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Biomedical subjects

N Tateishi

Publications and source records attributed to N Tateishi.

At least 37 records · Page 2Linked to original sources

Erythrocyte flow and elasticity of microvessels evaluated by marginal cell-free layer and flow resistance.

Flow dynamics of human erythrocytes was compared in elastic (E) and hardened (H) microvessels with inner diameters of 10-40 microns. The thickness of the marginal cell-free layer and the overall flow resistance were measured with a vascular bed isolated from rabbit mesentery (E vascular bed) as well as with a 4% paraformaldehyde-fixed bed (H vascular bed). 1) In both E and H microvessels, the thickness of the cell-free layer increased with increasing inner diameter of the microvessels and with decreasing hematocrit accompanied by an overall decrease in the flow resistance. The hematocrit-dependent change of the cell-free layer thickness was greater in the E microvessels than in the H microvessels. The flow resistance was always greater in the H vascular beds than in the E vascular beds. 2) With decreasing erythrocyte deformability induced by treatment with 2 mM diazenedicar-boxylic acid bis(N,N-dimethylamide), the thickness of the cell-free layer decreased at a low hematocrit in the E microvessels and at a high hematocrit in the H microvessels, although the flow resistance was increased in both vascular beds. 3) Dextran of 70,400 average molecular weight accelerated the formation of the cell-free layer by inducing erythrocyte aggregation. A drastic increase in the cell-free layer thickness at 2-4 g/dl of dextran in the E microvessels and at 1-2 g/dl of dextran in the H microvessels was accompanied by a significantly lower increase in the flow resistance. This study concludes that the elasticity of microvessels may play an important role for reducing the overall flow resistance of a vascular bed, which is modulated by the marginal cell-free layer, itself a function of the rheological properties of the erythrocytes.

Animals↗

Deformation of erythrocytes in microvessels and glass capillaries: effects of erythrocyte deformability.

OBJECTIVE: The deformation of erythrocytes in microvessels less than 15 microns in inner diameter was analyzed using a microvascular bed isolated from rabbit mesentery. The deformation was compared with that found in glass capillaries. METHODS: Human erythrocytes were perfused through two media: first, a microvascular-bed section isolated from rabbit mesentery; and second, a set of glass capillaries. Images of deformed erythrocytes were recorded on videotape under strobe light and analyzed with an image processor. The flow velocity of the erythrocytes was determined from the difference of their positions between video frames or by a dual-spot cross-correlation technique. Erythrocyte deformability was modified with diamide, diazene dicarboxylic acid bis[N,N-dimethylamide], by crosslinking spectrins. RESULTS: Symmetrical (parachute-like or slipper-like) deformation of erythrocytes was observed only in microvessels smaller than 13 microns in inner diameter. Erythrocytes in microvessels were less deformed than those in glass capillaries with corresponding diameters, and the marginal cell-free layer was narrower. The deformation increased by increasing the flow velocity of erythrocytes, and the cell-free layer became wider. Diamide-treated cells in microvessels were less deformed than normal cells and showed slightly narrower cell-free layers. Stronger stress in narrower microvessels induced further deformation of cells. CONCLUSIONS: Erythrocyte deformation in microvessels was essentially different from that in glass capillaries, and the effect of erythrocyte deformability on the flow dynamics of erythrocytes in microvessels was properly evaluated using an isolated microvascular bed.

Animals↗

Quantitative evaluation of flow dynamics of erythrocytes in microvessels: influence of erythrocyte aggregation.

Effects of erythrocyte aggregation on the flow dynamics of erythrocytes in microvessels were examined quantitatively by perfusing human erythrocytes suspended in isotonic medium containing various concentrations of dextran (70,400 avg mol wt, Dx-70) into a part of the microvascular bed isolated from rabbit mesentery. Thickness of the marginal cell-free layer was measured with an image analyzer, total flow resistance was determined on the basis of the perfusion pressure-volume flow relationship, and homogeneity of erythrocyte flow was evaluated by the power spectrum obtained by the fast Fourier transform of the light intensity change monitored on single microvessels. With increasing dextran concentration, suspension viscosity of erythrocytes at high shear rates increased linearly and thickness of the cell-free layer increased in a sigmoidal fashion. Flow resistance increased relatively little over the range of dextran concentrations in which the cell-free layer increased most rapidly. Furthermore, the flow pattern of erythrocytes in microvessels became inhomogeneous. In conclusion, the present study shows that Dx-70-induced erythrocyte aggregation results in increased flow resistance in the circulatory system, even through the widening of the cell-free layer tends to reduce the resistance and also results in inhomogeneous flow of erythrocytes in microvessels.

Animals↗

Simultaneous influence of erythrocyte deformability and macromolecules in the medium on erythrocyte aggregation: a kinetic study by a laser scattering technique.

The aggregation and sedimentation kinetics of human erythrocytes was studied by modifying the cellular properties and medium compositions simultaneously. Dextrans of average molecular weight 70400 and 494000 were used to provide suspending medium modifications, while diamide (diazene dicarboxylic acid bis(N,N-dimethylamide)) was used to alter the membrane structural properties. Laser scattering method was employed for this study, and it was compared with a kinetic method combined with a low-shear rheoscope and an image analyzer. From scattered light intensity profiles continuously obtained during aggregation of erythrocytes and sedimentation of the aggregates, characteristic kinetic parameters were computed. Kinetic parameters obtained from a phase of the one-dimensional aggregate formation and sedimentation corresponded well to the velocity of rouleaux formation obtained by the low-shear rheoscope technique. Dextrans accelerated the erythrocyte aggregation and the sedimentation, and diamide treatment suppressed the process by decreasing the erythrocyte deformability. The aggregating force by dextrans overcame the disaggregating force by the decreased deformability. However, the arrangement of erythrocytes as expressed in specific units for aggregates (i.e., rouleaux) became irregular by decreasing the erythrocyte deformability. In conclusion, the progression of erythrocyte aggregation and the structure of the aggregates were dependent on both erythrocyte properties and macromolecules in the medium.

Blood Sedimentation↗

Flow dynamics of erythrocytes in microvessels of isolated rabbit mesentery: cell-free layer and flow resistance.

A part of microvascular bed isolated from rabbit mesentery (composed of a few branches of superior mesenteric artery) was used for the study of the flow dynamics of erythrocytes in microvessels. The flow resistance in the microvascular bed was analyzed with respect to the thickness of a cell-free layer formed along the inner wall of vessels and the suspension viscosity of erythrocytes (in terms of hematocrit and erythrocyte deformability). The thickness of the cell-free layer increased with the increase in the inner diameter of microvessels. By lowering the hematocrit, the thickness increased and the flow resistance decreased. Meanwhile, by decreasing the erythrocyte deformability with diamide, the thickness decreased and the flow resistance increased. However, the thickness was not altered in microvessels less than 10 microns by the diamide treatment. The maximum inner diameter of microvessel required to induce parachute- and/or slipper-like deformation (at flow velocity of erythrocytes less than 2 mm s-1) was 13 microns for control cells and 6 microns for diamide-treated cells. The thickness of the cell-free layer could not be closely related to the flow resistance, while a good relationship applicable to both control and diamide-treated cells was obtained between the flow resistance and the suspension viscosity. The present results conclude that the flow resistance in the isolated microvascular bed is mainly provided by both hematocrit and erythrocyte deformability.

Animals↗

Induction of a 23 kDa stress protein by oxidative and sulfhydryl-reactive agents in mouse peritoneal macrophages.

The synthesis of 23 kDa protein was enhanced when mouse peritoneal macrophages were exposed to oxidative agents such as hydrogen peroxide and menadione, or to sulfhydryl-reactive agents such as diethylmaleate, cadmium chloride and sodium arsenite. After 11 h exposure to these agents the 23 kDa protein was one of the actively synthesized proteins in the macrophages. Under similar conditions the 34 kDa protein previously identified as heme oxygenase, was induced and its synthesis preceded that of the 23 kDa protein. Neither the 23 kDa or the 34 kDa protein was induced by hyperthermia. Conversely, the various oxidative and sulfhydryl-reactive agents employed here did not induce the major heat shock proteins in the macrophages. When the macrophages were activated by bacterial lipopolysaccharide or other stimulants, many proteins are known to be induced, however, the 23 kDa and 34 kDa proteins were not induced. The 34 kDa protein, i.e., heme oxygenase, has been found to be stress-induced in various types of cell, but not the 23 kDa protein. This suggests that the 23 kDa protein is a stress protein predominantly expressed in macrophages.

Animals↗

Rheological evaluation of X-ray irradiated blood.

CPD blood, in glass containers and polyvinyl chloride (PVC) bags, was X-irradiated with 50 Gy (n = 4), and then stored for up to 4 weeks. Rheological, hematological and biochemical changes of the red cells during storage were examined. Changes in blood viscosity and various properties of red cells (deformability, shape, hematological indices and concentrations of 2,3-diphosphoglycerate and nucleotides) during storage at 4 degrees C were not altered by the irradiation. The rheological, biochemical and morphological properties of red cells were as well preserved in PVC bags as in glass containers. No abnormal and/or cross-linked proteins in red-cell membranes were induced by the irradiation. Irradiation of PVC bags did not affect the above properties of red cells through any changes in the bag constituents. In conclusion, X-ray irradiation with 50 Gy to CPD blood in PVC bags does not affect the rheological functions of red cells.

Blood↗

GABAergic synaptic current in dissociated nucleus basalis of Meynert neurons of the rat.

gamma-Aminobutyric acid (GABA)-mediated spontaneous inhibitory postsynaptic currents (IPSCs) were recorded from dissociated rat nucleus basalis of Meynert neurons which still had their synaptic boutons attached. The membrane currents were recorded by the whole-cell patch-clamp technique. Elevated extracellular K+ concentration and the addition of the calcium ionophore, A23187, enhanced the amplitude and frequency of spontaneous IPSCs. Ryanodine and Ca(2+)-free external solution containing EGTA or BAPTA markedly decreased the spontaneous IPSC activities. Spontaneous IPSC activities were reversibly reduced by baclofen and increased by phaclofen, indicating that the GABAB receptor regulates the release of GABA from nerve terminals and acts as a negative autoreceptor.

Animals↗

ONO-4057, a novel, orally active leukotriene B4 antagonist: effects on LTB4-induced neutrophil functions.

ONO-4057(5-[2-(2-Carboxyethyl)-3-[6-(4-methoxyphenyl)-5E- hexenyl]oxyphenoxy]valeric acid), an orally active leukotriene B4(LTB4) antagonist, displaced the binding of [3H] LTB4 to the LTB4 receptor in human neutrophil (Ki = 3.7 +/- 0.9 nM). ONO-4057 inhibited the LTB4-induced rise in cytosolic free calcium (the concentration causing 50% inhibition (IC50) = 0.7 +/- 0.3 microM) and inhibited human neutrophil aggregation, chemotaxis or degranulation induced by LTB4 (IC50 = 3.0 +/- 0.1, 0.9 +/- 0.1 and 1.6 +/- 0.1 microM) without showing any agonist activity at concentration up to 30 microM. ONO-4057 did not inhibit fMLP or C5a-induced neutrophil activation at concentrations up to 30 microM. In the in vivo study, ONO-4057 given orally, prevented LTB4-induced transient neutropenia or intradermal neutrophil migration in guinea pig (the dose causing 50% efficacy (ED50) = 25.6mg/kg or 5.3mg/kg). Furthermore, ONO-4057 given topically, suppressed phorbol-12-myristate-13-acetate (PMA)-induced neutrophil infiltration in guinea pig ear (the effective dose = 1 mg/ear). These results indicate that ONO-4057 is a selective and orally active LTB4 antagonist and may be a potential candidate for the treatment of various inflammatory diseases.

Animals↗

A method for measuring the rate of oxygen release from single microvessels.

A system determining the rate of oxygen release from erythrocytes flowing in single microvessels was constructed with an inverted microscope by connecting 1) a scanning/grating spectrophotometer equipped with a photon-counting detector through a thin light guide, to obtain the visible absorption spectrum of a spot (5 microns in diameter) focused on a microvessel, 2) two photomultipliers (connected to a microcomputer via an analog-to-digital converter) through two light guides, to determine the flow velocity of erythrocytes by calculating the cross correlation between the light-intensity changes of two spots (3 microns in diameter, 5 microns apart from each other) focused on the microvessel, and 3) an image processor through a video camera, to estimate the diameter of microvessel from the digitized video images. The rate of oxygen release from single microvessels 7-25 microns in diameter in rat mesentery was measured under the superfusion of deoxygenated solution: 1) The maximal rate was obtained in capillaries, and the rate in arterial microvessels was larger than that in venous microvessels, when similar diameters were compared. 2) The rate was maximum at pH 7.0-7.2, and it decreased in more acidic and alkaline pH values. 3) The rate decreased with a decrease in temperature. The reliability of the measurement using the present apparatus was tested in detail.

Animals↗

Acetylcholine receptors in dissociated nucleus basalis of Meynert neurons of the rat.

Electrical and pharmacological properties of acetylcholine (ACh)-induced currents in neurons dissociated from the nucleus basalis of Meynert (nBM) of immature (2-week-old) rats were investigated with the whole-cell mode of the patch-clamp technique. At a holding potential (VH) of -50 mV, ACh (10(-4)M) evoked a transient inward current mimicked by nicotine (InACh), followed by a sustained outward current mimicked by carbamylcholine (ImACh). The KD values were 1.2 x 10(-4) M for ImACh and 8.7 x 10(-7) M for ImACh. The reversal potential of ImACh was close to EK. The ImACh was determined to be elicited via the M2 muscarinic receptor, based on the differences in sensitivity to muscarinic antagonists such as pirenzepine and AF-DX-116.

Acetylcholine↗

Modulatory action of cholinergic drugs on N-methyl-D-aspartate response in dissociated rat nucleus basalis of Meynert neurons.

The effect of cholinergic drugs on N-methyl-D-aspartate (NMDA)-activated responses in neurons dissociated freshly from the nucleus basalis of Meynert (nBM) of immature and mature rats were investigated with the whole-cell patch-clamp technique. The NMDA (10(-4) M)-evoked inward current (INMDA) consisted of transient peak and successive steady-state components. In mature neurons, the peak component was suppressed by pretreatment with acetylcholine (ACh, 10(-12)-10(-5) M) but facilitated by higher concentrations (greater than 10(-5) M). Regardless of the concentration, ACh had no effect on INMDA of immature neurons. The results suggest that ACh behaves as either inhibitory or excitatory modulator of glutaminergic transmission in nBM neurons of adult mammals depending on ACh concentration.

Acetylcholine↗

Lipid peroxidation in the liver of carcinogen-resistant rats.

Recently, we developed a new strain of rats that exhibit marked resistance to the hepatotoxic and carcinogenic actions of 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB) and some other carcinogens. In this work, we compared lipid peroxidation in the liver of these carcinogen-resistant (R) rats and the parental Donryu strain rats that are sensitive (S) to hazardous actions of these carcinogens. The liver microsomal fractions of the R group contained less amounts of polyunsaturated fatty acids. Microsomal lipid peroxidation in the presence of exogenous NADPH was much lower in R rats than in S rats. Liver microsomes of R rats were much less active than those of S rats also in producing 4-hydroxynonenal, carbonyl compounds and conjugated diene. The hepatic contents of ascorbic acid, glutathione, alpha-tocopherol and coenzyme Q in the R rats were similar to those in S rats. The activities of the free radical scavenger enzymes, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), in the two groups were also similar. Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are both thought to function in disposal of these cytotoxic aldehydes. The liver microsomal and mitochondrial ALDH activities of the two groups were similar. The ADH activity of the liver cytosolic fraction of R rats was nearly twice that of S rats, as measured with 4-hydroxynonenal as substrate. The higher ADH activity may explain the decreased lipid peroxidation in R rats at least partly, if this enzyme is involved in lipid peroxidation.

Alcohol Dehydrogenase↗

Effects of antagonists on N-methyl-D-aspartate response in acutely isolated nucleus tractus solitarii neurons of the rat.

The effect of antagonists on N-methyl-D-aspartate (NMDA)-induced response was investigated in isolated nucleus tractus solitarii (NTS) neurons freshly isolated from the rat using a conventional pathclamp technique. The NMDA-induced inward current consisted of an initial peak followed by a steady-state component. The competitive antagonists of NMDA receptor, D-2-amino-5-phosphonovalerate (APV), D-2-amino-4-phosphonoheptanoate (APH) and 3-3(2-carboxypiperazine-4-yl)propyl-1-phosphate (CPP), selectively suppressed the initial peak of NMDA-induced current more than the steady-state component at low concentrations. The non-competitive antagonists, MK-801, ketamine, Zn2+ and Mg2+, equally blocked both peak and steady-state components.

2-Amino-5-phosphonovalerate↗

Glycine-insensitive desensitization of N-methyl-D-aspartate receptors in acutely isolated mammalian central neurons.

Acutely isolated rat central neurons were recorded by whole-cell voltage-clamp and responses to a class of excitatory amino acid N-methyl-D-aspartate (NMDA) were examined. Rapid application of NMDA evoked inward current consisted of a fast initial peak followed by a sustained component. Glycine potentiated both initial and desensitized states of the NMDA response with identical concentration-dependence. The initial response, but not the sustained component, was abolished when low concentration of NMDA was pre-applied, and glycine could not reverse the desensitization. This evidence suggests that the NMDA receptor desensitization is sensitive to NMDA but not to glycine, and support the hypothesis that glycine initiates the activation of NMDA receptors rather than that glycine prevents desensitization at NMDA receptors in these cells.

Animals↗

Acetylcholine-activated ionic currents in parasympathetic neurons of bullfrog heart.

1. The electrical and pharmacologic properties of acetylcholine (ACh)-induced current (IACh) were studied in the parasympathetic neurons isolated from bullfrog heart with the use of the concentration-clamp technique, which allows intracellular perfusion and rapid change of external solution within 2 ms under the single-electrode voltage-clamp condition. 2. The IACh consisted of an initial transient peak component and a successive steady-state plateau component. Both currents increased in a sigmoidal fashion with increasing ACh concentration. The dissociation constant (Kd value) and the Hill coefficient for each component were 2.2 X 10(-5) M and 1.6, respectively. 3. In the K(+)-free solution, the reversal potential (EACh) of IACh was close to the Na+ equilibrium potential (ENa). The current-voltage (I-V) relation showed inward rectification at positive potentials. 4. Nicotine mimicked only the peak component of IACh. However both peak and steady-state components were blocked nonselectively by the nicotinic blockers d-tubocurarine and hexamethonium. 5. Carbamylcholine (CCh) mimicked the steady-state component of IACh. The steady-state component was selectively inhibited by atropine at concentrations 1,000 times lower than that required for inhibition of the peak component. The steady state was blocked equally by either pirenzepine (M1 blocker) or AF-DX-116 (M2 blocker). 6. It was concluded that the IACh consisted of a peak component having double exponential activation and inactivation, mediated through the nicotinic actions, and a steady-state component having no inactivation, mediated through the muscarinic action.

Acetylcholine↗

Proton-gated sodium current in parasympathetic ganglion cells of frog heart.

1. The proton-gated current was investigated in whole-cell configuration of the neurons isolated from bullfrog heart parasympathetic ganglia with the use of the "concentration-clamp" technique, which combines intracellular perfusion and extremely rapid exchange of external solution within 1-2 ms, under a single-electrode voltage-clamp condition. 2. In all isolated neurons, a "step" decrease in extracellular pH (pHo) induced a transient inward current that was followed by a complete inactivation within 1 s. 3. The proton-gated current increased in a sigmoidal fashion as pHo decreased. In the external solution containing 2 mM Ca2+, the threshold of current activation was at pHo 7.4, and the maximum response appeared at pHo 6.5-5.5. The dissociation constant (Kd) and Hill coefficient were 7.1 and 3.2, respectively. 4. The proton-gated current was reduced by decreasing the extracellular Na+ concentration. In the absence of K+, the current produced by reduced extracellular pH reversed at the Na+ equilibrium potential (ENa), indicating that the current was carried by Na+. 5. Kinetics of both the activation and inactivation phases of proton-induced current were single exponential. The time constants of activation (tau a) and inactivation (tau i) had no potential dependence but decreased slightly by decreasing pHo. 6. In the inactivation curve of the proton-induced current obtained by decreasing pHo from various conditioning pHos to 6.5, the half-maximum inactivation occurred at pHo 7.75. 7. The proton-gated current was suppressed as the extracellular Ca2+ concentration [( Ca2+]o) increased from 0.1 to 10 mM, and the half inhibition occurred at greater than 10 mM [Ca2+]o.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗