균성 Cellulase 유전자의 Cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Jung, Hee-Kyoung;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.311-317
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    • 2006
  • Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

Characterization of Extracellular Protease Secreted from Chryseobacterium sp. JK1 (Chryseobacterium sp. JK1이 분비하는 세포외 단백질분해효소 특성)

  • Lee, Yu-Kyong;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.78-82
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    • 2013
  • A novel Chryseobacterium sp. JK1 strain isolated from soil had been reported that this isolate produced large amount of extracellular protease at mesophilic temperature in previous study. The optimal temperature and pH of extracellular protease were $40^{\circ}C$ and 7.0, respectively, showing narrow range of optimal temperature and relatively broad activity from pH 6.0 to 9.0. In addition, the protease showed greatest activity against skim milk and lowest against bovine serum albumin (BSA). The protease strongly inhibited by ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA) or phenylmethylsulfonyl fluoride (PMSF), and addition of cation $Ag^+$ or $Cu^{2+}$, and slightly inhibited by $Al^{3+}$. No significant inhibition was found with pepstatin, and addition of cation, $K^+$, $Ca^{2+}$, $Na^+$, $Fe^{2+}$ or $Mg^{2+}$. On the contrary, protease was enhanced by addition of divalent cation $Mn^{2+}$ (5 mM). Zymography analysis of concentrated culture supernatant revealed two major bands at 67 and 145 kDa. These results suggest that Chryseobacterium sp. JK1 strain produced extracellular neutral serine proteases which could apply in food industry.

Bioluminescent Determination of Lactose Secretion: A Measure of the In Vitro Performance of Mammary Acini from Lactating Rats

  • Choi, B.H.;Stewart, K.W.;Davis, S.R.;Myung, K.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.2
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    • pp.274-278
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    • 2002
  • A culture system for lactating rat mammary acini was evaluated, where the primary indicator of performance was lactose secretion, measured by a sensitive bioluminescence assay. Lactose secretion was reduced by half (p<0.01) over the first 6 h of culture by overnight feed withdrawal (FW) from tissue donors but was sensitive to increased glucose concentration in the culture media (p<0.001) up to 30 mM. Lactose production of cells from fed donors over the first 6 h in culture in 30 mM glucose was 8.9 fmol/cell/h - a rate calculated to be about half that in vivo. No significant difference was shown in lactose secretion by cells from fed or FW rats over 6-24 h. Lactose secretion was 3.6 fmol/cell/h by cells from fed animals in 40 mM glucose concentration media over the 6-24 h culture period. Addition of insulin to the culture media had no effect on rates of lactose secretion while addition of prolactin and hydrocortisone, with or without insulin, significantly (p<0.001) decreased lactose production over both 0-6 h and 6-24 h culture periods. Lactose synthesis in vitro was significantly enhanced by aeration of the media during collagenase digestion of mammary tissue (p<0.05). No improvement in lactose secretion was effected by shaking of cells during culture, Matrigel coating of culture dishes or change in cell density over a range up to 2.5 million cells per ml.

The Effect of Additives on the Properties of Zn Electrode in Zn/AgO Secondary Battery (Zn/AgO Secondary Battery용 아연 양극의 성능에 미치는 첨가제의 영향)

  • Park, Kyung-Wha;Kim, Chang-Hwan;Moon, Kyung-Man
    • Journal of the Korean Electrochemical Society
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    • v.6 no.3
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    • pp.196-202
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    • 2003
  • We investigated the effect of both 4 types additives and $0.5wt\%\;Pb_3O_4$ which have been reported to show an improvement on the performance of Zinc anode. And Experimental methods such as corrosion potential measurement, potentiodynamic polorization test and charging-discharging cycle life test were carried out in $40 wt\%$ KOH with $Pb_3O_4(0.5, \;10\;&\;2.0wt\%)$ and 4 types additives $(0.4wt\%\;of\;Ca(OH)_2$, 0.025M of Citrate, Tartrate and Gluconate). Corrosion potential was shifted to high direction and also changed to high and low direction repeatedly with increasing of $Pb_3O_4$ quantity. However by adding $0.5wt\%\;Pb_3O_4$, corrosion potential shifted to low direction and showed stable condition. Furthermore it was well known that corrosion resistance was predominantly increased compared to no addition and improved charging-discharging property with adding additives. By SEM analysis, it was concluded that the morphology of surface in case of only $0.5wt\%\;Pb_3O_4$ addition was nearly the same as that of Tartrate additive and in the other additives such as $Ca(OH)_2$, Citrate, Tartrate and Gluconate, their morphologies showed dendrite growth. Eventually it was thought that the additive of Tartrate indicated comparatively good corrosion resistance effect as well as charging-discharging property improvement among those four types additives.

DNMT3a rs1550117 Polymorphism Association with Increased Risk of Helicobacter pylori Infection

  • Cao, Xue-Yuan;Jia, Zhi-Fang;Cao, Dong-Hui;Kong, Fei;Jin, Mei-Shan;Suo, Jian;Jiang, Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5713-5718
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    • 2013
  • Background: DNA methyltransferase-3a (DNMT3a) plays significant roles in embryogenesis and the generation of aberrant methylation in carcinogenesis. This study aimed to investigate associations between single nucleotide polymorphisms (SNPs) of the DNMT3a gene and risk of Helicobacter pylori infection, gastric atrophy and gastric cancer. Methods: The subjects comprised 447 patients with gastric cancer; 111 individuals with gastric atrophy and 961 healthy controls. Two SNPs (rs1550117 and rs13420827) of the DNMT3a gene were genotyped by Taqman assay. DNMT3a expression was analyzed in cancer tissues from 89 patients by tissue microarray technique. Odds ratio (ORs) and 95% confidence intervals were calculated by multivariate logistic regression. Results: Among healthy controls, risk of H.pylori infection was significantly higher in subjects with the rs1550117 AA genotype, compared to those with GG/AG genotypes of DNMT3a [OR=2.08, (95%CI: 1.02-4.32)]. However, no significant correlation was found between the two SNPs and risk of developing gastric atrophy or gastric cancer. In addition, no increase in DNMT3a expression was observed in the gastric cancer with H.pylori infection. Conclusions: This study revealed that DNMT3a rs1550117 polymorphism is significantly associated with an increased risk of H. pylori infection, but did not support any evidence for contributions of DNMT3a rs1550117 and rs13420827 to either gastric atrophy or gastric cancer. The biological roles of DNMT3a polymorphisms require further investigation.