• Title/Summary/Keyword: Small protein

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Oleoylethanolamide Exhibits GPR119-Dependent Inhibition of Osteoclast Function and GPR119-Independent Promotion of Osteoclast Apoptosis

  • Kim, Hyun-Ju;Lee, Dong-Kyo;Jin, Xian;Che, Xiangguo;Choi, Je-Yong
    • Molecules and Cells
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    • v.43 no.4
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    • pp.340-349
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    • 2020
  • Oleoylethanolamide (OEA), a bioactive lipid in bone, is known as an endogenous ligand for G protein-coupled receptor 119 (GPR119). Here, we explored the effects of OEA on osteoclast differentiation, function, and survival. While OEA inhibits osteoclast resorptive function by disrupting actin cytoskeleton, it does not affect receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. OEA attenuates osteoclast spreading, blocks actin ring formation, and eventually impairs bone resorption. Mechanistically, OEA inhibits Rac activation in response to macrophage colony-stimulating factor (M-CSF), but not RANKL. Furthermore, the OEA-mediated cytoskeletal disorganization is abrogated by GPR119 knockdown using small hairpin RNA (shRNA), indicating that GPR119 is pivotal for osteoclast cytoskeletal organization. In addition, OEA induces apoptosis in both control and GPR119 shRNA-transduced osteoclasts, suggesting that GPR119 is not required for osteoclast apoptosis. Collectively, our findings reveal that OEA has inhibitory effects on osteoclast function and survival of mature osteoclasts via GPR119-dependent and GPR119-independent pathways, respectively.

Chordoma - A Clinicopathologic Review of 4 Cases - (척삭종 - 4례의 임상 및 병리학적 검색 -)

  • Choi, Joon-Hyuk;Bae, Young-Kyung;Kim, Mi-Jin;Shin, Duk-Seop;Cho, Kil-Ho
    • Journal of Yeungnam Medical Science
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    • v.16 no.2
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    • pp.369-375
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    • 1999
  • Chordoma is a slow-growing malignant neoplasm arising from the remnants of the primitive notochord. It accounts for 1 to 4% of all malignant bone tumors. It occurs exclusively along the spinal axis. Authors experienced four cases of chordoma occurred in the sacrococcygeal region. There were two male and two female patients, with a mean age of 63.5 years(range, 57~75 years). Tissue was obtained by wide excision in two patients, by incisional biopsy in one patient and by needle biopsy in the other. Adjuvant radiation therapy was performed on all the patients after their biopsy. The mean diameter of the tumors was 7.6cm(range, 5.5 to 13.0cm). Grossly, tumor was multiobulated, soft and myxoid gelatinous mass. Microscopically, the tumor showed lobulated feature divided by fibrous septa within it. There were physaliphorous cells with vacuolated bubbly cytoplasm. And small uniform, round, and non-vacuolated tumor cells were also present. On immunohistochemical stain, all the cases were immunoreactive for cytokeratin, epithelial membrane antigen(EMA) and vimentin, respectively. One of the 4 cases was positive for S-100 protein. All the cases were negative for CEA.

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Purification and Characterization of HCV RNA-dependent RNA Polymerase from Korean Genotype 1b Isolate: Implications for Discovery of HCV Polymerase Inhibitors

  • Kim, Jeong-Min;Lee, Mi-Kyoung;Kim, Yong-Zu
    • Bulletin of the Korean Chemical Society
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    • v.26 no.2
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    • pp.285-291
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    • 2005
  • The nonstructural protein 5B (NS5B) of hepatitis C virus (HCV) is the viral RNA-dependent RNA polymerase (RdRp), which is the essential catalytic enzyme for the viral replication and is an appealing target for the development of new therapeutic agents against HCV infection. A small amount of serum from a single patient with hepatitis C was used to get the genome of a Korean HCV isolate. Sequence analysis of NS5B 1701 nucleotides showed the genotype of a Korean isolate to be subtype 1b. The soluble recombinant HCV NS5B polymerase lacking the C-terminal 24 amino acids was expressed and purified to homogeneity. With the highly purified NS5B protein, we established in vitro systems for RdRp activity to identify potential polymerase inhibitors. The rhodanine family compounds were found to be potent and specific inhibitors of NS5B from high throughput screening (HTS) assay utilizing the scintillation proximity assay (SPA) system. The binding mode of an inhibitor was analyzed by measuring various kinetic parameters. Lineweaver-Burk plots of the inhibitor suggested it binds not to the active site of NS5B polymerase, but to an allosteric site of the enzyme. The activity of NS5B in in vitro polymerase reactions with homopolymeric RNA requires interaction with multiple substrates that include a template/primer and ribonucleotide triphosphate. Steady-state kinetic parameter, such as Km, was determined for the ribonucleotide triphosphate. One of compounds found interacts directly with the viral polymerase and inhibits RNA synthesis in a manner noncompetitively with respect to UTP. Furthermore, we also investigated the ability of the compound to inhibit NS5B-directed viral RNA replication using the Huh7 cell-based HCV replicon system. The investigation is potentially very useful for the utility of such compounds as anti-hepatitic agents.

Biological, Physical and Cytological Properties of Pepper mottle virus-SNU1 and Its RT-PCR Detection

  • Han, Jung-Heon;Choi, Hong-Soo;Kim, Dong-Hwan;Lee, Hung-Rul;Kim, Byung-Dong
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.155-160
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    • 2006
  • A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

The Influence of Bcl-3 Expression on Cell Migration and Chemosensitivity of Gastric Cancer Cells via Regulating Hypoxia-Induced Protective Autophagy

  • Hu, Lin;Bai, Zhigang;Ma, Xuemei;Bai, Nan;Zhang, Zhongtao
    • Journal of Gastric Cancer
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    • v.20 no.1
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    • pp.95-105
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    • 2020
  • Purpose: Gastric cancer is a highly metastatic malignant tumor, often characterized by chemoresistance and high mortality. In the present study, we aimed to investigate the role of B-cell lymphoma 3 (Bcl-3) protein on cell migration and chemosensitivity of gastric cancer. Materials and Methods: The gastric cancer cell lines, AGS and NCI-N87, were used for the in vitro studies and the in vivo studies were performed using BALB/c nude mice. Western blotting, wound healing assay, Cell Counting Kit-8 assay, immunohistochemistry, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were used to evaluate the role of Bcl-3 in gastric cancer. Results: We found that the protein expression of hypoxia (HYP)-inducible factor-1α and Bcl-3 were markedly upregulated under hypoxic conditions in both AGS and NCI-N87 cells in a time-dependent manner. Interestingly, small interfering RNA-mediated knockdown of Bcl-3 expression affected the migration and chemosensitivity of the gastric cancer cells. AGS and NCI-N87 cells transfected with si-RNA-Bcl-3 (si-Bcl-3) showed significantly reduced migratory ability and increased chemosensitivity to oxaliplatin, 5-fluorouracil, and irinotecan. In addition, si-Bcl-3 restored the autophagy induced by HYP. Further, the protective role of si-Bcl-3 on the gastric cancer cells could be reversed by the autophagy inducer, rapamycin. Importantly, the in vivo xenograft tumor experiments showed similar results. Conclusions: Our present study reveals that Bcl-3 knockdown inhibits cell migration and chemoresistance of gastric cancer cells through restoring HYP-induced autophagy.

Molecular Cloning, Tissue Distribution and Expression of Porcine y+L Amino Acid Transporter-1

  • Zhi, Ai-min;Zhou, Xiang-yan;Zuo, Jian-jun;Zou, Shi-geng;Huang, Zhi-yi;Wang, Xiao-lan;Tao, Lin;Feng, Ding-yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.272-278
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    • 2010
  • In this study, we cloned, sequenced and characterized porcine y+L Amino Acid Transporter-1 (y+LAT1). By screening a translated EST database with the protein sequence of the human $y^{+}$LAT1 and by using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $y^{+}$LAT1 was isolated from porcine intestine RNA. It was 2,111 bp long, encoding a 511 amino acid trans-membrane glycoprotein composed of 12 transmembrane domains. The predicted amino acid sequence was found to be 91%, 90%, 87% and 87% identical to those of cattle, human, mouse and rat $y^{+}$LAT1 respectively. Real-time RT-PCR results indicated that the small intestine had the highest $y^{+}$LAT1 mRNA abundance and the lung had the lowest $y^{+}$LAT1 mRNA abundance. Baby hamster kidney (BHK) cells transfected with green fluorescent protein (GFP) tagged porcine $y^{+}$LAT1 cDNA indicated that the cellular localization of the gene product in BHK was on the plasma membrane.

Fodder Productivity and Growth Persistency of Three Local Cassava Varieties

  • Tung, C.M.;Liang, J.B.;Tan, S.L.;Ong, H.K.;Jelan, Z.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.9
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    • pp.1253-1259
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    • 2001
  • Three cassava varieties, namely MM 92 (MM), Black Twig (BT) and Local (LC), were arranged in a randomized complete block design to evaluate their dry fodder and crude protein (CP) productivity as well as growth persistency. Cassava plants grown in small plots of $5m{\times}10m$ at a planting distance of $25cm{\times}25cm$ were harvested every 6 weeks starting from 3 months after planting. Dry fodder yields of MM, BT and LC over the 8 harvests were 8.55, 8.01 and 6.15 t/ha, respectively. All varieties produced more leaves than stems with average leaf:stem ratios of 5, 5.9 and 4.8 for MM, BT and LC, respectively. In terms of CP production, MM was the highest yielder (272 kg/ha/harvest), followed by BT and LC (238 and 184 kg/ha/harvest, respectively). The total accumulative CP amounts over the 8 harvests were 2179, 1903 and 1474 kg/ha for MM, BT and LC, respectively. The mortality rates were 9.91, 14.01 and 13.98% for MM, BT and LC, respectively. Phosphorus content was more stable than potassium content during defoliation. MM, BT and LC had whole plant phosphorus contents of 0.41, 0.41 and 0.39%, respectively; whole plant potassium contents were 1.25, 1.38 and 1.20%.

Effects of different amylose to amylopectin ratios on rumen fermentation and development in fattening lambs

  • Zhao, Fangfang;Ren, Wen;Zhang, Aizhong;Jiang, Ning;Liu, Wen;Wang, Faming
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1611-1618
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    • 2018
  • Objective: The objective of this experiment was to examine the effects of different amylose/amylopectin ratios on rumen fermentation and development of fattening lambs. Methods: Forty-eight 7-day-old male Small-tailed Han sheep${\times}$Northeast fine wool sheep were randomly assigned to four treatments of dietary amylose/amylopectin ratios (0.12, 0.23, 0.24, and 0.48 in tapioca starch, corn starch, wheat starch and pea starch diets, respectively). Three lambs from each treatment were slaughtered at 21, 35, 56, and 77 days of age to determine the rumen fermentation and development. Results: Compared with tapioca starch diet, the pea starch diet significantly increased the concentration of ammonia nitrogen in the ruminal fluid of lambs but significantly decreased the bacterial protein content. At 56 and 77 d, the rumen propionate concentration tended to be greatest in the tapioca starch group than in other groups. The rumen butyrate concentration was the greatest in lambs fed on pea starch compared with those fed on other starch diets. Furthermore, the pea starch diet significantly stimulated rumen development by increasing the papillae height, width and surface area in the rumen ventral or dorsal locations in lambs. However, different amylose/amylopectin ratios diets did not significantly affect the feed intake, body weight, average daily gain, the relative weight and capacity of the rumen in lambs with increasing length of trial periods. Conclusion: Lambs early supplemented with a high amylose/amylopectin ratio diet had favourable morphological development of rumen epithelium, which was not conducive to bacterial protein synthesis.

Characteristics of Structure and Expression Pattern of ADSF/resistin Gene in Korean Native Cattle

  • Kang, Hye Kyeong;Park, Ji Ae;Seo, Kang Seok;Kim, Sang Hoon;Choi, Yun Jai;Moon, Yang Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.329-334
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    • 2006
  • Adipocyte-specific secretory factor (ADSF)/resistin, a hormone, is a small cysteine-rich protein secreted from adipose tissue and has been implicated in modulating adipogenesis in humans and rodents. The objective of this study was to clone a gene encoding ADSF/resistin and to characterize its function in Korean Native Cattle (Hanwoo). The coding sequence was 330 base pairs and it encoded a protein of 109 amino acids. An NCBI BLAST-search revealed the cloned cDNA fragment shared significant homology (82%) with the cDNA encoding the human ADSF/resistin. The nucleotide sequence homology of the Hanwoo sequence was 73% and 64% for the rat and mouse, respectively. A 654 bp ADSF/resistin gene promoter was cloned and putative binding sites of transcription factors were identified. Tissue distribution of ADSF mRNA was examined in liver, skeletal muscles (tenderloin, biceps femoris), subcutaneous fat, and perirenal fat by RT-PCR. ADSF mRNAs were detected in fat tissues but not in liver and muscles, suggesting that ADSF/resistin expression may be induced during adipogenesis. Although, the physiological function of ADSF/resistin in the cow remains to be determined, these data indicate ADSF is related to the adipocyte phenotype and may have a possibly regulatory role in adipocyte function.

Urban Particulate Matter-Induced Oxidative Damage Upon DNA, Protein, and Human Lung Epithelial Cell (A549): PM2.5 is More Damaging to the Biomolecules than PM10 Because of More Mobilized Transition Metals

  • Song, H-S;Chang, W-C;Bang, W-G;Kim, Y-S;Chung, N
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2002.10a
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    • pp.169-169
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    • 2002
  • The mobilizable amount of transition metals is a fraction of the total amount of the metal from urban particulate matter. Although the fraction is small, some metals (Fe, Cu) are the major participants in a reaction that generates reactive oxygen species (ROS), which can damage various biomolecules. Damaging effects of the metals can be measured by the single strand breakage (SSB) of X174 RFI DNA or the carbonyl formation of protein. In another study, we have shown that more metals are mobilized by PM2.5 than by PM10 in general. DNA SSB of >20% for PM2.5 and >15% for PM10 was observed in the presence of chelator (EDTA or citrate)/reductant (ascorbate), compared to the control (<3%) only with the chelator. The carbonyl formation by both PMs was very similar in the presence of the chelator, regardless of the kind of proteins. Compared to the control in the absence of chelator/reductant, 3.3 times and 4.9 times more carbonyl formation for PM2.5 and PM10, respectively, was obtained with BSA in the presence of chelator/reductant, showing that PM10 induced 33% more damage than PM2.5. However, 4.8 times and 1.9 times more carbonyl formation for PM2.5 and PM10, respectively, was observed with lysozyme in the presence of chelator/reductant, showing that PM2.5 induced 250% more damage than PM10. Although different proteins showed different sensitivities toward ROS, all these results indicate that the degrees of the oxidation of or damage to the biomolecules by the mobilized metals were higher with PM2.5 than with PM10. Therefore, it is expected that more metals mobilized from PM2.5 than from PM10, more damage to the biomolecules by PM2.5 than by PM10. We suggest that when the toxicity of the dust particle is considered, the particle size as well as the mobilizable fraction of the metal should be considered in place of the total amounts.

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