• Applied Biological Chemistry
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• 제35권4호
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• pp.276-280
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• 1992

HPLC에 의한 카테킨류 5성분의 정량분석법을 검토한 결과, 종래의 이동상을 acetonitril, acetic acid 및 methanol 수용액에서 acetontrile, N,N-dimethyl formamide, ethyl acetate의 0.06% 인산수용액으로 전환하였을 때 컬럼에서의 용출시간을 45분에서 28분으로 줄일 수 있었으며, 특히 (-)-epigallocatechin-3-gallate(EGCG)나 (-)-epicatechin gallate(ECG)의 peak를 종래의 넓은 형태에서 예리한 chromatogram을 얻으므로써 분석값의 오차를 최소화 할 수 있었고 머므름시간$(0.32{\sim}3.97%)$이나 peak 면적$(1.61{\sim}7.02%)$의 변동계수도 신뢰할 수 있는 값을 얻었다. 또한 시판되고 있는 수종의 녹차류와 홍차 및 대만산 오룡차를 분석한 결과 녹차류의 카테킨 함량은 $120.3{\sim}153.7\;mg/g$ 이었고 홍차는 녹차류의 1/4수준 이었다.

• 한국식품영양과학회지
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• 제20권3호
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• pp.260-265
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• 1991

Salmonella/microsome assay system을 이용하여 N -nitrosodimethylamine (NDMA)의 돌연변이유발성을 검토하는 효과적인 방법과 비타민 C가 NDMA자체의 돌연변이유발과 nitrite와 2급아민으로부터의 NDMA 생성에 미치는 영향을 연구하였다. NDMA를 활성화시키기 위한 S9중 Aroclor1254로 induce시킨 hamster S9이 가장 효과가 있었으며 Aroclor 1254나 phenobarbital로 induce시킨 rat 59 mix에 의하여서는 활성화가 약하였다. DMSO와 ethanol에 NDMA를 녹였을 때는 돌연변이유발실험에서 저해효과가 나타났으나 phosphate buffer(pH 7. 4), 증류수, 95% methanol, Tween 80 + water(l : 4)는 저해 효과가 없었다. 비타민 C는 이미 생성환 NDMA에 대해서는 돌연변이 유발저해 효과를 나타내지 않았지만 nitrite와 2급 아민(dimethylamine)으로 부터의 NDMA 생성은 크게 저해하였다. 반응물에 비타민C를 첨가한 경우 Salmonella typhimurium TAl00의 revertant 수가 spontaneous숫자 수준으로 감소되었으며 HPLC를 이용한 NDMA의 분석에서도 거의 95% 까지의 정량적인 감소를 관찰할 수 있었다.

• 대한화장품학회지
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• 제27권1호
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• pp.17-27
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• 2001

A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

• 한국식품위생안전성학회지
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• 제15권2호
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• pp.144-150
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• 2000

Phosphamidon의 가수분해속도상수는 25。C, pH 4, 7 및 9에서 각각 0.0020, 0.0022 및 0.0049이었고, 40。C에서 각각 0.0040, 0.0050및 0.0150으로 측정되었다 같은 pH에서 온도가 높을수록 촉진되었으며, pH 9의 염기성조건에서는 15。C상승에 반감기가 약 3배 정도 빨랐다. 또한 같은 온도에서 산성(pH 4)과 중성조건(pH 7)에 비해 염기성조건에서 가수분해반응이 약 2∼4배 정도 빨랐다. Profenofos의 가수분해속도상수는 25。C, pH 4, 7 및 9에서 각각 0.0022, 0.0047 및 0.0860이었고, 40。C에서 각각 0.0031, 0.0086 및 0.1245로 측정되었다. Profenofos의 가수분해는 phosphamidon과 마찬가지로 같은 pH에서 온도가 높을수록 촉진되었으며, 같은 온도에서 산성과 중성조건에 비해 염기성조건에서 가수분해반응이 약 15∼40배 정도 빨랐고 반감기가 모두 8시간 이내로 가수분해반응이 현저하게 일어났다. Profenofos의 가수분해속도가 phosphamidon보다 빠른 것을 알 수 있었다. 가수분해에 의한 분해산물을 확인하고자 GC/MS분석을 한 결과 phosphamidon의 분해생성물은 m/z=153의 O, O-dimethyl phosphate(DMP)와 m/z=149의 N, N-diethylchloro acetamide로 추정된다. Profenofos의 분해생성물은 m/z=208로 4-bromo 2-chloro phenol과 m/z=240으로 O-ethyl S-propyl phosphate로 추정된다.

• Toxicological Research
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• 제6권1호
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• pp.109-119
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• 1990

Ro09-0198, a cyclic peptide isolated from culture filtrates of Streptoverticillium griseoverticillatum, induced lysis of erythrocytes. Ro-09-0198-induced hemolysis was temperature-dependent and the sensitivity of hemolysis differed greatly among animal species. Preincubation of the peptide with phosphatidylethanolamine reduced the hemolytic activity, whereas other phospholipids present in erythrocytes in nature had no effect. A study of the structural requirements on phosphatidylethanolamine necessary for interaction with the peptide indicates that Ro09-0198 recognizes strictly a particular chemical structure of phosphatidylethanolamine: dialkylphosphoethanolamine as well as 1-acylglycerophosphoethanolamine showed the same inhibitory effct on hemolysis induced by Ro09-0198 as diacylphosphatidyl-ethanolamine, whereas phosphoethanolamine gave no inhibitory effect. Neither phosphatidyl-N-monomethylethanolamine nor alkylphosphopropanolamine had an inhibitory effect. Proton resonances of the peptide were observed in dimethyl sulfoxide solution in the presence of 1-dodecanoyl-sn-glycerophosphoethanolamine. This peptide caused permeability increase and aggregation of liposomes containing phosphatidylethanolamine. A glycerol backbone and a primary amino group of phosphatidylethanolamine are necessary for interaction with Ro09-0198 to cause membrane damage. Ro09-0198 induced a selective permeability change on liposomes. Glucose and umbelliferyl phosphate were effluxed significantly, but sucrose was only slightly permeable and inulin could not be released. Platelet aggregation and serotonin release simultaneously induced by Ro09-0198. Addition of peptide to rat platelet, loaded with the fluorescent $Ca^{++}$ chelator quin-2, caused immediate rise in cytosolic free $Ca^{++}$ to liposomal membrane containing phosphatidylethanolamine was observed dose dependently.

• 한국식품영양과학회지
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• 제22권6호
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• pp.815-822
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• 1993

This experiment was carried out to determine the separation condition of 1-dimethylaminonaphthalene-5-sulfony(Dansyl) derivatives of amino acids by reverse-phase high performance liquid chromatography with Nova-Pak C18 column. Determined solvent system was solvent A(200mA phosphate buffer pH 6.8 15%, acetonitrile 11%, water 74%) and solvent B(acetonitrile 65%, methanol 28%, water 7%). Linear gradient of solvent B was applied from 12% to 80% for 50min. Complete separation of 20 amino acids including asparagine and glutamine which constitute protein was achieved within 50min. As the detection limit was the range of picomole, the resolution power was excellent. Reproducibility of the retention time was less than mean $\pm$0.05min. According to the above optimum chromatographic conditions, the amino acid composition of some food and human blood was examined. The most affluent amino acid was alanine in human blood, aspartic acid and glutamic acid in soy sauce, alanine and threonine in soy milk and proline in milk and yoghurt.

• 전기화학회지
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• 제15권3호
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• pp.190-197
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• 2012

비닐에틸렌 카보네이트(VEC: vinyl ethylene carbonate)를 전해질 첨가제로 사용했을 때 하이브리드 커패시터(hybrid capacitors) 전극에서 나타나는 전기화학적 특성변화에 대해서 고찰하였다. 하이브리드 커패시터는 양극은 활성탄(AC : activated carbon) 음극은 리튬티타늄옥사이드(LTO: $Li_4Ti_5O_{12}$)를 사용하였고, 전해질로서는 에틸렌 카보네이트(EC: ethylene carbonate): 디메틸 카보네이트 (DMC: dimethyl carbonate) : 에틸메틸 카보네이트(EMC : ethyl methyl carbonate)를 사용하였고, 염으로 육불화인산리튬($LiPF_6$: lithium hexafluoro phosphate)을 사용하였다. 전극 표면의 산소관능기 그룹을 제거하고, 표면을 환원시킴으로써 전극에 안정성을 향상시킨다고 알려진 VEC의 첨가량에 따른 전기화학적 특성을 평가하였으며, 0.7%(부피비)의 VEC첨가시, 가장 우수한 전기화학적 특성을 얻을 수 있었다. 0.7% 이상 첨가하였을 경우, 오히려 부반응 증가로 전기화학적 성능이 감소하였다. X-ray photoelectron spectrocopy (XPS) 결과로부터 LTO 전극에서 VEC가 첨가되지 않은 전해질에 비해 LiF가 감소한 것을 확인 할 수 있었다. VEC가 첨가되지 않은 전해질은 2500 사이클 후, 43.2 %의 용량 유지를 나타냈지만, 최적화된 VEC 첨가를 통하여 82.7 %의 높은 용량을 유지하는 특성을 가진 하이브리드 커패시터를 얻을 수 있었다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1984년도 학술대회지
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• pp.257-275
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• 1984

Enzymatic browning is considered desirable in tea and tobacco processing but undesirable in many fruits processing at the present time. It is necessary to understand the nature of the enzyme, phenoloxidase, in order to control browning reactions, and extend its effects to formation of browning products as antioxidants in ginseng. Ginseng exhibits antioxidant activity when incorporated with turkey dark meat patties. The activity in red ginseng showed about two times stronger than white ginseng. One of the phenolic antioxidants from fresh, white and reprocessed white ginseng was identified as phenol 2.6 Bis(1.1 dimethyl ethyl) 4-methyl among several unknown compounds by GC/mass spectrometer. In red ginseng, no phenol 2.6 Bis (1.1 dimethyl ethyl) 4-methyl was detected, the compound may be polymerized by phenoloxidase and form some higher molecular compounds which may possess high antioxidant activity. Phenoloxidase isozymes in fresh Korean ginseng (panax ginseng C.A. Meyer) were extracted with phosphate buffer at pH 7.3. The isozymes were purified through ammonium sulfate fractionation, dialysis and chromatography on a DEAE-cellulose column. Two groups of phenoloxidase were shown to be present, one in the floating agglomerated group and the other in the precipitate. group from the 0.85 saturation ammonium sulfate. The DEAE-cellulose column chromatography, the phenoloxidase isozyme present in the precipitate appears as the first peak (I), and that in the agglomerate in the second peak (II). Isozyme I showed higher activity with catechin and catechal, and isozyme II showed higher activity with p-cresol. The isozyme showed two optimum pH activity one at pH 4.5 and the other at 8.5 with catechin as substrate. Korean ginseng phenoloxidase has high heat stability. When heated at $75^{\circ}C$ for 2 hours, its activity remained $90\%\;and\;80\%$ on phenoloxidase I and II respectively. Phenoloxidase I was most active on (+) catechin followed by p-cresal, catechol and epicatechin. Phenoloxidase II was most active on p-cresal followed by (+) catechin, catechol, p-coumanic acid and epicatechin. Sodium bisulfite, sodium cyanide, ascorbic acid glutachion in the oxidized form, sodium diethyl dithiocarbomate and ethylendiamine tetra acetate (EDTA) acted as inhibitors. Red ginseng color development was initiated by phenoloxidase and finished by a followed sun drying process. The antiaging activity of ginseng may be initiated by the antioxidant in the ginseng.

• 한국어병학회지
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• 제20권3호
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• pp.307-313
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• 2007

In this study, a QCM biosensor was made to detect Edwardsiella tarda (E. tarda) using a specific antibody. A 9 MHz AT-cut piezoelectric wafer layered with two gold electrodes of 5mm diameter had a reproducibility of 0.1 Hz in frequency response and was used as the transducer of the QCM biosensor. Self assembled layer (SAM) was conformed on a quartz crystal by treating with 3-mer-captopropionic acid (MPA) and activated with N-ethyl-N'-(3-dimethyl-aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The resulting NHS group was further converted to hydrazide by the reaction with hydrazine. Aldehyde group was introduced into the carbohydrate moiety of anti-E. tarda antibody by the reaction with periodic acid and was used to immobilise the antibody through the reaction with hydrazide group on the electrode surface. A baseline was established in the presence of phosphate-buffered saline (PBS) and a resonant frequency (F1) was measured. Sample was added to the sensor surface and second resonant frequency (F2) was measured after unbound substances were washed out with PBS several times. Finally, the frequency shift (ΔF) representing the mass change was calculated by subtracting F2 from F1. After adding the oxidized anti-E. tarda antibody to the electrode surface containing hydrazide group, frequency shift of 288.811.4 Hz (mean S.E) was observed, thus proving that considerable amount of antibody was immobilized. In the immunoassay test, the frequency shift of 1877.75 Hz, 580.67 Hz, 221.39 Hz, 7.671.83 Hz (mean S.E) were observed at doses of 1000, 500, 100, 50 g of bacterial cells, respectively. It was also demonstrated that the prepared sensor chip was stable enough to withstand repeated surface regeneration with 0.2 M Tris-glycine and 1 % DMSO, pH 2.3 more than ten times.

• 한국재료학회지
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• 제24권6호
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• pp.310-318
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• 2014

In this experiment, a highly porous scaffold of biphasic calcium phosphate (BCP) was prepared using the spongereplica method. The BCP scaffold was coated with 58S bioactive glass (BG) and sintered for a second time. The resulting scaffold was coated with gelatin (Gel) and cross-linked with [3-(3-dimethyl aminopropyl) carbodiimide] and N-Hydroxysuccinamide (EDC-NHS). The initial average pore size of the scaffold ranged from 300 to $700{\mu}m$, with more than 85 % porosity. The coating of BG and Gel had a significant effect on the scaffold-pore size, decreasing scaffold porosity while increasing mechanical strength. The material and surface properties were evaluated by means of several experiments involving scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffraction (XRD). Cytotoxicity was evaluated using MTT assay and confocal imaging of MC3T3-E1 pre-osteoblast cells cultured in vitro. Three types of scaffold (BCP, BCP-BG and BCP-BG-Gel) were implanted in a rat skull for in vivo evaluation. After 8 weeks of implantation, bone regeneration occurred in all three types of sample. Interestingly, regeneration was found to be greater (geometrically and physiologically) for neat BCP scaffolds than for two other kinds of composite scaffolds. However, the other two types of scaffolds were still better than the control (i.e., defect without treatment).