• Animal cells and systems
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• v.2 no.2
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• pp.223-227
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• 1998

We isolated two soluble forms of glutamate dehydrogenase isoproteins, GDH I and GDH II, from bovine brain. The regulation of GDH I and GDH II by phosphorylation and dephosphorylation has been examined in various conditions. There were dose- and time- dependent activation of the GDH isoproteins when phosphorylated by cAMP-dependent protein kinase. The phosphorylated GDH had 1.1 mol of covalently bound phosphate/mol of subunit and a 2-fold increased specific activity. The phosphorylated amino acid was identified as serine. When treated with alkaline phosphatase, the activities of the phosphorylated GDH isoproteins were reduced in dose and time dependent manner and returned to those of unphosphorylated enzymes. There were no significant differences between GDH I and GDH II in their sensitivities to the action of phosphorylation and dephosphorylation demonstrating that the microenvironmental structures of the phosphorylation site in GDH isoproteins are similar to each other, These results results suggest that the inter-conversion between less active form of brain GDH isoproteins and more active form is regulated by phosphorylation through cAMP-dependent protein kineses.

• Nutrition Research and Practice
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• v.18 no.1
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• pp.19-32
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• 2024

BACKGROUND/OBJECTIVES: Atherosclerosis is associated with increased inflammation in the visceral adipose tissue (VAT). Vitamin D has been reported to modulate the inflammatory responses of stromal vascular cells (SVCs) and adipocytes in adipose tissue, but the role of vitamin D in atherosclerosis biology is unclear. This study examined the effects of in vitro 1,25-dihydroxyvitamin D₃ (1,25[OH]₂D₃) treatment on the inflammatory responses of SVCs and adipocytes from atherosclerotic mice. MATERIALS/METHODS: C57BL/6J (B6) mice were divided randomly into 2 groups and fed a 10% kcal fat control diet (control group, CON) or 41% kcal fat, 0.21% cholesterol (high fat + cholesterol, HFC) diet (obese group, OB), and B6.129S7-Ldlr^tm1Her/J (Ldlr^-/-) mice were fed a HFC diet (obese with atherosclerosis group, OBA) for 16 weeks. SVCs and adipocytes isolated from VAT were pre-incubated with 1,25(OH)₂D₃ for 24 h and stimulated with lipopolysaccarides for the next 24 h. Proinflammatory cytokine production by adipocytes and SVCs, the immune cell population in SVCs, and the expression of the genes involved in the inflammatory signaling pathway in SVCs were determined. RESULTS: The numbers of total macrophages and SVCs per mouse were higher in OB and OBA groups than the CON group. The in vitro 1,25(OH)₂D₃ treatment significantly reduced macrophages/SVCs (%) in the OBA group. Consistent with this change, the production of interleukin-6 and monocyte chemoattractant protein 1 (MCP-1) by SVCs from the OBA group was decreased by 1,25(OH)₂D₃ treatment. The 1,25(OH)₂D₃ treatment significantly reduced the toll-like receptor 4 and dual-specificity protein phosphatase 1 (also known as mitogen-activated protein kinase phosphatase 1) mRNA levels in SVCs and MCP-1 production by adipocytes from all 3 groups. CONCLUSIONS: These findings suggest that vitamin D can attribute to the inhibition of the inflammatory response in VAT from atherosclerotic mice by reducing proinflammatory cytokine production.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.239-245
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• 2011

This study was conducted to determine the effects of excess vitamin A on alkaline phosphatase (ALP) activity, contents of calcium-binding protein (CaBP), bone gla-protein (BGP) in culture medium and CaBP mRNA expression in chicken osteoblasts in vitro. Osteoblastic cells in the tibia from 1-day-old Arbor Acre broiler chickens were isolated using enzyme digestion. The subconfluenced cells were divided into eight treatments with six replicates in each treatment and cultured in a medium containing either vehicle or different levels of vitamin A (0, 0.2, 0.6, 1.0, 2.0, 5.0, 10.0 and $20.0\;{\mu}g$/ml), and the control received an equivalent volume of ethanol. The incubation lasted 48 h. The results showed that vitamin A down-regulated ALP activity in the culture medium as well as CaBP mRNA expression of osteoblasts in a linear dose-dependent manner (p = 0.124 and p<0.10, respectively), and suppressed the contents of BGP and CaBP in the culture medium in a quadratic dose-dependent manner (p<0.05 and p<0.10, respectively) with increasing addition of vitamin A. The addition of 0-$0.2\;{\mu}g$/ml vitamin A to the culture medium increased ALP activity, BGP and CaBP contents as well as CaBP mRNA expression compared with other groups, but positive effects of vitamin A tended to be suppressed when vitamin A was increased to $1.0\;{\mu}g$/ml, and adverse effects occurred when vitamin A was increased to 10.0-$20.0\;{\mu}g$/ml. These results implied that there was a threshold level of vitamin A inclusion beyond which inhibitory effects occurred, and the mechanism by which overdose of vitamin A reduced bone growth in chickens was probably reduced osteoblastic cell activity, and inhibited expression of CaBP mRNA and CaBP secretion.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.665-670
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• 2002

The expediency of replacing cost prohibitive and often inaccessible traditional protein supplements prompted the monitoring of hematological parameters was carried out in female goats at 0, 30, 60 and 90 days post feeding. Rumen environment was (3), respectively fed supplements containing either a leaf meal mixture (LMTM) of Leucaena leucocephala-Morus alba-Tectona grandis (2:1:1) or traditional protein supplements groundnut cake (GNC) or soybean meal (SBM) and wheat straw as basal diet. The periodic monitoring of hematological parameters was carried out in female goats at 0, 30, 60 and 90 days post feeding. Rumen environment was studied in bucks in a $3{\times}3$ switch over design. Rumen liquor was collected at 0, 2, 4, 6 and 8 h post feeding after 4 weeks of feeding. The goats fed on LMTM or GNC had similar dry matter intake (g/kg $W^{0.75}$), which was significantly (p<0.05) higher than SBM. Except for packed cell volume (PCV), none of the blood biochemical constituents (Hemoglobin, serum glucose, total protein, serum albumin (A) and globulin(G), A:G ratio, alkaline phosphatase, transaminases) varied significantly due to replacement of 50% dietary protein by LMTM throughout the experiment. GNC group had significantly higher level of PCV than other treatments. However, the level of serum total protein (p<0.01) tended to increase from 60th day onwards irrespective of dietary treatments. The average rumen pH was significantly higher (p<0.001) on SBM followed by LMTM and GNC, respectively. Total volatile fatty acid (TVFA) production was comparable in goats given LMTM or GNC supplements, the corresponding values were significantly different (p<0.001) when compared with SBM. The ammonical-N, total-N and TCA-precipitable-N (mg/100 ml SRL) did not differ significantly among dietary treatments. It may be concluded that supplementing wheat straw with LMTM based concentrate had no adverse effect on voluntary intake, blood biochemical profile and rumen fermentation pattern of the goats.

• Molecules and Cells
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• v.24 no.2
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• pp.224-231
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• 2007

$H_2O_2$, as an example of oxidative stress, induces cardiac myocyte apoptosis. Bcl-2 family proteins are key regulators of the apoptotic response while their functions can be regulated by post-translational modifications including phosphorylation, dimerization or proteolytic cleavage. In this study, we examined the role of various protein kinases in regulating total BAD protein levels in adult rat cardiac myocytes undergoing apoptosis. Stimulation with 0.1 mM $H_2O_2$, which induces apoptosis, resulted in a marked down-regulation of BAD protein, which is attributed to cleavage by caspases since it can be restored in the presence of a general caspase inhibitor. Inhibition of PKC, p38-MAPK, ERK1/2 and PI-3-K did not influence the reduced BAD protein levels observed after stimulation with $H_2O_2$. On the contrary, inhibition of PKA or specifically $PKC{\delta}$ resulted in up-regulation of BAD. Decreased caspase 3 activity was observed in $H_2O_2$ treated cells after inhibition of PKA or $PKC{\delta}$ whereas inhibition of PKA also resulted in improved cell survival. Furthermore, addition of okadaic acid to inhibit selected phosphatases resulted in enhanced BAD cleavage. These data suggest that, during oxidative stress-induced cardiac myocyte apoptosis, there is a caspase-dependent down-regulation of BAD protein, which seems to be regulated by coordinated action of PKA, $PKC{\delta}$ and phosphatases.

• Toxicological Research
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• v.33 no.3
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• pp.219-224
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• 2017

Lipin1 was identified as a phosphatidate phosphatase enzyme, and it plays a key role in lipid metabolism. Since free radicals contribute to metabolic diseases in the liver, this study investigated the effects of free radicals on the regulation of Lipin1 expression in Huh7 and AML12 cells. Hydrogen peroxide induced mRNA and protein expression of Lipin1 in Huh7 cells, which was assayed by quantitative RT-PCR and immunoblotting, respectively. Induction of Lipin1 by hydrogen peroxide was confirmed in AML12 cells. Hydrogen peroxide treatment significantly increased expression of sterol regulatory element-binding protein (SREBP)-2, but not SREBP-1. Moreover, nuclear translocation of SREBP-2 was detected after hydrogen peroxide treatment. Hydrogen peroxide-induced Lipin1 or SREBP-2 expression was significantly reduced by N-acetyl-$\small{L}$-cysteine treatment, indicating that reactive oxygen species (ROS) were implicated in Lipin1 expression. Next, we investigated whether the hypoxic environments that cause endogenous ROS production in mitochondria in metabolic diseases affect the expression of Lipin1. Exposure to hypoxia also increased Lipin1 expression. In contrast, pretreatment with antioxidants attenuated hypoxia-induced Lipin1 expression. Collectively, our results show that ROS activate SREBP-2, which induces Lipin1 expression.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.821-838
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• 1999

On the basis of the evidences that electrical stimulation could enhance proliferation and differentiation of bone cells and promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on human periodontal ligament cells and gingival fibroblasts in vitro, which also have important roles in regeneration of periodontium, and to evaluate the potential of clinical application of electrical stimulation. Human periodontal ligament cells and gingival fibroblasts were primarily cultured from the root surface of extracted premolar and the adjacent gingiva without periodontal diseases. In control group, the cells ($5{\times}10^4$ cells/ml)were incubated only in Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In test groups, electrical stimulation was given at the current intensity of $0.25{\mu]A$(test group 1), $1.0{\mu}A$(test group 2), and $2.5{\mu}A$(test group 3) for 12 hours to the same culture media with the control group. After 12 hour exposure of electrical stimulation, the cells were incubated for 2 and a half days(60 hours), and then each group of cells was analyzed for cell proliferation, protein level, and activity of alkaline phosphatase. The results were as follows ; 1. The Rate of cell proliferation of every test group increased significantly in both periodontal ligament cells and gingival fibroblasts, and in periodontal ligament cells, test group 3 showed significantly increased proliferation compared to the other test groups(p<0.05). 2. In the protein levels, neither periodontal ligament cell nor gingival fibroblast showed statistically significant differences between control and test groups. 3. The activity of alkaline phosphatase in periodontal ligament cells increased significantly in all test groups(p<0.05), but there were no significant differences between 3 test groups. In gingival fibroblasts, the activity of alkaline phosphatase increased significantly only in test group 3(p<0.05). From the above results, it is concluded that electrical stimulation may have beneficial effects on the regeneration of destructed periodontal tissue in regard of the stimulation of periodontal ligament cells and gingival fibroblasts as well as electrically stimulated bone formation that has been known, and that electrical stimulation may have the potential of clinical application.

• Journal of Nutrition and Health
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• v.4 no.4
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• pp.15-23
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• 1971

On the assumption that the supplementation of certain nutrients or foods to the rice diet (low protein, low fat, and low vitamins) may decrease, to some extent, the degree of suffering from abnormal environments, such as vibration, noises, gases, dusts, smog etc. a series of experiments were started. As the first report the nutrition under vibration was studied in this experiment. Sixty (60) young growing male rats weighing about 65 grams were used, grouping to five (5) groups, twelve (12) rats each group. They were fed on the following five (5) experimental diets: rice diet (basal diet), rice diet+casein, rice diet+vitamins, rice $diet+{\alpha}-tocopherol$, and rice diet+ginseng powder (see the tables 1 and 2) for the period of 14 weeks experiment. During the experiment period the half number of the rats of each group were exposed to the three (3) hours vibration every day. The protective effect of each diet against the vibration may be summarized as follows. 1. The growth of rice diet group was impaired significantly under vibration, However, those of other groups (protein-supplemented, vitamin-supplemented, ${\alpha}-tocopherol-supplemented$ and ginseng-supplemented groups) were impaired much less compared with rice diet group. 2. The feed efficiency of the rice diet group was decreased significantly under vibration. It is estimated that the biological availability of nutrients was impaired under this environment. On the other hand, the feed efficiencies of protein supplemented, of vitamin supplemented, and of ginseng supplemented groups were not decreased under vibration, statistically. 3. There is tendency that the food spillages of vibration groups are higher than those of non-vibration groups. Especially it seems true in the case of rice diet group. The food spillage may be, to some extent, related with mental nervousness of animals. From the point that the food spillage of ginseng supplemented group is significantly lower than those of other groups it is thought ginseng acts some good role in protecting nervous system from suffering from vibration. 4. In all groups except protein supplemented group, liver fat of vibration group tends to be higher than that of non-vibration group. 5. It shows that, in general, the serum alkaline phosphatase activity of the vibration group is significantly higher than that of the non-vibration group. It seems that there may be, to some extent, corelation between the amount of liver fat and serum alkaline phosphatase activity. 6. There is tendency that, in rice diet group, the organs of vibration group are smaller than those of non-vibration group, especially lung is so. It is thought that this may be due to the poor growth of whole body size in vibration group.

• Journal of Periodontal and Implant Science
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• v.24 no.2
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• pp.303-320
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• 1994

Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

• Journal of Nutrition and Health
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• v.31 no.4
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• pp.716-728
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• 1998

To investigate the effect of dietary Ca levels on metabolic changes of Ca and skeleton in postmenopausal women, 10-month-old ovariectomized female rats were compared with 2 month old rats. The rats were fed either 0.2% or 1.2% Ca diets for 16 weeks. Food intake and weight gain as higher in rats fed high Ca diets and in ovariectomized rats. Apparent Ca absorption as higher, and Ca balance was lower in the low Ca groups. Vertebrae density was higher in old rats or those fed a high Ca diets. The old rats and ovariectomized rats showed decreased bone formation, increased bone resorption and kidney function deterioration resulting in increased urinary Ca excretion. Contradictory to the above observation, old rats and ovariectomized rats still showed higher bone mass and bone ash content. Therefore aging was not fully onging in 10-month-old rats. Bone weights, mineral contents, and mineral/wt ratio were lower in ovariectomized rats. Dietary Ca level did not affect urinary Ca excretion, urinary protein excretion, GFR, serum alkaline phosphatase, or urinary hydroxyporline excretion. This means that dietary Ca level did not influence kidney function or bone turnover. However Ca content and the ash content of femur, 4th vertebra, and scapula were increased in high Ca groups. Therefore, it is considered that decreased bone formation and accelerated bone resorption may account for the increased osteoporotic risk in women in menopause after middle age. However, Ca metabolism can be improved and bone components can be maintained if Ca is supplemented.