• Journal of Chest Surgery
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• v.35 no.12
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• pp.847-853
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• 2002

We previously published the data that proved the safety of retrograde cerebral perfusion for 120 minutes. At this time, we planned to check the neuron-specific enolase and S100-$\beta$ in serum and urine to assess the possibility of early detection of cerebral injury. Material and Method: We used pigs(Landrace species) weighing 35 kg and performed RCP for 120 minutes. After the weaning of cardiopulmonary bypass, we observed the pigs for another 120 minutes. Systemic arterial pressure, central venous pressure, and serum and urine levels of neuron-specific enolose (NSE) and S100$\beta$ protein were checked. Central venous pressure during RCP was maintained in the range of 20 to 25 mmHg. Result: Serum levels of NSE(ng/$m\ell$) were 0.67$\pm$0.18(induction of anesthesia), 0.53$\pm$0.47(soon after CPB), 0.44$\pm$0.27(20min alter CPB), 0.24$\pm$0.09(RCP 20min), 0.37$\pm$0.35(RCP 40min), 0.33$\pm$0.21 (RCP 60min), 0.37$\pm$0.22(RCP 80min), 0.41$\pm$0.23(RCP 100 min), 0.48$\pm$0.26(RCP 120min), 0.42$\pm$0.29(30min after rewarming), 0.35 $\pm$0.32(60min after rewarming, 0.42$\pm$0.37(CPBoff 30min), 0.47$\pm$0.34(CPBOff 60min), 0.47$\pm$0.28(CPBOff 90min), and 0.57$\pm$0.29(CPBOff 120min). There was no statistically significant difference in levels between before and after RCP(ANOVA, p＞0.05). Urine levels of NSE also showed no statistically significant difference in levels between before and after RCP. There was no correlation between urine and serum levels of NSE(Pearson correlation, p＞0.05). Serum levels of S100$\beta$ protein(ng/$m\ell$) during the same time frames were 0.14$\pm$0.08, 0.15$\pm$0.07, 0.22$\pm$0.15, 0.23$\pm$0.07, 0.28$\pm$0.10, 0.40$\pm$0.05, 0.47$\pm$0.03, 0.49$\pm$0.12, 0.43$\pm$0.11, 0.46$\pm$0.15, 0.62$\pm$0.17, 0.77$\pm$0.21, 0.78$\pm$0.23, 0.77$\pm$0.23, and 0.82$\pm$0.33. There was statistically significant difference in levels between before and after RCP(ANOVA, p＜0.05). Urine levels of NSE also showed statistically significant difference in levels between before and after RCP(ANOVA, p＜0.05). There was significant correlation between urine and serum levels of NSE(Pearson correlation, p＜0.05). Conclusion: The author observed the increase in serum and urine levels of S100$\beta$ after 120 minutes of RCP. Significant correlation between serum and urine levels was observed. The results were considered to be the fundamental data that could correlate this study with human-based study.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• Journal of Yeungnam Medical Science
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• v.9 no.2
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• pp.288-301
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• 1992

The objective of the present study was to identify the characteristics of phospholipase C (PLC) isozymes purified from bovine brain and to investigate their interrelationship with G protein. The purified PLC isozymes ${\beta}$, ${\gamma}$ and ${\delta}$ were obtained and the characteristics of PLC activity on various concentrations of free $Ca^{2+}$ were observed. The activity of PLC was increased with increasing $Ca^{2+}$ concentration and the activity PLC ${\delta}$ was increased higher in the presence of phosphatidyl choline(PC) than in the abscence of PC. For vesicle formation as the structure of cell membrane, cholic acid and deoxycholic acid as detergent on phosphatidylinositol bisphosphate($PIP_2$) substrate containing PC were used, and then the activity of PLC isozymes were increased with increasing concentration of cholate, from 0.2% to 1% and were increased slightly in deoxycholate. In the $PIP_2$ containing phospholipid and glycolipid as brain extract, the activity of PLC isozymes were checked in 0.2%-1% cholic acid. The activities of PLC isoyzmes were continuously increased up to 1% cholic acid. The quantitation of PLC isozymes from several bovine organs by radioimmunoassay was made. Brain was the most sufficient organ in terms of amount of PLC ${\beta}$and ${\delta}$. A large amount of PLC ${\delta}$ was existed in adrenal gland. The binding capacity of GTPrS and G protein was observed and other observations of the binding effect of GTPrS-G protein and PLC monoclonal Ab-Protein A from tissue homogenate with PLC were made. From the observation the binding capacity was revealed the range of 0.11%-1.49%. The effects of each type of G protein on the percent activity of purified PLC isozymes were observed. From the observation, activities of isozymes were increased in $Go{\alpha}$ & Gmix, and the activities of PLC ${\beta}$ and ${\delta}$ were increased in $G{\beta}{\gamma}$ and $Gi{\alpha}$. Activities of PLC ${\beta}$ and ${\gamma}$ were decreased in $Gt{\alpha}$ but PLC ${\delta}$ increased.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.765-785
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• 1999

Bone morphogenetic protein-2/4 (BMP-2/4) are members of Transforming Growth $Factor-{\beta}\;(TGF-{\beta})$ superfamily and they may differentiate the osteoprogenitor cell and induce formation of cartilage and bone in vivo. This study was performed to investigate the effects of bone morphogenetic protein-2/4 on the characteristics of rat periodontal ligament cells(RPDL) and rat calvaria cells(RCV). In the control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 20% fetal bovine serum, $100{\mu}/ml$ penicillin, $100{\mu}/ml$ streptomycin. In the experimental groups, recombinant human bone morphogenetic protein-2/4 (25ng, 100ng, 250ng/ml) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 2, 3, 5, 7th day, the amount of total protein synthesis and alkaline phosphatase activity at 2, 5, 7th day. And also, the calcified nodule was examed. The results were as follows ; 1 . Both RCV and RPDL cells in both control and experimental groups proliferated during the entire experimental period, but there is no stastically significant difference according to the BMP-2/4 concentration. 2 . Amount of total protein synthesis of both cells in both groups was steadily increased until 5th day, but all experimental groups were significantly different from the control group at 7th day. 3. Alkaline phosphatase activity of both cells in both groups was increased during the entire experiment period. In RCV cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 7th day. In RPDL cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 5th day, and all experimental groups were significantly different from the control group at 7th day. 4. In the both of the cultured Rat Periodontal ligament and calvaria cell treated with BMP-2/4 to compared with control group, it revealed more rapid cell polarization, cell aggregation and hyperchromatic stained on HE agent, and even though only 1 day treated with BMP-2/4 both RPDL and RCV showed more rapid cell reaction than control group. More sensivitve cell reaction of RCV were observed than RPDL in this experiment. From the above results, we could conclude that BMP-2/4 influenced the induction, proliferation and differentiation of bone forming cells

• Food Science and Biotechnology
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• v.15 no.2
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• pp.277-282
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• 2006

The aim of this study was to investigate the anti-inflammatory and anti-oxidant activity of Ligularia fischeri leaf extract on adjuvant induced arthritis in experimental mice. The oral administration of the L. fischeri leaf extract (LF), at doses of 100 and 200 mg/kg body weight once a day for 3 weeks, significantly reduced hindpaw swelling and the production of inflammatory cytokines (tumor necrosis factor(TNF)-${\alpha}$, interleukin(IL)-$1{\beta}$, and IL-6). Treatment with LF (100 mg/kg) also decreased the serum levels of triglyceride and low density lipoprotein(LDL)-cholesterol, and increased high density lipoprotein(HDL)-cholesterol contents compared with those of a control group. The induction of arthritis significantly increased oxidized proteins such as protein carbonyl, advanced oxidation protein products, and advanced glycation end-products in the lung, heart, and brain. Treatment with LF for 3 weeks reduced the levels of oxidized proteins. These results suggest that L. fischeri extract might be beneficial in the treatment of chronic inflammatory disorders.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.2
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• pp.257-264
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• 2019

Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to $500{\mu}g/mL$) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; ${\alpha}_{s2}$-casein increased 2-fold, ${\kappa}$-casein increased 5-fold, and the amount of unchanged ${\beta}$-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein ${\alpha}_{s2}$-casein, ${\kappa}$-casein, and ${\beta}$-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1832-1836
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• 2006

Mesorhizobium sp. LUK, which utilizes 3-amino-3-phenylpropionic acid as the sole source of nitrogen with high enantioselectivity (E(S)>100), was isolated using enrichment culture. The enzyme involved in the utilization of (S)-3-amino-3-phenylpropionic acid was confirmed to be a transaminase and was purified by 235-folds with a specific activity of 0.72 U/mg. The molecular weight of the purified protein was ca. 47 kDa and the active enzyme was determined as a dimer on gel filtration chromatography. The N-terminal sequence was obtained from the purified protein. Spontaneous decarboxylation of produced ${\beta}-keto$ acids was observed during the chiral resolution of 3-amino-3-phenylpropionic acid.

• Biomolecules & Therapeutics
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• v.25 no.6
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• pp.593-598
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• 2017

The $Na^+/H^+$ exchanger-1 (NHE-1) is a ubiquitously expressed pH-regulatory membrane protein that functions in the brain, heart, and other organs. It is increased by intracellular acidosis through the interaction of intracellular $H^+$ with an allosteric modifier site in the transport domain. In the previous study, we reported that glutamate-induced NHE-1 phosphorylation mediated by activation of protein kinase C-${\beta}$ (PKC-${\beta}$) in cultured neuron cells via extracellular signal-regulated kinases (ERK)/p90 ribosomal s6 kinases (p90RSK) pathway results in NHE-1 activation. However, whether glutamate stimulates NHE-1 activity solely by the allosteric mechanism remains elusive. Cultured primary cortical neuronal cells were subjected to intracellular acidosis by exposure to $100{\mu}M$ glutamate or 20 mM $NH_4Cl$. After the desired duration of intracellular acidosis, the phosphorylation and activation of PKC-${\beta}$, ERK1/2 and p90RSK were determined by Western blotting. We investigated whether the duration of intracellular acidosis is controlled by glutamate exposure time. The NHE-1 activation increased while intracellular acidosis sustained for >3 min. To determine if sustained intracellular acidosis induced NHE-1 phosphorylation, we examined phosphorylation of NHE-1 induced by intracellular acidosis by transient exposure to $NH_4Cl$. Sustained intracellular acidosis led to activation and phosphorylation of NHE-1. In addition, sustained intracellular acidosis also activated the PKC-${\beta}$, ERK1/2, and p90RSK in neuronal cells. We conclude that glutamate stimulates NHE-1 activity through sustained intracellular acidosis, which mediates NHE-1 phosphorylation regulated by PKC-${\beta}$/ERK1/2/p90RSK pathway in neuronal cells.

• Genomics & Informatics
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• v.11 no.4
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• pp.224-229
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• 2013

Receptor for advanced glycation endproducts (RAGE) is a multi-ligand receptor that is able to bind several different ligands, including advanced glycation endproducts, high-mobility group protein (B)1 (HMGB1), S-100 calcium-binding protein, amyloid-${\beta}$-protein, Mac-1, and phosphatidylserine. Its interaction is engaged in critical cellular processes, such as inflammation, proliferation, apoptosis, autophagy, and migration, and dysregulation of RAGE and its ligands leads to the development of numerous human diseases. In this review, we summarize the signaling pathways regulated by RAGE and its ligands identified up to date and demonstrate the effects of hyper-activation of RAGE signals on human diseases, focused mainly on renal disorders. Finally, we propose that RAGE and its ligands are the potential targets for the diagnosis, monitoring, and treatment of numerous renal diseases.

• Microbiology and Biotechnology Letters
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• v.9 no.4
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• pp.213-218
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• 1981

The molecular weight of the purified $\beta$-galactosidase of Penicillium sp. was estimated to be 130000 by both Sephadex G-200 gel filtration and SDS-polyacrylamide del electrophoresis. The SDS-electrophoresis gave two protein bands corresponding to the two molecular weights of 130000 and 70000. These results indicated that the enzyme consisted of two probably identical subunits which had a molecular weight of 70000. The optimum pH of the enzyme activity was 4.7 and maximum activity appeared at 5$0^{\circ}C$. The stable pH range for the enzyme was from 4.5 to 7.0. The purified $\beta$-galactosidase had no metal ion requirement for its activity or stability. The enzyme activity was inhibited by C $u^{++}$(1mM)and galactose (100mM). The hydrolysis of lactose in 5% lactose solution, pasteurized milk and 10% skim milk solution were 69.5%, 88.7% and 72.6% after 4 hr incubation at 5$0^{\circ}C$, when 10 units of $\beta$-glucosidase were used per $m\ell$ of the substrate solutions.s.