• Title/Summary/Keyword: p-conjugate.

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Application of Micro-BLISA in Serodiagnosis of Fascioliasis in Cattle (간질증의 효소면역학적 진단)

  • Lee, Jae-Gu;Baek, Byeong-Geol;Lee, Sang-Bok
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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    • pp.95-101
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    • 1985
  • Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

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Studies on the Biological Behaviors of Taxol Derivatives (Taxol 유도체들의 생물학적 거동에 관한 연구)

  • Awh, Ok-Doo;Yoo, Dae-Wung;Im, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.4
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    • pp.440-451
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    • 1997
  • This study was designed to prospect the $^{111}In$-labelled paclitaxel as tumor imaging agent. In order to provide a taxol molecule with a functional group which is able to chelate In-111, taxol-DTPA conjugate and 2'-hemisuccinyltaxol were synthesized by esterification of taxol at C-2'on C-13 carbon with DTPA anhydride and succinic anhydride, respectively. Synthesis yield of the taxol derivatives was 34% for taxol-DTPA and 80% for 2'-hemisuccinyltaxol. Cytotoxicity of the taxol derivatives were measured by MTT method toward cell lines HT29, B16, P388, and CT26. The cytotoxic activities of the taxol derivatives were maintained, although less active than taxol. Radiolabelling of the taxol derivatives were proceeded directly with $^{111}InCl_3$ or indirectly with $^{111}In$-citrate(ligand-exchange method). The ligand-exchange method was not suitable because some precipitates appeared during the reaction. On the contrary, by direct radiolabelling method, we were able to obtain taxol-DTPA-$^{111}In$ in 100% radiochemical yield. However, 2'-hemisuccinyltaxol was not labelled by both methods. Yield and radiochemical purity of the radiolabelled com-pound were determined by HPLC, paper chromatography and instant thin layer chromatography. Taxol-DTPA-$^{111}In$ was characterized to be hydrophilic by lipophilicity test, and nearly non-adhesive to HT29, B16, P388, and CT26 by cell binding affinity test. Binding affinity of the taxol-DTPA-$^{111}In$ complex to serum proteins was also examined by protein precipitation with 30% trichloroacetic acid. The results showed that 30% of the taxol-DTPA-$^{111}In$ complex binds with serum proteins.

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The Studies on the Change of Lipid Phosphorus and Influence of Irradiation with the Origination of Silkworm Eggs (Bombyx mori L.) (가잠난 발생과정에서 Lipid Phosphorus의 변동과 방사선영향에 관한 연구(제일보))

  • 김원경;임영우
    • Journal of Sericultural and Entomological Science
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    • v.13 no.1
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    • pp.31-34
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    • 1971
  • The pupa (7th. day after mounting) were irradiated for foully minutes with 1,000${\gamma}$-ray and the copulation was done under the various conditions when the pupa grew to be imagos. The silkworm eggs that the imagos laid were divided into many groups by five, fifteen, foully-five, ninety, seven-hundred, twenty and twenty-four hundred minutes eggs respectively and the lipid phosphorus in the each group of eggs was analyzed as follows. 1. The quantity of 928${\gamma}$/100mg dry powder (D. P.) was shown in the group of the fourt-five minutes eggs whose ovum nucleus and sperm nucleus were not conjugate in the control C (male and female). The number of the greatest quantity of 944${\gamma}$/100mg D.P. after the syngamy of ovum nucleus and sperm nucleus while the smallest quantity was 768${\gamma}$/100mg D.P. in the group of ninety minutes eggs. 2. Not much difference could be seen the copulation group of female irradiation and male control and the copulation group of male irradiation and female control compared to the both male and female control group, but the influence could be seen in the group of irradiated female pupas. 3. The great difference could be seen in the group of irradiated male and female pupa compared to the control group. The five minutes eggs showed the smallest quantity of 536${\gamma}$/100mg D.P. and the quantity of lipid phosphorus rapidly began to increase from the fifteen minutes eggs. The quantity amounted to 1,082${\gamma}$/100 mg D.P. in the case of seven hundred and twenty minutes eggs while the quantity decreased in the twenty-four hundred minutes eggs which showed 912${\gamma}$/100 mg D.P. The general result obtained in the present study was that the irradiated male and female pupa were greatly influenced before and after the syngamy of gamete.

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Liver Targetability and Pharmacokinetics of $[^3H]$-Methotrexate-Lactosaminated Bovine Serum Albumin Conjugates ($[^3H]$-메토트렉세이트-락토오스아미노화한 소 혈청 알부민 공유결합체의 간표적성 및 체내동태)

  • Kim, Chong-Kook;Lee, Woong-Doo;Park, Ho-Koon
    • YAKHAK HOEJI
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    • v.36 no.6
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    • pp.591-597
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    • 1992
  • The organ distribution of $[^3H]$-methotrexate-lactosaminated bovine serum albumin conjugates ($[^3H]$-MTX-LBSA) was investigated to examine their role as a liver-specific anticancer drug. Synthesis of lactosaminated bovine serum albumin(LBSA) with BSA, lactose and sodium cyanoborohydride through reductive amination was followed by its conjugation with methotrexate (MTX) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), thereby synthesizing [$[^3H]$-MTX-LBSA conjugates. Organ distribution and plasma elimination profiles were studied in male Wistar rats after intravenous injection of [$[^3H]$-MTX-LBSA conjugates. The fates of $[^3H]$-MTX and the $[^3H]$-MTX-BSA conjugates´fates were also investigated for comparison. The results showed that the plasma level of $[^3H]$-MTX-LBSA conjugates declined more rapidly than those of $[^3H]$-MTX-BSA and their liver concentration was significantly higher than those of other treatment (p<0.01). In addition, their uptake compared to the amount taken up by the liver (1 : 33.1 at 10 min, 1 : 24.1 at 120 min). All these suggested that MTX-LBSA conjugate is one of the drug delivery system (DDS) that is advanced in concentrating MTX in the liver and minimizing the renal toxicity of MTX.

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Effect of Boundary Layer Generated on the fin surfaces of a Compact Heat Exchanger on the Heat Transfer and Pressure Drop Characteristics (컴팩트형 열교환기의 핀 표면에서 발생하는 경계층이 열교환기의 전열 및 압력강하 특성의 변화에 미치는 영향에 관한 수치해석적 연구)

  • KIM Chul-Ho;Jung Ji-Yong
    • Journal of computational fluids engineering
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    • v.3 no.1
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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.

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Characterization of a Toxocara canis species-specific excretory-secretory antigen(TcES-57) and development of a double sandwich ELISA for diagnosis of visceral larva migrans

  • Iddawela, R.D.;Rajapakse, R.P.V.J.;Perera, N.A.N.D.;Agatsuma, Takeshi
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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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.

Human Immunodeficiency Virus-Infected T Cells Are Selectively Killed by Monoclonal Anti-gp120 Antibody Coupled to Pokeweed Antiviral Protein (섬자리공 유래 항바이러스 단백질과 항체 복합체를 이용한 HIV-1 감염세포의 선택적 제거)

  • Kang, Mi-Ran;Kim, Yoon-Kyu;Hong, Hyo-Jeong;Cho, Myung-Hwan;Shin, Hyung-Sik;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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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.

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Cloning of Chicken Microsomal Glutathione S-transferase 1 Gene (MGST1) and Identification of Its Different Splice Variants

  • Wang, X.-T.;Zhang, H.;Zhao, C.-J.;Li, J.-Y.;Xu, G.-Y.;Lian, L.-S.;Wu, C.-X.;Deng, Xuemei
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.155-161
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    • 2009
  • Mammal microsomal glutathione transferase 1 (MGST1) can conjugate many toxic or carcinogenic substances and depress oxidative stress. In this study, Chicken MGST1 and its variants were cloned for the first time and were composed of 956 or 944 nucleotides. The 12 nt deletion in the exon 2 did not alter the GT-AG rule and the ORFs for the two MGST1 variants were the same, which both comprised 465 nucletides and encoded a peptide with 155 amino acids. It was found that the two different splice variants identified using RT-PCR expressed in all three organs investigated of Dwarf Brown Chicken, namely liver, spleen and shell gland. Moreover, the expression level of MGST1 mRNA in the liver of Dwarf Brown chickens was the highest (p<0.01), and there were no significant differences between the spleen and the shell gland. These results provide a base for studying the biological function of Chicken MGST1.

Production of the Monoclonal Antibodies to the Escherichia coli Heat-Stable Enterotoxin (대장균의 내열성장독소 측정법개발을 위한 단세포군항체의 생산)

  • Chang, Woo-Hyun;Lee, Woo-Kon;Kim, Suck-Yong;Park, Jung-Bum
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.377-392
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    • 1987
  • Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.

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Rapid diagnosis of Pseudotuberclosis in yellowtail (Seriola quinqueradiata) by immunofluorescent antibody technique (면역형광항체법(免疫螢光抗體法)에 의(依)한 방어의 유결절증(類結節症) 신속(迅速) 진단(診斷))

  • Bang, Jong-Deuk;Jung, Sung-Hee;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.3 no.1
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    • pp.11-19
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    • 1990
  • Yellow tail (Seriola quinqueradiata) infected by Pasteurella piscicida have been occurred to mass mortality without showing apparent surface lesions in cage culture farms. In this case, it is necessary to consider countermeasure by rapid diagnosis of infected fish. The purpose of the present study was to investigate usefulness of the direct fluorescent antibody technique(FAT) for rapid diagnosis of pseudotuberclosis of cultured yellowtail caused by P. piscicida. Antibody produced by inoculating rabbit with formalin killed pseudotuberclosis bacteria antigen(strain KNP-2). Immunoglobulin-G(IgG) was purified from antisera by DEAE-cellulose column chromatography and conjugate with fluorescein isothiocyanate. Fluorescein-labeled antisera was purified by sephadex G-25 gel column chromatography. The fluorescein/protein molar ratio of labeled antisera was determined as 8.8-9.5. Diagnosis of cultured yellowtail was examined in cage culture farms which located in Tongyung, kyungnam from July to October 1990. The causative bacteria of pseudotuberclosis could be detected within two hours after the specimens were transferred to the laboratory for FAT, and it showed that FAT could be adapted as a rapid and accurate diagnostic method of pseudotuberclosis in yellowtail.

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