• Applied Biological Chemistry
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• v.36 no.6
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• pp.469-475
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• 1993

In order to elucidate the metabolism of the systemic insecticide, phosphamidon(2-chloro-2-diethylcarbamoyl-1-methylvinyl dimethyl phosphate), treated to pine trees against pine leaf gall midges (Thecodiplosis japonensis Uchida et Inouye), $[vinyl,\;carbonyl-^{14}C]$phosphamidon was implanted into the trunks of 10-year-old Korean red pine (Pinus densiflora Sieb. et Zucc.) and Japanese black pine (Pinus thunbergii Parl.), respectively. This chemical was degraded very quickly in pine trees after implanting, as confirmed by TLC/autoradiography of the extracts of pine needles. Phosphamidon metabolites in phosphate buffer extracts of pine needles include the major metabolite, ${\alpha}-chloroacetoacetic$ acid diethyl-amide, ${\alpha}-chloroacetoacetic$ acid ethylamide, 3-hydroxy-N,N-d iethylbutanamide, acetoacetamide, and trimethyl phosphate. The metabolism within pine trees is expected to be similar to this. Based on these findings, it is believed that the major pathway leading to the metabolites would be related to the P-O-vinyl hydrolysis of the chemical structure.

• The Korean Journal of Nuclear Medicine
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• v.28 no.1
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• pp.112-123
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• 1994

${\gamma}$-Glutamyltransferase (GGT: E.C. 2.3.2.2.) is a glycoprotein enzyme which is involved in glutathione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be associated with hepatocarcinogenesis. We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and protein A sepharose CL-4B column chromatography. Using these mAbs we preformed enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study the distribution of GGT isozyme in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kindey GGT. These mAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

• Animal cells and systems
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• v.4 no.1
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• pp.63-70
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• 2000

The distribution of receptor subtypes for endothelin (ET) in the kidney of the freshwater turtle, Amyda japonica, was examined by quantitative in vitro receptor autoradiography using iodinatd mammalian type ET-1 ($^125$/I-ET-1)as a radiolabeled ligand. Specific $^125$/I-ET-1 bindings were localized to renal tubules, renal arteries and ureter with binding densities of 111.21 $\pm$ 19.14, 182.13$\pm$10.57 and 219.46$\pm$12.83 amol/$mm^2$. respectively. Binding dissociation constants in renal tubules, renal arteries and ureter were 1.05 $\pm$ 0.63, 2.03 $\pm$0.56 and 1.70$\pm$0.47nM, respectively. Receptor subtypes for ET in the kidney were characterized by competition with BQ 123 and BQ 788 as specific antagonists for ET receptors, type A (ET$_A$ ), and type B (ET$_B$) subtypes, respectively. Specific $^125$/I-ET-1 bindings in renal arteries and ureter were potently inhibited by BQ 123 in a dose-dependent manner, whereas BQ 788 was not in competing for specific $^125$/I-ET-1 bindings in this structure. However, specific $^125$/I-ET-1 bindings in renal tubules were inhibited more potently by BQ 788. Therefore, these results indicate that specific ET receptors are localized in renal tubules, renal arteries and the ureter of the freshwater turtle. Results also suggest that the predominant ET receptor subtypes are like the ETA receptor in renal arteries and ureter, and like the ET/$_A$ receptor in the renal tubule.

• Journal of Sericultural and Entomological Science
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• v.28 no.1
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• pp.30-36
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• 1986

Polyacrylamide gel electrophoresis and autoradiography were applied to investigate the developmental profiles of the major hemplymph proteins (MHPs) and their biosynthesis. In addition, some biochemical methods were also used to isolate and purify the MHPs. The obtained results are summarized as follows. 1. MHP-a began to appear from the 2nd day of the fourth-instar larva while MHP-b and -c were detected first on the 1st day of the fifth-instar larva. All these proteins, however, showed a drastic increase in concentration at the 2nd day of the fifth-instar larva. 2. MHP-b and -c were synthesized in fat body on early day of the fifth-instar larva, but the possibility of MHP-a synthesis in fat body was excluded. 3. MHP-b was isolated and purified by heat-treatment (6$0^{\circ}C$), gel filtration on Sephadex G-100 and column chromatography on DEAE-cellulose and CM-cellulose. Purified MHP-b showed a single band on polyacrylamide gel- and SDS-polyacrylamide gel electrophoresis.

• Journal of Ginseng Research
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• v.47 no.1
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• pp.74-80
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• 2023

Background: Although many studies have evaluated the efficacy and pharmacokinetics of Korean Red Ginseng (KRG) components (Rg1, Rb1, Rg3, Rd, etc.), few have examined the in vivo pharmacokinetics of the radiolabeled components. This study investigated the pharmacokinetics of ginsenosides and their metabolite compound K (CK), 20(s)-protopanaxadiol (PPD), and 20(s)-protopanaxatriol (PPT) using radioisotopes in rat oral administration. Methods: Sprague-Dawley rats were dosed orally once with 10 mg/kg of the tritium(3H) radiolabeled samples, and then the blood was collected from the tail vein after 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24, 48, 96, and 168 h. Radioactivity in the organs, feces, urine, and carcass was determined using a liquid scintillation counter (LSC) and a bio-imaging analyzer system (BAS). Results and conclusion: After oral administration, as the ³H-labeled ginsenosides were converted to metabolites, C_max and half-life increased, and T_max decreased. Interestingly, Rb1 and CK showed similar values, and after a single oral administration of components, the cumulative excretion ratio of urine and feces was 88.9%-92.4%. Although most KRG components were excreted within 96-168 h of administration, small amounts of components were detected in almost all tissues and mainly distributed to the liver except for the digestive tract when observed through autoradiography. This study demonstrated that KRG components were distributed to various organs in the rats. Further studies could be conducted to prove the bioavailability and transmission of KRG components to confirm the mechanism of KRG efficacy.

• Journal of Sericultural and Entomological Science
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• v.37 no.1
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• pp.68-73
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• 1995

The toxic effect of Bacillus thuringiensis var, kurstaki $\delta$-endotoxin was determined in the midgut, fat body and Malpighian lobules from larvae of Hyphantria cunea using the au-toradiography and Western blot methods. Bt $\delta$-endotoxin crystals inhibited protein synthesis and elongation factor-2 activity of all tissues tested.

• Journal of Sericultural and Entomological Science
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• v.36 no.2
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• pp.119-123
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• 1994

By in vivo labelling of AMHP using[35S]-methionine, fat body culture and immunological analysis, it is proved that Bombyx adult fat body synthesizes 18K and 20K subunits of AMHP and releases them into haemolymph. Also these peptides are assembled to form native AMHP in the adult haemolymph.

• YAKHAK HOEJI
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• v.40 no.4
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• pp.442-448
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• 1996

Mitomycin C(MMC) has been used as an anticancer drug and behaves as an alkylating agent forming covalent cross-link between complementary strands of double strand DNA. The purpose of this research was to determine number of DNA adducts, formed in vivo by Mitomycin C, in mouse organs. DNAs from liver, lung, brain and pancreas were isolated and used for $^{32}P$-postlabelling. The labeled nucleotides were separated by 2D-TLC and subjected to autoradiography. Numbers of MMC-DNA adducts were 9,9,5,4 in liver, pancreas, lung and brain, respectively.

• The Journal of the Korean Society for Microbiology
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• v.3 no.1
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• pp.51-65
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• 1968

The site of the synthesis of Japanese encephalitis virus(JEV) in the actinomycin-treated and infecter PS Y15 cells(a porcine kidney stable cell line) was observed by the immunofluorescent antibody technique, acridine orange staining, and the autoradiographic analysis. In the parallel studies by immunofluorescent technique and acridine orange staining it the infected cells, Viral protein(as an antigen) and viral RNA were detected at the same site of cytoplasm. In the autoradiographic analysis, the cytoplasmic labeling of $^3H$-uridine was due to the synthesis of JEV-RNA, while the nucleolus and nucleus were not involved. In the autoradiographic studies on the secton of infected cells, the $^3H$-uridine was frequently incorporated around the cytoplasmic vacuoles. This localization of labeling agreed with the site of acridine orange positive granules. The results suggest that the syntheses of the viral RNA and viral protein occurred in the similar site of cytoplasm of the infected cells, and also the virus particles seem to be assembled in the sites of the viral RNA and protein syntheses.

• Journal of Microbiology and Biotechnology
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• v.2 no.2
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• pp.122-128
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• 1992

Post-translational modifications of the nucleocapsid protein of bovine coronavirus (Quebec strain) were investigated. Coronavirions were radiolabelled in vivo with inorganic $[^{32}P]$orthophosphate and analysed by SDS-PAGE, followed by autoradiography. A single polypeptide with a migration rate of 55 KDa was identified by metabolic phosphate labelling, demonstrating that the nucleocapsid protein of bovine coronavirus was a phosphoprotein. A gene encoding the nucleocapsid protein was inserted immediately downstream from the polyhedrin promoter of Autographa californica nuclear polyhedrosis baculovirus. Spodoptera frugiperda cells infected with this recombinant baculovirus synthesized a 55 KDa polypeptide, as demonstrated by immunoprecipitation with anti-nucleocapsid monoclonal antibody. The recombinant nucleocapsid protein synthesized in Spodoptera cells could also be labelled by $[^{32}P]$orthophosphate. Phosphoamino acid analysis showed that both serine and threonine residues were phosphorylated in authentic, as well as in recombinant nucleocapsid proteins, with a relative phosphorylation ratio of 7:3. Our studies demonstrated that the nucleocapsid protein of bovine coronavirus was a serine and threonine-phosphorylated protein and that Spodoptera insect cells were able to properly phosphorylate the relevant foreign proteins.