Search PubMed⌕ Search

Biomedical subjects

H Takagi

Publications and source records attributed to H Takagi.

At least 883 records · Page 49Linked to original sources

Role of interleukin-2-receptor-positive cells and natural killer cells on rejected liver grafts of rats.

We explored the subpopulation and distribution of infiltrating leukocytes in rejected liver grafts using an orthotopic liver transplantation model of rats. We obtained the following results: (1) OX08- and recombinant-interleukin 2-receptor-positive cells were numerously increased in rejected grafts. (2) There was an abnormal distribution of R73-positive OX08-positive OX19-negative cells in the sinusoidal area of rejected grafts. These results suggest that R73-positive OX08-positive OX19-negative cells (natural killer cells or T cells?) have a role in the immunologic reaction in the sinusoidal area of rejected liver grafts of rats.

Animals↗

Amino acid sequence of silkworm (Bombyx mori) hemolymph antitrypsin deduced from its cDNA nucleotide sequence: confirmation of its homology with serpins.

A cDNA clone of silkworm (Bombyx mori) larval hemolymph antitrypsin (sw-AT) has been isolated from a fat body cDNA library. The cDNA has an open reading frame which codes a 392-amino acid residue polypeptide comprising a 16-residue signal peptide and a 376-residue mature sw-AT of Mr 41,805. The reactive site of sw-AT for inhibition of bovine trypsin [Sasaki, T. et al. (1987) J. Biochem. 102, 433-441] was identified as Lys343-Val344. Alignment of the sw-AT amino acid sequence with those of 11 members of the serpin superfamily of proteins clearly confirmed the homology of sw-AT with serpins. The amino acid sequence of sw-AT is 56% identical with that of the proteinase inhibitor from a lepidopteron, Manduca sexta [Kanost, M.R. et al. (1989) J. Biol. Chem. 264, 965-972], but the sequence around the reactive site shows no homology and the inhibitory specificity for proteinases is very different.

Amino Acid Sequence↗

Synaptic and Nonsynaptic Localization of Benzodiazepine/GABAA Receptor/Cl- Channel Complex Using Monoclonal Antibodies in the Dorsal Lateral Geniculate Nucleus of the Cat.

The two monoclonal antibodies, bd-17 and bd-24, are specific for beta- and alpha-subunits of the GABAA/benzodiazepine receptor/chloride channel complex respectively. An abundance of both subunits has been revealed in the visual thalamus of the cat by light microscopic immunocytochemistry using these antibodies. The alpha-subunit specific antibody and electron microscopy were used to determine the subcellular distribution of immunoreactivity with respect to specific cell classes in the dorsal lateral geniculate nucleus. Immunoreactivity was always associated with membranes and the degree of immunoreactivity varied greatly between different types of cell as defined by: (i) immunoreactivity for GABA; (ii) soma area; (iii) presence or absence of cytoplasmic laminated bodies (CLB). GABA negative neurons with the smallest soma area showed the strongest immunoreactivity, mainly in the endoplasmic reticulum and also on the somatic plasma membrane. Cytoplasmic laminated bodies could be found in the majority of these neurons. Large GABA negative cells without CLBs were strongly immunoreactive on the plasma membrane of the soma and dendrites, but showed scant if any intracellular immunoreactivity. GABA-positive cells showed weak intracellular immunoreactivity but negligible if any immunoreactivity at the somatic and proximal dendritic plasma membrane. A similar reaction pattern was found in GABA negative cells which contained no CLBs and which constituted a medium sized cell population. It is suggested that the degree of intracellular receptor immunoreactivity is positively correlated with receptor turnover. The dendrites of projection cells, particularly outside the glomeruli, showed strong immunoreactivity on the plasma membrane. The synaptic junctions formed by many boutons (F terminals) establishing symmetrical synapses with dendrites of relay cells were immunopositive, but no immunoreactivity could be detected at the synapses established by the presynaptic dendrites of the local interneurons. Many axo-somatic F1 junctions were also immunoreactive. However, immunoreactivity for the receptor/channel complex was also widely distribution on nonsynaptic plasma membranes of somata and dendrites. Thus GABA may act at both synaptic and non-synaptic sites. Furthermore, the correlation of immunoreactivity for the GABAA receptor complex with previously published properties of physiologically identified cells suggests that the strongly immunoreactive, small, GABA negative cells with CLBs might correspond to the 'lagged' X-type cells, and the large GABA negative receptor outlined cells without CLBs might correspond to some of the Y-type neurons.

Journal Article↗

Functional reconstruction of purified Gi and Go with mu-opioid receptors in guinea pig striatal membranes pretreated with micromolar concentrations of N-ethylmaleimide.

Functional coupling between mu-opioid receptors and GTP-binding regulatory proteins (G proteins) was investigated in reconstituted membranes of the guinea pig striatum. Selective mu-opioid agonists stimulated low-Km GTPase in striatal membranes, in a Na(+)-dependent manner. The same mu-opioid agonist [( D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAGO)] caused no stimulation when the membranes were exposed to islet-activating protein (IAP; pertussis toxin). There was also no DAGO stimulation in preparations pretreated with a lower concentration (5 microM) of N-ethylmaleimide (NEM), which abolished the ADP-ribosylation of purified Gi (the G protein that mediates inhibition of adenylate cyclase) and Go (a G protein of unknown function purified from bovine brain) by IAP. In addition, as the NEM treatment caused no change in the mu-agonist binding, NEM could probably substitute for IAP in inactivating native G proteins, without exhibiting effects on the receptor binding in membranes. The mu-agonist stimulation of low-Km GTPase activity in NEM-treated membranes was recovered by reconstitution with purified Gi or Go. The mu-agonist stimulation of low-Km GTPase was additive when Gi and Go were simultaneously reconstituted in NEM-treated membranes in amounts of 0.5 pmol/assay, which was required for maximal recovery, in either reconstitution experiment. The present findings provide the first evidence that the mu-opioid receptor may exist in at least two different forms, separately coupled to Gi or Go.

Adenylate Cyclase Toxin↗

Conserved structures of cell wall protein genes among protein-producing Bacillus brevis strains.

Bacillus brevis HPD31 contains a surface (S)-layer protein, termed the HWP, which forms a hexagonal array in the cell wall. The 5' region of the HWP gene was isolated from a DNA library constructed in bacteriophage vector EMBL3 from a partial BamHI digest of the chromosomal DNA. The 3' region contained in a 2.7-kilobase BglII fragment of the DNA was cloned into Escherichia coli, using pUC118 as a vector. On the basis of the chemically determined N-terminal amino acid sequence, the HWP gene was found to encode a polypeptide consisting of 1,087 amino acid residues with a signal peptide of 53 or 23 amino acid residues. The deduced amino acid composition was similar to the chemical amino acid compositions of other S-layer proteins in the predominance of acidic relative to basic amino acids and in the very low content of sulfur-containing amino acids. The deduced amino acid sequence showed high homology (78%) with that of the middle wall protein of B. brevis 47. Furthermore, the multiple 5' ends of the HWP gene transcripts detected on S1 nuclease analysis closely resembled those of the middle wall protein gene transcripts. This complex structure was also conserved (greater than 85%) in the regulatory regions of two other cell wall protein genes isolated from B. brevis HPD52 and HP033, suggesting that the synthesis of the cell wall proteins is intricately regulated through a similar mechanism in protein-producing B. brevis.

Amino Acid Sequence↗

Adrenocorticotropin increases expression of c-fos and beta-actin genes in the rat adrenals.

It is widely accepted that expression of protooncogenes is coupled with cellular proliferation and differentiation. Since ACTH stimulates not only steroidogenesis but also cellular proliferation, we investigated whether ACTH affects the expression of c-fos, c-myc, and beta-actin genes. The effect of ACTH on adrenal glands was studied in hypophysectomized rats. Changes in the mRNA levels were studied by Northern and dot blot analyses. It was demonstrated that ACTH induces increases in mRNAs encoding c-fos and beta-actin in adrenal glands of hypophysectomized rats. When stimulated by ACTH (5 IU/100 g BW), the mRNA levels of both genes increase rapidly; the maximum levels are observed at 30 min for c-fos and 6 h for beta-actin. Both mRNAs declined to near-control levels by 6-24 h. The levels of mRNAs encoding cholesterol side-chain cleavage cytochrome P-450 and 21-hydroxylase cytochrome P-450 began to increase 3 and 12 h after ACTH administration, respectively. This increase continued for 24 h after ACTH treatment. Increases in total adrenal RNA and adrenal weight occurred slowly after ACTH treatment. On the other hand, the levels of c-myc mRNA were very low and were not increased by ACTH administration. These results suggest that increased expression of c-fos and beta-actin genes by ACTH may have important roles in mediating its action on adrenals.

Actins↗

[Physiological and pharmacological actions of a neuroactive dipeptide, kyotorphin, and its precursor, L-arginine, and clinical application].

This review describes: 1. Recent findings about the physiological and pharmacological actions of kyotorphin that was isolated from the brain. 2. Biosynthesis, distribution, release, uptake, enzymatic destruction of this peptide in the body. 3. L-Arginine was found to be a precursor of kyotorphin. 4. Clinical application of the precursor to the treatment of chronic pain.

Analgesics↗

Alteration in the expression of genes for cholesterol side-chain cleavage enzyme and 21-hydroxylase by hypophysectomy and ACTH administration in the rat adrenal.

The changes in steady-state levels of mRNA for cholesterol side-chain cleavage cytochrome P-450 (P-450scc) and steroid 21-hydroxylase cytochrome P-450 (P-450c21) caused by hypophysectomy and ACTH treatment were determined in rat adrenals. Hypophysectomy caused marked decreases in adrenal weight and total RNA per gland. Administration of ACTH resulted in increases in adrenal weight and total RNA. A significant correlation between the amount of RNA and adrenal weight was observed. Both P-450scc and P-450c21 mRNAs were decreased by hypophysectomy and increased by ACTH treatment. P-450scc mRNA decreased to 20% and P-450c21 mRNA to 76% of control values 1 day after hypophysectomy. ACTH caused a significant increase in P-450scc mRNA after 3 h. However, a significant increase in P-450c21 mRNA was observed 12 h after administration of ACTH. These results are concordant with previous studies in vitro utilizing cultured adrenocortical cells. Moreover, the induction of steady-state levels of P-450scc mRNA was faster than that observed by other investigators in studies in vitro. These results may indicate that integrity of the adrenal gland in vivo is important for the action of ACTH.

Adrenal Glands↗

Neurinoma of the oculomotor nerve--case report.

Only 10 cases of oculomotor nerve neurinoma have been reported previously. The authors report the rarity of a neurinoma originating from Schwann cells of the oculomotor nerve. The diagnosis was based on the initial sign of oculomotor nerve paresis without involvement of other cranial nerves, neuroradiological and surgical findings, and histological features of the specimen obtained at surgery.

Aged↗

Limited proteolysis of silkworm antitrypsin by several proteinases.

Silkworm antitrypsin (sw-AT) isolated from larval hemolymph was limitedly digested by Achromobacter lysylendopeptidase, alpha-chymotrypsin, subtilisin BPN', subtilisin Carlsberg, papain, or Pseudomonas elastase. Each proteinase could cleave specific site(s) around the reactive site identified for the reaction of sw-AT and bovine trypsin. Among these proteinases, only subtilisin BPN' was inhibited by sw-AT, although weakly. By the cleavable amino acid sequence in sw-AT, it was suggested that whether or not these proteinases were inhibited by sw-AT did not solely depend on their substrate specificities. The susceptibility to the attack of proteinase should indicate that this region is exposed on the molecular surface. The amino acid sequence in the COOH-terminal region slightly away from the reactive site in sw-AT had homology with that in the corresponding region of the serine proteinase inhibitor (serpin) group.

Amino Acid Sequence↗

[A case of CABG in a patient with chronic idiopathic thrombocytopenic purpura].

This is a report of the management of a patient with chronic idiopathic thrombocytopenic purpura (ITP) and ischemic heart disease. High-dose transvenous gamma-globulin therapy (400 mg/kg/day for five days) was started five days prior to operation, yielding an increase in platelet count from 5.8 X 10(4)/mm3 to 12.8 X 10(4)/mm3. Platelet transfusion was used at the end of cardiopulmonary bypass and on the first postoperative day. There were no problems with hemostasis during the operative procedure and the postoperative course was uneventful. We prefer high-dose transvenous gamma-globulin therapy for ITP prior to open heart surgery as a treatment of choice.

Aged↗

Effects of preceding ischemic time on the recovery course of energy metabolism in rat liver.

Effects of the duration of preceding ischemia on the recovery of liver energy metabolism after reperfusion were investigated. Liver ATP level was depleted after the first 30 min of ischemia, and the decrease remained steady thereafter. Recovery of ATP depended on the preceding ischemic time, i.e., 81.5%, 66.4% and 39.5% recovery of the control level were observed after 60 min of reperfusion following 30 min, 60 min and 120 min of ischemia, respectively. Ischemia-induced mitochondrial dysfunction depended on the duration of ischemia. Mitochondrial function was recovered fully after 60 min of reperfusion following both 30 min and 60 min of ischemia. However, deterioration of mitochondrial function did not recover significantly after 60 min of reperfusion following 120 min of ischemia. Similar decreases in adenylate energy charge were observed irrespective of the duration of ischemia, and it recovered fully after 60 min of reperfusion following 30 min, 60 min and 120 min of ischemia. These results suggest that not the energy charge but ATP level itself is a reliable marker of liver energy status.

Adenosine Triphosphate↗

[Subrenal capsule assay in normal mice immunosuppressed by cyclosporin A].

The subrenal capsule assay in normal mice has more advantages to evaluate sensitivities of anticancer agents than other assays in vivo. However, this method has difficulties because mice reject human tumor grafts by their immune reaction. We examined three methods of immunosuppression in mice; radiation, administration of cyclophosphamide and that of cyclosporin A. The administration of cyclosporin A suppressed host reactions most effectively among the three, and the human tumor grafts in subrenal capsules grew well. We also investigated the intervals of administration and the doses of cyclosporin A in order to evaluate drug response. The minimal dose of cyclosporin A which inhibited host reactions was revealed to be 50mg/kg/day administered subcutaneously to mice. Then the adequate dosage of each anticancer drug for this assay system was investigated so that the body weights of mice do not decrease more than 20%, and the following doses were determined; Mitomycin C: 5mg/kg; Cisplatinum: 4mg/kg; Cyclophosphamide: 100mg/kg and 5FU: 90 mg/kg.

Animals↗

[A basic study of cytotoxic effect of tumor necrosis factor-alpha on the hepatoma cell line, PLC/PRF/5].

In order to investigate the effect of tumor necrosis factor (recombinant TNF-alpha) on hepatoma, we assessed the antitumor activity of TNF-alpha on hepatoma cell line, PLC/PRF/5 in vitro, and combined effects of interferon-gamma (IFN-gamma), that is thought to increase TNF-receptors. TNF-receptors. TNF-alpha for itself, had no significant effect on DNA synthesis and viability, and the production of HBsAg from PLC/PRF/5. On the other hand, treatment of IFN-gamma- combined with TNF-alpha inhibited DNA synthesis and production of HBsAg. The decrease of viability of PLC/PRF/5 was also found in this combination treatment. According to these findings, antitumor activity of TNF-alpha had no effect on PLC/PRF/5 in itself, but was significantly enhanced when treated with IFN-gamma at the same time.

Carcinoma, Hepatocellular↗

[Studies on the effectiveness of adjuvant hepatic arterial chemotherapy after hepatectomy for primary or metastatic liver cancer].

From January 1980 to March 1990, 399 cases of primary liver cancer (hepatocellular carcinoma 357, cholangiocellular carcinoma 42) and 148 cases of metastatic liver cancer were treated in our hospital. Some 222 of H.C.C (hepatocellular carcinoma), 20 of C.C. (cholangiocellular carcinoma) and 42 of metastatic liver cancer were resected; 24 of H.C.C, 2 of C.C and 22 of metastatic cancer received adjuvant hepatic arterial chemotherapy, in which anti-cancer drugs were administered with oily contrast medium Lipiodol in hepatic artery. The relationship between operative findings and postoperative prognosis was studied in 168 resected H.C.C cases and risk factors for recurrence were determined. Risk factors are TW(+), which means that the cancer remains macroscopically within 1 cm of surgical margin; IM(+), which means intrahepatic metastasis exists; more than Vp2, which means tumor embolus exists in the second or more proximal branch of the portal vein; and Fc(-), which means lack of capsule formation. In 132 cases with the risk factors, the survival rate of 19 cases with adjuvant arterial chemotherapy was significantly higher than that of 113 cases without it. In the cases of liver metastasis of colon cancer, resection of metastases and adjuvant hepatic arterial chemotherapy improved the prognosis.

Antineoplastic Combined Chemotherapy Protocols↗

[Nucleotide metabolism in remnant rat liver after major hepatic resection].

Major hepatic resection is restricted in the presence of cirrhosis, because the cirrhotic liver is less able to regenerate. In the present study, to clarify what factors play a major role as inhibitors of the regeneration process, we focused on the changes in liver purine nucleotides and their catabolite levels in rats with cirrhosis. Decreases in adenine nucleotides and guanine nucleotides were observed after resection both in normal and thioacetamide-induced cirrhotic livers. These were prolonged in rats with cirrhosis. Tissue levels of hypoxanthine and xanthine increased both in the control group and in the cirrhotic group. The increase in xanthine level was remarkable in the cirrhotic group compared with the control group. These results indicate that xanthine oxidase activity is increased in cirrhotic liver after major hepatic resection. Xanthine oxidase catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid. Xanthine and uric acid are end metabolites of purine nucleotides. On the other hand, superoxide is generated in association with this reaction. Therefore, the disturbance in the energy metabolism and the increase in superoxide formation which are caused by the activation of xanthine oxidase might be involved in the regeneration process as inhibitory factors in cirrhotic rat liver.

Animals↗