• Title/Summary/Keyword: $eIF2{\alpha}$

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Reduced EGFR Level in eIF2α Phosphorylation-Deficient Hepatocytes Is Responsible for Susceptibility to Oxidative Stress

  • Kim, Mi-Jeong;Choi, Woo-Gyun;Ahn, Kyung-Ju;Chae, In Gyeong;Yu, Rina;Back, Sung Hoon
    • Molecules and Cells
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    • v.43 no.3
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    • pp.264-275
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    • 2020
  • Reactive oxygen species (ROS) play a significant role in intracellular signaling and regulation, particularly when they are maintained at physiologic levels. However, excess ROS can cause cell damage and induce cell death. We recently reported that eIF2α phosphorylation protects hepatocytes from oxidative stress and liver fibrosis induced by fructose metabolism. Here, we found that hepatocyte-specific eIF2α phosphorylation-deficient mice have significantly reduced expression of the epidermal growth factor receptor (EGFR) and altered EGFR-mediated signaling pathways. EGFR-mediated signaling pathways are important for cell proliferation, differentiation, and survival in many tissues and cell types. Therefore, we studied whether the reduced amount of EGFR is responsible for the eIF2α phosphorylation-deficient hepatocytes' vulnerability to oxidative stress. ROS such as hydrogen peroxide and superoxides induce both EGFR tyrosine phosphorylation and eIF2α phosphorylation. eIF2α phosphorylation-deficient primary hepatocytes, or EGFR knockdown cells, have decreased ROS scavenging ability compared to normal cells. Therefore, these cells are particularly susceptible to oxidative stress. However, overexpression of EGFR in these eIF2α phosphorylation-deficient primary hepatocytes increased ROS scavenging ability and alleviated ROS-mediated cell death. Therefore, we hypothesize that the reduced EGFR level in eIF2α phosphorylation-deficient hepatocytes is one of critical factors responsible for their susceptibility to oxidative stress.

ON A CLASS OF QUANTUM ALPHA-CONVEX FUNCTIONS

  • NOOR, KHALIDA INAYAT;BADAR, RIZWAN S.
    • Journal of applied mathematics & informatics
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    • v.36 no.5_6
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    • pp.567-574
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    • 2018
  • Let $f:f(z)=z+{\sum^{{\infty}}_{n=2}}a_nz^n$ be analytic in the open unit disc E. Then f is said to belong to the class $M_{\alpha}$ of alpha-convex functions, if it satisfies the condition ${\Re}\{(1-{{\alpha})}{\frac{zf^{\prime}(z)}{f(z)}}+{{\alpha}}{\frac{(zf^{\prime}(z))^{\prime})}{f^{\prime}(z)}}\}$ > 0, ($z{\in}E$). In this paper, we introduce and study q-analogue of the class $M_{\alpha}$ by using concepts of Quantum Analysis. It is shown that the functions in this new class $M(q,{\alpha})$ are q-starlike. A problem related to q-Bernardi operator is also investigated.

Purification and Characterization of the Functional Catalytic Domain of PKR-Like Endoplasmic Reticulum Kinase Expressed in Escherichia coli

  • Yun Jin-A;Chung Ho-Young;Kim Seong-Jun;Cho Hyun-Soo;Oh Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1453-1458
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    • 2006
  • PKR-like endoplasmic reticulum (ER) kinase (PERK) is a type I transmembrane ER-resident protein containing a cytoplasmic catalytic domain with a Ser/Thr kinase activity, which is most closely related to the eukaryotic translation initiation factor-$2{\alpha}$ ($eIF2{\alpha}$) kinase PKR involved in the antiviral defense pathway by interferon. We cloned and expressed the PERK C-terminal kinase domain (cPERK) in Escherichia coli. Like PERK activation in cells under ER stress, wild-type cPERK underwent autophosphorylation when overexpressed in E. coli, whereas the cPERK(K621M) with a methionine substitution for the lysine at amino acid 621 lost the autophosphorylation activity. The activated form cPERK which was purified to near homogeneity, formed an oligomer and was able to trans-phosphorylate specifically its cellular substrate $eIF2{\alpha}$. Two-dimensional phosphoamino acids analysis revealed that phosphorylation of cPERK occurs at the Ser and Thr residues. The functionally active recombinant cPERK, and its inactive mutant should be useful for the analysis of biochemical functions of PERK and for the determination of their three-dimensional structures.

SUBSTITUTION OPERATORS IN THE SPACES OF FUNCTIONS OF BOUNDED VARIATION BV2α(I)

  • Aziz, Wadie;Guerrero, Jose Atilio;Merentes, Nelson
    • Bulletin of the Korean Mathematical Society
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    • v.52 no.2
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    • pp.649-659
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    • 2015
  • The space $BV^2_{\alpha}(I)$ of all the real functions defined on interval $I=[a,b]{\subset}\mathbb{R}$, which are of bounded second ${\alpha}$-variation (in the sense De la Vall$\acute{e}$ Poussin) on I forms a Banach space. In this space we define an operator of substitution H generated by a function $h:I{\times}\mathbb{R}{\rightarrow}\mathbb{R}$, and prove, in particular, that if H maps $BV^2_{\alpha}(I)$ into itself and is globally Lipschitz or uniformly continuous, then h is an affine function with respect to the second variable.

Extracellular Production of Alpha-Interferon by Recombinant Escherichia coli : Part I. Construction of Expression Vectors (유전자 재조합 대장균을 사용한 Alpha-interferon의 생산과 분비: 제 1 부. 발현벡터의 제작)

  • 노갑수;최차용
    • KSBB Journal
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    • v.5 no.1
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    • pp.49-58
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    • 1990
  • We constructed hybrid plasmids to allow controlled and extracellular production of human alpha-interferon in Escherichia coli. The hybrid plassmids were constructed by transferring alpha-lFN gene from plasmid Hif-2h which has the alpha-lFN gene at PstI restriction site of pBR322, to plasmids pIN -IIIB3 and pIN-IIIC3 at restriction sites between HindIII and BamHI. Plasmids pIN-IIIB3 and pIN-IIIC3 carry E. coli lipoprotein promoter, lac promoter and operator in tandem. The plasmids also have lacl genes which encode for lac repressors, which allows controlled expression of genes cloned to the plasmids by using of inducer IPTG. Lipoprotein signal sequence is located just ahead of cloning sites of the plasmids, which helps cells to excrete or secrete cloned gene products. Plasmid pUC9 was used as a intermediate vector for transferring of alpha-lFN gene from Hif-2h to pIN vectors in order to solve the problem of different restriction sites between Hif-2h and pIN vectors.

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COMPLETE CONVERGENCE FOR ARRAY OF ROWWISE DEPENDENT RANDOM VARIABLES

  • Baek, Jong-Il;Park, Sung-Tae
    • Journal of applied mathematics & informatics
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    • v.27 no.3_4
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    • pp.829-842
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    • 2009
  • Let {$X_{ni}|1\;{\le}\;i\;{\le}\;n$, $n\;{\ge}\;1$} be an array of rowwise negatively associated random variables and let $\alpha$ > 1/2, 0 < p < 2 ${\alpha}p\;{\ge}\;1$. In this paper we discuss $n^{{\alpha}p-2}h(n)$ max $_{1\;{\le}\;k{\le}n}\;|\;{\sum}^k_{i=1}\;X_{ni}|/n^{\alpha}\;{\to}\;0$ completely as $n\;{\to}\;{\infty}$ under not necessarily identically distributed with a suitable conditions and h(x) > 0 is a slowly varying function as $x\;{\to}\;{\infty}$. In addition, we obtained that $n^{{\alpha}p-2}h(n)$ max $_{1\;{\le}\;k{\le}n}\;|\;{\sum}^k_{i=1}\;X_{ni}|/n^{\alpha}\;{\to}\;0$ completely as $n\;{\to}\;{\infty}$ if and only if $E|X_{11}|^ph(|X_{11}|^{1/\alpha})\;<\;{\infty}$ and $EX_{11}\;=\;0$ under identically distributed case and some corollaries are obtained.

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Search for extrasolar planets around K-giants: $\alpha$ Arietis - planet or surface features?

  • Lee, Byeong-Cheol;Mkrtichian, David E.;Han, In-Woo;Kim, Kang-Min;Park, Myeong-Gu
    • The Bulletin of The Korean Astronomical Society
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    • v.35 no.2
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    • pp.78.2-78.2
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    • 2010
  • We report the detection of a low-amplitude 380.8-day radial velocity (RV) variations in oscillating K2 III star ${\alpha}$ Ari (HD 12929). We do not found the correlation between RV variations and equivalent widths of chromospheric activity indicators ($H{\alpha}$ and CaII 8662 ${\AA}line$). The bisector analysis shows that bisector velocity span (BVS) and RV variations are not strongly correlated with each other. These result suggest that the RV variations could have been produced either by planetary companion or by the surface spots. If this RV variation is indeed caused by a planetary companion, an orbital solution with a period of P = 381 days, a semi-amplitude of K = 41 m/s, and an eccentricity of e = 0.25 fits the data best. Assuming a possible stellar mass of $M_{\bigstar} = 1.4-5.6 M\odot$, we estimate the minimum mass for the companion of m sini = 1.8-4.5 $M_{Jup}$ with an orbital semi-major axis of 1.2-1.9 AU. If confirmed, our finding gives a support to search for exoplanets around giant stars with multi-periodic oscillations.

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Purification of Recombinant Human Alpha-2a Interferon Without Using Monoclonal Antibodies

  • Kim, Dong Chung;Jin Jung
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.916-920
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    • 2002
  • This report describes a high-level expression of human alpha-2a interferon ($IFN{\alpha}-2a$) in Escherichia coli and its pilot scale purification by using a monoclonal antibody-independent chromatographic procedure that is based on anion-exchange, cation-exchange, hydrophobic interaction, and gel filtration. The recombinant E. coli produced much more $IFN{\alpha}-2a$ in a soluble form, when cultivated at low temperatures than at high-temperature fermentation. However, if the bacterial growth was taken into consideration, fermentation at $30^{\circ}C$ seemed optimal for the interferon production. By using our new protocol, we recovered approximately 160 mg of $IFN{\alpha}-2a$ with a specific activity of $3.59{\times}10^8$ IU/mg from 201 of the broth. The gel permeation chromatographic and SDS-PAGE indicated that the interferon preparation was purified to homogeneity and was of the correctly folded fast-migrating monomer.

Effect of Prostaglandin F2 Alpha on E-cadherin, N-cadherin and Cell Adhesion in Ovarian Luteal Theca Cells (난소의 황체협막세포에서 E-cadherin, N-cadherin과 세포부착에 미치는 Prostaglandin F2 Alpha의 영향)

  • Lee, Sang-Hee;Jung, Bae Dong;Lee, Seunghyung
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.360-369
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    • 2019
  • Cadherins are essential transmembrane proteins that promote cell-cell adhesion and maintain the corpus luteum structure in the ovary. This study examined the influence of prostaglandin F2 alpha ($PGF2{\alpha}$) on E-cadherin, N-cadherin, and adhesion in luteal theca cells (LTCs). The luteal cells were isolated from the mid-phase corpus luteum, and the LTCs were cultured separately from the luteal heterogeneous cells according to the morphology of the mesenchymal cells and to determine if steroidogenic and endothelial cells of LTCs, 3beta-hydroxysteroid dehydrogenase ($3{\beta}$-HSD), and vascular endothelial growth factor receptor 2 (VEGFR2) mRNA were used. The LTCs were then incubated in the culture medium supplemented with 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed $3{\beta}$-HSD mRNA expression in the LTC but no VEGF2R mRNA expression. The E-cadherin and N-cadherin proteins of the LTCs were damaged in the 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM $PGF2{\alpha}$ compared to the 0 mM $PGF2{\alpha}$ treatment groups (P<0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM $PGF2{\alpha}$ treatment group than in the 0 mM $PGF2{\alpha}$ treatment group (P<0.05). In conclusion, $PGF2{\alpha}$ affected the disruption of cadherin proteins and cell adhesion in LTCs. These results may help better understand the cadherin and adhesion mechanism during corpus luteum regression in the ovary.

Effects of Ojeoksangamibang on the Lipid Metabolism, Anti-oxidation and Concentration of Proinflammatory Cytokines in Rat Fed High Fat Diet (오적산가미방(五積散加味方)이 고지방식이 유도 비만쥐의 지질대사, 항산화계 및 전염증성 cytokine 생산에 미치는 영향)

  • Kong, In-Pyo;Park, Won-Hyung;Cha, Yun-Yeop
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.4
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    • pp.23-40
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    • 2011
  • Objectives: This study was designed to examine the effects of extracts of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) on the lipid lowering, anti-oxidation and concentration of proinflammatory cytokines and was investigated on hyperlipidemic rats. Methods: Male rats weighing $182.39{\pm}4.71g$ were fed high fat diet for 8 weeks and 36 rats(above 400 g) were divided into 4 groups. Each of 9 rats was divided a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats basal diet and administered an extract of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) extracts(100 mg/kg, 200mg/kg, 300 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflmmatory cytokines, anti-oxidative activity and $TNF-{\alpha}$, Apo-B, Apo-E and leptin gene expression. Results: 1. Concentration of plasma free fatty(FFA) showed no significant difference in all the treatment groups. Concentration of plasma triglyceride(TG) showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 2. Concentration of plasma total cholesterol showed a significant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma low density lipoprotein(LDL)-cholesterol showed a Significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma high density lipoprotein(HDL)-cholesterol showed a significant increment in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group. 3. Concentration of liver total cholesterol showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. Concentration of liver TG showed a significant decrement in all Ojeoksangamibang groups than that of control group. 4. Concentration of plasma and liver thiobarbituric acid reactive substance(TBARS) showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. 5. The values of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and catalase(CAT) activity showed a significant increment in all Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 6. The values of plasma aspartate aminotransferase(AST) and alanine aminotransferase(ALT) activity showed no significant different in all treatment group. 7. Concentration of plasma $interleukin(IL)-1{beta}$ showed no significant difference in all the treatment groups. Concentration of plasma IL-6 showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. Concentration of plasma tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ a siginifant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. However the concentration of plasma IL-10 in the 300 mg/kg Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant increment than that of control group. 9. In the analysis of reverse transcription-polymerase chain reaction(RT-PCR), gene expression of $TNF-{\alpha}$, Apo-B and Apo-E in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a lower expression than that of control group. However the gene expression of leptin showed no difference in the treatment groups. 10. The ratio of $TNF-{\alpha}$, Apo-B, and Apo-E per ${\beta}-actin$ expression in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant decrement than that of control group. However The ratio of leptin expression per ${\beta}-actin$ expression showed no significant difference among all the treatment groups. Conclusions: According to above results, in lowering lipid effect, anti-oxidation and control of pro-inflammatory cytokines production, Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) gives effect.