• Title/Summary/Keyword: Protein phosphatase 4

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Involvement of Phospholipase D in Norepinephrine Uptake in PC12 Cells

  • Rhee, Jong-Joo;Oh, Sae-Ock;Kim, Young-Rae;Park, Jong-Il;Park, Seung-Kiel
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.287-293
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    • 2009
  • Phospholipase D (PLD) is an enzyme hydrolyzing phosphatidylcholine to phosphatidic acid (PA) and choline. We investigated the involvement of PLD1 in the uptake of norepinephrine (NE) in PC12 cells, pheochromocytoma cells. NE uptake was specific in PC12 cells because nomifensine, a specific blocker of NE transporter, blocked NE uptake. Inhibition of PLD function in PC12 cells by the treatment of butanol suppressed the NE uptake. In contrast, overexpression of PLD1 in PC12 cells increased NE uptake efficiently. These results suggest that PLD activity is involved in NE uptake. We explored the action mechanism of PLD in NE uptake. PA phosphatase inhibitor, propranolol, blocks the formation of PKC activator diacylglycerol from PA. Propranolol treatment to PC12 cells blocked dramatically the uptake of NE. Specific PKC inhibitors, GF109203X and Ro31-8220, blocked NE uptake. Taken together, we suggest for the first time that PLD1 activity is involved in NE uptake via the activation of PKC.

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Hepatoprotective Effects of Waxy Brown Rice Fermented with Agrocybe cylindracea

  • Lee, Jong-Suk;Lee, Kyung-A;Kim, Han-Sup;Kang, Sung-Moo;Lee, Yoon-Joo;Yoon, Jae-Don;Chung, Min-Wook;Han, Gi-Dong;Lee, Jae-Sung
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.238-243
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    • 2006
  • The methanol extract of waxy brown rice fermented with Agrocybe cylindracea was prepared. The extract was then freeze dried and fed to rats at the level of 0.5, 1.0, 2.0 g/kg body weight for 14 days, followed by the treatment with carbon tetrachloride for three consecutive days to induce hepatotoxicity. After sacrificing the rats, the enzyme activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GTP) in serum was determined. Biochemical analysis on serum for albumin, total protein, triglyceride, and total as well as HDL-cholesterol were carried out along with a histopathological study of liver tissues. Based on these data, we suggest that the waxy brown rice cultured with A. cylindracea may exert hepatoprotective activity against hepatotoxicity caused by chemicals such as carbon tetrachloride.

Relationship of Somatic Cell Count, Physical, Chemical and Enzymatic Properties to the Bacterial Standard Plate Count in Different Breeds of Dairy Goats

  • Ying, Chingwen;Yang, Cheng-Bin;Hsu, Jih-Tay
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.4
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    • pp.554-559
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    • 2004
  • The objective of the present study was to investigate the accuracy of mastitis diagnostic indicators for different dairy goat breeds. Biweekly milk samples were collected from individual half mammary gland of seven Saanen and seven Alpine dairy goats in the period of 40 to 120 days in milk. With threshold value set at 2.8 and 3.1 for Alpine and Saanen dairy goats, respectively, log (SPC) offered good sensitivity (0.89, 0.93), specificity (0.88, 0.95), positive predictive value (0.75, 0.85) and negative predictive value (0.95, 0.98) as a mastitis diagnostic tool. The correlations of log (SPC) with milk yield, log (SCC), ALP, LDH, $Na^{+}$, $K^{+}$ and EC were significant in Saanen dairy goats (p<0.05), with the highest correlation coefficient (0.653) existing between log (SPC) and log (SCC). The correlations of log (SPC) with milk yield, milk fat, milk protein, log (SCC), $Na^{+}$, $K^{+}$, EC were significant in Alpine dairy goats (p<0.05), with the highest correlation coefficient (0.416) existing between log (SPC) and log (SCC). There were different best-fit regression equations with different multiple diagnostic indicators for Saanen and Alpine dairy goats. In conclusion, different breeds of dairy goats may have to adapt different mastitis diagnostic parameters for a better diagnosis.

Immunomodulating Activity of the Exopolymer from Submerged Mycelial Culture of Phellinus pini

  • Jeong, Sang-Chul;Cho, Sung-Pill;Yang, Byung-Keun;Jeong, Yong-Tae;Ra, Kyung-Soo;Song, Chi-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.15-21
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    • 2004
  • The immunomodulating activities and chemical characteristics of a water-soluble exopolymer from submerged mycelial culture of Phellinus pini were studied. Anticomplementary activity of this polymer was found to be $73.2\%$, and its activation system occurred through both classical and alternative pathways, where the classical pathway was detected to be the major one by crossed immunoelectrophoresis. Nitric oxide (NO) release ability and acid phosphatase activity of macrophage were increased by 1.6-fold ($100{\mu}g/ml$) and 3.4-fold ($500{\mu}g/ml$), respectively, and splenocyte proliferation in mixed lymphocyte reaction (MLR) was also increased by 2.6-fold ($200{\mu}g/ml$), compared to the control. The molecular weight of this polymer, determined by HPLC, was under 5 kDa. Total sugar and protein contents were 89.7 and 10.3%, respectively. Both sugar and amino acid compositions of the exopolymer were also analyzed.

Inhibitory Activity of Edible Plant Extracts on Proliferation of Human Umbilical Vein Endothelial Cells (HUVECs)

  • Song, Myoung-Chong;Kim, Sung-Hoon;Kwak, Ho-Young;Yang, Hye-Joung;Bang, Myun-Ho;Chung, In-Sik;Lee, Youn-Hyung;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.249-253
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    • 2007
  • Thirteen edible plants previously reported to show inhibitory activities on farnesyl protein transferase (FPTase) and phosphatase of the regenerating liver-3 (PRL-3) were evaluated for inhibitory activity on the proliferation of human umbilical vein endothelial cells (HUVECs). Four plant extracts, Oenothera erythrosepala, Perilla frutescens, Panicum miliaceum, and Quercus acutissima, significantly inhibited the proliferation of HUVECs induced by the basic fibroblast growth factor (bFGF) without cytotoxicity at 100 ${\mu}g/mL$. Myristica fragrans, Rosmarinus officinalis, and Syringa patula also showed inhibitory activity on the proliferation with only mild cytotoxicity.

Effect of Cynandione A of Cynanchi Wilfordii Radix in RANKL and Lipopolysaccharide-induced on Osteoclastogeneis in RAW 264.7 Cells (백하수오(Cynanchi Wilfordii Radix)의 Cynandione A가 RAW 264.7 세포에서 RANKL과 LPS로 유도된 파골세포형성에 대한 영향)

  • Hwang, Joon-Ho;Yi, Mi-Ran;Kang, Chang-Hee;Bu, Hee-Jung
    • Korean Journal of Pharmacognosy
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    • v.46 no.4
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    • pp.295-302
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    • 2015
  • Cynanchi wilfordii Radix roots have been utilized as traditional medicine for variety of diseases including diabetes mellitus, aging progression and scavenging free radicals, enhancing immunity, reducing high serum cholesterol, and anti-tumor activity. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of cynandione A on osteoclast cells. Thus, we was isolated cynandione A from Cynanchi wilfordii Radix roots and evaluated the effect of cynandione A on receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation. We found that cynandione A significantly inhibited osteoclast differentiation stimulated-RANKL in RAW 264.7 cells. Cynandione A conspicuously inhibited the mRNA and protein expression of matrix metallopeptidase 9 (MMP-9), tartrate-resistant acid phosphatase (TRAP) in cynandione A treated with RANKL. Taken together, our results demonstrated that Cynanchi Wilfordii Radix may be useful treatment option of bone-related disease such as osteoporosis leads to fracture of bone and rheumatoid arthritis.

Effect of Family Type on the Nutrient Intake and Nutritional Status in Elderly Women (가족형태에 따른 여자 노인의 영양소 섭취 및 영양 상태에 대한 연구)

  • 이현숙;장문정
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.4
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    • pp.934-941
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    • 1999
  • This study was performed to investigate the effect of family type on the nutrient intake and nutritional status in elderly women. Blood samples and anthropometric data of elderly women aged from 60 to 78 years were collected in 1996, who visited a health examination center in Seoul to screen their health status. They were divided into two groups by their family type, one was extended family(extended family elderly, n=37) and the other was independently living(independently living, n=29). Nutrients intake was measured by interview using a simplified questionaire. Albumin, cholesterol, triglyceride(TG), hemoglobin (Hb), hematocrit, Ca, Mg, and alkaline phosphatase activity of serum were analyzed. Intakes of energy, protein, carbohydrate, Fe, vitamin A, niacin, thiamin, riboflavin, and vitamin C were not different between the two groups. However, fat and Ca intakes were higher(p<0.05) in extended family elderly than indepen dently living elderly. There are no significant difference in hematocrit value and serum concentrations of Hb, TG, and Ca. Serum Mg and cholesterol levels tended to be higher in extended family elderly. However, serum albumin level was lower(p<0.05) in extended family elderly. These results show that the indepen dently living elderly women had intakes of daily energy, fat and Ca compared to the extended family elderly. Therefore, the nutritional education for the independently living elderly will be focused on the health risk resulted from less intakes of Ca and total energy.

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4-Hydroxynonenal Promotes Growth and Angiogenesis of Breast Cancer Cells through HIF-1α Stabilization

  • Li, Yao-Ping;Tian, Fu-Guo;Shi, Peng-Cheng;Guo, Ling-Yun;Wu, Hai-Ming;Chen, Run-Qi;Xue, Jin-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10151-10156
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    • 2015
  • 4-Hydroxynonenal (4-HNE) is a stable end product of lipid peroxidation, which has been shown to play an important role in cell signal transduction, while increasing cell growth and differentiation. 4-HNE could inhibit phosphatase and tensin homolog (PTEN) activity in hepatocytes and increased levels have been found in human invasive breast cancer. Here we report that 4-HNE increased the cell growth of breast cancer cells as revealed by colony formation assay. Moreover, vascular endothelial growth factor (VEGF) expression was elevated, while protein levels of hypoxia inducible factor 1 alpha (HIF-$1{\alpha}$) were up-regulated. Sirtuin-3 (SIRT3), a major mitochondria NAD+-dependent deacetylase, is reported to destabilize HIF-$1{\alpha}$. Here, 4-HNE could inhibit the deacetylase activity of SIRT3 by thiol-specific modification. We further demonstrated that the regulation by 4-HNE of levels of HIF-$1{\alpha}$ and VEGF depends on SIRT3. Consistent with this, 4-HNE could not increase the cell growth in SIRT3 knockdown breast cancer cells. Additionally, 4-HNE promoted angiogenesis and invasion of breast cancer cells in a SIRT3-dependent manner. In conclusion, we propose that 4-HNE promotes growth, invasion and angiogenesis of breast cancer cells through the SIRT3-HIF-$1{\alpha}$-VEGF axis.

A Case of Glycogen Storage Disease Type Ia Confirmed by Biopsy and Enzyme Assay (제Ia형 당원병 1례 (Glycogen Storage Disease , Type Ia))

  • Meen Sang-Ae;Rho Kwang-Sik;Kim Pyung-Kil;Jeong Hyeon-Joo;Park Young-Nyeon;Kim Myung-Joon;Kim Ji-Hong
    • Childhood Kidney Diseases
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    • v.2 no.1
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    • pp.77-81
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    • 1998
  • The author exprienced a case of glycogen storage disease type Ia(GSD-I) in an 18-year-old male patient who was admitted to our hospital due to proteinuria and hypertension. he was suspected to have GSD when 12 years old because of his family history of short stature and hepatomegaly. On admission, physical examination revealed short stature, heparomegaly, and The diagnosis of GSD-I was confirmed by compatible liver biopsy finding and enzyme assay which erealeddeficiency of glcose-6-phosphatase if hepatocyte. Sympromatic treatment was done using antihypertensive drugs and allopurinol with diet control. The authors report a case of glycogen storage disease type Ia completely confirmed by typical clinical manifestation, pathologic findings of the liver and the kidney, and the result of enzyme assay which revealed deficiency of glucose-6-phosphatase in hepatocytes with brief review fo related literatures.

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Establishment of Human Embryonic Stem Cells Derived from Frozen-Thawed Blastocysts (동결-융해된 인간 배반포기 배 유래의 배아 간(幹) 세포 배양)

  • Kim, Eun-Young;Nam, Hwa-Kyung;Lee, Keum-Sil;Park, Sae-Young;Park, Eun-Mi;Yoon, Ji-Yeon;Heo, Young-Tae;Cho, Hyun-Jung;Park, Se-Pill;Chung, Kil-Saeng;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.1
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    • pp.33-40
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    • 2001
  • Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

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