• Title/Summary/Keyword: Cellular metabolism

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Effects of Dietary Rice Bran Oil on Mitochondrial Respiration in M2-induced Bone Marrow-derived Macrophages (현미유가 생쥐의 골수로부터 M2로 유도한 대식세포의 미토콘드리아 호흡에 미치는 영향)

  • Lee, Sojung;Kim, Wooki
    • Food Engineering Progress
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    • v.22 no.4
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    • pp.353-357
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    • 2018
  • Previous studies have suggested that rice bran oil (RBO), an edible oil from the byproducts of rice milling, has anti-inflammatory effects in inflammation inducing macrophages, known as M1 subsets. Yet the effects of RBO on the counterpart M2 subsets, the "healing" macrophages, were poorly investigated to date. In this regard, recent studies on the molecular/cellular anti-inflammatory mechanisms of dietary components have demonstrated that mitochondrial respiration contributes to macrophage functioning. Therefore, the current study examined whether RBO regulates cytokine secretion by modulating mitochondrial metabolism in wound healing M2 subsets. Palm oil (PO), enriched with medium-chain fatty acids, served as a positive control. C57BL/6 mice were fed a diet containing either corn oil (CO), PO or RBO for 4 weeks, followed by purification of bone marrow-derived macrophages (BMDM) from their tibias and femurs. Cells were further polarized to M2-BMDM, and the expression of M2 marker (CD206) on cellular surfaces were not affected by dietary intervention. In addition, the secretion of anti-inflammatory cytokine (IL-10) in the culture supernatant was not affected by dietary lipids. Oxygen consumption rate, the indicator of mitochondrial respiration in M2-BMDM was not regulated by RBO intervention and PO treatment. Taken together, this study imply that RBO did not intervene both the regulation of inflammatory responses and mitochondrial respiration in M2 macrophages.

Effects of Gleditsiae Spina(GS) on Gene Expression of Human Breast Cancer Cells (조각자(皂角刺) 추출물이 인간 유래 유방암 세포의 유전자 발현에 미치는 영향)

  • Ban, Hye-Ran;Cho, Seong-Hee;Park, Kyung-Mi;Yang, Seung-Jeong
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.2
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    • pp.94-118
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    • 2009
  • Purpose: Gleditsiae spina (GS) has been used to treat patients with several diseases such as carbuncle, swelling and parasites. Recently GS is known to have anticancer activity in abdominal solid tumor, but the effects of GS on breast cancers is not clarified. For these reasons, we investigated effects of Gleditsiae spina (GS) on gene expression of human breast cancer cells. Methods: We investigated the effects of GS on proliferation of breast cancer cell line, MDA-MB-231. In addition, the genetic profile for the effect of GS on breast cancer cells was measured using microarray technique, and the functional analysis on these genes was conducted. Results: Total 1,434 genes were up-regulated and 2,483 genes down-regulated in the cells treated with GS. Genes induced or suppressed by GS were all mainly concerned with metabolic process, regulation of biological process and protein binding. The network of total protein interactions was measured using cytoscape program, and some key molecules that can be used for elucidation of therapeutical mechanism of medicine in future were identified. Conclusion: These results suggest possibility of GS as anti-cancer drug for breast cancer, and also suggest that related mechanisms are involved in regulation of intra-cellular metabolism in breast cancer cells.

Characterization of Rice Mutants with Enhanced Susceptibility to Rice Blast

  • Kim, Hye-Kyung;Lee, Sang-Kyu;Cho, Jung-Il;Lee, Sichul;An, Gynheung;Jwa, Nam-Soo;Kim, Byung-Ryun;Cho, Young-Chan;Han, Seong-Sook;Bhoo, Seong-Hee;Lee, Youn-Hyung;Hong, Yeon-Kyu;Yi, Gihwan;Park, Dae-Sup;Hahn, Tae-Ryong;Jeon, Jong-Seong
    • Molecules and Cells
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    • v.20 no.3
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    • pp.385-391
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    • 2005
  • As a first step towards identifying genes involving in the signal transduction pathways mediating rice blast resistance, we isolated 3 mutants lines that showed enhanced susceptibility to rice blast KJ105 (91-033) from a T-DNA insertion library of the japonica rice cultivar, Hwayeong. Since none of the susceptible phenotypes co-segregated with the T-DNA insertion we adapted a map-based cloning strategy to isolate the gene(s) responsible for the enhanced susceptibility of the Hwayeong mutants. A genetic mapping population was produced by crossing the resistant wild type Hwayeong with the susceptible cultivar, Nagdong. Chi-square analysis of the $F_2$ segregating population indicated that resistance in Hwayeong was controlled by a single major gene that we tentatively named Pi-hy. Randomly selected susceptible plants in the $F_2$ population were used to build an initial map of Pi-hy. The SSLP marker RM2265 on chromosome 2 was closely linked to resistance. High resolution mapping using 105 $F_2$ plants revealed that the resistance gene was tightly linked, or identical, to Pib, a resistance gene with a nucleotide binding sequence and leucine-rich repeats (NB-LRR) previously isolated. Sequence analysis of the Pib locus amplified from three susceptible mutants revealed lesions within this gene, demonstrating that the Pi-hy gene is Pib. The Pib mutations in 1D-22-10-13, 1D-54-16-8, and 1C-143-16-1 were, respectively, a missense mutation in the conserved NB domain 3, a nonsense mutation in the 5th LRR, and a nonsense mutation in the C terminus following the LRRs that causes a small deletion of the C terminus. These findings provide evidence that NB domain 3 and the C terminus are required for full activity of the plant R gene. They also suggest that alterations of the resistance gene can cause major differences in pathogen specificity by affecting interactions with an avirulence factor.

Identification of SNPs in Cellular Retinol Binding Protein 1 and Cellular Retinol Binding Protein 3 Genes and Their Associations with Laying Performance Traits in Erlang Mountainous Chicken

  • Wang, Yan;Xiao, Li-Hua;Zhao, Xiao-Ling;Liu, Yi-Ping;Zhu, Qing
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.8
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    • pp.1075-1081
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    • 2014
  • CRBP1 (cellular retinol binding protein 1) and CRBP3 (cellular retinol binding protein 3), are important components of the retinoid signaling pathway and take part in vitamin A absorption, transport and metabolism. Based on the role of vitamin A in chicken laying performance, we investigated the polymorphism of CRBP1 and CRBP3 genes in 349 chickens using single strand conformation polymorphism and DNA sequencing methods. Only one polymorphism was identified in the third intron of CRBP1, two polymorphisms were detected in CRBP3; they were located in the second intron and the third intron respectively. The association studies between these three SNPs and laying performance traits were performed in Erlang mountainous chicken. Notably, the SNP g.14604G>T of CRBP1 was shown to be significantly associated with body weight at first egg (BWFE), age at first egg (AFE), weight at first egg (WFE) and total number of eggs with 300 age (EN). The CRBP3 polymorphism g.934C>G was associated with AFE, and the g.1324A>G was associated with AFE and BWFE, but none of these polymorphisms were associated with egg quality traits. Haplotype combinations constructed on these two SNPs of CRBP3 gene were associated with BWFE and AFE. In particular, diplotype H2H2 had positive effect on AFE, BWFE, EN, and average egg-laying interval. We herein describe for the first time basic research on the polymorphism of chicken CRBP1 and CRBP3 genes that is predictive of genetic potential for laying performance in chicken.

Toxicogenomic Analysis and Identification of Estrogen Responsive Genes of Di (n-ethylhexyl) Phthalate in MCF-7 Cells

  • Kim, Youn-Jung;Yun, Hye-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.149-156
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    • 2005
  • Di (n-ethylhexyl) phthalate (DEHP) is thought to mimic estrogens in their action, and are called endocrine disrupting chemicals. DEHP is used in numerous consumer products, especially those made of flexible polyvinyl chloride and have been reported to be weakly estrogenic. In this study, DEHP were tested for estrogenic properties in vitro models and with microarray analysis. First, the E-screen assay was used to measure the proliferation of DEHP in MCF-7 cells, a human breast cancer cell line. DEHP induced an increase in MCF-7 cell proliferation at concentration of $10^{-4}M$. Second, we carried out a microarray analysis of MCF-7 cells treated with DEHP using human c-DNA microarray including 401 endocrine system related genes. Of the genes analyzed, 60 genes were identified showing significant changes in gene expression resulting from DEHP. Especially, 4 genes were repressed and 4 genes were induced by DEHP compared to $17{\beta}-estradiol$. Among these genes, trefoil factor 3 (intestinal), breast cancer 1, early onset and CYP1B1 are involved in estrogen metabolism and regulation. Therefore it suggests that these genes may be associated with estrogenic effect of the DEHP on transcriptional level. The rationale is that, as gene expression is a sensitive endpoint, alterations of these genes may act as useful biomarkers to define more precisely the nature and level of exposure to kinds of phthalates.

Screening of Differentially Expressed Genes by Desferrioxamine or Ferric Ammonium Citrate Treatment in HepG2 Cells

  • Park, Jong-Hwan;Lee, Hyun-Young;Roh, Soon-Chang;Kim, Hae-Yeong;Yang, Young-Mok
    • BMB Reports
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    • v.33 no.5
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    • pp.396-401
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    • 2000
  • A differential display method is used to identify novel genes whose expression is affected by treatment with ferric ammonium citrate (FAC) or desferrioxamine (DFO), an iron chelating agent in the human hepatoblastoma cell line (HepG2). These chemicals are known to deplete or increase the intracellular concentration of iron, respectively. Initially, we isolated seventeen genes whose expressions are down- or up regulated by the treatment of the chemicals, as well as their four differentially expressed genes that are designated as clone-1, -2, -3, and -4. These are further characterized by cDNA sequencing and Northern blot analysis. Through the cDNA sequencing, as well as comparing them to genes published using the NCBI BLAST program, we identified the sequence of the clone-1 that is up-regulated by the treatment of DFO. It is identical to the human insulin-like growth factor binding protein-1 (IGFBP-1). This suggests that the IGFBP-1 gene in the HepG2 cell is up-regulated by an iron depletion condition. Also, the expression of the clone-3 and -4 is up-regulated by FAC treatment and their eDNA sequences are identical to the human ferritin-fight chain and human NADH-dehydrogenase, respectively. However, the sequence of the clone-2 has no significant homology to any other known gene. Therefore, we suggest that changes of the cellular iron level in the HepG2 cell affects the transcription of cellular genes. This includes human IGFBP-1, ferritin-fight chain, and NADH-dehydrogenase. Regulation of these gene expressions may have an important role in cellular functions that are related to cellular iron metabolism.

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Choline-Lipid Release from Normal and Transformed Cells

  • Hong, Seong-Tshool;Jang, Yong-Suk;Park, Kie-In
    • BMB Reports
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    • v.29 no.1
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    • pp.73-80
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    • 1996
  • The effect of albumin on phosphatidylcholine (PC) metabolism in Hep-G2, 3T3-H.ras, and 3T3 cells pre-labelled with [Me-$^3H$]choline was studied. The [$^3H$]choline was more efficiently taken up and incorporated into cellular phospholipids in 3T3-H.ras cells than in Hep-G2 and 3T3 cells. In each of the three cell lines, most of the [$^3H$]choline metabolized into the phospholipids was incorporated into PC and only minor was incorporated into lysophosphatidylcholine (LPC). Bovine serum albumin stimulated the release of [$^3H$]LPC and [$^3H$]PC from each of the three cell lines pre-labelled with [$^3H$]choline. [$^3H$]PC was also released in the absence of albumin but [$^3H$]LPC was not. The efficiency of LPC secretion represented as the proportion of medium [$^3H$]LPC to cellular [$^3H$]choline lipid during a chase period is approximately 9 to 14 times greater in 3T3 cells compared with the transformed 3T3-H.ras and Hep-G2 cells. A similar comparison of published data for rat hepatocytes with Hep-G2 shows secretion to be 35~75 times greater from the rat hepatocytes than from Hep-G2. Also, PC secretion from 3T3 cells was 1.6 times more effective than from 3T3-H.ras, whereas rat hepatocytes secrete PC 2.8~3.8 times more effectively than does Hep-G2. The measurement of specific radioactivity of cellular PC in pre-labelled 3T3 cells showed it to be similar to that of the secreted PC. However, the specific radioactivity of secreted LPC was markedly lower than that of the cellular PC, which suggests that LPC is being secreted from a PC pool distinct from that used for PC secretion.

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Effect of Bevacizumab on Human Tenon's Fibroblasts Cultured from Primary and Recurrent Pterygium

  • Park, Young Min;Kim, Chi Dae;Lee, Jong Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.4
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    • pp.357-363
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    • 2015
  • The purpose of this study was to compare the inhibitory effect of bevacizumab on human Tenon's fibroblasts (HTFs) cultured from primary and recurrent pterygium. Cultured HTFs were exposed to 2.0, 5.0, 7.5, and 15.0 mg/mL concentration of bevacizumab for 24 hours. The 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase leakage assays were then performed to assess fibroblast metabolism and viability. The matrix metalloproteinase (MMP), procollagen type I C terminal propeptide (PIP), and laminin immunoassays were performed to examine extracellular matrix production. Changes in cellular morphology were examined by phase-contrast and transmission electron microscopy. Both metabolic activity and viability of primary and recurrent pterygium HTFs were inhibited by bevacizumab in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. Both types of HTFs had significant decreases in MMP-1, PIP, and laminin levels. Distinctly, the inhibitory effect of bevacizumab on MMP-1 level related with collagenase in primary pterygium HTFs was significantly higher than that of recurrent pterygium. Significant changes in cellular density and morphology both occurred at bevacizumab concentrations greater than 7.5 mg/mL. Only primary pterygium HTFs had a reduction in cellular density at a bevacizumab concentration of 5.0 mg/mL. Bevacizumab inhibits primary and recurrent pterygium HTFs in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. As the primary HTFs produces larger amounts of MMP-1 compared to recurrent HTFs, significant reduction in MMP-1 level in primary pterygium HTFs after exposure to bevacizumab is likely to be related to the faster cellular density changes in primary pterygium HTFs.

Peroxisome Proliferator-activated Receptor-γ Gene Polymorphisms are not associated with Osteonecrosis of the Femoral Head in the Korean Population

  • Kim, Tae-Ho;Hong, Jung Min;Park, Eui Kyun;Kim, Shin-Yoon
    • Molecules and Cells
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    • v.24 no.3
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    • pp.388-393
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    • 2007
  • Osteonecrosis of the femoral head (ONFH) is a multifactorial disease to which certain individuals are more at risk. Altered lipid metabolism is one of the major risk factors for osteonecrosis, especially corticosteroid therapy and alcoholism. Peroxisome Proliferator-Activated Receptor-${\gamma}$ ($PPAR{\gamma}$) plays a crucial role in differentiation of mesenchymal cells to adipocytes, lipid homeostasis, and bone metabolism. To investigate the possible association between $PPAR{\gamma}$ gene variants and susceptibility to ONFH, we genotyped three common polymorphisms (-796A > G, +34C > G[Pro12Ala], and +82466C > T[His477His]) in 448 ONFH patients and 336 control subjects. Genotypes, allele frequencies, and haplotypes of the polymorphisms in the complete set of patients as well as in subgroups by sex or etiology were not significantly different from those in the control group. This suggests that the examined polymorphisms and haplotypes of the $PPAR{\gamma}$ gene are unlikely to be associated with susceptibility to ONFH.

The Diversity of Lysine-Acetylated Proteins in Escherichia coli

  • Yu, Byung-Jo;Kim, Jung-Ae;Moon, Jeong-Hee;Ryu, Seong-Eon;Pan, Jae-Gu
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1529-1536
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    • 2008
  • Acetylation of lysine residues in proteins is a reversible and highly regulated posttranslational modification. However, it has not been systematically studied in prokaryotes. By affinity immunoseparation using an anti-acetyllysine antibody together with nano-HPLC/MS/MS, we identified 125 lysine-acetylated sites in 85 proteins among proteins derived from Escherichia coli. The lysine-acetylated proteins identified are involved in diverse cellular functions including protein synthesis, carbohydrate metabolism, the TCA cycle, nucleotide and amino acid metabolism, chaperones, and transcription. Interestingly, we found a higher level of acetylation during the stationary phase than in the exponential phase; proteins acetylated during the stationary phase were immediately deacetylated when the cells were transferred to fresh LB culture medium. These results demonstrate that lysine acetylation is abundant in E. coli and might be involved in modifying or regulating the activities of various enzymes involved in critical metabolic processes and the synthesis of building blocks in response to environmental changes.