• 제목/요약/키워드: CAT expression

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Change in Cationic Amino Acid Transport System and Effect of Lysine Pretreatment on Inflammatory State in Amyotrophic Lateral Sclerosis Cell Model

  • Latif, Sana;Kang, Young-Sook
    • Biomolecules & Therapeutics
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    • v.29 no.5
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    • pp.498-505
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    • 2021
  • Amyotrophic lateral sclerosis (ALS) is a lethal neurological disorder characterized by the deterioration of motor neurons. The aim of this study was to investigate alteration of cationic amino acid transporter (CAT-1) activity in the transport of lysine and the pretreatment effect of lysine on pro-inflammatory states in an amyotrophic lateral sclerosis cell line. The mRNA expression of cationic amino acid transporter 1 was lower in NSC-34/hSOD1G93A (MT) than the control cell line (WT), lysine transport is mediated by CAT-1 in NSC-34 cell lines. The uptake of [3H]L-lysine was Na+-independent, voltage-sensitive, and strongly inhibited by inhibitors and substrates of cationic amino acid transporter 1 (system y+). The transport process involved two saturable processes in both cell lines. In the MT cell line, at a high-affinity site, the affinity was 9.4-fold higher and capacity 24-fold lower than that in the WT; at a low-affinity site, the capacity was 2.3-fold lower than that in the WT cell line. Donepezil and verapamil competitively inhibited [3H]L-lysine uptake in the NSC-34 cell lines. Pretreatment with pro-inflammatory cytokines decreased the uptake of [3H]L-lysine and mRNA expression levels in both cell lines; however, the addition of L-lysine restored the transport activity in the MT cell lines. L-Lysine exhibited neuroprotective effects against pro-inflammatory states in the ALS disease model cell lines. In conclusion, studying the alteration in the expression of transporters and characteristics of lysine transport in ALS can lead to the development of new therapies for neurodegenerative diseases.

Gene Profile of Mesenchymal Stem Cell Induced by SAC or Hydrogen Peroxide (H2O2) (마늘성분 SAC 및 Hydrogen Peroxide에 의한 줄기세포의 유전자 발현 윤곽)

  • Park, Ran-Sook
    • The Korean Journal of Food And Nutrition
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    • v.25 no.4
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    • pp.863-870
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    • 2012
  • Though hydrogen peroxide ($H_2O_2$) causes a deleterious effect to cells with its reactive oxygen species resulting in cell death, S-allyl cysteine (SAC, a bioactive organosulfur compound of aged garlic extract) has been known to have a cytoprotective effect. Few reported profiles of gene expression of $H_2O_2$ and SAC treated human cord blood derived mesenchymal stem cells (MSC). This study revealed changes in the profile of twenty-one genes grouped by oxidative stress, antioxidant, cell death, anti-apoptosis and anti-aging by quantitative real time PCR. A concentration of $100{\mu}M$ of SAC or $50{\mu}M$ of $H_2O_2$ was applied to MSC which show moderate growth and apoptosis pattern. $H_2O_2$ treatment enhanced expression of eleven genes out of twenty-one genes compared with that of control group, on the contrary SAC suppressed expression of eighteen genes out of twenty-one genes except C ros oncogene. SAC decreased expression of oxidative stress genes such as SOD1, CAT and GPX. These results seemed consistent with reports which elucidated over-expression of NF-${\kappa}$B by $H_2O_2$, and suppression of it by SAC. This study will confer basic information for further experiments regarding the effects of SAC on gene levels.

Regulation of fpr Gene Encoding NADPH : Ferredoxin Oxidoreductase by the soxRS Locus in Escherichia coli

  • Koh, Young-Sang;Choih, Jenny;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.137-143
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    • 1996
  • We isolated a promoter inducible by paraquat, a superoxide-generating agent, from Escherichia coli using a promoter-probing plasmid pRS415. From sequence analysis we found out the promoter is for fpr ENCODING nadph : ferredoxin oxidoreductase. We constructed on operon fusion of lacZ gene with fpr promoter to monitor the expression of the gene in the single-copy state. LacZ expression generators, menadione and plumbagin, also induced the expression of .betha.-galactosidase in the fusion strain. On the other hand, no significant induction was observed by treatment with hydrogen peroxide, ethanol, and heat shock. Induction of .betha.-galactosidase was significantly reduced by introducing a .DELTA. sox 8 :: cat of soxS3 :: Tn10 mutation into the fusion strain, indicating that fpr gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis. Possible roles of fpr induction in superoxide stress were discussed.

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Enhanced Antioxidant Enzymes Are Associated with Reduced Hydrogen Peroxide in Barley Roots under Saline Stress

  • Kim, Sang-Yong;Lim, Jung-Hyun;Park, Myoung-Ryoul;Kim, Young-Jin;Park, Tae-Il;Seo, Yong-Won;Choi, Kyeong-Gu;Yun, Song-Joong
    • BMB Reports
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    • v.38 no.2
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    • pp.218-224
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    • 2005
  • Antioxidant enzymes are related to the resistance to various abiotic stresses including salinity. Barley is relatively tolerant to saline stress among crop plants, but little information is available on barley antioxidant enzymes under salinity stress. We investigated temporal and spatial responses of activities and isoform profiles of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), non-specific peroxidase (POX), and glutathione reductase (GR) to saline stress in barley seedlings treated with 200 mM NaCl for 0, 1, 2, 5 days, respectively. In the control plant, hydrogen peroxide content was about 2-fold higher in the root than in the shoot. Under saline stress, hydrogen peroxide content was decreased drastically by 70% at 2 d after NaCl treatment (DAT) in the root. In the leaf, however, the content was remained unchanged by 2 DAT and increased about 14 % at 5 DAT. In general, the activities of antioxidant enzymes were increased in the root and shoot under saline stress. But the increase was more significant and consistent in the root. The activities of SOD, CAT, APX, POX, and GR were increased significantly in the root within 1 DAT, and various elevated levels were maintained by 5 DAT. Among the antioxidant enzymes, CAT activity was increased the most drastically. The significant increase in the activities of SOD, CAT, APX, POX, and GR in the NaCl-stressed barley root was highly correlated with the increased expression of the constitutive isoforms as well as the induced ones. The hydrogen peroxide content in the root was most highly correlated with the CAT activity, indicating an increased role of CAT in hydrogen peroxide detoxification under salinity stress. In addition, the results suggest the significance of temporal and spatial regulation of each antioxidant isoform in determining the competence of the antioxidant capacity under saline stress.

Transcriptional Regulation of the VP16 Gene of Herpes Simplex Virus Type 1

  • Kwun, Hyun-Jin;Jun, Hong-Ki;Lee, Tae-Ho;Jang, Kyung-Lib
    • BMB Reports
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    • v.32 no.5
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    • pp.456-460
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    • 1999
  • The promoter of the HSV-1 VP16 gene contains binding sites for the cellular transcription factors such as USF, CTF, and Sp1, each of which affects basal level expression of the VP16 gene. Transcription of the VP16 gene was induced by viral immediate-early proteins, ICP0 and ICP4, in a synergistic manner but repressed by ICP22. To gain further insight into the role of ICP0 in the expression of the VP16 gene during virus infection, several mutants with deletions in each of their transcriptional regulatory elements were generated. According to transient gene expression assays of these mutants using the CAT gene as a reporter, the USF and CTF binding sites were necessary for efficient induction of the promoter in the presence of transfected ICP0 or during virus infection, whereas the Sp1 binding site had little effect on ICP0-mediated VP16 expression. These results indicate that the immediate early proteins of HSV-1 regulate expression of the VP16 gene during virus infection by modulating the activities of cellular transcription factors such as USF and CTF.

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Stress Responses of Olive Flounder (Paralichthys olivaceus) to Salinity Changes (염분변화에 따른 넙치(Paralichthys olivaceus)의 스트레스 반응)

  • Park, Hyung-Jun;Min, Byung-Hwa
    • Korean Journal of Ichthyology
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    • v.30 no.1
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    • pp.1-8
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    • 2018
  • We tried to determine the optimum salinity for a cultured of olive flounder (Paralichthys olivaceus) by investigating after exposing the fish at different salinity (10, 15, 20 and 25 psu) for 24 and 48 hours compared with control group (fish before transfer to experimental tank). As a control groups, we compared an analyzed with other experimental groups using olive flounder in natural sea water. Hematological parameters including hematocrit (Ht) and hemoglobin (Hb), cortisol and glucose, aspartate aminotransferase (AST) and alanine aminotransferase (ALT), $NH_3$, osmolality, total protein (TP), $Na^+$, $K^+$ and $Cl^-$ mostly exhibited significant changes at 10 and 15 psu groups compared with control groups for 24 and 48 hours exposed. Plasma SOD (superoxide dismutase) and CAT (catalase) activity also increased with experimental groups (10 and 15 psu) compared to the control groups. The expression of HSP70 mRNA was also higher at low-salinity (10 and 15 psu) than at control group. In particular, after 24 hours exposed, it expression to 15 psu groups showed a significant difference compared to the control group. However, after 48 hours exposed, it expression was higher in the 10 psu groups than the control. It is assumed that the changes in the hematological responses and hormone, homeostasis and metabolism were resulted in to protect fish body from stress. Based on these results, we are expected that it will be used as basic data for the culture of olive flounder prepared for low salinity.

Antioxidant Enzyme Activity and Anti-Adipogenic Effects of (-)-Epigallocatechin-3-Gallate in 3T3-L1 Cells ((-)-Epigallocatechin-3-Gallate의 3T3-L1 세포에서 항산화 효소 활성 및 지방세포 분화 억제 효과)

  • Kim, Younghwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.11
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    • pp.1293-1299
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    • 2017
  • Obesity contributes to the development of diseases, such as type II diabetes, hypertension, coronary heart disease, and cancer. In addition, oxidative stress caused by reactive oxygen species (ROS) is recognized widely as a contributing factor in the development of chronic diseases. This study was examined the antioxidant and anti-adipogenic activities of epigallocatechin-3-gallate (EGCG) in 3T3-L1 preadipocytes. 3T3-L1 cells were differentiated with or without EGCG for 6 days. The production of glutathione (GSH) and the activities of the antioxidant enzymes, such as glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD) were measured. EGCG inhibited significantly the lipid accumulation and the expression of adipogenic specific proteins including CCAAT/enhancer binding protein ${\alpha}$ and adipocyte fatty acid binding protein. The production of intracellular ROS was decreased significantly by EGCG in 3T3-L1 cells. EGCG increased the GSH production and the activities of GPx, GR, CAT, and SOD. Moreover, EGCG increased the protein expression of glutamate-cysteine ligase and heme oxygenase-1 in 3T3-L1 cells. These results suggest that EGCG increased the activity and expression of antioxidant enzymes and suppressed the lipid accumulation in 3T3-L1 cells. Therefore, the use of phytochemicals that can maintain the GSH redox balance in adipose tissue could be promising for reducing obesity.

Effect of Transgenic Genotype on Transgene Expression in Mud Loach (Misgurnus mizoIepis): I. Copy Number-Dependent Expression in Gynogenetically Derived Homozygous Transgenics

  • Nam Yoon Kwon;Noh Jae Koo;Kim Dong Soo
    • Fisheries and Aquatic Sciences
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    • v.4 no.1
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    • pp.39-46
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    • 2001
  • To examine the effect of copy number-dependent transgenic genotype on the expression of foreign gene, stable hemizygous and homozygous transgenic breeding line was established using artificial parthenogenesis. For this purpose, induced diploid gynogenetic transgenesis was optimized in mud loach (Misgurnus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to $4,050\;ergs/mm^2$ The UV-irradiated sperm were inseminated into eggs from recessive color strain (yellow) or heterozygous transgenic mud loach containing CAT gene. Cold shock at $2^{\circ}C$ for 60 min, 5 min post fertilization successfully restored the diploidy of eggs inseminated with UV-irradiated sperm. Restoration to diploidy was confirmed by flow cytometry and gynogenetic status was verified by examining maternal exclusive inheritance of multi-locus DNA fingerprints, body color and transgenic marker. Putative isogenic transgenic fish clearly showed homozygous status at trans gene locus based on Southern blot hybridization and progeny testing. Further, such homozygous gynogenetic diploids revealed the increased levels of transgene expression, when compared to those of heterozygous (hemizygous) transgenic fish.

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Molecular Characterization and Expression Analysis of Adrenergic Receptor Beta 2 (ADRB2) Gene before and after Exercise in the Horse

  • Cho, Hyun-Woo;Shin, Sangsu;Song, Ki-Duk;Park, Jeong-Woong;Choi, Jae-Young;Lee, Hak-Kyo;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.686-690
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    • 2015
  • The adrenergic receptor beta 2 (ADRB2) plays a role in various physiological responses of the muscle to exercise, such as contraction and relaxation. Given its important role in muscle function, we investigated the structure of the horse ADRB2 gene and its expression pattern after exercise to determine if it can serve as a putative biomarker for recovery. Evolutionary analyses using synonymous and non-synonymous mutation ratios, were compared with other species (human, chimpanzee, mouse, rat, cow, pig, chicken, dog, and cat), and revealed the occurrence of positive selection in the horse ADRB2 gene. In addition, expression analyses by quantitative polymerase chain reaction exhibited ubiquitous distribution of horse ADRB2 in various tissues including lung, skeletal muscle, kidney, thyroid, appendix, colon, spinal cord and heart, with the highest expression observed in the lung. The expression of ADRB2 in skeletal muscle was significantly up-regulated about four folds 30 minutes post-exercise compared to pre-exercise. The expression level of ADRB2 in leukocytes, which could be collected with convenience compared with other tissues in horse, increased until 60 min after exercise but decreased afterward until 120 min, suggesting the ADRB2 expression levels in leukocytes could be a useful biomarker to check the early recovery status of horse after exercise. In conclusion, we identified horse ADRB2 gene and analyzed expression profiles in various tissues. Additionally, analysis of ADBR2 gene expression in leukocytes could be a useful biomarker useful for evaluation of early recovery status after exercise in racing horses.

Effects of Ixeris dentata ext. on Lowering Lipid and Anti-oxidation (씀바귀의 지질강하 및 항산화효과)

  • Lee, Eun
    • Korean Journal of Plant Resources
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    • v.24 no.1
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    • pp.55-60
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    • 2011
  • To investigate effects of Ixeris dentata EtOH ext. on lowering lipid levels and anti-oxidation activity, hyperlipidemic rats were treated with Ixeris dentata EtOH ext. and lipid levels and anti-oxdation activity were measured. The Ixeris dentata EtOH ext. groups showed low concentration of plasma FFA, plasma triglyceride, plasma total cholesterol, and plasma LDL-cholesterol compared to control group. However, concentration of plasma HDLcholesterol was not significantly different among all the treatment groups. The Ixeris dentata EtOH ext. groups showed lower level of liver total cholesterol, liver triglyceride, plasma TBARS, and liver TBARS than those of control group. The Ixeris dentata EtOH ext. groups also showed higher level of GSH-Px activity, SOD activity, and CAT activity than those of control group. Moreover, the Ext. showed lower level of TNF-${\alpha}$, Apo-B, Apo-E, and leptin expression than those of control group. The results in this study shows that the Ixeris dentata EtOH ext. have positive effect in lowering lipid level, and anti-oxidative activity.