• Applied Chemistry for Engineering
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• v.29 no.4
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• pp.452-460
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• 2018

This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.625-634
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• 2011

Green tea seed coat (GTSC) was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether (PE), ethyl acetate (EtOAC) and butanol (BuOH). The EtOAC fraction showed the highest level in total phenol contents and the lowest level in nitric oxide (NO) production in LPS-stimulated RAW264.7 macrophage cell. Thus, this study was carried out to investigate the anti-inflammatory and its mechanisms of GTSC EtOAC fraction in LPS-stimulated RAW264.7 macrophage cell. GTSC EtOAC fraction contained EGC ($1146.48{\pm}11.01\;{\mu}g/g$), tannic acid ($966.99{\pm}32.24\;{\mu}g/g$), EC ($70.88{\pm}4.39\;{\mu}g/g$), gallic acid ($947.61{\pm}1.03\;{\mu}g/g$), caffeic acid ($37.69{\pm}1.46\;{\mu}g/g$), ECG ($35.46{\pm}3.19\;{\mu}g/g$), and EGCG ($15.53{\pm}0.09\;{\mu}g/g$) when analyzed by HPLC. NO production was significantly (p<0.05) suppressed in a dose-dependent manner with an $IC_{50}$ of $80.11\;{\mu}g$/mL. Also prostaglandin $E_2$ level was also inhibited in a dose-dependent manner. Moreover, iNOS protein expression was suppressed in dose-dependent manner but COX-2 gene expression was not affected. Total antioxidant capacity and glutathione (GSH) levels were enhanced more than the LPS-control. Expressions of antioxidative enzymes including catalase, GSH-reductase and Mn-SOD were elevated compared to LPS-control. Nuclear p65 level was decreased in the GTSC EtOAC fraction in a dose-dependent manner. These results indicate that GTSC EtOAC fraction inhibit oxidative stress and inflammatory responses through elevated GSH levels, antioxidative enzymes expressions and suppression of iNOS expression via NF-${\kappa}B$ down-regulation.

• Journal of Applied Biological Chemistry
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• v.51 no.1
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• pp.36-44
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• 2008

Organic forms of arsenic (As) were determined through fractionation procedure of soil organic matter (SOM) in soil, sediments and mine tailing samples from the Myungbong, Dongil, and Okdong mining areas of southern Korea. An alkaline extraction method was applied to soil samples followed by the fractionation procedures of SOM by the DAX-8 and XAD-4 resin adsorption method. Major fraction of organic As species (42% to 98%) was found in acid-soluble fraction, whereas minor fraction (0.1 % to 67.8%) was present in the humic-associated As. In acid-soluble fractions, the transphillic- and hydrophilic-associated As were dominant in addition to As binding with humic and fulvic SOM. Arsenic binding was the strongest between pH 6 to 8 and reduced to about 70% at both low and high pH regions. The amount of both transphillic and hydrophillic associated As was less changed than humic and fulvic-associated As, in both low and high pH regions. This apparently indicates that As has stronger affinity towards hydrophillic rather than hydrophobic organics. From the experimental observation of As-binding SOM in natural soil, the ligand exchange model may be a feasible explanation of transphillic and hydrophillic affinity of As.

• KSBB Journal
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• v.24 no.1
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• pp.59-64
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• 2009

The callus formation from inferior leaf of Aloe saponaria was induced in M & S medium supplemented with 10-30 ${\mu}M$ NAA (${\alpha}$-naphthalene acetic acid) and 3-7 ${\mu}M$ kinetin under incubation in the dark at $25^{\circ}C$ for 6 weeks. The hot water extract ($100^{\circ}C$, 24 hrs) from cultured callus was obtained and the components analysis for the extract were examined to determine the callus can synthesized the bioactive component such as Aloe polysaccharide. The freeze dried extract contained the sugar of 53.2%, protein of 7.3%, ash of 18.5% and water of 21% (w/w). Two fractions (Fr-I and Fr-II) were obtained by Sepharose CL-4B gel permeation chromatography and Fr-I, major fraction was further purified with dialysis. From sugar analysis by TLC and GC, the purified Fr-I fraction consisted of glucose (77.6%), galactose (17.7%), mannose (4.7%, w/w) and uronic acid (trace). The molecular weight of purified Fr-I fraction determined by GPC was about 110 kDa.

• Biomolecules & Therapeutics
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• v.6 no.4
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• pp.378-382
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• 1998

In the previous screening on antidiabetic effect of Lycii fructus by glucose transport method using $N_2$-STZ diabeted rat model, each extracts showed the potent antidiabetic activity. We obtained three compounds isolated from the water fraction, EtOAc fraction and n-BuOH fraction of Lycii fructus in the present work and their structures were identified as 1-carboxy-N,N,N-trimethylmethanaminium hydroxide inner salt, 2,4(1H,3H)-pyrimidinedione and 3,3',4',5,7-pentahydroxyflavone-3-rutinoside . Among the constituents separated from Lycii fructus, 2,4(IH,3H)-pyrimidinedione, 3,3',4',5,7-pentahydroxyflavone-3-rutinoside and ascorbic acid were shown a remarkable antidiabetic effect.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.3
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• pp.705-709
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• 2005

The purpose of this research was to investigate the effects of Coix Lachryma-Jobi L. var. Ma-yuen Stapf. on the proliferation of L1210 cells and immune cells. The hexane fraction of Coix Lachryma-Jobi decreased the proliferation of L1210 cells and induced DNA fragmentation of L1210 cells. Also, the fraction decreased the cell viability of murine thymocytes and splenocytes, but increased the phagocytic activity of murine peritoneal macrophages. In addition, fatty acids and fatty acid methyl esters decreased the proliferation of L1210 cells and induced DNA fragmentation of L1210 cells. The main components of the fraction are fatty acid and their derivatives. These results indicate that Coix Lachryma-Jobi has a cytotoxicity on tumor cells and immune cells.

• Toxicological Research
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• v.30 no.1
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• pp.33-38
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• 2014

The human cytochrome P450 2J2 catalyzes an epoxygenase reaction to oxidize various fatty acids including arachidonic acid. In this study, three recombinant enzyme constructs of P450 2J2 were heterologously expressed in Escherichia coli and their P450 proteins were successfully purified using a $Ni^{2+}$-NTA affinity column. Deletion of 34 amino acid residues in N-terminus of P450 2J2 enzyme (2J2-D) produced the soluble enzyme located in the cytosolic fraction. The enzymatic analysis of this truncated protein indicated the typical spectral characteristics and functional properties of P450 2J2 enzyme. P450 2J2-D enzymes from soluble fraction catalyzed the oxidation reaction of terfenadine to the hydroxylated product. However, P450 2J2-D enzymes from membrane fraction did not support the P450 oxidation reaction although it displayed the characteristic CO-binding spectrum of P450. Our finding of these features in the N-terminal modified P450 2J2 enzyme could help understand the biological functions and the metabolic roles of P450 2J2 enzyme and make the crystallographic analysis of the P450 2J2 structure feasible for future studies.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.472-477
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• 1996

Natural human epidermal growth factor (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C$_{18}$ HPLC system. Following characterization by Western blot analysis and double immu- nodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporatioin of 5-bromo-2'-deoxy uridine (BrdU), a non-radioactive alternative for [$^{3}$H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1 - 10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others. Based on the results from the HPLC analysis spiked with recombinant human epidermal growth factor (rhEGF) and amino acid sequencing, we concluded that the second fraction was nhEGF and the other three fractions were the derivatives of nhEGF. In addition, the proportion of nhEGF was approximately 46% is compared with that of the other three derivatives.

• Advances in Traditional Medicine
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• v.6 no.2
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• pp.86-95
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• 2006

The ethanol extract of the Crotalaria juncea seeds, which showed promising antispermatogenic and antiandrogenic activities in albino mice, was taken up further for the isolation of the active fractions present in it. Two fractions that were obtained from thin layer chromatography were subjected for testing to know their antispermatogenic and antiandrogenic activities. After preliminary trials the fraction I showed maximum antifertility activity at the dose level of 200 mg/kg body weight when administered orally to the rats for 50 days. The fraction I was found to affect spermatogenesis as well as the endocrine functions of the testis as indicated by gravimetric, histopathological and biochemical changes. Further this fraction has caused degenerative changes in the seminiferous tubules and Leydig cells of the testis. The accessory reproductive organs like epididymis, seminal vesicles, vas deferens, prostrate, Cowper's gland and Levator Ani muscle showed significant malfunction. Cauda epididymal sperm count and sperm motility were reduced significantly. The treatment has also resulted in increase in the cholesterol level and alkaline phosphatase activity, and decrease in protein, glycogen, sialic acid contents and acid phosphatase activity in testis. It is noteworthy that RIA studies have shown significant reduction in serum FSH, LH and testosterone. Scanning electron microscopic observations revealed abnormalities in sperm structure.

• Food Science of Animal Resources
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• v.18 no.2
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• pp.185-191
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• 1998

Human milk and bovine milk in normal stage were fractionated four parts : whey, skimmilk membrane, and casein pellet. The specific activity (nmole / mim / mg protein) and distribution ratio(%) of suborganella marker enzymes in each separated milk fraction were determined. Especially, neutral $Ca^{2+}$-ATPase, acid $Ca^{2+}$-ATPase, NADH-cytochrome C reductase, and acid phosphatase were higher in human milk. However, both $Ca^{2+}$-ATPases were not detected in all fractions of bovine milk. On the other hand, 5'-nucleotidase, phosphodiesterase I, alkaline phosphatase, and $\gamma$-glutamyl transpeptidase activities in bovine milk were higher than in human milk. Most of the marker enzymes were highly distributed in cream fraction of either human milk or bovine milk, and their specific activities were high to 24 fold from 3 fold when compared with that of whole milk. These results suggest that marker enzymes in mammary epitherial cell are transfered into cream fraction by the membrane rearrangement, and different biochemical reaction between human and bovine exists for milk secretion in mammary gland.