• Title/Summary/Keyword: E2 protein

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Study on Biochemical Pollutant Markers for Diagnosis of Marine Pollution VI. Changes in Cholinesterase Activity of Flounder (Pleuronichthys cornutus) in the Yellow Sea (해양오염의 진단을 위한 생화학적 오염지표에 관한 연구 VI. 황해산 도다리 (Pleuronichthys cornutus)의 콜린에스테라아제 활성의 변화)

  • CHOI Jin-Ho;KIM Dong-Woo;PARK Chung-Kil;YANG Dong Beom
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.4
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    • pp.614-619
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    • 1997
  • This study was designed to investigate the biochemical pollutant markers for diagnosis of marine pollutions by changes in cholinesterase activity of the flounder (Pleuronichthys cornutus) in the Yellow Sea of Korea. Acetylchotinesterase (AChE) activities in brain and muscle of wild flounders in the Yellow Sea were significantly lower $(20\~30\%\;and\;10\~40\%,\;respectively)$ than those of wild flounder in Pohang (control) of the last Sea. Butyrylcholinesterase (BChE) activities in brain and muscle of wild flounders in the Yellow Sea were significantly lower $(10\~30\%\;and\;35\~45\%,\;respectively)$ than those of wild flounder in Pohang of the East Sea. lactate dehydrogenase (LDH) activities in serum of wild flounders in the yellow Sea were significantly $(about\;30\%)$ lower than those of wild flounder in Pohang of the East Sea. These results suggest that AChE and BChE activities in brain and muscle of wild flounders of the Yellow Sea may be used as the most effective mean in a biochemical marker for diagnosis of pollutant effects by organophosphorus pesticides.

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Curcumin Induces Downregulation of E2F4 Expression and Apoptotic Cell Death in H CT116 Human Colon Cancer Cells; Involvement of Reactive Oxygen Species

  • Kim, Kyung-Chan;Lee, Chu-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.391-397
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    • 2010
  • E2F transcription factors and their target genes have been known to play an important role in cell growth control. We found that curcumin, a polyphenolic phytochemical isolated from the plant Curcuma longa, markedly suppressed E2F4 expression in HCT116 colon cancer cells. Hydrogen peroxide was also found to decrease E2F4 protein level, indicating the involvement of reactive oxygen species (ROS) in curucmin-induced downregulation of E2F4 expression. Involvement of ROS in E2F4 downregulation in response to curcumin was confirmed by the result that pretreatment of cells with N-acetylcystein (NAC) before exposure of curcumin almost completely blocked the reduction of E2F4 expression at the protein as well as mRNA level. Anti-proliferative effect of curcumin was also suppressed by NAC which is consistent to previous reports showing curcumin-superoxide production and induction of poly (ADP-ribose) polymerase (PARP) cleavage as well as apoptosis. Expression of several genes, cyclin A, p21, and p27, which has been shown to be regulated in E2F4-dependent manner and involved in the cell cycle progression was also affected by curcumin. Moreover, decreased (cyclin A) and increased (p21 and p27) expression of these E2F4 downstream genes by curcumin was restored by pretreatment of cells with NAC and E2F4 overexpression which is induced by doxycycline. In addition, E2F4 overexpression was observed to partially ameliorate curcumin-induced growth inhibition by cell viability assay. Taken together, we found curcumin-induced ROS down-regulation of E2F4 expression and modulation of E2F4 target genes which finally lead to the apoptotic cell death in HCT116 colon cancer cells, suggesting that E2F4 appears to be a novel determinant of curcumin-induced cytotoxicity.

Effects of the Dietary Protein and Energy Levels on Growth in Fat Cod (Hexagrammos otakii Jordan et Starks) (사료의 단백질 및 에너지 함량이 쥐노래미 성장에 미치는 영향)

  • LEE Jong Kwan;LEE Sang-Min
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.4
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    • pp.464-473
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    • 1996
  • Two-month feeding experiment was conducted to investigate the optimum dietary protein level and energy to protein ratio in fat cod (Hexagrammos otakii Jordan et STARKS). The fish averaging 29 g were fed with one of the isocaloric diets containing 30, 40, 50 or $60\%$ of protein, or with one of the isoproteic diets containing 9, 10, 11 or 12 of available energy/protein (E/P) ratio. Weight gain and feed efficiency increased significantly with dietary protein level up to $50\%$, then decreased with $60\%$ protein diet (P<0.05). Daily protein intake increased significantly with dietary protein level, whereas protein efficiency ratio decreased with dietary protein level (P<0.05). Second order polynomial regression analyses of percent weight gain and daily protein intake may indicate that the adequate dietary protein level is $45\%$ and daily protein requirement per 100g fish is 1.5g for maximal growth. Weight gain, feed efficiency and protein efficiency from fish led the diet containing 12 of E/P ratio were significantly higher than those from fish fed the other diets (P<0.05). Daily feed or protein intake from fish fed the diet containing 12 of E/P ratio was significantly lower than those from fish fed the other diets (P<0.05). Daily lipid intake increased significantly with dietary E/P ratio (P<0.05).

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Effects of Amifostine on Apoptosis, Cell Cycle and Cytoprotection of Human Colon Cancer Cell Lines

  • Eun Ju Lee
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.287-295
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    • 2023
  • Amifostine was developed to protect cells, but it is known to induce cytotoxicity and apoptosis, and the exact mechanism is unknown. In this study, we investigated how the DNA mismatch repair (MMR) system interacts with p53 to prevent apoptosis, cell cycle arrest, and cytoprotective effects induced by amifostine. HCT116 colon cancer cells sublines HCT116/p53+,HCT116/p53+, HCT116/p53-, HCT116/E6 and HCT116+ch3/E6 cells were used for evaluation. Amifostine induced G1 arrest and increased toxicity two-fold in p53- cells regardless of MMR expression. Both G1 cell cycle arrest and induction of p53 protein peaked at 24 h after the start of amifostine exposure. Both G1 cell cycle arrest and induction of p53 protein peaked at 24 h after the start of amifostine exposure. Amifostine induced the expression of p21 protein in both p53+ and p53- cells. As for apoptosis, compared to p53- cells, p53+ cells showed 3.5~4.2 times resistance to amifostine-induced apoptosis. HCT116+E6 with both p53 and MMR loss showed maximum apoptosis at 48 h, and HCT116+ch3/E6HCT116+ch3/E6 with p53 loss showed maximum apoptosis at 24 h. As a result, it was confirmed through in vitro experiments that amifostine-induced G1 cell cycle arrest and apoptosis are mediated through a pathway dependent on MMR and p53 protein.

Optimal Conditions for the Expression of Glycoprotein E2 of Classical Swine Fever Virus using Baculovirus in Insect Cells

  • Bae, Sung Min;Lee, Seung Hee;Kwak, Won Suk;Ahn, Yong Oh;Shin, Tae Young;Woo, Soo Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.207-213
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    • 2014
  • The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein $E^{rns}$ (E0), E1 and E2. Among them, E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. In this study, to determine the optimal expression conditions of glycoprotein E2 using baculovirus system, we investigated the influence of insect cells and media to the expression of recombinant E2. Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZA DNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21 cells was first observed after 3 days and reached a maximum on the 5th day after infection. Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity of infection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) were tested, High-Five cells showed the highest production. In addition, four different serum-free and serum-supplemented media, respectively, were tested for the expression of glycoprotein E2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided the best conditions for protein production and the BV yield.

Metabolic Elasticity and Induction of Heat Shock Protein 70 in Labeo rohita Acclimated to Three Temperatures

  • Das, T.;Pal, A.K.;Chakraborty, S.K.;Manush, S.M.;Chatterjee, N.;Apte, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.7
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    • pp.1033-1039
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    • 2006
  • The metabolic response of Labeo rohita to thermal acclimation was assessed. Advanced fingerlings of L. rohita (average weight $31{\pm}1.4g$) were acclimated to 31, 33 and $36^{\circ}C$ compared with ambient temperatures ($26^{\circ}C$) for 30 days and different enzymes associated with stress response were estimated. Glycolytic enzyme-Lactate dehydrogenase, (LDH, E.C.1.1.1.27), TCA cycle enzyme-Malate dehydrogenase (MDH, E.C.1.1.1.37), Protein metabolizing enzymes-Aspartate amino transferase (AST, E.C.2.6.1.1) and Alanine amino transferase (ALT, E.C.2.6.1.2) of liver, gill and muscle, Gluconeogenic enzymes-Fructose 1,6 Bi phosphatase (FBPase, E.C. 3.1.3.11) and Glucose 6 phosphatase (G6Pase, E.C. 3.1.3.9) of liver and kidney were significantly (p<0.05) different with increasing acclimation temperatures. Heat Shock Protein-70 (HSP-70) was expressed in increasing intensity at 31, 33 and $36^{\circ}C$ but was not expressed at $26^{\circ}C$. Results suggest that higher acclimation temperatures enhance metabolism and L. rohita maintains homeostasis between $26-36^{\circ}C$ via an acclimation episode. Such adaptation appears to be facilitated by resorting to gluconeogenic and glycogenolytic pathways for energy mobilization and induction of HSPs.

Daidzein Modulations of Apolipoprotein B and Fatty Acid Synthase mRNA Expression in Chick Liver Vary Depending on Dietary Protein Levels

  • Choi, Jinho;Song, Jungmin;Choi, Yeon-Mi;Jang, Dong-Ju;Kim, Eunmi;Kim, Inho;Chee, Kew-Mahn
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.236-244
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    • 2006
  • This study was designed to determine the effects of daidzein (DE) on hepatic lipid metabolism in chicks fed with low protein (LP) diet based on casein. In experiment 1, the male chicks were fed with one of the three levels of dietary protein containing 10.95%, 21.9% and 43.8% protein content for 2 days. In experiment 2, the chicks were fed one of the three levels of protein with or without DE at 1,000 mg/kg diet for 2 days. Experiment 3 was conducted to compare DE (LP+DE) with estradiol (LP+E2) in chicks fed with LP diet for 7 days. Plasma lipid profiles, hepatic lipid profiles, activities of hepatic malic enzyme and isocitrate dehydrogenase (ICDH) were measured. Transcriptions of hepatic fatty acid synthase, apolipoprotein-B (APO-B), and fructose bisphosphatase mRNA were measured by RT-PCR. Increasing dietary protein levels markedly decreased the concentrations of plasma triglycerides, hepatic total lipids, hepatic TG, and the mRNA transcriptions while the increased dietary protein levels increased hepatic ICDH activities in experiment 1. In experiment 2, the effects of dietary protein levels on blood and hepatic lipid content were more prominent than those of the additional DE. Interestingly, plasma TG levels were affected by DE supplementation (p<0.05). In experiment 3, DE inhibited APO-B mRNA expressions and stimulated the accumulation of lipid in the liver through mechanisms different from E2. In this study, we demonstrate that DE has beneficial effects on blood lipid profiles, but that it inhibits APO-B mRNA transcription and aggravates the fatty liver induced by LP diet in chicks.

Prediction of hub genes of Alzheimer's disease using a protein interaction network and functional enrichment analysis

  • Wee, Jia Jin;Kumar, Suresh
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.39.1-39.8
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    • 2020
  • Alzheimer's disease (AD) is a chronic, progressive brain disorder that slowly destroys affected individuals' memory and reasoning faculties, and consequently, their ability to perform the simplest tasks. This study investigated the hub genes of AD. Proteins interact with other proteins and non-protein molecules, and these interactions play an important role in understanding protein function. Computational methods are useful for understanding biological problems, in particular, network analyses of protein-protein interactions. Through a protein network analysis, we identified the following top 10 hub genes associated with AD: PTGER3, C3AR1, NPY, ADCY2, CXCL12, CCR5, MTNR1A, CNR2, GRM2, and CXCL8. Through gene enrichment, it was identified that most gene functions could be classified as integral to the plasma membrane, G-protein coupled receptor activity, and cell communication under gene ontology, as well as involvement in signal transduction pathways. Based on the convergent functional genomics ranking, the prioritized genes were NPY, CXCL12, CCR5, and CNR2.

Effects of maternal undernutrition during late pregnancy on the regulatory factors involved in growth and development in ovine fetal perirenal brown adipose tissue

  • Yang, Huan;Ma, Chi;Zi, Yang;Zhang, Min;Liu, Yingchun;Wu, Kaifeng;Gao, Feng
    • Animal Bioscience
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    • v.35 no.7
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    • pp.1010-1020
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    • 2022
  • Objective: The experiment was conducted to evaluate the effects of maternal undernutrition during late pregnancy on the expressions of genes involved in growth and development in ovine fetal perirenal brown adipose tissue (BAT). Methods: Eighteen ewes with singleton fetuses were allocated to three groups at day 90 of pregnancy: restricted group 1 (RG1, 0.33 MJ metabolisable energy [ME]/kg body weight [BW]0.75/d, n = 6), restricted group 2 (RG2, 0.18 MJ ME/kg BW0.75/d, n = 6), and a control group (CG, ad libitum, 0.67 MJ ME/kg BW0.75/d, n = 6). The fetuses were removed at day 140 of pregnancy. All data were analyzed by using the analysis of variance procedure. Results: The perirenal fat weight (p = 0.0077) and perirenal fat growth rate (p = 0.0074) were reduced in RG2 compared to CG. In fetal perirenal BAT, the protein level of uncoupling protein 1 (UCP1) (p = 0.0001) was lower in RG1 and RG2 compared with CG and UCP1 mRNA expression (p = 0.0265) was decreased in RG2. The protein level of myogenic factor 5 (Myf5) was also decreased in RG2 (p = 0.0001). In addition, mRNA expressions of CyclinA (p = 0.0109), CyclinB (p = 0.0019), CyclinD (p = 0.0015), cyclin-dependent kinase 1 (CDK1) (p = 0.0001), E2F transcription factor 1 (E2F1) (p = 0.0323), E2F4 (p = 0.0101), and E2F5 (p = 0.0018) were lower in RG1 and RG2. There were decreased protein expression of peroxisome proliferator-activated receptor-γ (PPARγ) (p = 0.0043) and mRNA expression of CCAAT/enhancer-binding protein-α (C/EBPα) (p = 0.0307) in RG2 and decreased PPARγ mRNA expression (p = 0.0008) and C/EBPα protein expression (p = 0.0015) in both RG2 and RG1. Furthermore, mRNA expression of bone morphogenetic protein 4 (BMP4) (p = 0.0083) and BMP7 (p = 0.0330) decreased in RG2 and peroxisome proliferator-activated receptor co-activator-1α (PGC-1α) reduced in RG2 and RG1. Conclusion: Our observations support that repression of regulatory factors promoting differentiation and development results in the inhibition of BAT maturation in fetal perirenal fat during late pregnancy with maternal undernutrition.

Effects of Dietary Conjugated Linoleic Acid (CLA) on Antioxidant System in the Liver of Chronically Ethanol-Treated Rats (식이에 첨가한 Conjugated Linoleic Acid (CLA)가 만성적으로 알코올을 섭취한 쥐에서 간조직의 항산화 체계에 미치는 영향)

  • Kim, Se-Na;Kim, Min-Seok;Park, Hyun-Suh
    • Journal of Nutrition and Health
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    • v.40 no.2
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    • pp.105-110
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    • 2007
  • The study was designed to observe antioxidant activities of conjugated linoleic acid (CLA) by determining antioxidant enzyme protein levels [cytochrome P4502 El (CYP2E1), Copper, Zinc-superoxide dismutase (CuZn-SOD), glutathione peroxidase (CSH-Px), glutathione S-transferase (GST)] by Western blot analysis and the levels of ${\alpha}$-tocopherol and 2-thiobarbituric acid reactive substances (TBARS) in the liver of chronically ethanol-treated rats. Sixty Sprague Dawley male rats were divided into 3 groups (Control, EtOH, EtOH+CLA). All rats were fed Lieber-DeCarli liquid diet for 4 weeks by pair-feeding against the EtOH group. The liquid diet was supplemented with 1.77g CLA mixture per kg diet in the EtOH+CLA group. Isocaloric maltose dextrin was added in replace of 50g ethanol (36%kcal) for the Control group. Ethanol ingestion significantly increased the levels of CYP2E1 protein and TBARS, but significantly reduced CuZn-SOD protein level and increased GST protein level. There was no significant effect on the level of GSH-Px protein and ${\alpha}$-tocopherol in the liver by ethanol. CLA supplementation with ethanol significantly increased the levels of CuZn-SOD, GSH-Px and GST and also significantly attenuated TBARS level, whereas there was no significant effect on the levels of CYP2E1 protein and ${\alpha}$-tocopherol by CLA. Overall, the CLA supplemented to ethanol could significantly increase the levels of CuZn-SOD, GSH-Px and GST proteins and reduce the level of TBARS in the liver of chronically ethanol-treated rats.