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H Kon

Publications and source records attributed to H Kon.

At least 55 records · Page 3Linked to original sources
PubMed1988

A flow EPR study of deformation and orientation characteristics of erythrocyte ghosts: effects of lysing and resealing conditions.

The effects of various conditions in lysing and resealing the red cell membrane on the degree of ghost deformation and orientation in flow are investigated using the flow EPR and spin-label method. The relatively low deformability of the standard ghost, which is lysed and resealed, respectively, in hypotonic and isotonic NaCl-Tris buffer, is markedly enhanced by the presence of Mg-ATP, chlorpromazine, or Ca2+ ion during resealing. The effect is concentration dependent, and there is an optimal level for each treatment. Chlorpromazine and Ca2+ are also effective when added to the resealed ghosts. Mg2+ ion shows an opposite effect reducing the ghost deformability in flow at all concentrations. An isotonic lysis in NH4HCO3 solution with less osmotic stress substantially raises ghost deformability above that of the standard ghosts. These results are interpreted on the basis of a misalignment between the bilayer leaflets that is probably brought about during hypotonic lysis and its recovery to the nearly normal bilayer state by the agents used during or after resealing. The novel finding of deformability enhancing effect of calcium is assumed to be caused by the electrostatic expansion of the inner layer relative to the outer leaflet. The explanations are supported by the resealed ghost shapes observed before and after the treatments; shape recovery from the monoconcave spheroid toward biconcave discoid is observed in most cases concomitantly with improvements of flow characteristics.

Adenosine Triphosphate↗
PubMed1988

A flow EPR study of deformation and orientation characteristics of erythrocyte ghosts: a possible effect of an altered state of cytoskeletal network.

Using the flow EPR technique, we investigated the resealed ghost deformability in shear flow and the effects of the altered state of cytoskeletal network induced by hypotonic incubation of ghosts. Isotonically resealed ghosts in the presence of Mg-ATP, in which alteration of cytoskeletal network is not effected, have smooth biconcave discoid shapes, and show a flow orientation and deformation behavior similar to that of erythrocytes, except that higher viscosities are required to induce the same degrees of deformation and orientation as in erythrocytes. The flow behavior of resealed ghosts is Mg-ATP dependent, and the shape of the ghosts resealed without Mg-ATP is echinocytic. In contrast, the ghosts resealed by hypotonic incubation show a markedly reduced deformability even with Mg-ATP present. Nonreducing, nondenaturing polyacrylamide gel electrophoresis (PAGE) of the low ionic strength extracts from hypotonically resealed ghosts reveals a shift of the spectrin tetramer-dimer equilibrium toward the dimers. In the maleimide spin-labeled ghosts, the ratios of the weakly immobilized to the strongly immobilized EPR intensities are larger in hypotonically resealed ghosts than in the isotonically resealed ghosts, indicating an enhanced mobility in the spectrin structure in the former. Photomicrographs of hypotonically resealed ghosts show slightly stomatocytic transformations. These data suggest that the shape and the deformability loss in hypotonically resealed ghosts are related to an alteration of the spectrin tetramer-dimer equilibrium in the membrane. Thus, the shift of the equilibrium is likely to affect the regulation of the membrane deformability both in normal and pathological cells such as hereditary elliptocytes.

Adenosine Triphosphate↗
PubMed1988

[Effect of gonadectomy on the onset of diabetic syndrome in the female WBN/Kob rats].

Almost all the male animals of WBN/Kob rat strain show the diabetic syndrome whereas none of the female animals develop such diseased conditions even at elder age. We investigated the effect of sex hormones on the onset of diabetic syndrome of this rat strain by comparing the results of body weight gain and various clinical tests such as glucose tolerance, serum biochemistry and histopathology of spayed female rats with those of non-treated and sham-operated female animals kept until 88-week-old. Non-treated and sham-operated female animals had shown no abnormal result even at 88-week-old. Spayed female animals began to reveal glucosuria associated with polydipsia and polyuria from 72-week-old, and gradually developed emaciation and cataract. Increased body weight gain, impaired glucose tolerance and lasting hyperglycemia were observed prior to the onset of the symptoms. Pancreatic changes consisted of atrophy of acinar tissue and atrophy or disappearance of islet tissue attributable to clinical data also were detected in spayed female animals. These diabetic syndrome and pancreatic change were analogous to those of aged male WBN/Kob rats but the onset of spayed females was delayed and less severe. Present results suggest that female sex hormones are protective from the onset of diabetic syndrome of WBN/Kob rats.

Animals↗
PubMed1987

Low-temperature electron paramagnetic resonance study of the ferricytochrome c-cardiolipin complex.

The electron paramagnetic resonance spectrum of the ferricytochrome c complex with cardiolipin was observed at temperatures below 20 K. For the low-spin iron(III) heme system complexed with the negatively charged lipid, the tetragonal and rhombic ligand field parameters (delta/lambda = 3.58, V/lambda = 1.82) differ significantly from those (delta/lambda = 2.53, V/lambda = 1.49) of the free ferricytochrome c sample. The g values of the complex (gx = 1.54 +/- 0.02, gy = 2.26 +/- 0.01, gz = 3.02 +/- 0.01) are compared to the values for free ferricytochrome c (gx = 1.25 +/- 0.02, gy = 2.25 +/- 0.01, gz = 3.04 +/- 0.01). Spectral alterations are interpreted in terms of the ligand field changes induced within the heme group by association with the negatively charged phosphoglyceride.

Cardiolipins↗
PubMed1987

The copper-induced deformability loss and echinocyte formation in human erythrocytes: an electron paramagnetic resonance study.

The effects of incubating human erythrocytes with CuSO4 under prelytic conditions were investigated to clarify the cause of the decreased whole-cell deformability in flow, using electron paramagnetic resonance and the spin label methods. The effect of Cu(II) on erythrocytes was manifested by the dose-dependent echinocyte formation as well as by the decreased whole-cell deformability in flow. The deformability loss was attributed in part to the altered rheological characteristics due to echinocytosis. By using various reagents capable of reversing the effects of Cu(II) on morphology and deformability loss, it was shown that at least two types of damage sites can be distinguished. They include (1) the Cu(II) coordination sites which can be dissociated by chelating reagents and (2) the sites involving sulfhydryl groups in which disulfide bonds and probably Cu(II)-bridged strong coordination bonds are formed. The latter can be reversed by sulfhydryl reagents such as dithioerythritol, but not by EDTA or penicillamine. Pretreatment of erythrocytes with iodoacetate eliminates the variance in susceptibility to Cu(II) among donors and generally enhances the effects of Cu(II) by lowering the cellular glutathione level. A possible relation between the nature of the damage sites and the cellular glutathione level is discussed.

Adult↗
PubMed1987

Spin label study of erythrocyte deformability. Ca2+-induced loss of deformability and the effects of stomatocytogenic reagents on the deformability loss in human erythrocytes in shear flow.

The Ca2+-induced loss of deformability in human erythrocytes and the recovery of the lost deformability by stomatocytogenic reagents were investigated by means of a new flow electron paramagnetic resonance (EPR) spin label method, which provides information on deformation and orientation characteristics of spin labeled erythrocytes in shear flow. The Ca2+-induced loss of deformability is attributed mainly to the increase in intracellular viscosity resulting from efflux of intracellular potassium ions and water (Gardos effect). Partial recovery of the lost deformability is demonstrated in the presence of stomatocytogenic reagents, such as chlorpromazine, trifluoperazine, W-7, and calmidazolium (R24571). The recovery can not be explained solely by suppression of the Gardos effect due to the reagents. Incorporation of an optimal amount of the reagents into the membrane appears to compensate for the membrane modification due to Ca2+ ions to restore a part of the lost deformability.

Calcimycin↗
PubMed1986

Cold shock hemolysis in human erythrocytes studied by spin probe method and freeze-fracture electron microscopy.

When human erythrocytes are osmotically stressed or chemically treated, they hemolyze on cooling below 10 degrees C (called cold shock). We have studied the effects of osmotic stress and cooling on the state of membrane by the spin-probe method and freeze-fracture electron microscopy. At room temperature, the membrane fluidity detected by 12-doxyl stearate spin probe showed a steady decrease with osmolality in hypertonic NaCl solutions up to 900 mOsm/kg, above which it remained unchanged. In hypertonic sucrose solutions, the electron paramagnetic resonance spectra showed an additional pair of absorptions, indicating development of regions, in the membrane, further immobilized than in NaCl solutions. Mobility of a cholesterol analogue probe, androstane, did not show change by hypertonicity, but the spectral intensity dropped at 1,200 mOsm/kg, probably due to formation of loose aggregates in the cholesterol phase. On cooling the osmotically stressed cells in NaCl solution, the isotropic rotational correlation time vs. inverse temperature plot of 12-doxyl stearate probe exhibited a step-wise discontinuity at approximately 10 degrees C, suggestive of a drastic transition in the state of the membrane. At about the same temperature, the freeze-fracture pattern of osmotically stressed cells revealed the development of large wrinkles and aggregation of membrane particles, in contrast to the case of the cells in isotonicity. Significance of these findings in understanding cold shock hemolysis is discussed.

Cold Temperature↗
PubMed1985

Effect of the presence of hardened erythrocytes on deformation-orientation characteristics of normal erythrocytes in shear flow studied by the spin label method.

The effect of the presence of hardened red blood cells (HRBC) in a mixed suspension on the deformation-orientation characteristics of normal cells in flow is experimentally probed by the electron spin resonance (ESR) spin label method, using a phosphatidylcholine label which does not transfer between the cells. The average deformation-orientation of the normal cells is generally suppressed by the presence of HRBC to different degrees, depending upon the shape and the way the HRBC are prepared. The effects are qualitatively explained by disturbance of laminar shear flow due to the random tumbling of the HRBC.

Blood Flow Velocity↗
PubMed1985

Study of the effect of varying hematocrit on free deformation and orientation of erythrocytes in flow.

The electron paramagnetic resonance (EPR) spin label method was used to investigate the effect of varying hematocrit on the deformation and orientation behavior of erythrocytes in shear flow. The relative EPR spectral change due to flow, which we use as a measure of the average deformation and orientation of erythrocytes, was observed as a function of the hematocrit. The profile generally shows a rising and a declining phase with the maximum in-between. The position of the maximum with respect to the hematocrit and the level of the spectral change are influenced by the suspending medium viscosity, osmolarity, and depend upon modifications of the red cell properties such as the internal viscosity, area-to-volume ratio and membrane rigidity. Results show that there is an upper limit of free deformation and orientation of the cell for a given hematocrit value. A possible role of the cell-cell interaction is discussed in restricting the space around a cell which is required for free deformation and orientation. A significance of the findings is that the actual deformation and orientation of an ensemble of cells which give rise to the EPR spectral change in flow is determined not only by the single cell deformability but also by the way the cells interact with each other under a given fluid dynamic condition.

Blood Viscosity↗
PubMed1984

Changes in cytochrome P-450 iron spin state as a function of castration, testosterone or estrogen treatment, vigorous exercise, or starvation.

3-Methylcholanthrene treatment induced large increases in high-spin P-450 form(s) in intact microsomal membranes from New Zealand White rabbit or C57BL/6N mouse liver but not in rabbit lung, kidney or intestine. Testosterone enhanced the high-spin/low-spin ratio of P-450 in female C57BL/6N liver to the same degree as 3-methylcholanthrene treatment; there was no additive increase in the ratio with treatment of testosterone plus 3-methylcholanthrene. The same effect was seen in ovariectomized females. In the liver of normal or castrated C57BL/6N mice, estradiol-17 beta pretreatment caused no significant change in the high-spin/low-spin ratio. Vigorous homogenization of the microsomes can decrease the high-spin/low-spin ratio, more so in control than in 3-methylcholanthrene-induced C57BL/6N microsomes. Vigorous exercise (20-min swim) and fasting for 2 days significantly enhanced the high-spin/low-spin ratio in C57BL/6N liver. Temperature studies showed that the low-spin electron paramagnetic resonance signal intensity, as a function of absolute temperature in the range of 16 to 70 degrees K, was consistent with a simple Boltzmann dependence within the Kramers doublet giving rise to the microwave transition. We conclude that, within the limits of measurement, the low-spin state acts like an isolated state and is not in thermal equilibrium with the high-spin form of P-450.

Animals↗
PubMed1984

Spin-label studies of erythrocyte deformability. IV. Relation of electron spin resonance spectral change with deformation and orientation of erythrocytes in shear flow.

Electron spin resonance (ESR) spectra of spin-labeled human erythrocytes in shear flow are simulated to derive semi-empirical relations of the ESR spectral change with deformation and orientation of the cells by using a modified theoretical model developed for deformation and orientation of liquid drops. The six observed spectra at different shear stress values were simultaneously simulated by adjusting only two parameters. One parameter can be related to the ratio of the internal to the external viscosity, and the other to the elastic property of the cell membrane. From these results we have derived a semi-empirical relationship between the average deformation index or the orientation angle with a spectral measure, which characterizes the spectral shape change induced by shear stress. Thus, it becomes possible to obtain improved quantitative information on the rheological behavior of red blood cells by using the spin-label ESR method.

Electron Spin Resonance Spectroscopy↗
PubMed1983

Spin label study of erythrocyte deformability. III. Further characterizations of electron spin resonance spectral change in shear flow.

It is demonstrated that the change in the spin label ESR spectrum induced by shear flow reflects the whole cell deformation as a function of the cell surface area-to-volume ratio (s/v), the morphology and intracellular viscosity. Since the effect of the change in the membrane mechanical property on the ESR spectrum has been described previously, the spin label ESR spectrum is now shown to contain full information concerning the whole cell deformability which is determined by the major intrinsic and extrinsic properties of the red blood cells. The result of microphotographic observations shows also that the cells in the flow are elongated and aligned approximately along the flow direction to an increasing extent as the cells flow near the surface of the flat channel walls. Thus, the entire observation confirms the view that the ESR spectral difference-shear rate profile is closely related to the elongation ratio shear rate characteristics obtained by other (optical) methods.

Blood Viscosity↗
PubMed1982

A spin-label study of the correlation between stomatocyte formation and membrane fluidization of erythrocytes.

Change in the membrane fluidity of human erythrocytes on transformation to stomatocytes was observed by ESR spectroscopy using 12-doxyl stearic acid or its methyl ester as a probe. When the transformation to stomatocytes was induced by four qualitatively different methods, i.e. (a) addition of cationic amphiphilic agents such as chloropromazine, tetracaine, chloroquine or primaquine, (b) addition of Triton X-100, a non-ionic detergent, (c) lowering the pH, and (d) depleting membrane cholesterol, membrane fluidization was always observed. This indicates the existence of a close correlation between stomatocyte formation and increase in the membrane fluidity. Furthermore, since the stomatocytes fixed by diamide treatment exhibited membrane fluidization only in the presence of the reagent, the enhanced membrane fluidity was a direct consequence of the reagent interacting with, and changing the state of, the lipid bilayer itself, and not through the influence of some structural alteration of spectrin. These results provide experimental support for the theoretical prediction made by Brailsford et al. [J. theoret. Biol. 86, 531 (1980)]. Plausible mechanisms for the discocyte-stomatocyte transformation are discussed.

Chlorpromazine↗