• Title/Summary/Keyword: Extracellular protein

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Pulmonary Function Test and Body Composition Analysis in Obese Children (비만 소아에서 폐기능 검사와 체성분 분석에 대한 연구)

  • Shin, Jee Seon;Park, Ji Hye;Kim, Ji Young;Kim, Su Jung;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.48 no.6
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    • pp.588-593
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    • 2005
  • Purpose : Obesity is associated with disturbances of ventilatory functions in adults. But few studies have evaluated the pulmonary complications of obesity in the pediatric population. The purpose of this study is to clarify the effects of obesity on pulmonary function and body composition in obese children. Methods : Forty seven obese children whose ages ranged from nine to twelve years were evaluated for their body composition(intracellular fluid, extracellular fluid, protein mass, mineral mass, soft lean mass, fat mass, percent body fat, fat distribution) by bioelectrical impedance analysis. Hemoglobin, serum glucose, aspartate aminotransferase(AST), alanine aminotransferase(ALT), total cholesterol and triglycerides were measured. Pulmonary function test was performed by spirometer. Results : Intracellular fluid, protein mass, fat mass, percent body fat and fat distribution were significantly higher in severely obese children with an obesity index of more than 150 percent compared with those with an index of less than 150 percent. Peak expiratory flow rate(PEFR) was significantly lower in severely obese children with obesity index of more than 150 percent compared with those with less than 150 percent($241.7{\pm}14.6L/sec$ vs $276.8{\pm}64.3L/sec$). PEFR, forced expiratory flow 25 percent($FEF_{25}$), mid expiratory flow rate(MEFR), forced expiratory flow 50 percent($FEF_{50}$), forced expiratory volume in 1st second($FEV_1$) and forced vital capacity(FVC) were decreased in 37.0 percent, 14.8 percent, 14.8 percent, 11.1 percent, 3.7 percent and 3.7 percent of obese children, respectively. Conclusion : PEFR was significantly decreased in obese children. Pulmonary function test must be performed in severely obese children and more extended study is needed in other age groups.

Relationship between water and coffee intake on Health-related Physical Fitness and obesity of women's university student (여대생의 건강체력과 비만에 대한 물과 커피 섭취량과의 관계)

  • Lee, Lu-Ry;Ko, You-Sun
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.4
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    • pp.649-658
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    • 2020
  • The purpose of this study is to verify the difference in health and obesity according to water and coffee intake. The study subject is 900 female college students. Physical fitness factors measured in this study were physical strength(grip strength, back muscle strength), muscle endurance (sit-up), Flexibility(Sit & reach) and cardiopulmonary endurance(harvard step). The factors of body composition (intracellular fluid, extracellular fluid, protein, mineral, body water content, muscle mass, lean body mass, body fat mass) and obesity (BMI, body fat percentage, abdominal fat percentage, obesity) were measured with the Inbody equipment. SPSS program was used for all data analysis. Group differences were verified by two-way ANOVA and one-way ANOVA. The results are as follows. There was no interaction effect between water and coffee intake on health and obesity. Therefore, the difference of the dependent variables according to each independent variable was verified. There was no significant difference in physical fitness according to water intake and coffee intake, but there was a significant difference in body composition (intracellular fluid, protein, minerals, body water, muscle mass and lean body mass). Water drinking group more than 7 cups had significantly higher body composition than the 1-4 cups water drinking group. In the case of coffee, the body composition of the 3 cup intake group was significantly higher than that of the none intake group. Obesity was significantly higher than the none intake and 1-2 cup intake groups. Based on the study results, water intake was not related to obesity, but coffee was related. For further studies, it would be better to consider gender and age differences in drinking various beverages such as water and coffee and that would be used to suggest a desirable way of water intake according to individual characteristics of the better health.

Removal of Chromium (VI) by Escherichia coli Cells Expressing Cytoplasmic or Surface-Displayed ChrB: a Comparative Study

  • Zhou, Xiaofeng;Li, Jianghui;Wang, Weilong;Yang, Fan;Fan, Bingqian;Zhang, Chenlu;Ren, Xiaojun;Liang, Feng;Cheng, Rong;Jiang, Fengying;Zhou, Huaibin;Yang, Juanjuan;Tan, Guoqiang;Lyu, Jianxin;Wang, Wu
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.996-1004
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    • 2020
  • Various genetically engineered microorganisms have been developed for the removal of heavy metal contaminants. Metal biosorption by whole-cell biosorbents can be enhanced by overproduction of metal-binding proteins/peptides in the cytoplasm or on the cell surface. However, few studies have compared the biosorption capacity of whole cells expressing intracellular or surface-displayed metal-adsorbing proteins. In this study, several constructs were prepared for expressing intracellular and surface-displayed Ochrobactrum tritici 5bvl1 ChrB in Escherichia coli BL21(DE3) cells. E. coli cells expressing surface-displayed ChrB removed more Cr(VI) from aqueous solutions than cells with cytoplasmic ChrB under the same conditions. However, intracellular ChrB was less susceptible to variation in extracellular conditions (pH and ionic strength), and more effectively removed Cr(VI) from industrial wastewater than the surface-displayed ChrB at low pH (<3). An adsorption-desorption experiment demonstrated that compared with intracellular accumulation, cell-surface adsorption is reversible, which allows easy desorption of the adsorbed metal ions and regeneration of the bioadsorbent. In addition, an intrinsic ChrB protein fluorescence assay suggested that pH and salinity may influence the Cr(VI) adsorption capacity of ChrB-expressing E. coli cells by modulating the ChrB protein conformation. Although the characteristics of ChrB may not be universal for all metal-binding proteins, our study provides new insights into different engineering strategies for whole-cell biosorbents for removing heavy metals from industrial effluents.

Effects of Mifepristone and Tamoxifen on Calcium Modulation in DU-145 Prostate Cancer Cells (DU-145 전립선 암세포에 있어서 mifepristone과 tamoxifen이 칼슘조절에 미치는 영향)

  • Kim, Yeo-Reum;Kim, Byeong-Gee
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1324-1331
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    • 2010
  • Mifepristone (MIF) and Tamoxifen (TAM) have been used in the treatment of prostate cancer and breast cancer for more than a decade. MIF can induce apoptosis in both AR-positive and negative prostate cancer cells. Because of its pleiotropic ligand-receptor properties, TAM exerts cytotoxic activity in estrogen (ER)-positive and various ER.negative cancer cells. However, the molecular mechanisms of these two substances are not yet clear. In the present work, we report that the cytotoxic effects of MIF and TAM are due to the modulation of intracellular $Ca^{2+}$ level in DU-145, androgen-insensitive cells. When the cells were treated with micromolar concentrations of either MIF or TAM, the growth and viability were significantly decreased in a dose- and time-dependent manner. The apoptosis induced by MIF or TAM was further proved and analyzed by confocal laser scanning microscopy (CLSM) and fluorescence-activated cell sorting (FACS). In the cells cultivated in a normal 1.5 mM $Ca^{2+}$ medium, both MIF and TAM also induced an increase of the intracellular $Ca^{2+}$ level in a dose-dependent fashion. Since a change in calcium level could not be found in cells of the $Ca^{2+}$-free medium, the increase of intracellular $Ca^{2+}$ level might be due to an increase in extracellular calcium uptake. Our results show that the apoptotic effect was more prominent in TAM treatment compared to MIF treatment in DU-145 cells. The above findings might be due to the difference in the uppermost pathways of apoptosis induced by either MIF or TAM. When we checked the level of procaspase-8 activation, TAM showed minor level of activation, as opposed to MIF, which exerted strong activation. In both treatments, the levels of anti-apoptotic protein Bcl-2 decreased, and pro-apoptotic protein Bax level increased more than 2-fold. The activation of caspase-3, a key protease enzyme in the downstream pathway of apoptosis, was much higher in the cells treated with TAM, compared to the MIF treatment. The overall apoptotic activity shown in the present work was closely related to intracellular $Ca^{2+}$ concentration levels. Therefore, the cytotoxic activity induced by MIF and TAM might have been due to intracellular calcium modulation.

The Anti-inflammatory Effect of Skipjack Tuna (Katsuwonus pelamis) Oil in LPS-induced RAW 264.7 Cells and Mouse Models (LPS 유도 RAW 264.7 세포와 마우스 모델에서 참치(Katsuwonus pelamis) 유의 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Ji-Hye;Bae, Nan-Young;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.45-55
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    • 2015
  • This study was carried out to demonstrate the anti-inflammatory effect of tuna oil (TO) using LPS-induced inflammation responses and mouse models. First, nitric oxide (NO) and pro-inflammatory cytokines levels were suppressed up to 50% with increasing concentrations of TO without causing any cytotoxicity. Also, the expression of a variety of proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and nuclear factor kappa B (NF-κB), was suppressed in a dosedependent manner by treatment with TO. Furthermore, TO also inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 protein kinase (p38). Moreover, in in vivo testing the formation of ear edema was reduced at the highest dose tested compared to that in the control, and a reduction of ear thickness and the number of mast cells was observed in histological analysis. In acute toxicity test, no mortalities occurred in mice administrated 5,000 mg/kg body weight of TO over a two-week observation period. Our results suggest that TO has a considerable anti-inflammatory property through the suppression of inflammatory mediator productions and that it could prove to be useful as a potential anti-inflammatory therapeutic material.

Secretory Expression System of Xylose Reductase (GRE3) for Optimal Production of Xylitol (Xylitol 생산에 최적화된 xylose reductase (GRE3)의 분비발현 시스템)

  • Jung, Hoe-Myung;Kim, Jae-Woon;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1376-1382
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    • 2016
  • Xylitol is widely used in the food and medical industry. It is produced by the reduction of xylose (lignocellulosic biomass) in the Saccharomyces cerevisiae strain, which is considered genetically safe. In this study, the expression system of the GRE3 (YHR104W) gene that encodes xylose reductase was constructed to efficiently produce xylitol in the S. cerevisiae strain, and the secretory production of xylose reductase was investigated. To select a suitable promoter for the expression of the GRE3 gene, pGMF-GRE3 and pAMF-GRE3 plasmid with GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter for secretory production. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$, and $SEY2102{\Delta}trp1$/pGMF- GRE3 and $SEY2102{\Delta}trp1$/pAMF-GRE3 transformants were selected. In the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, the total activity of xylose reductase reached 0.34 unit/mg-protein when NADPH was used as a cofactor; this activity was 1.5 fold higher than that in $SEY2102{\Delta}trp1$/pAMF-GRE3 with ADH1 as the promoter. The secretion efficiency was 91% in both strains, indicating that most of the recombinant xylose reductase was efficiently secreted in the extracellular fraction. In a baffled flask culture of the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, 12.1 g/l of xylitol was produced from 20 g/l of xylose, and ~83% of the consumed xylose was reduced to xylitol.

Dietary Brown Seaweed Modulates Nutrient Metabolism, Anti-oxidant System and Immune Response in Broiler Chicks Injected LPS i.p. (사료 중 미역은 복강내 LPS 주입 육계 병아리에서 영양소 대사, 항산화계 및 면역 반응을 조정한다)

  • Park, In-Kyung;Im, Jin-Taek;Choi, Do-Yul;Koh, Tae-Song
    • Journal of Animal Science and Technology
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    • v.50 no.2
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    • pp.185-198
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    • 2008
  • Influences of dietary brown seaweed(BSW) on the nutrient metabolism, anti-oxidant enzyme activity and cell-mediated immune response were studied in broiler chicks activated acute phase response. 72 Hatched male broiler chicks(Ross) were divided into 12 pens, 6 heads per pen, and fed the BSW 0.0% (Basal) or 2.0% diet, respectively, and injected with the Salmonella typhimurium lipopoly saccharide(LPS) for activation of the acute phase response three times at 8, 10 and 12 d of age. During 4 wks of experimental feeding, growth performance of broiler chicks was not affected by dietary BSW and the acute phase response. Compared with control birds, the acute phase response did not affect the daily weight gain in birds fed BSW 2.0% diet, decreased nitrogen balance(NB) or metabolizable energy(ME) utilization per metabolic body size(kg0.75), and enhanced activities of peroxidase or extracellular SOD(EcSOD), tumor necrosis factor-alpha and ovotransferrin in plasma and MnSOD and CuZnSOD in erythrocyte cytosol. Compared to BSW 0.0% diet, 2.0% diet enhanced protein retention(NB) per kg0.75 regardless the acute phase response, did not affect uric acid nitrogen excretion(UAN) per kg0.75 in birds during the acute phase response, decreased(p<0.05) the UAN excretion per kg0.75 in control birds. And BSW 2.0% diet also decreased(p<0.05) plasma peroxide level and erythrocyte peroxidase or MnSOD activity but increased plasma peroxidase and EcSOD activity and interleukin-1 activity secreted from LPS-stimulated PBMC in 4 week broiler chicks.

Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

Lack of the Association between Microsatellite Polymorphism in Toll-like Receptor 2 Gene and Development of COPD (Toll-like Receptor 2 유전자의 Microsatellite 유전자 다형성과 만성폐쇄성폐질환 발생과의 연관성 결여)

  • Lee, Hee Seok;Lee, Hye Won;Kim, Deog Kyeom;Ko, Dong Seok;Park, Gun Min;Hwang, Yong Il;Lee, Sang-Min;Yoo, Chul Gyu;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yim, Jae-Joon
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.367-374
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    • 2005
  • Background : The fact that only 10-20% of chronic cigarette smokers develop chronic obstructive pulmonary disease (COPD) reflects the presence of genetic factors associated with the susceptibility to COPD. Recently, it was reported that the surfactant protein A increases the secretion of matrix metalloprotease 9, which degrades extracellular matrices of the lung, through a Toll-like receptor 2 (TLR2). In this context, possible role of TLR2 in the pathogenesis of COPD was postulated, and a functional dinucleotide repeat polymorphism in intron II of TLR2 was evaluated for any association with COPD. Method : Male patients with COPD and male smokers with a normal pulmonary function were enrolled in this study. The number of Guanine-Thymine repeats in intron II of the TLR2 gene were counted. Because the distributions of the repeats were trimodal, the alleles were classified into three subclasses, 12-16 repeats: short (S) alleles; 17-22 repeats: medium length (M) alleles; and 23-27 repeats: long (L) alleles. Result : 125 male patients with COPD and 144 age- and gender-matched blood donors with a normal lung function were enrolled. There were no differences in the distribution of each allele subclass (S, M and L) between the COPD and control group (p=0.75). The frequencies of the genotypes with and without each allele subclass in the COPD and control group were similar. Conclusion : A microsatellite polymorphism in intron II of TLR2 gene was not associated with the development of COPD in Koreans.

A novel cold-active lipase from Psychrobacter sp. ArcL13: gene identification, expression in E. coli, refolding, and characterization (새로운 Psychrobacter sp. ArcL13 유래 저온활성 지질분해효소 : 유전자 분리동정, 대장균에서의 발현, refolding 및 특성 연구)

  • Koo, Bon-Hun;Moon, Byung-Hern;Shin, Jong-Suh;Yim, Joung-Han
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.192-201
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    • 2016
  • Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.