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Biomedical subjects

A Tamura

Publications and source records attributed to A Tamura.

At least 451 records · Page 25Linked to original sources

Trans-bilayer movement of added phosphatidylcholine and lysophosphatidylcholine species with various acyl chain lengths in plasma membrane of intact human erythrocytes.

14C-Labeled phosphatidylcholine (PC) and lysophosphatidylcholine (lysoPC) species with two homologous saturated acyl chains and of a saturated acyl chain of various lengths, respectively, were each incorporated into the outer leaflet of the membrane lipid bilayer of intact human erythrocytes, and the transbilayer movement into the inner leaflet during incubation at 37 degrees C of the lipid-loaded erythrocytes was followed. The labeled PC and lysoPC molecules present in the outer leaflet were extracted with egg-yolk PC liposome suspension and BSA solution, respectively, and the amount which moved into the inner leaflet during the incubation was measured by determining the residual amount of the labeled lipid in the membrane. Translocation of lysoPC molecules was also measured by assaying the decrease in the amount of the added labeled lysoPC in the membrane during the incubation on the basis of the previously reported fact that lysoPC molecules are all converted metabolically to PC or glycerylphosphorylcholine plus fatty acid as soon as they are translocated from the outer to the inner leaflet. Every lipid tested showed significant transbilayer movement during the course of the incubation for up to 10 h. With the C8, C10, and C12 species of PC the rate of the transbilayer movement increases with decreasing acyl chain length. The same is true with the C14, C16, and C18-lysoPC species.

Animals↗

Analysis of polypeptide composition and antigenic components of Rickettsia tsutsugamushi by polyacrylamide gel electrophoresis and immunoblotting.

Polyacrylamide gel electrophoresis of lysates of purified Rickettsia tsutsugamushi revealed as many as 30 polypeptide bands, including major bands corresponding to molecular sizes of 70, 60, 54 to 56, and 46 to 47 kilodaltons. Compared with the polypeptide composition of the rickettsiae of Gilliam, Karp, and Kato strains and a newly isolated Shimokoshi strain, the major polypeptide in the Kato strain (54-56K) and in the Karp strain (46-47K) migrated a little faster and slower, respectively, than the corresponding polypeptides in the other strains. The largest major polypeptide (54-56K) was digestible by the treatment of intact rickettsiae with trypsin and variable in content in separate preparations, suggesting that the polypeptide exists on the rickettsial surface and is easily degraded during the handling of these microorganisms. Several surface polypeptides of rickettsiae, including the 54-56K and 46-47K polypeptides, were detected by radioiodination of intact rickettsiae followed by polyacrylamide gel electrophoresis of the lysate; however, the 70K and 60K polypeptides were not labeled. Immunoblotting experiments with hyperimmune sera prepared in guinea pigs against each strain demonstrated that the 70K, 54-56K, and 46-47K polypeptides showed antigenic activities. The 54-56K polypeptide appeared to be strain specific, whereas the 70K and 46-47K polypeptides cross-reacted with the heterologous antisera.

Animals↗

Ultrastructure of cerebral arteries following experimental subarachnoid haemorrhage.

Scanning and transmission electron microscopy have been used to examine the ultrastructure of cerebral arteries taken from dogs up to nine days following the injection of autologous blood into the suprachiasmatic cistern and using meticulous perfusion-fixation technique. No ultrastructural changes in the cerebral arteries were noted either at sites of radiologically demonstrated arterial constriction or elsewhere. The only abnormality noted was the presence of some leucocytes and macrophages in the subarachnoid space surrounding the arteries. These results are discussed in relation to changes in cerebrovascular reactivity that occur at this stage following subarachnoid haemorrhage.

Animals↗

Mechanisms affecting peplomycin sensitivity of Chinese hamster cell lines.

Chinese hamster lung cell line V79 was ca. 13 times more resistant to peplomycin (PEP), and 6 times more resistant to bleomycin (BLM)-A2 than Chinese hamster ovary (CHO) cell line. The natural resistance of V79 cells to PEP or BLM was attributed to higher levels of BLM hydrolase activity and lower cellular uptake of the antibiotic. The sensitivity to PEP of a mutant clone CHO/O-2 T-1 was similar to that of CHO. A hybrid clone of CHO/O-2 T-1 X V79 showed an intermediate sensitivity to PEP between those of both parental cell lines, suggesting that the gene responsible for the natural resistance to PEP appears codominantly in the hybrid. The BLM hydrolase activity of the hybrid was also found intermediate between those of both parental cells. Mutant clones CHO/O-2 T-5 and CHO/O-2 T-6 were 8.3-9.0 times more sensitive to PEP than CHO cells. Hybrid clones CHO/O-2 T-5 X V79 and CHO/O-2 T-6 X V79 displayed PEP sensitivity similar to that of V79, suggesting that the gene responsible for the PEP supersensitivity (PEPss) behaves recessively in the hybrids. Both PEPss clones showed levels of BLM hydrolase and cellular uptake of [3H]PEP similar to the parental CHO cells, suggesting that the PEPss is due to neither BLM hydrolase nor cellular uptake of the antibiotic. Increased PEP-induced DNA cleavage and decreased DNA repair in the PEPss clones were demonstrated by alkaline sucrose density gradient sedimentation method. The results suggest that the PEPss of these mutant clones is attributed to decreased DNA-repairing activity and/or increased DNA-breaking activity.

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↗

Analysis of the mode of antigen presentation required for triggering of T cell proliferation by using various azobenzenearsonate-tyrosine derivatives.

Various azobenzenearsonate-tyrosine (ABA-Tyr) derivatives were synthesized by modifying amino and carboxyl groups at the alpha-carbon of tyrosine, with preservation of most of the ABA-Tyr moiety (ABA plus hydroxyphenyl portion of tyrosine). These derivatives were tested for the ability to stimulate ABA-L-Tyr specific T cell lines derived from B10.BR and B10.S mice. ABA-acetyltyramine, ABA-hydroxyphenylpropionic acid (ABA-PPr), and ABA-propylphenol, which lack either the carboxyl or amino group or both, could not induce T cell proliferation. The lack of stimulation by these derivatives was not due to their cytotoxic effects. A similar pattern of proliferation was obtained on stimulating lymph node T cells from B10.BR and B10.S mice primed with ABA-L-Tyr. Some differences were observed, however, between B10.BR and B10.S mice. ABA-L-Tyr-specific T cells from B10.BR mice could not respond well to ABA-D-Tyr in contrast to B10.S T cells. Furthermore, B10.BR mice primed with ABA-acetyltyramine or ABA-PPr in complete Freund's adjuvant could not induce ABA-L-Tyr-reactive T cells, whereas T cells from B10.S mice primed with these derivatives could proliferate in the presence of ABA-L-Tyr. The differences between B10.BR and B10.S mice were further investigated by using (B10.S X B10.BR)F1 mice. T cells from ABA-L-Tyr-immunized F1 mice responded poorly to ABA-D-Tyr when presented with B10.BR antigen-presenting cells (APC), but responded well when presented with B10.S APC. Similarly, T cells from ABA-PPr-primed F1 mice did not proliferate to ABA-L-Tyr in the presence of B10.BR APC, but could proliferate in the presence of B10.S APC. Our results clearly indicate that the presence of charged groups at the alpha-carbon of tyrosine plays a critical role in the triggering of ABA-L-Tyr-specific T cell proliferation. The significance of these results is discussed.

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[Changes in regional cerebral catecholamines following middle cerebral artery occlusion in the rat].

Ischemic brain injury affects the content and metabolism of brain monomines. Our aim was to know the time course of changes in regional cerebral catecholamines during focal cerebral ischemia, and whether focal cerebral ischemia may affect the metabolism of catecholamines in distant area of the brain. Methods Fifty-five rats were subjected to occlusion of the middle cerebral artery (MCA) on the olfactory tract, under halothane anesthesia. Fourteen animals were sham-operated group. Animals were decapitated at 1/2, 1,2,3,6,12 and 24 hours post-occlusion (PO), respectively. The brains were removed, and the brain structures dissected out include bilateral corpus striatum, cerebral cortex (MCA territory) and cerebellar hemisphere. Catecholamines were extracted by alumina procedure, and determined by high-performance liquid chromatography with electrochemical detection. Results Dopamine (DA) contents, in ipsilateral corpus striatum and cerebral cortex to the ischemia, decreased at 1 hour PO, and reached, at 6 hours PO, to 40% of control value in corpus striatum and 30% in cerebral cortex, respectively. After 6 hours PO, DA remained low. Norepinephrine (NE) content in the ipsilateral corpus striatum gradually reduced and reached to 60% of control value at 24 hours PO. NE in the ipsilateral cerebral cortex decreased to 50% of control at 1 hour PO, and thereafter remained reduced. In the contralateral corpus striatum and cerebral cortex, either DA or NE showed no significant changes, except 1/2 hour PO. NE contents in bilateral cerebral cortex showed a transient increase at 1/2 hour PO. Cerebellar NE content, bilaterally, reduced slowly to 70% of control at 24 hours PO.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Shape change of human erythrocytes induced by phosphatidylcholine and lysophosphatidylcholine species with various acyl chain lengths.

Intact human erythrocytes were treated, under non-haemolytic conditions at 37 degrees C, with synthetic phosphatidylcholine which has homologous, saturated acyl chains of 8-18 even-numbered carbon atoms (C8-C18-PC) or with lysophosphatidylcholine which has a saturated acyl chain of 8-18 carbon atoms (C8-C18-lysoPC). The C8-C14-PC and C12-C18-lysoPC species were rapidly incorporated into the erythrocytes and induced a shape change of the crenation (echinocyte formation) type. The site of the incorporation was found to be most probably on the outer leaflet of the membrane lipid bilayer. The extent of the shape change was dependent on the amount of each lipid incorporated. When the same amount of a PC or lysoPC species was incorporated into the membrane, about the same extent of crenation was induced, independent of acyl chain length. However, C16-PC, C18-PC, C8-lysoPC and C10-lysoPC, which were not incorporated into the erythrocytes, did not induce any shape change. It is therefore suggested that the hydrophobic moiety of these amphiphilic lipids may greatly contribute to their transfer from the outer medium into the erythrocyte membrane, but do not influence so much the perturbation of the membrane lipid bilayer which may be responsible for induction of the shape change.

Erythrocyte Membrane↗

Delayed neuronal death in the rat hippocampus following transient forebrain ischemia.

An unusual, slowly progressing neuronal damage has been reported to occur in the gerbil hippocampus following ischemia (Kirino 1982). Delayed neuronal death following ischemia has also been noticed in the rat four-vessel occlusion model (Pulsinelli et al. 1982). By light microscopy this slow neuronal injury in the rat was not different from the previously known neuronal ischemic cell change. This report lead us to the question as to whether neurons in the rat hippocampus are damaged rapidly following an initial latent period or deteriorate slowly and progressively until they display overt changes. To clarify this point, observation was done on the hippocampal CA1 sector of the rat following ischemia. Rats were subjected to four-vessel occlusion, and those which developed ischemic symptoms were perfusion-fixed. Although the change appeared very slowly and lacked microvacuolation of the cytoplasm, neuronal alteration was practically not different from classical ischemic cell change. By electron microscopy, however, the change was detectable when the neurons still appeared intact by light microscopy. An increase in the membranous organelles and deposition of dark substances were the initial manifestations. It seemed that the CA1 neurons deteriorated very slowly and progressively, and that they retained partial viability in the initial phase of the change. In spite of the difference in light-microscopic findings, the mechanisms underlying delayed neuronal death in the rat and gerbil hippocampus seemed to be identical.

Animals↗

Isolation of Rickettsia tsutsugamushi antigenically different from Kato, Karp, and Gilliam strains from patients.

Rickettsia tsutsugamushi strains from three recent patients of Tsutsugamushi disease in Niigata Prefecture were isolated primarily in mice and then in L cell cultures. By this procedure, low virulent strains to mice, as well as high virulent ones, could be isolated and cultivated serially in L cell cultures, suggesting the usefulness of L cells for isolation of this species of rickettsia. Each newly isolated strain was identified as a member of R. tsutsugamushi from the results of cross immunological tests and morphological observation. On the other hand, it was recognized that one of these rickettsiae showed immunological properties distinguishable from the prototype strains of Kato, Karp, and Gilliam by the cross complement fixation test, and also had low virulence in mice.

Animals↗