• 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.

• 한국전산유체공학회지
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• 제3권1호
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• pp.82-88
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• 1998

As a par of a project related to the development of the design algorithm of a compact heat exchanger for the application of the electronic home appliances, the effect of the discreteness of the airflow boundary generated on the cooling fin surface on the heat transfer and pressure drop characteristics of the heat exchanger was studied numerically. In general, there are two critical design parameters seriously considered in the design of the heat exchanger; heat transfer rate(Q) and pressure drop coefficient(C/sub p/). Even though the higher heat transfer rate with lower pressure drop characteristics is required in a design of the heat exchanger, it is not an easy job to satisfy both conditions at the same time because these two parameters are phenomenally inversely proportional. To control the boundary layer thickness and its length along the streamline, the surface of the flat fin was modified to accelerate the heat transfer rate on the fin surface. To understand the effect of the discreted fin size(S/sub w/) and its location(S/sub h/) on the performance of the heat exchanger in the airflow field, the flat fin was modified as shown in Fig. 1. From this study, it was found that the smaller and more number of slits on the fin surface showed the higher energy diffusion rate. It means that the discreteness of the boundary layer is quite important on the heat transfer rate of the heat exchanger. On the other hand, if the fin surface configuration is very complex than needed, higher static pressure drop occurs than required in a system and it may be a reason of the induced aerodynamic noise in the heat exchanger.

• Biomolecules & Therapeutics
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• 제11권3호
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• pp.163-168
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• 2003

Pharmacokinetics of eupatilin (an active components of $Stillen^{\circledR}$, a new antigastritic agent) were investigated after both intravenous and oral administration at a dose of 30mg/kg to rats. After intravenous administration, the plasma concentrations of unchanged eupatilin declined rapidly with a mean terminal half-life of 0.101 h. Eupatilin was eliminated fast in rats; the total body clearance was 121 mL/min/kg. Eupatilin was mainly metabolized in rats; the percentage of intravenous dose of eupatilin excreted in 24 h urine and feces as unchanged eupatilin was only 2.5 and 0.919％, respectively. Eupatilin was mainly metabolized to form its glucuronide conjugate; after intravenous administration, 15.9 and 51.7％ of intravenous dose was excreted in 24 h urine and feces, respectively, as eupatilin plus its glucuronide. After oral administration, the absolute bioavailability was only 3.86％ based on $AUC_{0-24h}$ of eupatilin plus its glucuronide. Approximately 68.5％ of oral dose was not absorbed from the entire gastrointestinal tract. Therefore, it could be concluded that the superior effect of eupatilin in experimental animal models of gastric ulcer and inflammatory bowel disease after oral administration could be due to the local action of eupatilin. Further pharmacokinetic studies to elucidate the local action of eupatilin are required.

• Parasites, Hosts and Diseases
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• 제45권1호
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• pp.19-26
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• 2007

This study describes the isolation of a Toxocara canis species-specific excretory-secretory(ES) antigen and the development of an enzyme-linked immunosorbent assay(ELISA) based on this antigen. Analysis of the ES antigens of T. canis, Toxocara vitulorum, Ascaris lumbricoides and Necator americanus larval antigen was performed by SDS-PAGE followed by western blotting. A 57 kDa T. canis-specific antibody fraction(TcES-57) was identified by western blotting and labelling with anti-Toxocara antibodies(from experimental rabbits and human patients) and tracing with anti-human or anti-rabbit peroxidase conjugate. No protein fraction of 57 kDa was detected in ES or larval antigens collected from T. canis, T. vitulorum, A. lumbricoides and N. americanus. Using TcES-57, a specific anti-serum was produced in rabbits and a double sandwich ELISA was developed. This test was validated using known seropositive sera from toxocariasis patients, sera from A. lumbricoides or N. americanus patients, and 50 serum samples from cats. These tests revealed that TcES-57 antigen is specific to T. canis infection and does not cross react with sera of other related infections. Thus, ELISA based on TcES-57 antigen was proven to be an effective tool in the diagnosis of toxocariasis and studies on the role of T. canis in the epidemiology of human toxocariasis.

• 한국강구조학회 논문집
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• 제11권2호통권39호
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• pp.213-222
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• 1999

ALFD방법은 연속합성판형교의 과재하중과 최대하중에 대한 강도검토에 있어서 실제적인 거동을 나타낼 수 있는 탄소성해석법이다. 주어진 하중으로 인한 최대 정, 부모멘트단면에서의 항복은 활하중이 통과한 후에 잔류하는 소성회전을 일으킨다. 또한 제조시의 잔류응력으로 인하여 이론적인 항복모멘트 이하에서도 소성변형을 일으킨다. 이러한 국부항복은 다음에 작용되는 과재하중하에서 탄성화되어 정의 자생모멘트를 유발한다. 본 연구에서는 지점과 최대 정모멘트 단면에서의 단위소성회전각으로 인한 자생모멘트를 공액보법과 3연모멘트법에 의하여 구하였고, 9개의 설계경간을 지점단면을 감소시켜가면서 본 연구에서 개발한 전산프로그램에 의하여 연속관계와 모멘트-비탄성회전각 실험곡선과 일치하는 자생모멘트를 구하였다. 또한 한국도로교시방서에 준하여 비조밀단면을 갖는 3경간 연속합성판형교의 평가를 수행하였다.

• 대한바이러스학회지
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• 제28권4호
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• pp.383-391
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• 1998

A murine monoclonal antibody (mAb) specific for the envelope glycoprotein gp120 of human immunodeficiency virus type-I (HIV -1) was chemically coupled to pokeweed antiviral protein (PAP) from Phytolacca americana. The immunotoxin was purified by FPLC using S200 colum. The purified immunotoxin efficiently bound to HIV-infected T cells as evidenced by fluorescenceactivated cell sorter analysis. The immunotoxin selectively killed human T lymphoid lines infected with $HIV-1_{IIIB}$ at less than 250 pM of the immunotoxin cells, while PAP or mAb alone did not have any significant effect on infected cells. The uninfected control T cell lines were not affected. Human cells infected with HIV-2 or other HIV-1 strains were not killed, suggesting that the killing depends completely on the antibody used for coupling. These in vitro results suggest that the PAP-mAb conjugate may be used to selectively remove cells expressing viral antigens from individuals infected with HIV.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.109-109
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• 1997

Inhibitory effect on DNA topoisomerase-I, rate of glutathione conjugation and cytotoxicity of naphthoquinone derivatives were correlated. During 5 min exposure of the derivatives to glutathione (GSH), it was found that 14% of 5,8-dimethoxy-1,4-naphthoquinone(DMNQ) was converted into a GSH-conjugate, whereas 5,8-dihydroxy-1,4-naphthoquinone(DHNQ) did not interact with GSH, implying that DMNQ exerted higher electrophilicity than DHNQ. However, DHNQ (IC$\_$50/, 0.15 ${\mu}$M) showed stronger cytotoxicity in L1210 cells than DMNQ(IC$\_$50/, 0.45 ${\mu}$M). The stronger cytotoxicity of DHNQ, compared to DMNQ, could be ascribed to more rapid redox cycling. Both naphthoquinones (IC$\_$50/, 60-65 ${\mu}$M) exhibiting about the same inhibitory effect on DNA topoisomerase-I were more potent than 1,4-naphthoquinone(1,4-NQ, IC$\_$50/, 134 ${\mu}$M). Thus, 5,8-oxy groups in the structure seem to be important for the inhibition of the enzyme. DMNQ showed a broader dose range while maintaining a good antitumor activity against S-180 fluid tumor. For these reasons, DMNQ was taken as useful pharmacophore for structural modification. Introduction of 1-hydroxyalkyl groups at C-2 of DMNQ lowered all of the activities mentioned above, while acetylation of 1-hydroxyalkyl moiety enhanced the activities by 4-5 times. Introduction of the same side chains at C-6 exhibited stronger activities than 2-substituted ones. Based on these results it was suggested that the quinonoid moiety in 6-substituted DMNQ was more exposed to cellular nucleophiles such as DNA, thiols of enzymes and so on. The synthesis of DHNQ or DMNQ derivatives are going on, and the corelationship between structure-activity will be discussed.

• Applied Biological Chemistry
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• 제36권1호
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• pp.7-10
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• 1993

ELISA법을 zearalenone 생성균주 검색에 응용하였다. 먼저 zearalenone에 대한 항체를 500배 희석한 후 microtiter well에 $125\;{\mu}l$씩 주입하여 $40^{\circ}C$에서 overnight시켜 coating하고, 시료용액과 enzyme을 $37^{\circ}C$에서 30분간 반응시켰다. 배양후 washing buffer로 6회 세척한 다음 plate에 2,2'-azino-di-3-ethyl-benzthiazoline sulfonic acid(ABTS) 용액을 $100\;{\mu}l$씩 첨가하여 15분간 발색시킨 다음 반응 정지액 $100\;{\mu}l$씩을 가해 반응을 정지시키고 ELISA Reader로서 흡광도(410 nm)를 측정하였다. 그 결과 zearalenone 생성이 확인된 19 균주 중 분리균 R-5, C-46 및 S-134가 50 ng/ml 이상의 zearalenone을 생성하였다.

• 한국측량학회지
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• 제14권1호
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• pp.9-16
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• 1996

본 연구에서는 상호표정과 사진기준점측량을 위한 새로운 매칭방법을 제안하였다. 이 방법은 수작업에 의한 점이사와 관측을 대신하여 디지탈 항공사진에서 연결점과 접합점으로 사용할 수 있는 공액점들을 매칭한다. 제안된 매칭방법에는 다음과 같은 세 가지의 독특한 단계가 포함되어 있다. 첫째 단계는 interest point를 찾고 이들을 매칭후보점으로 사용한다. 둘째 단계는 중복찰영된 서상에서 표준상관계수법에 의해 양방향으로(왼쪽에서 오른쪽으로 오른쪽에서 왼쪽으로) 매칭을 실시한다. 세째 단계는 양매칭방향에서 일치성이 있는 쌍들을 선택한다. 제안된 매칭방법에 기초한 컴퓨터 프로그램을 개발하고, 전면지상기준점측량을 실시한 디지탈 항공사진을 이용하여 제안된 방법을 사용하였을 때의 신뢰성과 위치정확도를 조사하기 위한 시험을 하였다. 시험 결과 제안된 매칭방법은 다른 매칭방법보다 불량하게 매칭된 쌍들을 제거하는 데 더 효과적이었으며 , 따라서 이 방법을 상호표정 및 사진기준점측량에 사용할 경우 매우 매우 신뢰성이 높다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권6호
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• pp.743-749
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• 2004

The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.