• Title/Summary/Keyword: Glutamine

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Effect of Heat Shock Protein 70 on Inducible Nitric Oxide Synthase during Sepsis in Rats (패혈증에서 Heat Shock Protein 70의 과도발현이 Inducible Nitric Oxide Synthase의 발현에 미치는 효과)

  • Lee, Yong Keun;Shin, Hyo-Keun;Kwon, Woon Yong;Suh, Gil Joon;Youn, Yeo Kyu
    • Journal of Trauma and Injury
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    • v.21 no.1
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    • pp.59-65
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    • 2008
  • Purpose: The aim of this study was to evaluate the effect of overexpression of heat shock protein 70 (HSP70) on the expression of inducible nitric oxide synthase and on the concentration of nitric oxide and to determine the mechanism for the relationship between HSP70 and inducible nitric oxide synthase (iNOS) in sepsis. Methods: Experiments were performed on male Sprague-Dawley rats, and sepsis was induced by using cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 h after initiation of sepsis. We acquired serum and lung tissues from the rats 12 h or 24 h after initiation of sepsis. We analyzed the concentration of nitric oxide, the expression of HSP70 in the lung, and the gene expression of iNOS in the lung. Results: In CLP+GLN, glutamine given after initiation of sepsis enhanced the expression of HSP70 in the lung at 12 h (CLP+GLN vs. CLP:: $47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, p = 0.004). In CLP+GLN, glutamine attenuated the expression of iNOS mRNA in the lung at 12 h (CLP+GLN vs. CLP: $4167.17{\pm}951.59$ vs. $5513.73{\pm}1051.60$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $9,437.65{\pm}2,521.07$ vs. $18,740.27{\pm}8,241.20$, p = 0.016) and reduced the concentration of nitric oxide in serum at 12 h (CLP+GLN vs. CLP: $0.86{\pm}0.48$ vs. $3.82{\pm}2.53{\mu}mol/L$, p = 0.016) and 24 h (CLP+GLN vs. CLP: $0.39{\pm}0.25$ vs. $1.85{\pm}1.70{\mu}mol/L$, p = 0.025). Conclusion: The overexpression of HSP70 induced by the administration of glutamine in sepsis attenuated the gene expression of iNOS and reduced the concentration of nitric oxide.

Effects of L-glutamine supplementation on degradation rate and rumen fermentation characteristics in vitro

  • Suh, Jung-Keun;Nejad, Jalil Ghassemi;Lee, Yoon-Seok;Kong, Hong-Sik;Lee, Jae-Sung;Lee, Hong-Gu
    • Animal Bioscience
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    • v.35 no.3
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    • pp.422-433
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    • 2022
  • Objective: Two follow-up studies (exp. 1 and 2) were conducted to determine the effects of L-glutamine (L-Gln) supplementation on degradation and rumen fermentation characteristics in vitro. Methods: First, rumen liquor from three cannulated cows was used to test L-Gln (50 mM) degradation rate and ammonia-N production at 6, 12, 24, 36, and 48 h after incubation (exp. 1). Second, rumen liquor from two cannulated steers was used to assess the effects of five levels of L-Gln including 0% (control), 0.5%, 1%, 2%, and 3% at 0, 3, 6, 12, 24, 36, and 48 h after incubation on fermentation characteristics, gas production, and degradability of nutrients (exp. 2). Results: In exp. 1, L-Gln degradation rate and ammonia-N concentrations increased over time (p<0.001). In exp. 2, pH was reduced significantly as incubation time elapsed (p<0.001). Total gas production tended to increase in all groups as incubation time increased. Acetate and propionate tended to increase by increasing glutamine (Gln) levels, whereas levels of total volatile fatty acids (VFAs) were the highest in 0.5% and 3% Gln groups (p<0.001). The branched-chain VFA showed both linear and quadratic effects showing the lowest values in the 1% Gln group particularly after 6 h incubation (p<0.001). L-Gln increased crude protein degradability (p<0.001), showing the highest degradability in the 0.5% Gln group regardless of incubation time (p<0.05). Degradability of acid detergent fiber and neutral detergent fiber showed a similar pattern showing the highest values in 0.5% Gln group (p<0.10). Conclusion: Although L-Gln showed no toxicity when it was supplemented at high dosages (2% to 3% of DM), 0.5% L-Gln demonstrated the positive effects on main factors including VFAs production in-vitro. The results of this study need to be verified in further in-vivo study.

Functional Assessments of Spodpotera Cell-expressed Human Erythrocyte-type Glucose Transport Protein with a Site-directed Mutagenesis

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.119-122
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    • 2008
  • The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. In order to exploit this, the effects of substitution at the highly conserved residue glutamine 282 of the human erythrocyte-type glucose transporter have been examined by in vitro site-directed mutagenesis. The modified human transport protein has been expressed in Spodoptera frugiperda 21 cells by using the recombinant baculovirus AcNPV-GTL. To assess the functional integrity of the expressed transporter, measurements of the transport inhibitor cytochalasin B binding were performed, involving the membranes prepared from 4 days post infection with no virus, with wild-type virus or AcNPV-GTL virus. Data obtained showed that there was little or no D-glucose-inhibitable binding in cells infected with the wild type or no virus. Only the recombinant virus infected cells exhibited specific binding, which is inhibitable by D- but not by L-glucose. However, there was a notable reduction in the affinity for the potent inhibitor cytochalasin B when binding measurements of AcNPV-GTL were compared with those of AcNPV-GT, which has no substitution. It is thus suggested that although the modified and unmodified human transporters differed slightly in their affinity for cytochalasin B, the glutamine substitution did not interfere the heterologous expression of the human transporter in the insect cells.

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A Study on Nitrogen Compounds and the Vicissitude of Free Amino Acids in Mong-bean Saute (녹두(綠豆)지짐에 있어서의 함질소화합물(含窒素化合物) 특(特)히 유리(遊離) 아미노산(酸)의 소장(消長)에 관(關)하여)

  • Kang, Young-Hee;Lee, Jung-Ock;Chung, Ha-Boon;Lee, Ki-Yull
    • Journal of Nutrition and Health
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    • v.4 no.1
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    • pp.63-67
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    • 1971
  • The nitrogen content and free amino acids were determined from the saute of mong bean paste which is one of the important protein sources on Korean diet. During the saute' process of mong-bean paste, valine, r-aminobutyrate, glutamine, arginine, methionine, and unknown acids were lost, but, proline, lysine, homoserine, and tyrosine were detected.

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Analysis of amino acids in the liver of Hamster treated with Dimethylnitrosamine (Dimethylnitrosamine이 투여된 Hamster 간 속의 아미노산 분석)

  • Kim, Soo Gyung;Jung, Ha Seung;Park, Taek Kyu
    • Analytical Science and Technology
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    • v.7 no.2
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    • pp.165-171
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    • 1994
  • The effect of the chemical carcinogen dimetylnitrosamine(DMN) on the composition of amino acids of the liver in hamsters orally administered with DMN was studied by using the reversed phase high performance liquid chromatography technique. In the liver, the concentration of aspartic acid, glycine, glutamine, histidine, proline, tyrosine and leucine were increased ca. 2-fold of those observed in liver of control group, valine and tryptophan were increased ca. 3-fold, phenylalanine was markedly increased ca. 10-fold, whereas the concentration of threonine was decreased, serine, alanine, arginine, methionine, isoleucine and lysine were unchanged, respectively.

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Effects of Low-Serum Medium and Various Culture Additives on Production of Recombinant Human Erythropoietin in CHO Cell Cultures (CHO 세포 배양을 통한 Recombinant Human Erythropoietin의 생산에서 저혈청 배지와 배양 첨가물질이 미치는 영향)

  • Lee, Kyung-Sun;Cha, Hyun-Myoung;Lim, Jin-Hyuk;Kim, Dong-Il
    • KSBB Journal
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    • v.32 no.2
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    • pp.90-95
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    • 2017
  • Mammalian cell cultures have been used extensively to produce proteins for therapeutic agent because of their ability to perform post-translational modification including glycosylation. To produce recombinant protein, many factors and parameter are considered such as media composition, host cell type, and culture process. In this study, recombinant human erythropoietin (rhEPO) producing cell line was established by using glutamine synthetase system. To reduce serum concentration in media, we compared direct adaptation with step adaptation. Cell growth was faster in step adaptation. In low-level serum media, there were insufficient glucose for cell growth. Thus, we added glucose in low-level serum media from 2 g/L to 4.5 g/L. Titer of rhEPO was higher than other conditions at 4.5 g/L of glucose. Additionally, N-methyl-D-aspartate (NMDA), 13-cis-retinal, and pluronic F-68 (PF-68) were added to enhance productivity in CHO cell cultures. In conclusion, we applied CHO cell producing rhEPO to low-level of serum in media using step-adaptation. Also, we confirmed positive effect of NMDA, 13-cis-retinal, and PF-68.

A METHOD OF MUCOSA CULTURE (구강점막의 배양에 관한 연구)

  • Choi, Byung-Ho;Yoo, Jae-Ha
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.17 no.4
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    • pp.331-336
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    • 1995
  • To use cultured mucosa as a graft of full thickness, our laboratory has been involved in the development of techniques to grow epidermis together with connective tissue. Human oral mucosa was obtained at dental surgery. Under sterile conditions the tissues were cut into explants of 0.1 $cm^2$ which were placed in the center of 24 well tissue culture dishes and incubated in a growth medium. The growth medium used for epithelial was MEM(Minimum Essential Medium) supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide, glutamine (0.292 g/l), epidermal growth factor (40 ug/ml), cholera toxin (30 ng/ml), hydrocortisone (2 ug/ml), insulin (40 ug/ml) and transferin (5 ug/ml). The medium for stratification of epithelial cells was MEM supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide and glutamine (0.292 g/l). The medium used for fibroblasts was MEM supplemented with 10% fetal calf serum. With the three types of media used alternatively, a mucosa composed of epidermis and connective tissue was obtained after 3 weeks of culture.

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