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Biomedical subjects

K Terao

Publications and source records attributed to K Terao.

At least 163 records · Page 9Linked to original sources

The effects of naturally occurring bisdihydrofuran ring-containing mycotoxins on cultured chick embryonal hepatocytes.

The cytotoxicity of 16 naturally occurring mycotoxins which contain the bisdihydrofuran ring was evaluated by electron-microscopy and radioautography in a primary culture of chick embryonal hepatocytes. A series of nucleolar changes occurred, including the appearance of microsegregation (stage I), concentration of nucleolar structure (stage II), formation of macrosegregation, fragmentation of nucleoli, and the appearance of homogeneously granular nucleoli (stage III). These nucleolar changes correlate well with the inhibition of RNA synthesis calculated from the results of radioautography. The compounds containing a 2,3-double bond in the bisdihydrofuran ring were most toxic. However, it was clear that there was a remarkable difference between mycotoxins of angular type (group A) and those of linear type (group C). Therefore, not only the active site in the molecule but also the shape of the molecule may play an important role in the cytotoxicity of these mycotoxins.

Aflatoxins↗

Cross-linking of L5 protein to 5 S RNA in rat liver 60-S subunits by ultraviolet irradiation.

After rat liver 60-S ribosomal subunits were irradiated with ultraviolet light at 254 nm, they were treated with EDTA and then subjected to sucrose density-gradient centrifugation to isolate 5 S RNA-protein complex. When 5 S RNA-protein was analyzed by SDS-acrylamide gel electrophoresis which dissociated noncovalent 5 S RNA-protein, two protein bands were observed. The one showed a slower mobility than the protein band (L5) of 5 S RNA-protein from non-irradiated 60 S subunit and the other showed the same mobility as L5 protein. Since the former band was shown to be specific to ultraviolet-irradiation, it was considered as cross-linked 5 S RNA-protein. After the two protein bands were iodinated with 125 I, labeled protein was extracted and treated with RNAase. Thereafter, it was analyzed by two-dimensional acrylamide gel electrophoresis, followed by autoradiography. The results indicate that the protein component of cross-linked 5 S RNA-protein is L5 protein (ribosomal protein; these proteins are designated according to the proposed uniform nomenclature. The correlation between that and our nomenclature was reported by McConkey et al. (1979) Mol. Gen. Genet. 169, 1-6. They also confirm the results previously reported (Terao, K., Takahashi, Y. and O(gata, K. (1975) Biochim. Biophys. Acta 402, 230-237).

Animals↗

Identification of neighbouring protein pairs in the rat liver 40-S ribosomal subunits cross-linked with dimethyl suberimidate.

(1) The 40-S ribosomal subunits of rat liver were treated with a bifunctional cross-linking reagent, dimethyl suberimidate. Cross-linked protein-protein dimers were separated by two-dimensional acrylamide gel electrophoresis. The stained cross-linked complexes within the gel were radioiodinated without the elution of proteins from the gel and were cloven into the original monomeric protein constituents by ammonolysis. The proteins in each dimer were finally identified by two-dimensional acrylamide gel electrophoresis of the cloven monomeric proteins, followed by radioautography of the stained gel. (2) The molecular weights of cross-linked complexes were determined by SDS-polyacrylamide gel electrophoresis and were compared with those of their constituent proteins. (3) The following dimers were proposed from these results: S3-S12 (S3 or S3a-S11), S4-S12 (S3b-S11, S5-S7 (S4-S6), S5-S22 (S4-S23 or S24), S6-S8 (S5-S7), S8-S16 (S7-S18), S17-S21 (S16--S19) and S22A-S22B (S23-S24), designated according to our numbering system [1]. The designations according to the proposed uniform nomenclature [2] are described in parentheses.

Acetates↗

Stimulation by aminoacyl-tRNA of the GTPase and ATPase activities of rat liver 5S RNA protein particles in the presence of EF-2.

The GTP- and ATP-hydrolyzing activities of the rat liver 5S RNA-L5 (according to the proposed common nomenclature (1) protein complex designated as 5S RNP were stimulated by pig liver elongation factor 2 (EF-2) plus aminoacyl-tRNA, both of which were required for the stimulation. The stimulative effect of aminoacyl-tRNA on GTP hydrolysis by the complete system containing 5S RNP and EF-2 was dependent on the concentration of aminoacyl-tRNA. Aminoacyl-tRNA also stimulated ATP hydrolysis by the complete system but did not stimulate the ATP hydrolysis by 5S RNP alone or EF-2 alone. While deacylated tRNA stimulated the ATPase activity of the complete system, it had no effect on the GTPase activity. Preincubated tRNA lacking the 3'-terminal CCA moiety had little effect on the GTPase and ATPase activities of the complete system. Fusidic acid inhibited the GTPase and the ATPase activities of the complete system with and without aminoacyl-tRNA, although the extent of the inhibition was larger in the presence of aminoacyl-tRNA. These results suggest that 5S RNP may be a component of the GTPase center of rat liver 60S subunits.

Adenosine Triphosphatases↗

Identification of neighboring protein pairs in rat liver 60S ribosomal subunits cross-linked with dimethyl suberimidate or dimethyl 3,3'-dithiobispropionimidate.

Protein-protein neighbors in rat liver 60S ribosomal subunits were investigated by using two bifunctional imidoesters; dimethyl suberimidate (DMS) and dimethyl 3,3'-dithiobispropionimidate (DTP). 1. Complexes cross-linked with DMS were separated by two-dimensional acrylamide/urea gel electrophoresis. Each complex in the gel was labeled with 125I (1) and was cleaved into the original monomeric protein constituents of ammonolysis. The products were analyzed by two-dimensional acrylamide/urea gel electrophoresis, followed by radioautography of the stained gel. Eight protein pairs are proposed according to our numbering system (2): L1-L3(L3-L5), L2-L21 (L4-L26), L5-L13 (L7/L7a-L15), L5-L34 L7/L7a-L36), L6-L34 (L8-L36), L6-L32 (L8-L35), L12-L32 (L13/L13a-L35), and L18-L27 (L18-L27/L27a). The designations according to the proposed uniform nomenclature (3) are given in parentheses. 2. Complexes cross-linked with DTP were analyzed by acrylamide/SDS diagonal gel electrophoresis (4), and the molecular weights of the complexes and their components were determined. The monomer components of cross-linked pairs were labeled with 125I in the gel, and identified by two-dimensional acrylamide/urea gel electrophoresis followed by radioautography. Six pairs are proposed; L1-L5 (L3-L7/L7a), L1-L6 (L3-L8). L5-L19 (L7-/L7a-L21/L23/L23a), L13-L15 (L15-L14), L13-L16 (L15-L19), and L23-L27 (L30-L27/L27a). 3. Two pairs which were formed by protein-protein interaction without the use of cross-linking reagents were identified as L2-L4 (L4-L6) and L4-L30 (L6-L29).

Animals↗

Proteins of small subunits of rat liver ribosomes that interact with poly(U). II. Cross-links between poly(U) and ribosomal proteins in 40 S subunits induced by UV irradiation.

(1) When rat liver 40 S ribosomal proteins in 6 M urea were were mixed with poly(U) at an appropriate ratio, a precipitate was formed which was also insoluble in the sample solution for two-dimensional acrylamide gel electrophoresis. Analyses by two-dimensional acrylamide gel electrophoresis showed that S7 and S10 proteins (according to our numbering system) had disappeared selectively from the fraction soluble in 6 M urea. These two proteins were present in the fraction insoluble in 6 M urea, and became soluble in the sample solution after treating it with RNase. The results suggest that S7 and S10 proteins have strong affinities for poly(U). When rat liver 40 S subunits were incubated with poly(U), similar results were obtained. (2) After incubation of 40 S subunits with [3H]poly(U) and then with unlabeled poly(U), UV irradiation cross-linked poly(U) to the protein moiety of the 40 S subunit. When the protein fraction insoluble in the sample solution for two-dimensional electrophoresis was prepared from 40 S subunits cross-linked to poly(U) and then subjected to two-dimensional acrylamide gel electrophoresis after RNase treatment, S7 and S10 proteins were detected on the gel. In addition to the S7 protein spot, a triangular area spreading from the spot to the origin contained radioactivity. The results suggest that poly(U) is cross-linked to S7 protein and oligo(U) fragments bound to S7 protein affect its electrophoretic mobility. (3) Ribosomal proteins were prepared from 40 S subunits cross-linked to carrier-free [3H]poly(U) and analyzed by three-dimensional acrylamide gel electrophoresis (Terao, K. & Ogata, K. (1975) Biochim. Biophys. Acta 402, 214--229) after RNase treatment. It was found that S7, S6, and S15 proteins are cross-linked to poly(U). From the results of the present and preceding experiments it is concluded that S7 is the poly(U)-binding protein. The possibility that other proteins in 40 S ribosomal subunits interact with poly(U) is discussed.

Animals↗

Mixed cell agglutination reaction of ABO substances in the saliva and determination of human-type ABO blood groups in the cynomolgus monkey.

For detection of ABO substances in the saliva of the cynomolgus monkey, the mixed cell agglutination reaction (MCAR) gave specific and clear results with a very small amount of saliva. Anti-A and anti-B antibodies in the sera of the same species showed the clearest hemagglutination by the saline agglutination method. The combined use of both methods was demonstrated to be easily and accurately applicable to the determination of human-type ABO blood groups of the cynomolgus monkey.

ABO Blood-Group System↗

The effects of trichothecene toxins on the Bursa of Fabricius in day-old chicks.

The effects of T-2 toxin, Fusarenon X (FX), and Nivalenol (NV) on the bursa of Fabricius in the day-old chick were examined. After injections of 5 mg/kg of the mycotoxins into the residual yolk sac, cellular injury was limited at first to the smaller epithelial cells with coarse microvilli, which were located in the central portion of the follicle-associated epithelium. Subsequently necrosis spread out to the periphery. Degeneration and necrosis followed in the lymphoid cells in the lymphoid follicles. The other epithelial components in the follicle were relatively resistant to the mycotoxins. Both FX and NV were less potent than T-2 toxin, although the effects on the bursa of Fabricius were essentially the same. These findings suggest that the follicle-associated epithelium is clearly distinguished from other epithelial components in the bursa of Fabricius in day-old chicks.

Animals↗

The different effects of N-ethylmaleimide and iodoacetamide on the activity of rat liver 60S subunits for peptide bond elongation.

The activity of 60S subunits of rat liver ribosomes in poly(U)-dependent polyphenylalanine synthesis was inhibited by incubation with N-ethylmaleimide. However, when 60S subunits were incubated with iodoacetamide, their activity decreased only slightly. Furthermore, iodoacetamide-pretreated 60S subunits became insensitive to N-ethylmaleimide. Similar results were obtained for the activity of EF-2-dependent GTPase of 60S subunits. As a whole, the labeling patterns of ribosomal proteins on two-dimensional gel electrophoresis were similar for 60S subunits labeled with both 14C-labeled sulfhydryl reagents, although the extent of labeling of some proteins was somewhat different. These results indicate that the SH groups in the 60S subunits are not directly involved in the activities of the subunits described above.

Animals↗

Mesotheliomas induced by sterigmatocystin in Wistar rats.

Male Wistar rats were given repeated injections of sterigmatocystin (stg) and related compounds into the peritoneal cavity once a week for 23 weeks. Only stg alone produced mesothelioma in 20 of 40 rats given a total dose of 20 approximately 25 mg, while no mesotheliomas were found in rats injected O-acetyl-stg (AcO-stg) and dihydro-stg. There were two histological types in the mesothelioma induced by stg; the epithelioid and mesenchymal types. There was little loose connective tissue between the epithelioid cell tumors, whereas abundant dense collagen fibers were found between the cell nests of the mesenchymal cell types of mesothelioma. The hepatocellular carcinomas were found in a rat treated with stg and in five animals given AcO-stg. The fact that mesotheliomas could be induced in high incidence by direct intraperitoneal application of stg suggests a potent carcinogenicity of stg. Although both dihydro-stg and stg formed fine needle-like crystals when injected into the peritoneal cavity, dihydro-stg could not produce any mesothelioma. Dihydro-stg, reduction product of stg, has no double bond in the terminal furan ring. Therefore, the carcinogenicity of stg may be related more closely with the presence of a double bond in the terminal furan ring of stg than the physical form in the peritoneal cavity. AcO-stg may easily be absorbed from the peritoneum, so that it could not persist in the peritoneium for sufficient duration for carcinogenesis.

Animals↗

Lysis of halophilic Vibrio alginolyticus and Vibrio costicolus induced by chaotropic anions.

High concentration (1.0 M) of KSCN, but not of NaSCN, induced lysis of slightly halophilic Vibrio alginolyticus and moderately halophilic Vibrio costicolus, and the decrease in absorbance of the cell suspension was complete after 30 min at 25 degrees C. Replacement of K+ with Na+ effectively prevented the lysis by SCN-.K+ salts of NO3-, Br- and I-, however, induced no significant lysis. In electron micrographs, a prolonged exposure of the cells of V. alginolyticus to 1.0 M KSCN displaced the nucleoplasm to maintain close contact with the cell membranes. After 40 min of interaction, 50% of the cellular protein, 96% of RNA and 94% of DNA were recovered in the lysed cells. In contrast to lysis in hypotonic conditions, the lysis induced by KSCN is due mainly to a partial release of protein from the cells. V. costicolus was more susceptible to SCN- than V. alginolyticus, whereas nonhalophilic Escherichia coli was resistant to 1.0 M KSCN. Thus, lysis by SCN- is characteristic of halophilic bacteria and cell membranes of more halophilic bacteria are more susceptible to chaotropic anions. The protective effect of Na+ observed here was considered to be manifested by specific interactions of Na+ with components of cell membranes, thereby rendering their structures resistant to the action of chaotropic anions.

Bacteriolysis↗

The effects of some carcinogenic nitroso compounds on the rat liver nucleolus.

The hepatocyte nucleoli of rats undergo a variety of specific and nonspecific alterations after the administration of 8 nitroso compounds and 5 corresponding non-nitroso compounds. After the oral administration of dimethylnitrosamine and diethylnitrosamine, and unusual segregation of the nucleolus was encountered in the cells with karyorrhexis. Most characteristic of the segregation are the scattered ribosome-like granules at the outermost layer of the altered nucleolus. Frequently microspherules with halos 150 nm in diameter are found in all rats treated with nitroso compounds, which have been reported to be carcinogenic in the organs other than the liver. With the exception of butylurea and butylamine no such specific microspherules were found in the nucleoli after the administration of corresponding non-nitroso compounds.

Animals↗

HeLa cell-tumor in nude mice and its response to antitumor agents.

HeLa cells were heterotransplanted to nude mice and their response to some of antitumor agents was investigated. HeLa cell-tumor readily grew in nude mice and no regression was observed. Metastases to the lung and other organs were noticed in some of the animals. Histopathological examination revealed that the tumor retained the original characteristics of human epidermoid carcinoma. Standardization of HeLa cell-tumor in nude mice for the screening and evaluation of antitumor chemotherapeutics was attempted. Marked inhibition of tumor growth was observed with lower doses of Mitomycin-C, 5-fluorouracil, Adriamycin, and Bleomycin. The tumor regression was observed with high doses of Mitomycin-C, Adriamycin, and 5-fluorouracil. However, cyclophosphamide, cytosine arabinoside, and daunorubicin had little effect on the tumor growth. Complete regression was not obtained with any of the test agents and active regrowth took place even with the most effective compound. Considerable variation in the effect on tumor growth was observed among the test compounds, while histopathological findings were much alike; few mitotic figures, vacuolization, and pyknosis were main changes in tumor cells, and large foci of necrosis and hemorrhage were present in the degenerative areas. The regrowth was initiated around the capillaries in the necrotic tumor tissue.

Animals↗