• Title/Summary/Keyword: Glucose starvation

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Generation of Transgenic Mice Overexpression Mouse RESISTIN

  • J. R. Chun;S. J. Song;J. T. Do;K. S. Chung;Lee, H. T.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.99-99
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    • 2002
  • The hormone resistin is associated with typeII diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downregulates a resistin which is induced during adipocyte differentiation. But the connection between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin cDNA and to generate transgenic mice overexpressing mouse resistin gene. The 555 bp of mouse resistin was amplified from mob cDNAS by polymerase chain reaction (PCR) and cloned into pCR$\^$(R)/ 2.1 TOPO T-vector. Mouse resistin mRNA on the basis of Genbank sequence (acession no. AF323080). Then, the PCR product was cloned into pTargeT$\^$TM/ mammalian expression vector that has pCMV promoter and chimeric intron. Restriction enzyme analysis with BamH I and Not I was carried out to determine an orientation of the insert in the vector. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$\^$TM/-mus/resistin by digestion of Bgl II, and has used for microinjection into pronuclei of one cell embryos. The microinjected embryos were transfered to pseudopregnant foster-mother. Mouse resistin expression was detected in transgenic F1 mice by Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR). Resistin gene expression mouse has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, the abdominal fat pads were isolated from each 24h starvation and re-feeding after fasting group mice that were assessed by RT-PCR analysis. In fasting group mice, resistin expression was higher than that of re-feeding group mice. This result suggests that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes mellitus caused by insulin resistance of resistin.

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Stem Cell Properties of Gastric Cancer Stem-Like Cells under Stress Conditions Are Regulated via the c-Fos/UCH-L3/β-Catenin Axis

  • Jae Hyeong Lee;Sang-Ah Park;Il-Geun Park;Bo Kyung Yoon;Jung-Shin Lee;Ji Min Lee
    • Molecules and Cells
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    • v.46 no.8
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    • pp.476-485
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    • 2023
  • Gastric cancer stem-like cells (GCSCs) possess stem cell properties, such as self-renewal and tumorigenicity, which are known to induce high chemoresistance and metastasis. These characteristics of GCSCs are further enhanced by autophagy, worsening the prognosis of patients. Currently, the mechanisms involved in the induction of stemness in GCSCs during autophagy remain unclear. In this study, we compared the cellular responses of GCSCs with those of gastric cancer intestinal cells (GCICs) whose stemness is not induced by autophagy. In response to glucose starvation, the levels of β-catenin and stemness-related genes were upregulated in GCSCs, while the levels of β-catenin declined in GCICs. The pattern of deubiquitinase ubiquitin C-terminal hydrolase-L3 (UCH-L3) expression in GCSCs and GCICs was similar to that of β-catenin expression depending on glucose deprivation. We also observed that inhibition of UCH-L3 activity reduced β-catenin protein levels. The interaction between UCH-L3 and β-catenin proteins was confirmed, and it reduced the ubiquitination of β-catenin. Our results suggest that UCH-L3 induces the stabilization of β-catenin, which is required to promote stemness during autophagy activation. Also, UCH-L3 expression was regulated by c-Fos, and the levels of c-Fos increased in response to autophagy activation. In summary, our findings suggest that the inhibition of UCH-L3 during nutrient deprivation could suppress stress resistance of GCSCs and increase the survival rates of gastric cancer patients.

Effect of Fasting and Refeeding on Growth and Blood Chemistry in Juvenile Olive Flounder Paralichthys olivaceus L.

  • Cho, Sung-Hwoan
    • Journal of Aquaculture
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    • v.22 no.1
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    • pp.11-15
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    • 2009
  • Effect of fasting and refeeding on growth and blood chemistry of juvenile olive flounder Paralichthys olivaceus L. was investigated when fish achieved compensatory growth. Fish were fed the experimental diet for 6 days a week. Five treatments in triplicate were prepared: C, S1, S2, S3 and S4. Fish in the control group (C) were hand-fed to apparent satiation twice a day. Fish in treatments of S1, S2, S3 and S4 experienced 1, 2, 3 and 4 weeks of starvation and were then hand-fed to satiation twice daily during the remaining 7, 6, 5 and 4 weeks of the experiment, respectively. Weight gain of fish in C, S1 and S2 were higher than those of fish in S3 and S4. A significant difference in plasma total protein, glucose, triglyceride, $T_3$ and $T_4$ was observed in between starved and refed fish for the rest periods of the feeding trial. Plasma total protein and $T_3$ of flounder decreased with week of fasting and following correlationships were obtained; Y (Total protein) = -0.13X (week of fasting) + 1.54, $R^2=0.9792$ and $Y(T_3)=-11.48X$ (week of fasting) + 79.57, $R^2=0.8822$, respectively.

Phosphate and Carbon Source Regulation of Alkaline Phosphatase and Phospholipase in Vibrio vulnificus

  • Oh, Wan-Seok;Im, Young-Sun;Yeon, Kyu-Yong;Yoon, Young-Jun;Kim, Jung-Wan
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.311-317
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    • 2007
  • In this study, the effects of phosphate concentration and carbon source on the patterns of alkaline phosphatase (APase) and phospholipase (PLase) expression in Vibrio vulnificus ATCC 29307 were assessed under various conditions. The activities of these enzymes were repressed by excess phosphate (4 mM) in the culture medium, but this repression was reversed upon the onset of phosphate starvation in low phosphate defined medium (LPDM) containing 0.2 mM of phosphate at approximately the end of the exponential growth phase. The expressions of the two enzymes were also influenced by different carbon sources, including glucose, fructose, maltose, glycerol, and sodium acetate at different levels. The APase activity was derepressed most profoundly in LPDM containing fructose as a sole carbon source. However, the repression/derepression of the enzyme by phosphate was not observed in media containing glycerol or sodium acetate. In LPDM-glycerol or sodium acetate, the growth rate was quite low. The highest levels of PLase activity were detected in LPDM-sodium acetate, followed by LPDM-fructose. PLase was not fully repressed by high phosphate concentrations when sodium acetate was utilized as the sole carbon source. These results showed that multiple regulatory systems, including the phosphate regulon, may perform a function in the expression of both or either APase and PLC, in the broader context of the survival of V. vulnificus.

Aspartate-glutamate carrier 2 (citrin): a role in glucose and amino acid metabolism in the liver

  • Milan Holecek
    • BMB Reports
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    • v.56 no.7
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    • pp.385-391
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    • 2023
  • Aspartate-glutamate carrier 2 (AGC2, citrin) is a mitochondrial carrier expressed in the liver that transports aspartate from mitochondria into the cytosol in exchange for glutamate. The AGC2 is the main component of the malate-aspartate shuttle (MAS) that ensures indirect transport of NADH produced in the cytosol during glycolysis, lactate oxidation to pyruvate, and ethanol oxidation to acetaldehyde into mitochondria. Through MAS, AGC2 is necessary to maintain intracellular redox balance, mitochondrial respiration, and ATP synthesis. Through elevated cytosolic Ca2+ level, the AGC2 is stimulated by catecholamines and glucagon during starvation, exercise, and muscle wasting disorders. In these conditions, AGC2 increases aspartate input to the urea cycle, where aspartate is a source of one of two nitrogen atoms in the urea molecule (the other is ammonia), and a substrate for the synthesis of fumarate that is gradually converted to oxaloacetate, the starting substrate for gluconeogenesis. Furthermore, aspartate is a substrate for the synthesis of asparagine, nucleotides, and proteins. It is concluded that AGC2 plays a fundamental role in the compartmentalization of aspartate and glutamate metabolism and linkage of the reactions of MAS, glycolysis, gluconeogenesis, amino acid catabolism, urea cycle, protein synthesis, and cell proliferation. Targeting of AGC genes may represent a new therapeutic strategy to fight cancer.

Effect of Various Fat and Oil Types on Weight Gain and Liver Metabolism in Fasting or Normal Diet Rats (불포화도가 다른 유지가 단식과 정상식이 흰쥐의 체중 및 간대사에 미치는 영향)

  • 서화중;박현희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.1
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    • pp.19-26
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    • 1993
  • In this study rats in fasting or fed protein free restricted diet including only fat showed much lowered level of serum cholesterol and triglyceride accompanied by utmost weight loss and high level of blood urea nitrogen indicated the tissue degradation, especially in liver with signs of damage or necrosis of hepatic parenchymal cell leading to elevated glutamic pyruvate transaminase value and to death. Rats fed only perilla oil in starvation or as fat source in normal diet dropped down the level of serum cholesterol and triglyceride compared to beef tallow fed rat. But with evidence of glutamic pyruvate transaminase values which was significantly elevated long term ingestion of perilla oil is likely to cause the lesion or any damage of hepatic function.

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Effect of Osmotic Pressure on hCTLA-lg Production in Transgenic Rice Cell Suspension Cultures (형질전환된 벼세포 배양에 있어서 삼투압 조절에 따른 hCTLA4-lg 생산성 변화)

  • Choi Sung-Hun;Lee Song-Jae;Hong Seok-Mi;Cho Ji-Suk;Kim Dong-Il
    • KSBB Journal
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    • v.20 no.4
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    • pp.278-284
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    • 2005
  • An immunosuppressive agent, human cytotoxic T lymphocyte antigen 4 (hCTLA4), is used for the prevention of graft rejection and treatment of autoimmune diseases. hCTLA4-Ig, a CTLA4-immunoglobulin fusion protein, was produced and secreted from transgenic rice cell suspension cultures using rice a-amylase (RAmy3D) expression system. In this system, hCTLA4-Ig expression was regulated metabolically by sugar starvation. For the purpose of improving hCTLA4-Ig production, the effects of osmotic pressure was investigated in suspension cultures of transgenic rice cells. The highest production level was achieved at 40 mM sorbitol $(140\;mOsm{\cdot}kg^{-1}\;H_2O)$. Using the medium with 8 mM glucose, the level of hCTLA4-Ig in the medium reached 45.3 mg/L. By adjusting the osmotic pressure of induction medium, it was found that the hCTLA4-Ig production could be increased up to 2.1-fold compared with that in batch culture.

Carbon Source-Dependent Regulation of the Schizosaccharomyces pombe pbh1 Gene

  • Kim, Su-Jung;Cho, Nam-Chul;Ryu, In-Wang;Kim, Kyung-Hoon;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.689-693
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    • 2006
  • Pbh1, from the fission yeast Schizosaccharomyces pombe, is a baculoviral inhibitor of apoptosis (IAP) repeat (BIR) domain-containing protein. Its unique encoding gene was previously found to be regulated by nitric oxide and nitrogen starvation. In the current work, the Pbh1-lacZ fusion gene was used to elucidate the transcriptional regulation of the pbh1 gene under various carbon sources. When fermentable carbon sources, such as glucose (at a low concentration of 0.2 %), sucrose (2.0 %) and lactose (2.0 %), were the sole carbon source, the synthesis of $\beta$-galactosidase from the Pbh1-lacZ fusion gene was reasonably enhanced. However, the induction by these fermentable carbon sources was abolished in the Pap1-negative S. pombe cells, implying that this type of induction of the pbh1 gene is mediated by Pap1. Ethanol (2.0%), a nonfermentable carbon source, was also able to enhance the synthesis of $\beta$-galactosidase from the fusion gene in wild-type cells but not in Pap1-negative cells. The results indicate that the S. pombe pbh1 gene is up-regulated under metabolic oxidative stress in a Pap1-dependent manner.

Studies on the effects of PUERARIAE RADIX(葛根) and ARTEMISIAE HERBA(茵蔯) on experimental liver damages induced by alcohol, d-galactosamine and $CCl_4$ (인진(茵蔯)과 갈근(葛根)이 d-galactosamine, 급성(急性) alcohol중독(中毒) 및 $CCl_4$중독(中毒) 백서(白鼠)의 간손상(肝損傷)에 미치는 영향(影響))

  • Woo, Hong-Jung;Lee, Jang-Hoon;Kim, Young-Chul
    • The Journal of Korean Medicine
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    • v.18 no.1
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    • pp.411-429
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    • 1997
  • Recent survey shows that chronic liver disease such as chronic hepatitis, liver cirrhosis and hepatoma is the third leading causes for death in Korea. In oriental medicine, viral hepatitis is related to Hwangdal(黃疸) and alcoholic liver disease is related to Joosang(酒傷). ARTEMISIAE HERBA and PUERARIAE RADIX have long been used in treating those symptoms. This study was done to evaluate the effect of AR1EMISIAE HERBA and PUERARIAE RADIX on viral and alcoholic hepatitis. ARTEMISIAE HERBA and PUERARIAE RADIX were decocted respectively with water and followed by vaccum evaporation. The solution was diluted to adequate concentration. Sprague-Dawley rats were used in this experiment. Each group was given PUERARIAE RADIX or ARTEMISIAE HERBA solution orally and CCl4, d-galactosamine or alcohol was given orally 30 minutes later. After 24 hours of starvation, blood samples were taken to check serum GOT, GPT, LDH and ALP activities, TC, TG, glucose and BUN levels. These results show that ARTEMISIAE HERBA has better effect on liver injury induced by d-Galactosamine than PUERARIAE RADIX and that both ARTEMISIAE HERBA and PUERARlAE RADIX have good effect on acute alcoholic liver disease while in the liver injury induced by $CCl_{4}$, PUERARIAE RADIX has better inhibitory effect on serum AST, ALT and ALP levels and ARTEMISIAE HERBA has better inhibitory effect on serum total cholesterol and triglyceride. And the result that high concentration group has better effect shows these effects are concentration-dependent. Further study on the mechanism of these herbs is still required.

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Cloning and Expression Analysis of a Chitinase Gene Crchi1 from the Mycoparasitic Fungus Clonostachys rosea (syn. Gliocladium roseum)

  • Gan, Zhongwei;Yang, Jinkui;Tao, Nan;Yu, Zefen;Zhang, Ke-Qin
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.422-430
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    • 2007
  • Clonostachys rosea (syn. Gliocladium roseum) is a well-known biocontrol agent and widely distributed around the world. In this study, an endochitinase gene Crchi1 was isolated from the mycoparasitic fungus C. rosea using the DNA walking strategy. The Crchi1 ORF is 1,746 bp long and interrupted by three introns. The cloned gene Crchi1 encodes 426 amino acid residues and shares a high degree of similarity with other chitinases from entomopathogenic and mycoparasitic fungi. Several putative binding sites for transcriptional regulation of Crchi1 in response to carbon (5'-SYGGRG-3') and nitrogen (5'-GATA-3') were identified in the upstream of Crchi1. Expression of Crchi1 gene in different carbon sources was analyzed using real-time PCR (RT-PCR). We found that the Crchi1 expression was suppressed by glucose but strongly stimulated by chitin or solubilized components of the cell wall from Rhizoctonia solani. Phylogenetic analysis of chitinases from entomopathogenic and mycoparasitic fungi suggests that these chitinases have probably evolved from a common ancestor.