• Biotechnology and Bioprocess Engineering:BBE
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• 제10권3호
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• pp.218-224
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• 2005

Surface modification of glutaraldehyde fixed bovine pericardium (GFBP) was success­fully carried out with hyaluronic acid (HA) derivatives. At first, HA was chemically modified with adipic dihydrazide (ADH) to introduce hydrazide functional group into the carboxyl group of HA backbone. Then, GFBP was surface modified by grafting HA-ADH to the free aldehyde groups on the tissue and the subsequent HA-ADH hydrogel coating. HA-ADH hydrogels could be prepared through selective crosslinking at low pH between hydrazide groups of HA-ADH and crosslinkers containing succinimmidyl moieties with minimized protein denaturation. When HA­ADH hydrogels were prepared at low pH of 4.8 in the presence of erythropoietin (EPO) as a model protein, EPO release was continued up to $85\%$ of total amount of loaded EPO for 4 days. To the contrary, only $30\%$ of EPO was released from HA-ADH hydrogels prepared at pH=7.4, which might be due to the denaturation of EPO during the crosslinking reaction. Because the carboxyl groups on the glucuronic acid residues are recognition sites for HA degradation by hyaluronidase, the HA-ADH hydrogels degraded more slowly than HA hydrogels prepared by the crosslinking reaction of divinyl sulfone with hydroxyl groups of HA. Following a two-week subcutaneous implantation in osteopontin-null mice, clinically significant levels of calcification were observed for the positive controls without any surface modification. However, the calcification of surface modified GFBP with HA-ADH and HA-ADH hydrogels was drastically reduced by more than $85\%$ of the positive controls. The anti-calcification effect of HA surface modification was also confirmed by microscopic analysis of explanted tissue after staining with Alizarin Red S for calcium, which followed the trend as observed with calcium quantification.

• 한국환경복원기술학회지
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• 제7권5호
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• pp.94-99
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• 2004

This study was carried out to find out what is the optimum conditions for RAPD of Zelkova serata. We changes the factors what affect to PCR band patterns, as a result, we established the optimum conditions as follows; template DNA 100mg, Primer 0.25uM, dNTP 100mM, Taq polymerase 1.0u, and total reaction volume was filled up to 10uL with distilled water. As the amount of primers went higher, PCR reaction rates were lowered. This reason was cause by exhaustion of primers during initial reaction. The amount of dNTP didn't showed noticable differtations between the range, but the optimum amount was 100mM for efficiency. Taq polymerase 1.0 unit was the best in the range. As the concentration of polymerase were increased, many non-specific bands were appeared, In primer selection, most Openron Random Primers are amplified in this experiment. The primers GC contents were 60, and set A, B, C, D, E, X were tested. Thermal cycler(ASTEC PC808, Japan) condition was, $95^{\circ}C$, 5min, initial denaturation, $94^{\circ}C$, 20sec, denaturation, $37^{\circ}C$, 40sec, annealing, $72^{\circ}C$, 1min, extention, 45cycle repeated and final extention $72^{\circ}C$10min.

• BMB Reports
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• 제30권5호
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• pp.326-331
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• 1997

To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

• Asian Journal of Atmospheric Environment
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• 제6권1호
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• pp.33-40
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• 2012

In this study, we characterized the physical form of allergenic Cry j 1 in the urban atmosphere. Through an immunofluorescence antibody method, we showed that allergenic Cry j 1 exists as fine particles (${\leq}1.1{\mu}m$). To determine Cry j 1 concentrations and its particle size distribution, we used the ELISA method to confirm that most Cry j 1 exists as fine particles in the urban atmosphere and is found at high concentrations on fine day next to rainy day. Furthermore, we evaluated Cry j 1 denaturation by using the Biacore J system based on the surface plasmon resonence (SPR) principle and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We showed that the dissociation constant ($K_D$) of Cry j 1 that has been exposed to urban polluted air is lower ($1.76{\times}10^{-14}$ M) than that of Cry j 1 ($1.32{\times}10^{-9}-3.37{\times}10^{-9}$ M) of original pollen grains that has not been exposed to air pollutants. Cry j 1 turns into low molecular weight proteins by reacting with various acidic solutions. In sum, we showed that allergenic Cry j 1 exists as fine particles that can deposit in the lower respiratory tract. This finding clarifies the relationship between Japanese cedar pollinosis and air pollutants.

• Journal of Pharmaceutical Investigation
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• 제39권6호
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• pp.423-429
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• 2009

Liposomes have been used as one of the efficient carriers for drug delivery. In this study, anionic liposomes of which surface was modified by using both electrostaic interaction between anionic liposomes and cationically charged BSA molecules at lower pH than isoelectric point (pI) of BSA and denaturation of the BSA-coated liposomes by thermal treatment. The thermally denatured BSA-coated liposomes (DBAL) had mean particle diameter of 125.2${\pm}$1.7 nm and zeta potential value of -22.4${\pm}$4.5 mV. Loading efficiency of model drug, doxorubicin (DOX), into liposomes was 83.0${\pm}$2.6%. Results of in vitro stability study of DBAL in blood plasma showed that the mean particle diameter of DBAL 400 did not increase in blood plasma and adsorption of plasma protein was much less than plain or anionic liposomes. Intracellular uptake of DBAL 400 evaluated by confocal microscopy observation was higher than that of PEG liposomes.

• 대한한방소아과학회지
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• 제11권1호
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• pp.249-273
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• 1997

Hyunggyeyungyotang has been used for treatment of sinusitis and otitis media in oriental medicine since ancient times. It is reported that Hyunggyeyungyotang has good effects on inflammatory and allergic diseases of otorhinolaryngology in clinical medicine. Kamihyunggyeyungyotang was made by adding several herbs to Hyunggyeyungyotang which has such good effects. To investigate the effects of Hyunggyeyungyotang and Kamihyunggyeyungyotang on inflammatory, algesic and allergic diseases, the author examined the analgesic effect by acetic acid reaction, studied the anti-inflammatory effect through the experiments of the protein thermo-denaturation and circumscribed edema. Besides researched the anti-allergic effect through the vascular permeability response to Chemical Mediator and the delayed type hypersensitivity response to Picryl Chloride. The obtained results were as follows ; 1. In the analgesic effect of Hyunggyeyungyotang and Kamihyunggyeyungyotang extract by acetic acid method, both of the sample groups showed the analgesia, but didn't show useful effect. 2. In the anti-inflammatory effect on the protein thermo-denaturation, the sample groups revealed the inhibitory effect in proportion to concentration as compared with the control group. 3. In the inhibitory action on circumscribed edema induced by Caraggeenin, both of Hyunggyeyungyotang and Kamihyunggyeyungyotang administration showed the significant effect after 4 hours in comparison to the control group. 4. In the delayed type hypersensitivity response to Picryl Chloride, both of the sample groups revealed the significant effects. 5. Both of the sample groups decreased the vascular permeability induced by Histamine in comparison with the control group, but the significancy was admitted in only Hyunggyeyungyotang administration. According to above results, Hyunggyeyungyotang and Kamihyunggyeyungyotang are considered to be used for treament of the inflammatory diseases including sinusitis.

• 한국식품영양과학회지
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• 제19권4호
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• pp.349-355
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• 1990

Actomyosine prepared from Yellowtail fish(seriola quinqueradiata) were stored at $0^{\circ}C$(ice-cooling) -3.5$^{\circ}C$(partial freeaing) and -2$0^{\circ}C$(freezing) Another actomyosin samples were prepared from the fish previously stored at the temperatures for a week as the maximum .Remaining activity of {{{{ {Ca }^{2+ } }}}}-and {{{{ {Mg }^{2+ } }}}}- dependent adenosine triphosphatase(ATPase) activity was measured fronm the actomyosin preparations. Specific activity of {{{{ {Mg }^{2+ } }}}}-ATPase of actomy-osin before storagew was 0.253$\mu$ mole pi/min/mg of protein and it was 1.5 times higher than that of {{{{ {Ca }^{2+ } }}}} -ATPase. The enzyme activities were markedly decreased during early period of storage. However no significant differences in the enzyme activity were revealed among the samples stored at different temperature. The enzyme of actomyosin prepared from the fish previously stored at the temperatures for a week revealed an acitivity of 2-3 times higher than that of freezing. Apparent denaturation constant of {{{{ {Mg }^{2+ } }}}} -ATPase of actomyosin was between 0.810-1.139 per day and it was about 1.5 times hgiher than that of {{{{ {Ca }^{2+ } }}}} -ATPase. But the constant of {{{{ {Mg }^{2+ } }}}} ATPase of actomyosin extracted from the fist stored for a week at each temperature was between 0.176-0.356 per day. This constant was 4 times higher than that of {{{{ {Ca }^{2+ } }}}}- ATPase in frozen stored fish. It was presumed from these results that denaturation of ATPase is largely accorded to the structural changes of actomyosin.

• 한국수산과학회지
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• 제6권3_4호
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• pp.101-111
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• 1973

생 전복을 액체질소동결, 송풍동결, 반송풍동결, 정지공기동결의 네가지 동결방법으로 동결하여 $-20^{\circ}C$에서 3개월간 저장하여 동결방법에 의한 전복근육의 물성적 변화, 단백질의 변성 등을 검토하여 다음과 같은 결과를 얻었다. 1) 생전복의 사후경직은 다른 어류보다 빨랐다. 2) 압측시험치, 전단시험치는 액체질소동결을 제외하고는 동결방법에 의한 차는 없었고, 저장기간이 길어짐에 따라 약간 감소하였다. 3) 염가용성질소는 동결저장기간중 용출량이 완만하게 감소하였다. 4) 유리수의 유출량은 패주와 다리 사이에는 차가 있었고, 저장기간중 패주는 유리수가 증가하는 경향이었다. 5) Texture의 Panel test 결과 저장기간을 통하여 Texture가 감소하는 경향이었으며, Texture의 변화는 유리수의 변화(P<0.01), 단백질의 변성(P<0.05), 압축시험치의 변화(P<0.05)와 유의의 상관관계가 있었다. 본 논문을 지도하여 주신 동경수산대학 교수 전중 화부 박사 그리고 논문의 작성에 조언해 주신 부산 수산대학 식품공학과 이응호 박사님께 감사의 뜻을 표한다.

• Journal of Plant Biology
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• 제39권1호
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• pp.57-61
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• 1996

Thermal denaturation, reassociation kinectics 및 핵부피 측정 방법으로 고추 게놈의 염기 조성, kinetic component 및 게놈 크기를 조사하였다. 변성온도(Tm)에 근거한 고추 게놈의 염기 조성은 37% (G＋C)였으며, Cot 커브에 근거한 고추 게놈은 카피수가 10,754, 178 및 1이고 kinetic complexity가 $5.6{\times}10^{3}\;bp,\;1.9{\times}10^{6}\;bp\;및\;1.9{\times}10^{8}\;bp$인 3개의 성분이 게놈의 4.2%, 26% 및 65%를 차지하고 있었다. 이로부터 계산한 고추 게놈의 크기는 $1.25{\times}10^{9}\;bp/C$인데, 이는 핵부피($62.4\;\mu\textrm{m}^3/C$)로부터 계산한 $4.053{\times}10^{9}\;bp/C$의 약 33%에 해당한다.

• Journal of Dairy Science and Biotechnology
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• 제35권4호
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• pp.270-285
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• 2017

Among milk proteins, caseins are not subjected to chemical changes during heat treatment of milk; however, whey proteins are partially denatured following heat treatment. The degree of whey protein denaturation by heat treatment is decreased in the order of high temperature short time (HTST) > low temperature long time (LTLT) > direct-ultra-high temperature (UHT) > indirect-UHT. As a result of heat treatment, several changes, including variations in milk nitrogen, interactions between beta-lactoglobulin and k-casein, variations in calcium sulfate and casein micelle size, and delay of milk coagulation by chymosin action, were observed. Lysine, an important essential amino acid found in milk, was partially inactivated during heat treatment. Therefore, the available amount of lysine decreased slightly (1~4% decrease) after heat treatment, However, the influence of heat treatment on the nutritional value of milk was negligible. Nutritional value and nitrogen balance did not differ significantly between UHT and LTLT in milk. In conclusion, our results showed that heat treatment of milk did not alter protein quality. Whey proteins denatured to a limited extent during the heat treatment process, and the nutritional value and protein quality were unaffected by heat treatment.