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Biomedical subjects

M Miyazaki

Publications and source records attributed to M Miyazaki.

At least 739 records · Page 41Linked to original sources

A histopathologic study of 102 cases of intrahepatic cholangiocarcinoma: histologic classification and modes of spreading.

In our survey of 102 cases of intrahepatic cholangiocarcinoma, 71 peripheral type cases and 31 hilar type cases were observed, the former being prone to metastasize to remote organs and lymph nodes when compared with the latter. These cases were histologically classified into nine histologic types according to the predominant features and rearranged into three groups (well-differentiated, less-differentiated, and uncommon). Although most of these cases were mucin-producing adenocarcinomas with variable grades of differentiation (92 of 102; 90%), several uncommon types were also encountered (ten of 102; 10%), such as adenosquamous, squamous, mucinous, or anaplastic carcinoma. Remote organ metastases were observed more frequently in the less-differentiated group than in the well-differentiated group. In regard to intrahepatic tumor spreading, expansion via sinusoidal spaces (93%), vascular (52%) or lymphatic (18%) involvement, perineural invasion (16%), replacing growth in the bile duct (12%), and permeation in the portal connective tissue (19%) were observed. The cholangiocarcinomas with the vascular involvement presented a higher tendency of intrahepatic as well as extrahepatic metastasis. For correct diagnosis and treatment of cholangiocarcinoma, it seems to be important to possess some knowledge of these histologic types, including their variations and patterns of intrahepatic spread.

Adenoma, Bile Duct↗

Qualitative morphogenesis of synaptic glomeruli in the rat cerebellum.

The synaptic glomeruli of rat cerebellum was investigated at postnatal ages of 1 to 7, 9 and 15 days in an attempt to formulate a morphogenic analysis for synaptic junctional development, using the E-PTA stain for synapses. The synaptic junctions, which are composed of dense projection, intercleft density and postsynaptic band in synaptic glomeruli, were observed at a postnatal age of 6 days. Two types of synaptic junctions, symmetrical and asymmetrical, were recognized.

Animals↗

Peptide elongation factor 1 from yeasts: purification and biochemical characterization of peptide elongation factors 1 alpha and 1 beta (gamma) from Saccharomyces carlsbergensis and Schizosaccharomyces pombe.

Cytoplasmic elongation factor 1 alpha (EF-1 alpha) [corrected] was purified to homogeneity in high yield from the two different yeasts Saccharomyces carlsbergensis (S. carls.) and Schizosaccharomyces pombe (S. pombe). The purification was easily achieved by CM-Sephadex column chromatography of the breakthrough fractions from DEAE-Sephadex chromatography of cell-free extracts. The basic proteins have a molecular weight of 47,000 for the S. carls. factor and of 49,000 for the S. pombe factor. While the purified yeast EF-1 alpha s function analogously to other eukaryotic factors and the E. coli EF-Tu in Phe-tRNA binding and polyphenylalanine synthesis, the yeast factor unusually hydrolyzed GTP on yeast ribosomes upon addition of Phe-tRNA in the absence of poly(U) as mRNA. This novelty is probably owing to the yeast ribosomes, which are assumed to lack elongation factor 3-equivalent component(s). Trypsin and chymotrypsin selectively cleaved the two yeast factors to generate resistant fragments with the same molecular weight of 43,000 (by trypsin) and of 44,000 (by chymotrypsin), respectively. Those cleavage sites were characteristically protected by the presence of several ligands bound to EF-1 alpha such as GDP, GTP, and aminoacyl-tRNA. Based on the sequence analysis of the fragments generated by the two proteases, the partial amino acid sequence of the S. carls. EF-1 alpha was deduced to be in accordance with the N-terminal region covering positions (1) to 94 and two Lys residues at the C-terminal end of the predicted total sequence of the Saccharomyces cerevisiae (S. cerev.) factor derived from DNA analysis, except for a few N-terminal residues, confirming the predicted S. cerev. sequence at the protein level. EF-1 beta and EF-1 beta gamma were isolated and highly purified as biologically active entities from the two yeasts. EF-1 beta s from the two yeasts have the same molecular weight of 27,000, whereas component gamma of the S. carls. EF-1 beta gamma showed a higher molecular weight (47,000) than that of the S. pombe factor (40,000). It was also shown that a stoichiometric complex was formed between EF-1 alpha and EF-1 beta gamma from S. pombe. Furthermore, a considerable amount of Phe-tRNA binding activity was distributed in the EF-1H (probably EF-1 alpha beta gamma) fraction from freshly prepared cell-free extracts of yeast.

Amino Acid Sequence↗

Characterization of the ATPase and GTPase activities of elongation factor 3 (EF-3) purified from yeasts.

Three steps of chromatography of a post-ribosomal supernatant fraction have provided a highly purified preparation of peptide elongation factor 3 (EF-3) with a molecular weight of 125,000 from the typical budding yeast Saccharomyces carlsbergensis and of the factor with a molecular weight of 120,000 from the fission yeast Schizosaccharomyces pombe. Both of the proteins consist of a single peptide chain. The purified factors fulfilled the requirement for polyphenylalanine synthesis on yeast ribosomes and exhibited strong ATPase and GTPase activities dependent on yeast ribosomes. The activity profiles of the nucleotidases dependent on pH and salt concentration and the inhibition studies indicated that the ATPase and GTPase activities of EF-3 were displayed by the same active site with a wide substrate specificity, showing the highest activity with ATP. Those experiments also revealed that the ATPase and GTPase of EF-3 were characteristically different from the GTPases of EF-1 alpha and EF-2. Both Km and kcat of EF-3 for ATP (Km = 0.12 mM and Kcat = 610 mol/mol/min) and GTP (Km = 0.20 mM and kcat = 390 mol/mol/min) are much higher than those of the GTPases of EF-1 alpha and EF-2. Inactivation experiments and studies on the ATP effect led us to conclude that this ATPase activity was an essential requirement for the functional role of EF-3 and therefore, in addition to the GTPases of EF-1 alpha and EF-2, the third nucleoside triphosphate hydrolyzing step by the ATPase of EF-3 was necessary for the yeast peptide elongation cycle.

Adenosine Triphosphatases↗

Role of yeast peptide elongation factor 3 (EF-3) at the AA-tRNA binding step.

The stimulatory effect of peptide elongation factor 3 (EF-3), which is uniquely required for the yeast elongation cycle, on the step of binding of aminoacyl-tRNA (AA-tRNA) to ribosomes has been investigated in detail. Yeast EF-1 alpha apparently functions in a stoichiometric manner in the binding reaction of AA-tRNA to the ribosomes. The addition of EF-3 and ATP to this binding system strikingly stimulated the binding reaction, and the stimulated reaction proceeded catalytically with respect to both EF-1 alpha and EF-3, accompanied by ATP hydrolysis, indicating that EF-3 stimulated the AA-tRNA binding reaction by releasing EF-1 alpha from the ribosomal complex, thus recycling it. This binding stimulation by EF-3 was in many respects distinct from that by EF-1 beta gamma. The idea that EF-3 may participate in the regeneration of GTP from ATP and the formed GDP, as indicated by the findings that the addition of EF-3 along with ATP allowed the AA-tRNA binding and Phe polymerization reactions to proceed even in the presence of GDP in place of GTP, was not verified by the results of direct measurement of [32P]GTP formation from [gamma-32P]ATP and GDP under various conditions. Examination of the stability of the bound AA-tRNA disclosed the different binding states of AA-tRNA on ribosomes between in the cases of the complexes formed with EF-1 alpha alone, or factor-independently, and with EF-1 alpha and EF-3.(ABSTRACT TRUNCATED AT 250 WORDS)

Fungal Proteins↗

The yeast peptide elongation factor 3 (EF-3) carries an active site for ATP hydrolysis which can interact with various nucleoside triphosphates in the absence of ribosomes.

ATP (GTP) hydrolysis was clearly demonstrated by using at most 16 pmol of yeast peptide elongation factor 3 (EF-3) in the absence of ribosomes. However, the highly active yeast ribosomes (up to 48 pmol) displayed virtually no ATPase (or GTPase) activity in the absence of EF-3. Several lines of evidence indicated that both the catalytic and binding sites of the ATPase reside in the elongation factor itself, not on the ribosomes. The patterns of protection by various nucleoside triphosphates against tryptic digestion of EF-3, reflecting the wide substrate specificity of the ATPase, confirmed that the active center of the endogenous ATPase is located on the factor itself and not on contaminants. The intrinsic activity was stimulated up to two orders of magnitude by the presence of the yeast ribosomes fully active in polyphenylalanine synthesis. The activation was achieved by enhancing the catalytic activity (kcat) to a much greater extent than the binding affinity (Km). On the other hand, the ribosome-activated ATPase activity was revealed to inherit its wide substrate specificity from the intrinsic property of EF-3, which shows an affinity to various XTPs, including pyrimidine- and purine-nucleoside triphosphates, irrespective of 2'-hydroxylation of the sugar moiety. From experiments on protection against tryptic digestion, we determined that intricate conformational changes of the factor molecule occur upon interaction with the substrate XTP and ribosomes.

Adenosine Triphosphatases↗

Leakey virus: a new hantavirus isolated from Mus musculus in the United States.

A hantavirus, designated Leakey virus, was isolated from a Mus musculus captured in Real County, Texas, U.S.A. in August 1986. Virus-specific fluorescence was first detected 13 days after inoculation of Vero-E6 cells with spleen tissue from the seropositive M. musculus. Ultrastructurally, the new isolate resembled other hantaviruses. Leakey virus induced a fatal meningoencephalitis in infant Fischer rats, with viral antigen detectable in brain, lung, liver, kidney and spleen. Serum dilution, plaque reduction neutralization tests indicated that Leakey virus was antigenically distinct from Hantaan, Seoul, Puumala and Prospect Hill viruses, and therefore constitutes a new serotype.

Animals↗

Suppression of the pressor effect of centrally administered thyrotropin-releasing hormone under halothane, pentobarbital and flunitrazepam anaesthesia.

Intracerebroventricular (i.c.v.) administration of thyrotropin-releasing hormone (TRH) caused an increase in blood pressure (BP) and heart rate (HR) in conscious rats. The pressor effect was greatly diminished by adrenalectomy as well as after pretreatment with phentolamine, an alpha-receptor antagonist or with mecamylamine, a ganglion blocker, suggesting that centrally administered TRH increases BP mainly by stimulating sympathetic activity. Under halothane (0.8%), pentobarbital (33 mg/kg, i.p.) and flunitrazepam (0.8 mg/kg, i.v.) anaesthesia, the pressor effect of TRH was almost completely blocked. The increase in BP induced by peripheral alpha-receptor stimulation with phenylephrine was not affected by the anaesthetics at these doses. Pretreatment with atropine (50 micrograms, i.c.v.) significantly reduced the pressor effect of TRH. Intracerebroventricularly administered haloperidol and bicuculline also partially diminished the increase in BP produced by TRH, while other neurotransmitter blockers such as phentolamine, propranolol and naloxone did not. These results indicate that the anaesthetics at the doses employed interfere with the central neuronal pathway(s), probably cholinergic pathways, through which TRH exerts its pressor effect.

Animals↗

Functional and histological changes in mesenteric arteries and aortas from monkeys fed a high cholesterol diet.

Treatment of Japanese monkeys for 8 months with a high fat, high cholesterol diet produced atherosclerotic lesions in the aorta and mesenteric arteries, such as fatty dots, streaks and plaques, intimal thickening with accumulation of spindle-shaped cells and macrophages and endothelial cell flattening. Contractile responses of mesenteric arteries from control and atherosclerotic monkeys to electrical stimulation of adrenergic nerves, norepinephrine and angiotensin II did not differ, whereas contractions caused by serotonin in the atherosclerotic monkey arteries were significantly greater. Ketanserin and cinanserin suppressed the serotonin-induced contraction. Relaxations caused by adenosine and K+ (5 mM) were moderately attenuated in atherosclerotic monkey mesenteric arteries, and those by acetylcholine were reduced only slightly or not affected in the arteries or aortas. Relaxations of control and atherosclerotic arteries in response to nitroglycerin, isoproterenol and prostaglandin I2 did not differ. The relaxant response to K+ was reversed to a contraction by ouabain. Acetylcholine-induced relaxations were dependent on the endothelium and suppressed by atropine. Diet-induced atherosclerosis appears to potentiate contractions mediated via serotonergic 5-HT2 receptors and to attenuate relaxations possibly caused by activation of the electrogenic Na+ pump in the smooth muscle cell membrane. Endothelium-dependent relaxations via muscarinic receptors would not evidently be affected in mesenteric arteries and aortas from atherosclerotic Japanese monkeys.

Animals↗

A method for rapid preparation of single-cell suspensions from rat hepatocyte primary cultures on collagen substratum and the mechanism of cell dissociation.

A method has been developed for the rapid preparation of single-cell suspensions from rat hepatocyte primary cultures on collagen substratum. Hepatocytes were adequately dissociated into single cells when the cultures were first treated with a combination of trypsin and ethylenediaminetetraacetic acid (EDTA) and then with collagenase. However, when the order was reversed, hepatocytes were inadequately dispersed. The possible mechanism of cell dissociation is discussed on the basis of the experimental data obtained.

Animals↗

Decreased albumin secretion in serum-free primary cultures of adult rat hepatocytes during proliferation induced by epidermal growth factor and insulin.

Upon addition of epidermal growth factor (EGF, 0.1 microgram/ml) and insulin (0.1 microM), adult rat hepatocytes proliferated and increased 120-134% in number in serum-free primary culture. However, in the absence of the growth factors, hepatocytes decreased in number with time. The average albumin secretion per cell was much lower in the proliferating cultures than in the non-proliferating cultures. The results suggest that albumin production in hepatocytes decreases during cell proliferation.

Albumins↗

[Treatment of delayed brain injury after pituitary irradiation].

Treatment for delayed brain injury after pituitary irradiation is discussed. Six cases with delayed brain injury were treated with a combination of dexamethasone or betamethasone, with heparin, glycerol, dextran 40 and some vasodilators. Two cases with temporal lobe syndrome were treated in the early stages of brain injury for a period of over 12 months were almost completely cured, another two cases with chiasma syndrome were treated in the relatively late stages, showed a partial improvement. One case which was irradiated 120 GY during 13 years did not improve. The final case treated with steroids for a short period also resulted in failure and the patient underwent an operation for the removal of the necrotic mass three years after the radiotherapy. Steroid therapy started in the early stages of brain injury after irradiation for over the 12 months is thought to be effective. Heparin therapy was also effective in one out of three cases, but in one of the cases subarachnoid hemorrhage from a traumatic aneurysm occurred during the therapy. In an acute phase, showing edematous change of the injured brain, the administration of glycerol is also thought to be useful. But the effectiveness of the other medicines containing some vasodilators was obscure or doubtful. We propose the following: (1) A meticulous observation is essential for the patients who received high doses of irradiation to diagnose brain injury in the early reversible stage. (2) Steroids should be given immediately in this reversible stage of brain injury before the irreversible "necrosis" occurs. (3) Steroids should be maintained for a long period over 12 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenoma↗

Role of vascular angiotensin converting enzyme in hypertension.

The possible role of vascular angiotensin converting enzyme (ACE) in the maintenance of one-kidney, one clip (1-K,1C) hypertensive rats was studied in comparison with age-matched, one-kidney (1-K) normotensive rats. Mean blood pressure was elevated after partial occlusion of the left renal artery with unilateral nephrectomy, and the high blood pressure persisted for at least 11 weeks, whereas no significant changes in mean blood pressure were observed in 1-K rats. Plasma and vascular renin activities and plasma ACE activity did not differ between the two groups of rats, both 5 and 11 weeks after operation. In contrast, ACE activity in lung and mesenteric arteries in 1-K,1C hypertensive rats was significantly elevated 11 weeks after operation, but not 5 weeks after operation, compared with the matched 1-K normotensive rats. In addition, the decreases in mean blood pressure induced by ACE inhibition were significantly greater in 1-K,1C rats than those in 1-K rats. These results indicate that the elevation of vascular ACE activity may play an important role in the maintenance of high blood pressure and may result in hypotension in response to ACE inhibitors in the chronic stage of 1-K,1C hypertension.

Angiotensin-Converting Enzyme Inhibitors↗

[The effect of naproxen on fever following transcatheter arterial embolization of hepatic tumors].

In the cases of trans-catheter arterial embolization (TAE) for hepatic malignancies, naproxen has been evaluated for its antipyretic effect. Each patient not given naproxen manifested a remarkable and prolonged fever despite treatment by antibiotics. In those who received an administration of naproxen, however, a significant suppression of fever was achieved (P 0.01-0.001). No remarkable influence was found on the white blood cell counts, the serum GOT and LDH levels by the naproxen. Similarly, no untoward effect due to naproxen was found on the ulcerogenicity in the gastro-duodenum. Thus, for hepatic malignancies, naproxen could be useful in the symptomatic treatment of a fever following a TAE.

Adult↗