• YAKHAK HOEJI
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• v.41 no.4
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• pp.484-491
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• 1997

The effect of prolonged treatment with Mori Folium column fraction(MFCF) on intestinal ${\alpha}$-glycosidase activity has been studied in mice made hyperglycemic and hyperinsulinemic by feeding a high carbohydrate-containing diet. Mice were treated for 10 week with or without MFCF, added to the high carbohydrate-containing diet at 50mg/100g food. While MFCF had no effect on body weight, it prevented the rise in glycemia and insulinemia. Maltase, sucrase and lactase activities were measured in intestinal homogenates of proximal, middle and distal segments of jejunoileum. Following 10 week of MFCF administration, MFCF significantly induced all three enzyme activities especially at middle and distal segments. The mechanisms responsible for these changes and their potential biochemical implications remain to be determined.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.72-80
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• 2000

Lipopolysaccharides (LPS) of Pasteurella multocida (P multocida) and several Gram-negative bacterial pathogens were analyzed by methanolysis, trifluoroacetylation and gas chromatography (GC) on a fused-silica capillary column. The GC analysis indicated that LPS prepared from a strain of P multocida by phenol-water (PW) or trichloroacetic acid (TCA) extraction were quite different in chemical composition. However, LPS prepared from Salmonella enteritidis by the two extraction methods were very similar. PW-LPS extracts from different Pasteurella strains of a serotype had essentially identical GC patterns. Endotoxic LPS extracted from 16 different serotypes of P multocida by PW or by phenol-chloroform-petroleum ether procedures yielded chromatograms indicating similar composition of the fatty acid moieties but minor differences in carbohydrate content. When the chemical composition of endotoxic LPS extracted from several Gram-negative bacteria (P multocida, Pasteurella hacmolytica, Haemophilus somnus, Actinobacillus ligniersii, Brucella abortus, Treponema hyodysenteriae, Escherichia coli, Bacteriodes fragilis, Salmonella abortus equi and Salmonella enteritidis) were examined, each bacteria showed a unique GC pattern. The carbohydrate constituents in LPS of various Gram-negative bacteria were quite variable not only in the O-specific polysaccharides but also in the core polysaccharides. The LPS of closely related bacteria shared more fatty acid constituents with each other than with unrelated bacteria.

• Archives of Pharmacal Research
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• v.21 no.5
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• pp.521-526
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• 1998

The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

• Applied Biological Chemistry
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• v.36 no.4
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• pp.310-314
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• 1993

High performance liquid chromatographic method using a TSK-gel amide 80 column and isocratic elution with acetonitrile-water (63 :35 ;v/v) mixture was used for the separation and the quantitation of fructo (GF2-GF7)- and inulo-oligosaccharides (F2-F4). Retention time of each standard carbohydrate was highly reproducible. Standardization curves obtained by plotting the peak areas against the amounts of each carbohydrate showed very high coefficient of determination$({\ge}0.9884)$ and similar slopes, and a wide range of y-intercepts. Our results suggest the use of each Pure oligosaccharide for its own standardization curve instead of using a certain carbohydrate as an internal standard.

• Korean Journal of Food Science and Technology
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• v.28 no.1
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• pp.99-104
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• 1996

Carbohydrate-free caseinomacropeptide (CMP) was isolated from the sweet whey powder by a precipitation method using 12% trichloroacetic acid. The yield of carbohydrate-free CMP was 2.7 g from 100 g sweet whey powder. The electrophoretic pattern and the amino acid analysis of CMP showed that isolated CMP was quite pure, indicating the precipitation with 12% trichloroacetic acid was very effective for isolating carbohydrate-free CMP from the sweet whey powder. Trypsin, covalently immobilized on pore glass beads by carbodiimide (EDC) method, was 20mg per 1g glass beads. CMP was almost completely hydrolyzed by soluble trypsin in 24hr, but not by immobilized trypsin. The tryptic hydrolysates were fractionated on a Bio-Gel P 4 column $(1.5{\times}120\;cm)$and separated peptides were tested for their capacities to inhibit platelet aggregation using a aggregometer. The hydrolysate obtained from CMP after 24hr digestion by immobilized trypsin showed the highest activity.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.521-529
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• 1996

In an effort to obtain basic data of carbohydrate metabolism during spermiogenesis of the sexually-matured Jindo dog, the glycogen distribution in the testis was investigated by light and transmission electron microscopy. Periodic acid thiocarbohydrazide silver proteinate physical development(PA-TCH-SP-PD) staining method provided better results in the detection of glycogen granules from Sertoli cells and germ cells than the periodic acid schiff(PAS) staining method did. Pre-treatment of the tissue sections with ${\alpha}$-amylase elicited a significant decrease in PA-TCH-SP-PD stained granules, which suggested that the stained granules were of glycogen origin. High concentration of the glycogen granules were observed in the Sertoli cells, especially in its column, sheet-like processes, club-like processes, and tubular processes. The glycogen granules were unevenly distributed in some Sertoli cell columns. These results strongly indicated that the Sertoli cells of Jindo dogs showed vigorous activity of carbohydrate metabolism.

• Journal of the Korean Society of Food Science and Nutrition
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• v.11 no.3
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• pp.81-86
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• 1982

Ginseng saponins separated from Panax ginseng C.A. Meyer were analyzed by high-performance liquid chromatography using a carbohydrate analysis column. The effect on the resolution and retention time of each ginsenosides, as well as the addition effect of n-butanol on a acetonitrile/water system, was examined using various proportions of acetonitrile/water system (80/20-94/6) which have been used with typical solvent of carbohydrate analysis column. The retention time of each ginsenosides was greatly affected by the compositions and mixture proportions of the mobile phase and also markedly increased as the proprotion of acetonitrile in mobile phase increased. It was proved that acetonitrile/water system (80/20) and acetonitrile/water/n-butanol system (86/14/10) were very effective mobile phases for diol and triol sapoin analyses, respectively. According to the result obtained by this method, the PT/PD ratios of white and red ginseng saponins were 0.401 % and 0.561 %, respectively. Red ginseng increased PT/PD ratio, compared with white ginseng. This is attributed to the change of saponin pattern by processing method and the change of PT/PD ratio would greatly influence on the biochemical and pharmacological effects of ginseng and its products.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.519-526
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• 2003

Pancreatic lipase and ${\alpha}$-amylase inhibitory activities in purified extracts of pumpkin and job's tear were studied. Pancreatic lipase inhibitory activity was determined by measuring the rate of releasing oleic acid from triolein, and ${\alpha}$-amylase inhibitory activity was determined by iodometric method. The extracts of pumpkin and job's tear were purified using silica gel and C-18 gel column chromatographies. Treatment of pumpkin extract $(120\;{\mu}g/mL)$ in 3T3-L1 preadipocyte decreased differentiation about 95% and blocked accumulation of lipid. Body weights of rats fed high-fat diet containing dietary supplement decreased about 13% as compared with those fed only highfat diet. These results revealed dietary supplement is a good obesity-reducing material for decreasing lipid and carbohydrate absorptions.

• Journal of Ginseng Research
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• v.35 no.1
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• pp.31-38
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• 2011

In order to distinguish the cultivation area of Panax ginseng, principal component analysis (PCA) using quantitative and qualitative data acquired from HPLC was carried out. A new HPLC method coupled with evaporative light scattering detection (HPLC-ELSD) was developed for the simultaneous quantification of ten major ginsenosides, namely $Rh_1$, $Rg_2$, $Rg_3$, $Rg_1$, Rf, Re, Rd, $Rb_2$, Rc, and $Rb_1$ in the root of P. ginseng C. A. Meyer. Simultaneous separations of these ten ginsenosides were achieved on a carbohydrate analytical column. The mobile phase consisted of acetonitrile-water-isopropanol, and acetonitrile-water-isopropanol using a gradient elution. Distinct differences in qualitative and quantitative characteristics for ginsenosides were found between the ginseng roots produced in two different Korean cultivation areas, Ganghwa and Punggi. The ginsenoside profiles obtained via HPLC analysis were subjected to PCA. PCA score plots using two principal components (PCs) showed good separation for the ginseng roots cultivated in Ganghwa and Punggi. PC1 influenced the separation, capturing 43.6% of the variance, while PC2 affected differentiation, explaining 18.0% of the variance. The highest contribution components were ginsenoside $Rg_3$ for PC1 and ginsenoside Rf for PC2. Particularly, the PCA score plot for the small ginseng roots of six-year old, each of which was light than 147 g fresh weight, showed more distinct discrimination. PC1 influenced the separation between different sample sets, capturing 51.8% of the variance, while PC2 affected differentiation, also explaining 28.0% of the variance. The highest contribution component was ginsenoside Rf for PC1 and ginsenoside $Rg_2$ for PC2. In conclusion, the HPLC-ELSD method using a carbohydrate column allowed for the simultaneous quantification of ten major ginsenosides, and PCA analysis of the ginsenoside peaks shown on the HPLC chromatogram would be a very acceptable strategy for discrimination of the cultivation area of ginseng roots.

• Biomedical Science Letters
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• v.18 no.2
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• pp.96-103
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• 2012

Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.