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Biomedical subjects

H Orii

Publications and source records attributed to H Orii.

At least 37 records · Page 2Linked to original sources

A new type of plasmid from a wild isolate of Dictyostelium species: the existence of closely situated long inverted repeats.

A circular plasmid having high copy number was found in a wild isolate of Dictyostelium species. Gel electrophoresis, electron microscopy and Southern blot hybridization revealed that the plasmid, named pDG1, is 4.5 Kb (1.5 micron) in size with closely situated long inverted repeats. The plasmid seems to be located in the nuclei. It was not a derivative of ribosomal DNA. The possible correlation of the plasmid with the putative intermediate DNA of retrotransposon DIRS-1 found in Dictyostelium discoideum is discussed.

DNA Restriction Enzymes↗

[Focal cerebral infarction in the rat: II. Neuropathological study and local cerebral blood flow pattern].

We have recently reported that middle cerebral artery (MCA) occlusion in the rat produces a uniform pattern of cerebral ischemia in an acute phase. This study was done to determine if this model is also useful for quantitative evaluation of infarction size in a chronic phase. [Methods] Sprague-Dawley rats were anesthetized with halothane and left MCA was occluded via transretro-orbital approach. The following studies were done. Neuropathological study was done one week after MCA occlusion. After perfusion fixation, the brain was cut into 6 coronal slices and stained sections were examined. Local cerebral blood flow patterns were observed by 14C-iodoantipyrine autoradiographic technique 1, 2, and 5 days after the occlusion. [Results] Neuropathological studies invariably showed infarct in the cortex and the lateral part of the basal ganlia. The ratio of the infarct to the total areas of both hemispheres in 6 coronal sections was 14.05 +/- 2.66% (Mean +/- SD) in MCA occluded animals (N = 14) and 0.59 +/- 0.46% in sham operated animals (N = 12). Relative to the contralateral hemisphere, marked reduction in CBF was seen in the territory of the MCA and moderate reductions were also seen in the surrounding areas. The same pattern of increased CBF as previously reported was also seen in the ipsilateral substantia nigra and globus pallidus 1, 2, and 5 days after the occlusion. These results indicate the usefulness of this chronic focal cerebral infarction model in the evaluation of infarction.

Animals↗

[Regional blood flow of experimental brain tumors with special reference to effects of chemotherapy and radiotherapy].

Regional blood flow and capillary permeability in the experimental brain tumors and their surrounding brain tissue of rats were measured with quantitative 14C-antipyrine and 14C-alpha-aminoisobutyric acid (AIB) autoradiographic method. The pharmacokinetic implications with respect to drug delivery to tumor tissue and the effect of ionizing irradiation were discussed in these physiological measurement. A suspension of 1 X 10(4) rat glioma cells (E239 RG 12) was stereotactically implanted into the right basal ganglia of CD-Fisher rats, and spherical brain tumors developed 10-17 days after implantation with a diameter of 1--5 mm. Autoradiographic investigations were performed for rats with small tumors (1--2 mm in diameter) and large tumors (4--5 mm in diameter). The uniform blood flow (91.7 +/- 13.1 ml/100 g/min: mean +/- S.E.) was observed in small tumors with the patchy low flow area (56.7 +/- 12.5 ml/100 g/min) surrounding the tumor. In large tumors, the blood flow was markedly decreased in the central part of the tumor (28.3 +/- 2.4 ml/100 g/min) with a ring shaped high flow area in the peripheral part (59.3 +/- 5.9 ml/100 mg/min). The blood flow in the brain adjacent to the tumor (30.5 +/- 2.5 ml/100 g/min) was lower than that in the peripheral part of the tumor. The uptake of 14C-AIB was quite similar to that of 14C-antipyrine suggesting the smaller permeability in the central part of the tumor. Neuropathological studies did not reveal necrotic foci, but viable cells in these areas.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The role of ferritin in the intracellular distribution of gallium 67.

The binding of gallium 67 or iron 59 to ferritin in vitro was investigated using equilibrium dialysis. Gallium 67 did not bind to apo-ferritin until the protein was transformed into ferritin in the presence of iron citrate. Apotransferrin inhibited the binding of 67Ga to ferritin, especially in the presence of sodium bicarbonate and citrate, thus indicating that 67Ga has not gained access to ferritin from its complex with transferrin. Similar inhibition was observed for ferritin-59Fe. The release of 59Fe from its transferrin complex was enhanced by ATP, citrate, or ascorbic acid, while these reagents did not stimulate the dissociation of 67Ga from its transferrin complex. On the other hand, 67Ga injected intravenously in vivo was not found in the ferritin fractions of rat liver, kidney, and tumor. The difference between experimental results in vivo and in vitro supports the hypothesis that 67Ga in the cytoplasm is not labile enough to be bound to ferritin. We have indicated a significant role of ferritin in distinguishing between 67Ga and 59Fe in the cell, and provided some clues to interpret the chemical forms of 67Ga in the cytoplasm.

Animals↗

A novel cyclic AMP metabolism exhibited by giant cells and its possible role in the sexual development of Dictyostelium discoideum.

In Dictyostelium discoideum cyclic AMP (cAMP) metabolism during macrocyst development, i.e., the sexual cycle of this organism, and in giant cells, i.e., fusion products from opposite mating-type cells, was investigated. The pattern of change in cAMP levels during macrocyst development differed considerably from that observed during fruiting-body formation, i.e., the asexual cycle. Giant cells produced and excreted considerable amounts of cAMP. Adenylate cyclase activity catalyzing cAMP production in giant cells was comparable to that of unfused cells. However, the activity of membrane-bound phosphodiesterase in giant cells was extremely low, and no extracellular phosphodiesterase was excreted. A phosphodiesterase inhibitory protein was secreted in excess by giant cells.

3',5'-Cyclic-AMP Phosphodiesterases↗

[Binding of 67 Ga and 59 Fe to ferritin or transferrin].

The bindings of 67Ga and 59Fe to ferritin or transferrin in vitro has been investigated. Affinity constants have been measured using the equilibrium dialysis, and the results have been obtained as follows: 1 Apo-ferritin could not bind to 67Ga until it was transformed into ferritin in presence of Fe-citrate. On the contrary, the affinity of 67Ga to ferritin was reduced when Fe was released from ferritin; thus indicating that Fe-core has been required for the binding of 67Ga to ferritin. 2 Binding of 67Ga to ferritin was inhibited with apo-transferrin, and this was also shown in the case of 59Fe. In the presence of NaHCO3 or citrate, more remarkable inhibitions were observed. NaHCO3 or citrate was found to give a synergistic effect on the binding of 67Ga to transferrin, as well as Fe-transferrin. Therefore, both 67Ga and 59Fe could not bind to ferritin in the state of 67Ga- or 59Fe-transferrin. 3 The release of 59Fe from 59Fe-transferrin was enhanced with adenosine triphosphate (ATP), citrate, or ascorbic acid, while any of these reagents did not affect the release of 67Ga from 67Ga-transferrin. The comparison of 59Fe and 67Ga through their bindings to ferritin or transferrin has suggested one of points to distinguish 67Ga from 59Fe in the cell.

Apoferritins↗

Changes in membrane surface properties of hepatic peroxisomes of rats under several conditions as determined by partition in aqueous polymer two-phase systems.

Changes in membrane surface properties of hepatic peroxisomes of rats under several conditions were observed by aqueous polymer two-phase systems, which contained 6% (w/w) dextran T 500, 6% (w/w) polyethyleneglycol 4000, 250 mmol sucrose/kg and various concentrations of sodium phosphate buffer. The partition of peroxisomes into the upper phase depended to a large extent on their membrane surface charge. The cross-points of peroxisomes shifted from 5.55 to 5.25 and 5.2 after the administration of clofibrate and aspirin for 2 weeks, respectively, although that of alloxan-diabetic rat peroxisomes was not altered. The hydrophobic properties of peroxisomes, examined by means of a partition containing polyethyleneglycol monostearate, were altered by diabetes and starvation, but no change occurred in rats treated with clofibrate or aspirin. In the liver of rats fed a high-fat diet, the partition of peroxisomes was the same as that of the control. These findings indicate that hypolipidemic drugs such as clofibrate and aspirin induce the proliferation of peroxisomes and lead to the alteration of the surface charge of peroxisomal membranes. Diabetes or fasting lead to an alteration mainly of the hydrophobic properties. Both changes are probably due to alteration of content and/or composition of the proteins and the phospholipids in peroxisomal membrane under the conditions used.

Animals↗

[Tumor affinity and DNA interactions of 57Co-bleomycin (author's transl)].

Cobalt-57-bleomycin (BLM) has been proven to be the most stable and useful tumor-diagnostic agent among several radiolabelled BLMs. However, the considerably long half life of 57Co causes troubles in handling and preclude its extensive uses. In our previous work, BLM was proved to form two geometrical isomers, in chelating with Co. In this paper, we compared the tumor affinity of two isomers, to make an improvement of this drug's merit. We investigated biodistribution in tumor-bearing mice and DNA binding properties by fluorescence quenching technique and DNA melting study. A comparison of the data obtained suggests that isomerism affects the tumor affinity of the drug. Both tumor accumulation in tumor-bearing mice and stability of DNA binding of type I isomer were higher than those of type II. If a certain suitable radionuclide is inserted into cold Co-BLM type I isomer, this agent will be a greatly useful tumor-imaging radiopharmaceutical.

Animals↗

DNA interaction with 57Co-bleomycin.

Tumor-diagnostic 57Co-bleomycin is a mixture of two isomers: types I and II. Interaction between these and DNA was studied by fluorescence spectrometry and thermal denaturation. The fluorescence study indicated that cobalt chelation resulted in a remarkable increase in the apparent DNA-bleomycin association constant and a slight increase in bleomycin-DNA binding; a remarkable difference was observed between the two isomers. In the thermal denaturation study, the difference of DNA binding behavior was also observed. The tumor affinity of these isomers was slightly different, and type I isomer showed higher tumor affinity than type II. These results indicate that cobalt chelation to bleomycin enhances DNA-bleomycin binding and its DNA binding stability, and these mechanisms, although not fully understood, appear to underly the difference in tumor affinity of cobalt-bleomycin isomers.

Animals↗

The mechanism of adsorption of Ga-67 citrate to cultured cells.

Gallium adsorption was investigated at various pH values, with and without chelating agents or buffers. Evidence was obtained that Ga-67 citrate was adsorbed to cells in a polymeric form at a certain pH. Adsorption of Ga-67 to cells and to a glass surface reached a maximum at pH 4.5. When Ga-67 solutions were centrifuged to separate the precipitate, the highest precipitated radioactivity was found at the stated pH. With the addition of chelating agents which interact strongly with gallium, the adsorption to either cells or a glass surface was stoichiometrically inhibited. No such inhibition occurred, however, when counter ions unbound to gallium were added. The amount of Ga-67 released from the cell depended on the pH, suggesting that most of the gallium is deposited on the cell surface. Our results support Glickson's conclusion that gallium citrate forms a polymer at a certain acidic pH, which results in an increased cellular uptake by unknown mechanisms, presumably by pynocytosis.

Adsorption↗