• Biomolecules & Therapeutics
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• v.23 no.5
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• pp.486-492
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• 2015

Drug metabolism mostly occurs in the liver. Cytochrome P450 (CYP) is a drug-metabolizing enzyme that is responsible for many important drug metabolism reactions. Recently, the US FDA and EU EMA have suggested that CYP enzyme induction can be measured by both enzymatic activity and mRNA expression. However, these experiments are time-consuming and their interassay variability can lead to misinterpretations of the results. To resolve these problems and establish a more powerful method to measure CYP induction, we determined CYP induction by using luminescent assay. Luminescent CYP assays link CYP enzyme activity to firefly luciferase luminescence technology. In this study, we measured the induction of CYP isozymes (1A2, 2B6, 2C9, and 3A4) in cryopreserved human hepatocytes (HMC424, 478, and 493) using a luminometer. We then examined the potential induction abilities (unknown so far) of mesalazine, a drug for colitis, and mosapride citrate, which is used as an antispasmodic drug. The results showed that mesalazine promotes CYP2B6 and 3A4 activities, while mosapride citrate promotes CYP1A2, 2B6, and 3A4 activities. Luminescent CYP assays offer rapid and safe advantages over LC-MS/MS and qRT-PCR methods. Furthermore, luminescent CYP assays decrease the interference between the optical properties of the test compound and the CYP substrates. Therefore, luminescent CYP assays are less labor intensive, rapid, and can be used as robust tools for high-throughput CYP screening during early drug discovery.

• Applied Biological Chemistry
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• v.47 no.4
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• pp.379-383
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• 2004

For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

• Applied Microscopy
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• v.29 no.4
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• pp.437-450
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• 1999

Bile canaliculi are the structure delivering bile secreted by hepatocytes into the bile passage. Bile secretion is mainly controlled by the cytoskeletal elements, mainly of actin in the microvilli, pericanalicular web. Most studies on the bile secretion have been done in viva situation, however, to control the various parameters in vitro culture system seem to be more useful. To set up an in vitro experimental system, the investigator isolated hepatocytes with an enzymatic method using a mixture of collagenase and hyaluronidase from normal Sprague-Dawley rat liver and cultured. Isolated hepatocytes were round and formed cords in culture. Microvilli covered the whole surface of hepatocytes. Bile canaliculi were formed between hepatocytes and were characterized by the presence of microvilli of various lengths and shapes mainly arising from small surface mounds. Actin filament core in the microvilli and pericanalicular actin web were incomplete. After cytochalasin D treatment, cultured hepatocytes were round but the surface were irregular with surfacen blebs, folds and grooves. Microvilli on the surface were scarce. Bile canaliculi were markedly dilated often with the detached junctional complexes. Bile canaliculi lacks microvilli almost completely and extended into the pericanalirular cytoplasm showing complex vacuolar and tubular structures by transmission electron mciroscopy. Pericanalicular actin web, intermediate filaments were hardly identified. Subsurface actin filaments were scattered scarcely under the cell membranes. These results suggest that hepatocytes isolated from rats can survive and form bile canaliculi in culture and the actin filaments are involved in the formation and/or maintenance of the bile canaliculi.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.8
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• pp.5492-5500
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• 2015

Low-density lipoprotein cholesterol (LDL-C) is a major modifiable risk factor for cardio- cerebrovascular disease. In clinical practice, however, it is primarily calculated using the Friedewald formula as a cost-effective method. The aim of this study was to compare Friedewald-estimated and directly measured LDL-C values and assess the concordance in guideline LDL-C risk classification between the two methods. The data were derived from the 2009 and 2010 Korea National Health and Nutrition Survey (KNHANES). Analysis was done for 4,319 subjects with lipid panels-total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), directly measured LDL-C using an enzymatic homogeneous assay, and triglycerides (TG). For subjects with TG lower than 400 mg/dL, Friedewald-estimated and directly measured LDL-C were highly correlated (r = 0.958, p < 0.001) and overall concordance was 82.7%. As TG increased, overall concordance decreased. Overall concordance was 85.4% at TG lower than 150 mg/dL; 78.2% at TG of 150-199 mg/dL; and 71.4% at TG of 200-399 mg/dL. The Friedewld equation tended to overestimate LDL-C when TG are of < 150 mg/dL; however, underestimate LDL-C when TG are of ${\geq}150mg/dL$. As a result, Friedewald estimation misclassified 382 subjects (9.1%) in a higher category versus 348 subjects (8.3%) in a lower category. Our findings suggest that overestimation of LDL-C by the Friedewald formula can be a great problem as well as underestimation.

• Nutrition Research and Practice
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• v.11 no.2
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• pp.105-113
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• 2017

BACKGROUND/OBJECTIVES: A high-fat diet (HFD) induces obesity, which is a major risk factor for cardiovascular disease and cancer, while a calorie-restricted diet can extend life span by reducing the risk of these diseases. It is known that health effects of diet are partially conveyed through epigenetic mechanism including DNA methylation. In this study, we investigated the genome-wide hepatic DNA methylation to identify the epigenetic effects of HFD-induced obesity. MATERIALS AND METHODS: Seven-week-old male C57BL/6 mice were fed control diet (CD), calorie-restricted control diet (CRCD), or HFD for 16 weeks (after one week of acclimation to the control diet). Food intake, body weight, and liver weight were measured. Hepatic triacylglycerol and cholesterol levels were determined using enzymatic colorimetric methods. Changes in genome-wide DNA methylation were determined by a DNA methylation microarray method combined with methylated DNA immunoprecipitation. The level of transcription of individual genes was measured by real-time PCR. RESULTS: The DNA methylation statuses of genes in biological networks related to lipid metabolism and hepatic steatosis were influenced by HFD-induced obesity. In HFD group, a proinflammatory Casp1 (Caspase 1) gene had hypomethylated CpG sites at the 1.5-kb upstream region of its transcription start site (TSS), and its mRNA level was higher compared with that in CD group. Additionally, an energy metabolism-associated gene Ndufb9 (NADH dehydrogenase 1 beta subcomplex 9) in HFD group had hypermethylated CpG sites at the 2.6-kb downstream region of its TSS, and its mRNA level was lower compared with that in CRCD group. CONCLUSIONS: HFD alters DNA methylation profiles in genes associated with liver lipid metabolism and hepatic steatosis. The methylation statuses of Casp1 and Ndufb9 were particularly influenced by the HFD. The expression of these genes in HFD differed significantly compared with CD and CRCD, respectively, suggesting that the expressions of Casp1 and Ndufb9 in liver were regulated by their methylation statuses.

• The Korean Journal of Food And Nutrition
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• v.28 no.6
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• pp.1026-1032
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• 2015

This study attempted to find an efficient method for the preparation of high-purity galactooligosaccharides (HP-GOS) using ${\beta}$-galactosidase and yeast fermentation. GOS prepared using Lactozym 3000L showed the greatest enhancement in total GOS of the six ${\beta}$-galatosidases tested. GOS alone achieved 51% conversion of initial lactose. GOS production was enhanced by fermentation with commercial yeast (Saccharomyces cerevisiae); its concentration reached 71% after 36h fermentation with 8% yeast. Component sugar analysis with HPLC indicated that HP-GOS fermented with S. cerevisiae showed significantly increased levels of 4'/6'-galactosyllactose and total GOS as well as a significantly decreased glucose level. HP-GOS facilitated the growth of Lactobacillus sp. (L. acidophilus and L. casei) and Bifidobacterium sp. (B. longum and B. bifidum). In sum, high-purity GOS has been successfully produced through both an enzymatic process and yeast fermentation. GOS encourages the growth of bacteria such as Lactobacillus and Bifidobacterium that may be beneficial to human gastrointestinal health.

• Korean Journal of Food Science and Technology
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• v.27 no.1
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• pp.86-91
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• 1995

In order to expand the utility of alginic acid in the food industry, we have investigated to prepare low viscous alginic acid using a method for degradation of alginic acid with the enzyme system of Vibrio sp. AL-145. The enzyme showed maximum activity at pH 8.0 and $37^{\circ}C$, and was stable in the pH range 7.5 to 8.5 and at temperature up to $30^{\circ}C$, and 0.5M NaCl needed for the enzyme activity. The viscosity of alginic acid decreased with the reaction time courses regardless of alginic acid and enzyme concentration, and 90% of viscosity decreased with 60 min of reaction time, but the changes of reducing sugar not exhibited. The soluble concentration of partially degradated alginic acid(PDA) in water was about 10%(w/v), and adsorption capacity of $Ca^{2+}$ ion increased with increasing the concentration of PDA. The alcohol concentration on precipitation of PDA was higher than Na-alginic acid.

• Journal of Digital Convergence
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• v.14 no.1
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• pp.253-262
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• 2016

Yeonsan Ogae has been known as supporting health and high efficacy of treatment. In recent days, as the efficacy of functional peptides has known, the optimization of oligo peptides production and its characteristics from Ogae viscera has been performed. Response surface method was used to perform the optimizaion of enzyme hydrolysis. The range of processes was temperature (40, 50 and $60^{\circ}C$), pH(6.0, 7.0 and 8.0), and enzyme(1, 2 and 3%). The degree of hydrolysis, amono acids, molecular weight of products were analyzed. The optimum process of enzyme hydrolysis were determined as temperature $58^{\circ}C$, pH 7.5, and enzyme concetration 3%. At optimum conditions, the degree of hydrolysis after 2 h reaction was 75-80%. The total amino acids of amino acid and were 386.15 mg/100 g and 155.26 mg/100 g, respectively. The molecular weight of products by using Maldi-TOF was ranged from 300 to 1,000 Da.

• Journal of Life Science
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• v.19 no.8
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• pp.1088-1092
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• 2009

This study was conducted to determine an estimated value and change in TDF (total dietary fiber) binding major minerals (Ca, P, K Mg) of soybean sprouts depending on cultivation periods of 1, 3 or 5 days, and parts of cotyledon and hypocotyl. The compositions of TDF binding major minerals in soybean sprouts were evaluated by the enzymatic-gravimetric method developed by Prosky and adopted by AOAC. The content of TDF binding P ranged between 915.28 mg and 1037.82 mg per 100 g cotyledon. Average contents of Ca, K, and Mg in cotyledon ranged between 541.67 mg and 634.34 mg, 180.91 mg and 253.98 mg, 231.90 mg and 301.01 mg, respectively, on dry matter basis. The average contents of TDF binding Ca in hypocotyl per 100 g were between 454.36 mg and 540.33 mg, and other major minerals contents were between 149.24 mg and 186.21 mg for P, 164.17 mg and 182.78 mg for K, and 152.53 mg and 161.22 mg for Mg, respectively. The proportional changes of Ca, an estimated value in cotyledon, ranged between 5.58% and 30.98%, 47.18% and 59.10% for P, 95.24% and 95.50% for K, and 58.45% and 68.64% for Mg, respectively, based on dry matter. An estimated value of Ca, P, K, and Mg in hypocotyl ranged between 6.71% and 13.52%, 91.03% and 91.76%, 96.91% and 98.92%, and 61.03% and 66.37%, respectively, on dry matter basis.

• Journal of Life Science
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• v.23 no.4
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• pp.569-577
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• 2013

Soy sauce is a traditional fermented seasoning in several oriental countries, such as Korea and Japan, and recently it has been reported to have biological activities. In Korean soy sauce, soybeans and wheat are the two main raw materials. Polysaccharides that originate from the cell wall of soybeans are resistant to enzymatic hydrolysis. These polysaccharides remain in the soy sauce even after fermentation and are termed Kanjang polysaccharides (KPS). In this study, polysaccharides were obtained from dialysate of different soy sauces labeled as A~T and manufactured by fermentation or the acid-hydrolyzate method. We investigated anti-inflammatory activities by examining the effects of these KPS on proinflammatory cytokine release and mRNA expression in mast cells. Histamine and ${\beta}$-hexosaminidase release were strongly decreased by the KPS treatment in RBL-2H3 cells. Treatment with KPS clearly reduced mRNA expression and the release of the proinflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha (TNF-${\alpha}$) in PMACI-stimulated HMC-1 cells. In particular, KPS derived from fermented Kanjang products showed a significant anti-inflammation effect on mast cells compared to the acid-hydrolyzed Kanjang products. This study suggests that KPS appear to be effective in suppressing allergic inflammatory reactions.