• Title/Summary/Keyword: Protein phosphatase

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Effects of Scytosiphon lomentaria on osteoblastic proliferation and differentiation of MC3T3-E1 cells

  • Park, Mi Hwa;Kim, Seoyeon;Cheon, Jihyeon;Lee, Juyeong;Kim, Bo Kyung;Lee, Sang-Hyeon;Kong, Changsuk;Kim, Yuck Yong;Kim, Mihyang
    • Nutrition Research and Practice
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    • v.10 no.2
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    • pp.148-153
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    • 2016
  • BACKGROUND/OBJECTIVES: Bone formation and bone resorption continuously occur in bone tissue to prevent the accumulation of old bone, this being called bone remodeling. Osteoblasts especially play a crucial role in bone formation through the differentiation and proliferation. Therefore, in this study, we investigated the effects of Scytosiphon lomentaria extract (SLE) on osteoblastic proliferation and differentiation in MC3T3-E1 cells. MATERIALS/METHODS: A cell proliferation assay, alkaline phosphatase (ALP) activity assay, alizarin red staining and protein expression analysis of osteoblastic genes were carried out to assess the osteoblastic proliferation and differentiation. RESULTS: The results indicated that treatment of SLE promoted the proliferation of MC3T3-E1 cells and improved ALP activity. And, SLE treatment significantly promoted mineralized nodule formation compared with control. In addition, cells treated with SLE significantly upregulated protein expression of ALP, type 1 collagen, bone morphogenetic protein 2, runt-related transcription factor 2, osterix, and osteoprotegerin. CONCLUSIONS: The results demonstrate that SLE promote differentiation inducement and proliferation of osteoblasts and, therefore may help to elucidate the transcriptional mechanism of bone formation and possibly lead to the development of bone-forming drugs.

Effects of Anthropometric Indices and Nutrients Intake on Bone Mineral Density and Biochemical Markers of Bone Metabolism in Korean Premenopausal and Postmenopausal Women (폐경 전$\cdot$후 여성의 신체 계측치와 영양소 섭취량이 골밀도 및 골밀대사 지표에 미치는 영향)

  • Joo Eun-Jung
    • Journal of the East Asian Society of Dietary Life
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    • v.15 no.5
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    • pp.531-541
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    • 2005
  • This study investigated the influence of anthropometric data and nutrient intake on bone mineral density(BMD) and biochemical markers of bone metabolism The mean age of 21 premenopausal women were 47.0 years and that of 41 postmenopausal women whose menopausal age was 49.46 years were 60.56 years. The waist and WHR of postmenopausal women were significantly higher than those of premenopausal ones. The animal protein intake of premenopausal and postmenopausal women were 38.5 and 21.03 g which comprised 54.35 and $31.84\%$ of total protein intake, respectively. The calcium intake of premenopausal and postmenopausal women were 446.45 and 546.97mg which was 63.78 and $78.14\%$ of Korean RDA, respectively. The ALP(Alkaline phosphatase) of premenopausal women was 65.81 U/L, which was significantly lower than that(90.24 U/L) of postmenopausal women (p<0.01). BMD of lumbar spine of premenopausal women was correlated significantly with body weight(r=0.690, p<0.01), waist(r=0.682, p<0.01), WHR(r=0.672, p<0.01), BMI(r=0.559, p<0.01), and body fat(r=0.457, p<0.01). Urinary Ca/creatinine ratio of the premenopausal women was negatively correlated with plant protein(r=-0.529, p<0.05) and plant calcium(r=-0.579, p<0.05). BMD of lumbar spine of postmenopausal women showed positive correlation with lean body mass(r=0.469, p<0.01) and body weight(r=0.383, p<0.05). Urinary Ca/creatinine ratio for the postmenopausal women was positively correlated with ALP(r=0.404, p<0.01) and urinary Na/creatinine ratio(r=0.389, p<0.05). In conclusion, it is necessary to maintain adequate body weight and to increase calcium intake for the premenopausal women. It is also important to increase muscle mass and reduce salt intake for the postmenopausal women.

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Purification and biological activity of recombinant human bone morphogenetic protein-2 produced by E. coli expression system (E. coli 발현 시스템에 의해 생산된 recombinant human bone morphogenetic protein-2의 정제와 생물학적 활성)

  • Choi, Kyung-Hee;Moon, Keumok;Kim, Soo-Hong;Yun, Jeong-Ho;Jang, Kyung-Lib;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
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    • v.38 no.1
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    • pp.41-50
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    • 2008
  • Purpose: Bone morphogenetic protein-2(BMP-2) has been shown to possess significant osteoinducitve potential. There have been attempts to overcome a limitation of mass production, and economical efficiency of BMP. The aim of this study was to produce recombinant human BMP-2(rhBMP-2) from E. coli in a large scale and evaluate its biological activity. Materials and Methods: The E.coli strain BL21(DE3) was used as a host for rhBMP-2 production. Dimerized rhBMP-2 was purified by affinity chromatography using Heparin column. To determine the physicochemical properties of the rhBMP-2 expressed in E. coli, we examined the HPLC profile and performed Western blot analysis. The effect of the purified rhBMP-2 dimer on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and representing morphological change using C2C12 cell. Results: E. coli was genetically engineered to produce rhBMP-2 in a non-active aggregated form. We have established a method which involves refolding and purifying a folded rhBMP-2 dimer from non-active aggregates. The purified rhBMP-2 homodimer was characterized by SDS-PAGE as molecular weight of about 28kDa and eluted at 34% acetonitrile, 13.27 min(retention time) in the HPLC profile and detected at Western blot. The purified rhBMP-2 dimer stimulated ALP activity and induced the transformation from myogenic differentiation to osteogenic differentiation. Conclusion: rhBMP-2 was produced in E. coli using genetic engineering. The purified rhBMP-2 dimer stimulated ALP activity and induced the osteogenic differentiation of C2C12 cells.

Novel Genetic Associations Between Lung Cancer and Indoor Radon Exposure

  • Choi, Jung Ran;Koh, Sang-Baek;Park, Seong Yong;Kim, Hye Run;Lee, Hyojin;Kang, Dae Ryong
    • Journal of Cancer Prevention
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    • v.22 no.4
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    • pp.234-240
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    • 2017
  • Background: Lung cancer is the leading cause of cancer-related death worldwide, for which smoking is considered as the primary risk factor. The present study was conducted to determine whether genetic alterations induced by radon exposure are associated with the susceptible risk of lung cancer in never smokers. Methods: To accurately identify mutations within individual tumors, next generation sequencing was conduct for 19 pairs of lung cancer tissue. The associations of germline and somatic variations with radon exposure were visualized using OncoPrint and heatmap graphs. Bioinformatic analysis was performed using various tools. Results: Alterations in several genes were implicated in lung cancer resulting from exposure to radon indoors, namely those in epidermal growth factor receptor (EGFR), tumor protein p53 (TP53), NK2 homeobox 1 (NKX2.1), phosphatase and tensin homolog (PTEN), chromodomain helicase DNA binding protein 7 (CHD7), discoidin domain receptor tyrosine kinase 2 (DDR2), lysine methyltransferase 2C (MLL3), chromodomain helicase DNA binding protein 5 (CHD5), FAT atypical cadherin 1 (FAT1), and dual specificity phosphatase 27 (putative) (DUSP27). Conclusions: While these genes might regulate the carcinogenic pathways of radioactivity, further analysis is needed to determine whether the genes are indeed completely responsible for causing lung cancer in never smokers exposed to residential radon.

Regulation of Hepatic Gluconeogenesis by Nuclear Receptor Coactivator 6

  • Oh, Gyun-Sik;Kim, Si-Ryong;Lee, Eun-Sook;Yoon, Jin;Shin, Min-Kyung;Ryu, Hyeon Kyoung;Kim, Dong Seop;Kim, Seung-Whan
    • Molecules and Cells
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    • v.45 no.4
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    • pp.180-192
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    • 2022
  • Nuclear receptor coactivator 6 (NCOA6) is a transcriptional coactivator of nuclear receptors and other transcription factors. A general Ncoa6 knockout mouse was previously shown to be embryonic lethal, but we here generated liver-specific Ncoa6 knockout (Ncoa6 LKO) mice to investigate the metabolic function of NCOA6 in the liver. These Ncoa6 LKO mice exhibited similar blood glucose and insulin levels to wild type but showed improvements in glucose tolerance, insulin sensitivity, and pyruvate tolerance. The decrease in glucose production from pyruvate in these LKO mice was consistent with the abrogation of the fasting-stimulated induction of gluconeogenic genes, phosphoenolpyruvate carboxykinase 1 (Pck1) and glucose-6-phosphatase (G6pc). The forskolin-stimulated inductions of Pck1 and G6pc were also dramatically reduced in primary hepatocytes isolated from Ncoa6 LKO mice, whereas the expression levels of other gluconeogenic gene regulators, including cAMP response element binding protein (Creb), forkhead box protein O1 and peroxisome proliferator-activated receptor γ coactivator 1α, were unaltered in the LKO mouse livers. CREB phosphorylation via fasting or forskolin stimulation was normal in the livers and primary hepatocytes of the LKO mice. Notably, it was observed that CREB interacts with NCOA6. The transcriptional activity of CREB was found to be enhanced by NCOA6 in the context of Pck1 and G6pc promoters. NCOA6-dependent augmentation was abolished in cAMP response element (CRE) mutant promoters of the Pck1 and G6pc genes. Our present results suggest that NCOA6 regulates hepatic gluconeogenesis by modulating glucagon/cAMP-dependent gluconeogenic gene transcription through an interaction with CREB.

Comparison of Isozyme Patterns among Varieties of Ginseng. Panax spp. (인삼품종간 Isozyme pattern 비교)

  • Son, Eung-Ryong;Park, Won-Mok;Lee, Yong-Se;Ahn, Sang-Deug;Chun, Seong-Ryong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.4
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    • pp.350-355
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    • 1984
  • The present researches were carried out to classify the species of ginseng by electrophoretic methods with isozyme patterns of LAP. esterase, GOT, phosphatase, peroxidase and proteins. All variants of Korean and Japanese ginseng had identical band patterns of the investigated enzymes in roots as well as in seeds. However, American ginseng had different patterns from those of Korean or Japanese.

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Association of Killer Cell Ig-like Receptor (KIR) with an Adaptor Protein Shc

  • Cho, Hyun-Il;Chwae, Yong-Joon;Park, Sang-Myun;Kim, Jong-Sun
    • IMMUNE NETWORK
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    • v.6 no.2
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    • pp.67-75
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    • 2006
  • Background: Cytotoxic function of killer cells is inhibited by specific recognition of class I MHC molecules on target cells by inhibitory killer Ig-like receptors (KIR) expressed on NK cells and some cytotoxic T cells. The inhibitory effect of KIR is accomplished by recruitment of SH2-containing protein tyrosine phosphatase (SHP) to the phosphotyrosine residues in the cytoplasmic tail. Methods: By in vitro coprecipitation experiments and transfection analysis, we investigated the association of KIR with an adaptor protein Shc in Jurkat T cells. Results: The cytoplasmic tail of KIR appeared to associate with an adaptor protein Shc in Jurkat T celilysates. Similar in vitro experiments showed that phosphorylated KIR cytoplasmic tail bound SHP-1 and Shc in Jurkat T cell lysates. The association of KIR with Shc was further confirmed by transfection analysis in 293T cells. Interestingly, however, Shc appeared to be replaced by SHP-2 upon engagement of KIR in 293T cells. Conclusion: Our data indicate that KIR associate with an adaptor protein Shc in Jurkat T cells, and suggest that KIR might have an additional role which is mediated by this adaptor protein.

Some of the Food Color Additives Are Potent Inhibitors of Human Protein Tyrosine Phosphatases

  • Shrestha, Suja;Bhattarai, Bharat Raj;Lee, Keun-Hyeung;Cho, Hyeong-Jin
    • Bulletin of the Korean Chemical Society
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    • v.27 no.10
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    • pp.1567-1571
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    • 2006
  • Synthetic color additives approved for general food use are sixteen in European Union, seven in U. S. A. and twelve in Japan. Twelve food dyes were examined for their inhibitory potency against human protein tyrosine phosphatases (PTPases). Half of the food colorants inhibited PTPases significantly and three of them were potent inhibitors with low micromolar IC50 values. Also examined were the synthetic dyes structurally similar but not allowed in food. Some of them were potent inhibitors of PTPases. Considering the importance of PTPases in cellular signal transduction, inhibition of PTPases by food colorants might cause harmful effects in human health.

Relationship between Nutritionally-related Blood Metabolites and Gastrointestinal Parasites in Nguni Goats of South Africa

  • Gwaze, F. Rumosa;Chimonyo, M.;Dzama, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1190-1197
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    • 2010
  • The objective of the study was to determine the relationship between faecal egg counts and nutritionally-related blood metabolites in Nguni goats of South Africa. Body weights, body condition scores (BCS), FAMACHA scores, faecal and blood samples were collected from 96 Nguni castrates. Faecal samples were analysed using the modified McMaster technique for nematodes and the sedimentation method for trematodes. Blood was analysed for packed cell volume (PCV), glucose, cholesterol, total protein, albumin, urea and creatinine. Season had an effect on glucose, globulin, total protein, creatinine, PCV and faecal egg counts (FEC). Globulin, PCV, creatinine and FEC were significantly higher in the wet season compared to the dry season. A quadratic relationship existed between faecal egg count loads and BCS whilst negative linear relationships were observed between faecal egg counts and creatinine, albumin and cholesterol levels of Nguni goats.

Effect of Estrogen and Dietary Protein Level on Ca and Skeletal Metabolism in Ovariectomized Rats (난소절제쥐에서 Estrogen을 투여하였을 때 식이 단백질 수준이 Ca 및 골격 대사에 미치는 영향)

  • 김화영
    • Journal of Nutrition and Health
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    • v.28 no.4
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    • pp.298-308
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    • 1995
  • To investigate the effect of estrogen and dietary protein level on Ca metabolism, female rats were undergone ovariectomy or sham-operation. Ovariectomized rate were divided into either estrogen-or vehicle-treated groups. Each treatment group was again divided into 40%-casein(H) or 10%-casein(L) diet groups. All experimental diets contained 0.2% Ca, 0.4% P and fed to rats for 8 weeks. Apparant Ca absorption and Ca balance were not affected by dietary protein level and ovariectomy, however they were increased by estrogen injection and this effect was even higher in low protein groups. Urinary Ca excretion were higher in high protein groups. GFR was not affected by dietary protein level, ovariectomy, or by estrogen injection. Urinary protein excretion was higher in high protein groups, which implies that the kidney funtion was deteriorated by high protein diet, and this may account partly for the higher urinary Ca in high protein groups. Ovariectomy or estrogen treatment had no effect on urinary protein excretion. Urinary hydroxyproline was higher in ovariectomized rats and increased in high protein grous. Elevated value of ovarictomized rats was lowered by estrogen injection, especially in low protein group. Alkaline phosphatase tended to increase in ovariectomized groups and lowered with estrogen treatment, but this difference was not statistically significant. Serum PTH was not affected by ovariectomy and dietary protein level. Therefore the increased hydroxproline excretion does not seem to be attributed to PTH. Dietary protein level, ovariectomy and estrogen treatment did not affect the weights and components of femur, scapular, and 4th vertebra. Ash/wt ratio of femur was, however, lower in ovariectomized rats and increased with estrogen treatment. Therefore, among the bones studied, femur seemed to be the most vulnerable. The results of this study shows that estrogen treatment may alleviate or reduce bone loss in postmenopausal women somewhat, especially for those people with low protein diet.

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