Tomiyasu Koyama

Membrane Viscosity Correlates with α1-Adrenergic Signal Transduction of the Aged Rat Cerebral Cortex

Abstract: We investigated, using adult (2‐month‐old) and senescent (12‐ and 24‐month‐old) rats, the effects of aging on the relationship between the α1‐adrenergic coupling system and the membrane viscosity of the cerebral cortex. There was no age‐related difference in the KD values of [3H]prazosin binding on the membranes. The Bmax values of [3H]prazosin binding were reduced with advanced age. Norepinephrine induced formation of 3H‐labeled inositol phosphates (3H‐IPs) in the slices increased with advanced age. The EC50 values for norepinephrine to stimulate the formation of 3H‐IPs at advanced age were lower than that at adult age. The cholesterol content in membranes increased with advanced age. No changes in the phospholipid content in membranes were observed with advanced age. Concomitantly, an increase of the molar ratio of cholesterol to phospholipids was observed with advanced age. The membrane viscosity as measured by 1,6 diphenyl‐1,3,5‐hexatriene increased with advanced age. These results indicate that the altered cholesterol content and/or viscosity in cortical membranes of the aged rat may account for the loss of α1‐adrenergic receptor density and/or compensatory changes in the receptor‐phospholipase C coupling system.

Red Blood Cell Deformability in Renal Failure

Red blood cell deformability was measured in 74 cases of renal failure and diabetic nephropathy by means of a modified nuclepore membrane filter method. Low red blood cell deformability was observed in a certain proportion of the patients. In cases of renal failure only weak correlation was found between reduced red blood cell deformability and BUN as well as between reduced red blood cell deformability and creatinine level, while simultaneous changes in these variable were observed in several patients. The reduction in red blood cell deformability in cases of diabetic nephropathy slightly exceeded that in cases of renal failure and correlated with an increment in HbA1c content. The effects of uremic toxins were unclear in in vitro tests. The red blood cell deformability was impaired in nephrectomized rabbits.

Improved filtration method for red cell deformability measurement

A method which enables reliable measurements of red cell deformability in whole blood to be performed is presented. It is based on the Nuclepore filtration method described byReidet al. (1976). Its reliability was much improved, obtaining an accuracy of within 5% by a technique to remove air bubbles trapped in the filter pores, which caused the poor reproducibility observed before. Simple analyses of the flow characteristics of whole blood and the haematocrit dependence of whole blood passage time were given to eliminate the effect of transient pore blockings of white cells and the contribution of different haematocrit in the whole blood passage time. A mean pore passage time of single red cells is obtainable as a quantitative index of red cell deformability. Furthermore, a good utilisation was achieved by making full use of electrically operated valves and aspiration pumps. It is possible to repeat measurements at intervals of 3 min. This improved filtration method will be useful for diagnostic purposes since it can avoid artefacts caused by artificial treatments added on blood samples and time changes after blood sampling.

The capillarity of left ventricular tissue of rats subjected to coronary artery occlusion

OBJECTIVE The effects of chronic coronary occlusion on the capillarity of left ventricular tissue in rat heart. METHODS The heart was exposed through a left-sided thoracotomy in ethylether-anesthetized and ventilated rats. The left coronary artery was occluded with a thin suture and the thorax was closed. Thirty days after the surgery the heart was removed and cross-sections were made for morphometric analyses. RESULTS The cross-sectional area of cardiomyocytes increased markedly in the subepi- and subendocardia with a concomitant increase in the capillary to myocyte ratio. The total capillary density increased in the non-ischemic zone. The balance between the increase in capillary density and myocyte hypertrophy was assessed from the capillary domain area (CDA). CDAs were significantly smaller for the venular capillary portion in the subendocardial non-ischemic zone. However, CDAs in the transition zone around the necrosis were significantly increased in the venular capillary portion in the subendocardium and in all capillary portions in the subepicardium compared with those in the non-ischemic zone. Immunohistological staining for basic fibroblast growth factor (bFGF) revealed punctate bFGF distribution in both the non-ischemic and transition zones of ventricular tissues 30 days after occlusion, suggesting a persistent stimulation for capillary angiogenesis. CONCLUSION The remodelling of the capillary bed is not uniform throughout the cardiac tissue in coronary-occluded rat hearts. CDAs decreased in the non-ischemic zone but increased in the transition zone. The rate of capillary angiogenesis failed to parallel the development of hypertrophy in the transition zone over 30 days, as could be seen from the significant reduction in the percentage of venular capillary portions.

Oxygen transport to tissue X

Hyperthermia is a developing modelity for the treatment of cancer. This therapy is occasionally used by itself, however, usually it is used as an adjuvate with chemo or radiation therapy. The mechanism for this treatment is based on, the fact that cancer cells are heated preferentially by heat application due to lower vascularity in the tumor tissue as compared with the surrounding normal tissue and that, when used with radiation therapy or chemo therapy, higher oxygen partial pressure in the tumor results in increased tumor cell damage. Appropriate mathematical models and their real time prediction of oxygen and temperature profiles could be very helpful in achieving optimal results via hyperthermia and to avoid possible danger which might occur during the treatment. Because of the complexity and the heterogeneous nature of physiological system, it is necessary to include heterogeneous properties in the mathematical models for them to be useful for biomedicai calculations. Of course, it is much more difficult to sorve mathematically the heterogeneous system than the homogeneous one. In this paper, the importance of the implementation of heterogeneities in the heat and mass transport for biological system mathematical modelling is discussed. Results of a three dimensional computer simulation of mass and heat transfer in tumor tissue with different capillary geometries during hyperthermia are demonstrated. The method used for the computer simulation is a deterministic/ probabilistic technique, Williford-Bruley calculational strategy.

Blood flow velocity in pulmonary microvessels of bullfrog

Flow velocity in the pulmonary microvessels of the exposed lung of bullfrogs was measured by means of a laser Doppler microscope of an oblique backward mode, together with a signal-analyzing system having a time sharing circuit triggered by the R-wave of the ECG. By these means, measurements of the changes of flow velocity contour in the cardiac cycle were made. Flow velocity was clearly pulsatile in response to cardiac cycles in all microvessels including capillaries. Flow velocities in the arteriole and venule consistently decreased for a short period after the R-wave (84 +/- 33 msec (mean +/- SD) in the arteriole and 130 +/- 31 msec in the venule, respectively) and rapidly increased up to a maximicronm value. The mean flow velocities in arterioles (diameter 50 +/- 17 micron) and venules (39 +/- 9 micronm) were 2.29 +/- 0.32 and 2.30 +/- 0.27 mm/sec. The amplitudes of pulsatile flow in these vessels were 0.83 +/- 0.31 and 0.63 +/- 0.16 mm/sec, respectively. In the capillary the times from the R-wave to the minimicronm and maximum values were variable. In some cases the velocity gradually increased without first decreasing and the increase sharply accelerated a certain time after the R-wave. The mean velocity in the pulmonary capillary and the amplitude of the pulsatile flow ere 1.78 +/- 0.31 and 0.37 +/- 0.12 mm/sec, resepctively. The ratios of the pulsatile amplitude to the mean velocity in the pulmonary capillary, venule and arteriole averaged 0.21, and 0.36, respectively.

Different enzyme activities in coronary capillary endothelial cells.

Differential distributions of alkaline phosphatase (AP) and dipeptydylpeptidase IV (DPPIV) were studied in coronary microvascular endothelial cells. Endothelial cells were obtained by the perfusion of coronary vessels with 0.1% trypsin PBS solution and cultured in uncoated culture dishes. Staining of cultured endothelial cells with AP- and DPPIV-sensitive reagents revealed blue or red staining, respectively. Most colonies showed cells of only one color, blue or red, even at the fifth passage. AP-sensitive cells, which were originally elongated, shortened and widened, proliferating to form monolayer colonies of cobble stone-like cells. AP-stainability became weak with repeated passages. DPPIV-sensitive endothelial cells remained elongated even after repeated passages. The cell shape and stainability seemed to be coupled and maintained through the five passages studied.

Reduced deformability of erythrocytes exposed to hypercapnia.

The effect of hypercapnia on the deformability of erythrocytes was studied by means of a nuclepore membrane filter method. A decrement of the deformability by 20–40% was observed when

Viscosity and order in erythrocyte membranes studied with nanosecond fluorometry

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