• Title/Summary/Keyword: Small protein

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Analysis of TIMP-2 and Vimentin Protein Expression and Epigenetic Reprogramming in Cloned Bovine Placentae

  • Kim, Hong-Rye;Han, Rong-Xun;Lee, Hye-Ran;Yoon, Jong-Taek;Cheong, Hee-Tae;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.97-102
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    • 2007
  • The objective of this study was to analyzed pattern of proteins expression abnormally in cloned bovine placenta. TIMP-2 protein whose function is related to extracellular matrix degradation and tissue remodeling processes was one of differentially up-regulated proteins in SCNT placenta. And one of down-regulated protein in SCNT placenta was identified as vimentin protein that is presumed to stabilize the architecture of the cytoplasm. The expression patterns of these proteins were validated by Western blotting. To evaluate how regulatory loci. of TIMP-2 and vimentin genes was programmed reprogramming in cloned placenta. the status of DNA methylation in the promoter region of TIMP-2 and vimentin genes was analyzed by sodium Bisulfite mapping. The DNA methylation results showed that there was not difference in methylation pattern of TIMP-2 and vimentin loci between cloned and normal placenta. Histone H3 acetylation state of the nucleosome was analyzed in the cloned placental and normal placenta by Western blotting. A small portion of the protein lysates were subjected to Western blotting with the antibodies against anti acetyl-Histone H3. Overall histone H3 acetylation state of SCNT placenta was significantly higher than those of normal placenta cells. It is postulated that cloned placenta at the end of gestation seems to be unusual in function and morphology of placenta via improper expression of TIMP-2 and vimentin by abnormal acetylation states of cloned genome.

Effects Protein Quality, Energy Restriction and Subsequent Rehabilitation on Growth in Young Rats (단백질 급원의 변화와 열량제한 및 식이회복이 흰쥐의 성장발달에 미치는 영향)

  • Kim, Wha-Young;Chung, Sook-Hyun
    • Journal of Nutrition and Health
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    • v.16 no.4
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    • pp.263-272
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    • 1983
  • Effects of dietary protein qualify, energy restriction, and subsequent rehabilitation were studied in the weanling rats. Rats were devided into the six experimental diet groups. Rats in AC (casein- containing diet ), AS (soyprotein-containing diet), and AG(gluten-containing diet) group consumed their diets ad libitum, and rats in PC, PS, PG groups were pair-fed in 60% of the of amounts eaten by corresponding rats in the AC, AS, AG groups, respectively. After 3 weeks of nutritional restriction, all rats were switched to the AC diet for 14 days. At the end of restriction period, body weight, weight gain, food intake, and FER were higher in casein- fed rats than other groups of rats. The differences were, however, disappeared with rehabilitation diet, except body weights which were not fully recovered until the end of experiment. The weights of liver, gastrocnemius muscle, kidney, small intestine, spleen, and lung were the highest in the casein group, and the lowest in gluten group at the end of restriction period. The recovery with rehabilitation diet were differ in different organs. The muscle and liver seemed to be the most affected organs by dietary protein quality and energy restriction. There were no differences in brain weights among the experimental groups during the restriction period, however, after 2 weeks of rehabilitation, rats in AC group showed lower brain weight compared to AC and AS groups, and the brain weight of PC group was lower than AC group. Protein contents in liver and muscle showed the same trends as their weights. Gluten-fed rats showed low serum protein concentration, but recovered fully with rehabilitation diet for 3 days.

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Full-length cDNA, Expression Pattern and Association Analysis of the Porcine FHL3 Gene

  • Zuo, Bo;Xiong, YuanZhu;Yang, Hua;Wang, Jun
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1473-1477
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    • 2007
  • Four-and-a-half LIM-only protein 3 (FHL3) is a member of the LIM protein superfamily and can participate in mediating protein-protein interaction by binding one another through their LIM domains. In this study, the 5'- and 3'- cDNA ends were characterized by RACE (Rapid Amplification of the cDNA Ends) methodology in combination with in silico cloning based on the partial cDNA sequence obtained. Bioinformatics analysis showed FHL3 protein contained four LIM domains and four LIM zinc-binding domains. In silico mapping assigned this gene to the gene cluster MTF1-INPP5B-SF3A3-FHL3-CGI-94 on pig chromosome 6 where several QTL affecting intramuscular fat and eye muscle area had previously been identified. Transcription of the FHL3 gene was detected in spleen, liver, kidney, small intestine, skeletal muscle, fat and stomach, with the greatest expression in skeletal muscle. The A/G polymorphism in exon II was significantly associated with birth weight, average daily gain before weaning, drip loss rate, water holding capacity and intramuscular fat in a Landrace-derived pig population. Together, the present study provided the useful information for further studies to determine the roles of FHL3 gene in the regulation of skeletal muscle cell growth and differentiation in pigs.

Targeted Efficacy of Dihydroartemisinin for Translationally Controlled Protein Expression in a Lung Cancer Model

  • Liu, Lian-Ke;Wu, Heng-Fang;Guo, Zhi-Rui;Chen, Xiang-Jian;Yang, Di;Shu, Yong-Qian;Zhang, Ji-Nan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2511-2515
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    • 2014
  • Objective: Lung cancer is one of the malignant tumors with greatest morbidity and mortality around the world. The keys to targeted therapy are discovery of lung cancer biomarkers to facilitate improvement of survival and quality of life for the patients with lung cancer. Translationally controlled tumor protein (TCTP) is one of the most overexpressed proteins in human lung cancer cells by comparison to the normal cells, suggesting that it might be a good biomarker for lung cancer. Materials and Methods: In the present study, the targeted efficacy of dihydroartemisinin (DHA) on TCTP expression in the A549 lung cancer cell model was explored. Results and Conclusions: DHA could inhibit A549 lung cancer cell proliferation, and simultaneously up-regulate the expression of TCTP mRNA, but down-regulate its protein expression in A549 cells. In addition, it promoted TCTP protein secretion. Therefore, TCTP might be used as a potential biomarker and therapeutic target for non-small cell lung cancers.

Euchromatin histone methyltransferase II (EHMT2) regulates the expression of ras-related GTP binding C (RRAGC) protein

  • Hwang, Supyong;Kim, Soyoung;Kim, Kyungkon;Yeom, Jeonghun;Park, Sojung;Kim, Inki
    • BMB Reports
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    • v.53 no.11
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    • pp.576-581
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    • 2020
  • Dimethylation of the histone H3 protein at lysine residue 9 (H3K9) is mediated by euchromatin histone methyltransferase II (EHMT2) and results in transcriptional repression of target genes. Recently, chemical inhibition of EHMT2 was shown to induce various physiological outcomes, including endoplasmic reticulum stress-associated genes transcription in cancer cells. To identify genes that are transcriptionally repressed by EHMT2 during apoptosis, and cell stress responses, we screened genes that are upregulated by BIX-01294, a chemical inhibitor of EHMT2. RNA sequencing analyses revealed 77 genes that were upregulated by BIX-01294 in all four hepatic cell carcinoma (HCC) cell lines. These included genes that have been implicated in apoptosis, the unfolded protein response (UPR), and others. Among these genes, the one encoding the stress-response protein Ras-related GTPase C (RRAGC) was upregulated in all BIX-01294-treated HCC cell lines. We confirmed the regulatory roles of EHMT2 in RRAGC expression in HCC cell lines using proteomic analyses, chromatin immune precipitation (ChIP) assay, and small guide RNA-mediated loss-of-function experiments. Upregulation of RRAGC was limited by the reactive oxygen species (ROS) scavenger N-acetyl cysteine (NAC), suggesting that ROS are involved in EHMT2-mediated transcriptional regulation of stress-response genes in HCC cells. Finally, combined treatment of cells with BIX-01294 and 5-Aza-cytidine induced greater upregulation of RRAGC protein expression. These findings suggest that EHMT2 suppresses expression of the RRAGC gene in a ROS-dependent manner and imply that EHMT2 is a key regulator of stress-responsive gene expression in liver cancer cells.

Molecular Analyses of the Metallothionein Gene Family in Rice (Oryza sativa L.)

  • Zhou, Gongke;Xu, Yufeng;Li, Ji;Yang, Lingyan;Liu, Jin-Yuan
    • BMB Reports
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    • v.39 no.5
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    • pp.595-606
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    • 2006
  • Metallothioneins are a group of low molecular mass and cysteine-rich metal-binding proteins, ubiquitously found in most living organisms. They play an important role in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting against intracellular oxidative damages. Analysis of complete rice genome sequences revealed eleven genes encoding putative metallothionein (OsMT), indicating that OsMTs constitute a small gene family in rice. Expression profiling revealed that each member of the OsMT gene family differs not only in sequence but also in their tissue expression patterns, suggesting that these isoforms may have different functions they perform in specific tissues. On the basis of OsMT structural and phylogenetic analysis, the OsMT family was classified as two classes and class I was subdivided into four types. Additionally, in this paper we also present a complete overview of this family, describing the gene structure, genome localization, upstream regulatory element, and exon/intron organization of each member in order to provide valuable insight into this OsMT gene family.

Food Protein-induced Enterocolitis Syndrome: an Update on Clinical Approaches and Its Pathophysiology (식품 단백질 유발성 장염 증후군: 임상적 접근과 병태생리의 최신 지견)

  • Hwang, Jin-Bok
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.10 no.2
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    • pp.117-128
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    • 2007
  • Food protein-induced enterocolitis syndrome (FPIES) is a non-IgE mediated hypersensitivity disorder, which is associated with mainly gastrointestinal symptoms and has a delayed onset. The vomiting and/or diarrheal symptoms of FPIES typically begin in the first month of life in association with a failure to thrive, metabolic acidosis, and shock. Therefore, the differential diagnosis of FPIES and neonatal or infantile sepsis-like illnesses or gastroenteritis is difficult. The early recognition of indexes of suspicion for FPIES may help in the diagnosis and treatment of this disorder. The diagnosis of FPIES is generally made through clinical practice and food-specific IgE test findings are typically negative in this condition. Therefore, oral cow's milk challenge (OCC) remains the valid diagnostic standard for FPIES. An investigation of positive OCC outcomes helps to find out a diagnostic algorithm of criteria of a positive challenge in FPIES. Moreover, it has not been clearly determined in infantile FPIES when $1^{st}$ follow up-oral food challenge (FU-OFC) should be performed, with what kind of food protein (e.g., cow's milk, soy), and how much protein should be administered. Hence, to prevent the risk of inappropriate FU-OFC or accidental exposure and achieve appropriate dietary management, it is necessary to identify tolerance rates to major foods under the careful follow up of infantile FPIES patients. On the other hand, small intestinal enteropathy with villous atrophy is observed in FPIES and this enteropathy seems to be in part induced by both of epithelial apoptosis and intercellular junctional complex breakdown. The purpose of this report is to introduce an update on diagnostic and therapeutic approaches in FPIES and suggest the possible histopathological evidences in this disorder.

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Comparison of the Efficiency of Absorbed Nitrogen Use from Different Protein Sources in Diets Having Similar Amino Acid Balance

  • Lee, K.U.;Boyd, R.D.;Austic, R.E.;Ross, D.A.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.6
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    • pp.725-731
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    • 1998
  • Nine crossbred female pigs fitted with the bladder catheters were used to investigate the effects of dietary protein form on the efficiency of absorbed nitrogen for nitrogen retention in growing pigs. Combinations of the main protein sources were corn-soybean meal (CSM; slow + slow absorption rate form), corn-hydrolyzed casein (CAS; slow + rapid absorption rate form) and corn-porcine plasma (CPL; slow + intermediate absorption rate form). All experimental diets were formulated to be isonitrogenous (CP 11%) and isocaloric (3.5 Mcal/kg) and synthetic amino acids were added to the diet as required to maintain an equivalent amino acid profile among diets. Fecal digestibility of nitrogen was not different among treatments (p > 0.10). Ingested nitrogen was absorbed with an apparent efficiency of 82% to 84%. Mean nitrogen retention in pigs fed the CSM diet was as high as for pigs fed the CPL diet (0.74 g N/kg $BW^{0.75}$ per d), which was higher than the N retention rate in pigs fed CAS diet (0.68 g/kg $BW^{0.75}$ per d; P < 0.05). Apparent biological values (ABV = 100 ${\times}$ N retention/absorbed nitrogen) were 63.3%, 58.0% and 61.6% for CSM, CAS, and CPL groups, respectively (p < 0.05). There was no difference in mean energy digestibility among treatments. The efficiency of absorbed lysine utilization was significantly different among treatments (p < 0.05). Pigs fed the CAS diet were inferior to counterparts on the other diets in utilizing absorbed lysine. The ratios of free (and small peptide-bound) to protein-bound amino acids in CSM diet differed considerably from the CAS diet. This may affect the efficiency of amino acids utilization for nitrogen retention if hydrolyzed and intact amino acid pools reach the blood at different times.

Involvement of GRP78 in the Resistance of Ovarian Carcinoma Cells to Paclitaxel

  • Zhang, Li-Ying;Li, Pei-Ling;Xu, Aili;Zhang, Xin-Chen
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3517-3522
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    • 2015
  • Background: Glucose regulated protein 78 (GRP78) is a type of molecular chaperone. It is a possible candidate protein that contributes to development of drug resistance. We first examined the involvement of GRP78 in chemotherapy-resistance in human ovarian cancer cell. Materials and Methods: The expression of GRP78 mRNA and protein were examined by RT-PCR and western blotting, respectively, in human ovarian cancer cells line (HO-8910). Sensitivity of HO-8910 to paclitaxel was determined with methyl thiazolyl tetrazolium (MTT). Suppression of GRP78 expression was performed using specific small-interfering RNA (siRNA) in HO-8910 cells, and cell apoptosis was assessed by flow cytometry. Statistical analysis was performed using the SPSS 15.0 statistical package. Results: HO-8910 cells, with high basal levels of GRP78, exhibited low sensitivity to paclitaxel. The mRNA and protein levels of GRP78 were dramatically decreased at 24h, 48h and 72h after transfection and the sensitivity to paclitaxel was increased when the GRP78 gene was disturbed by specific siRNA transfection. Conclusions: The results suggested that high GRP78 expression might be one of the molecular mechanisms causing resistance to paclitaxel, and therefore siRNA of GRP78 may be useful in tumor-specific gene therapy for ovarian cancer.

Effect of Protein and Protein Hydrolysate on Nitrogen Metabolism in Rats with Gastric Ulcer Induced by Restraint and Water-Immersion Stress (단백질과 단백질 가수분해물이 침수 속박 스트레스로 유도된 위 궤양 흰쥐의 질소대사에 미치는 영향)

  • 김창임
    • Journal of Nutrition and Health
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    • v.28 no.4
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    • pp.291-297
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    • 1995
  • This study aimed to verify the nutritional and curative effects of protein hydroysate in rats model with gastric ulcer induced by restraint and water-immersion stress. Sprague-Dawley, famale rats weighing approximtely 200g were forced in 5$\times$5$\times$15cm plexiglas cage. The restraint and water immersion stress was carried at 20$\pm$2$^{\circ}C$ for 8-hours. After stress 4 kinds of diets(10% casein, 20% casein, 10% casein hydrolysate, 20% casein hydrolysate) were given for 5 days. In the gastric ulcer rats model, the growth, gastric emptying rate, trypsin activity in gastrointestinal content, plasma total protein, albumin, $\alpha$-amino-N, UUN, creatinine and hydroxyproline of the urine and nitrogen retention were analyzed for nutritional effects of dietary nitrogen levels(10%, 20%) and sources (casein, casein hydrolysate). The results were as follows ; In gastric ulcer rats model, severeness of ulcer, plasma protein, gastric emptying rate, nitrogen retention rate were not different between 20% casein-fed group and 20% casein hydrolysatefed group. But 10% casein hydrolysate-fed group had more curative group. The casein hydrolysate diet-fed group was lower trysin activity in small intestianl content than the casein-fed group, at both casein level(10%, 20%). Finally at 20% levels, there was no difference between casein and casein hydrolysate diet, but 10% level, casein hydrolysate diet was more curative of ulcer than casein diet in gastric ulcer rat model. The results of this study provide useful information concerning diet therapy for the patients with gastrointestinal diseases and the field of enteral diet materials.

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