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H Kuroda

Publications and source records attributed to H Kuroda.

At least 361 records · Page 20Linked to original sources

Origin of the membrane potential in plasmodial droplets of Physarum polycephalum. Evidence for an electrogenic pump.

Spherical droplets, derived from Physarum plasmodia by incubation in 10 mM caffeine, seemed to be an excellent system for electrophysiological studies because they were large (less than or equal to 300 micrometer in diameter) and because they tolerated intracellular electrodes filled with 3 M KCl and 10 mM EDTA for a few hours. Intact plasmodia, by contrast, gave valid records for only a few minutes. Under standard conditions ([K+]o = 1 mM, [Na+]o = 5 mM, [Ca++]0 = 0.5 mM, [Mg++]o = 2 mM, and [Cl-]o = 6 mM at pH 7.0), the potential difference across droplet membranes was -80 to -120mV, interior negative. The membrane potential was only slightly sensitive to concentration changes for the above-mentioned ions, and was far negative to the equilibrium diffusion potentials calculated from the known internal contents of K, Na, Ca, Mg, and CL (29.4, 1.6, 3.7, 6.5, and 27.8 mmol/kg, respectively). Variations of external pH did have a strong influence on the membrane potential, yielding a slope of 59 mV/pH between pH 6.5 and 5.5. In this pH range, however, the equilibrium potential for H+ (assuming 6.2 less than or equal to pHi less than or equal to 7.0) was greater than 75 mV positive to the observed membrane potential. Membrane potential was directly responsive to metabolic events, being lowered by potassium cyanide, and by cooling from 25 to 12 degrees C. This ensemble of results strongly indicates that the major component of membrane potential in plasmodial droplets of Physarum is generated by an electrogenic ion pump, probably one extruding H+ ions.

Antimetabolites↗

Nine cases with marked retention of indocyanine green test and normal sulfobromophthalein test without abnormal liver histology: constitutional indocyanine green excretory defect.

Nine patients revealed marked retention in the indocyanine green (ICG) test and the normal sulfobromophthalein (BSP) test. The results of liver function tests in these cases were within normal limits with the exception of the ICG test. The ICG plasma disappearance rate ranged from 0.017 to 0.025, whereas that of BSP ranged from 0.058 to 0.126. The transfer rate of ICG from plasma to liver markedly decreased on two-compartmental analysis of the ICG decay curve. A transient delay of the ICG plasma disappearance (step formation) on the decay curve was observed over 20 to 25 minutes after an injection in all cases after repeated observation. Binding of ICG to plasma proteins in these cases failed to demonstrate a significant difference from normal subjects. The fasting serum bile acid levels in two cases and postprandial serum bile acid level in one case were normal. Light microscopic findings of the liver revealed normal histology. Electron microscopic findings showed increase of lipofuscin-like lysosomes, modification and paracrystalline-like array of mitochondria, and an increase of reticulum fiber in Disse's space. It is suggested that these cases represent a new type of dye excretory disorder of the liver with impaired hepatic uptake of ICG.

Adenocarcinoma↗

Fluorescent staining of microfilaments with heavy meromyosin labeled with N-(7-dimethylamino-4-methylcoumarinyl) maleimide.

For fluorescent staining of microfilaments in cells, heavy meromyosin (HMM) or subfragment-1 (S-1) was labeled with a novel thiol-directed fluorescent dye, N-(7-dimethylamino-4-methylcoumarinyl) maleimide (DACM), instead of the usual dyes, such as fluorescein-isothiocyanate (FITC). DACM-labeled HMM or S-1 gave characteristic fluorescence patterns to a variety of cell types similar to those reported with the use of FITC-labeled HMM or S-1 or with immunofluorescence techniques using anti-actin antibody. The fluorescence of DACM was fairly photoresistant as compared with FITC, so that HMM or S-1 required only 1 mol of the dye per myosin head. Consequently, F-actin need not be used to preserve the actin binding activity of the myosin fragments when labeling with the dye.

Animals↗

Calcium accumulation in vacuoles of Physarum polycephalum following starvation.

The plasmodium of Physarum polycephalum contained 15.3 mmol Ca/kg fresh weight of sample, 11.8 mmol Mg/kg, 24.5 mmol K/kg and 1.4 mmol Na/kg. When the plasmodium was starved of food, the Ca content increased gradually up to 71.9 mmol/kg during 5 days of starvation. The concentration of other elements changed only slightly. The endoplasm contained 23.0 mmol Ca/kg, 12.6 mmol Mg/kg, 26.6 mmol K/kg and 1.7 mmol Na/kg, but these contents changed only slightly during starvation. The Ca, Mg, K and Na contents of the slime and the soluble fraction were also determined. In order to clarify where the accumulated Ca was localized, Ca in the plasmodium was precipitated with potassium pyroantimonate and examined by electron microscopy. In the starved plasmodium, the vacuoles which contained the electron-opaque precipitates and were located in the ectoplasm increased in number, compared with the unstarved plasmodium. At the same time the large electron-opaque granules in the extracellular slime increased in number. The electron-opaque precipitates were identified as Ca pyroantimonate by its susceptibility to removal by chelation with ethyleneglycol bis (beta-aminoethyl ether) N, N, N', N'-tetraacetic acid (EGTA) and X-ray microprobe analysis.

Calcium↗

Induction of phenylalanine ammonia-lyase and increase in phenolics in lettuce leaves in relation to the development of russet spotting caused by ethylene.

Russet spotting (RS), consisting of numerous small brown spots on the midrib of head lettuce (Lactuca sativa), is a physiological disorder induced by exposure to ethylene. In leaves suffering RS, the increase in spotting was accompanied by a parallel increase in the amount of phenolic compounds. Of these, chlorogenic acid and isochlorogenic acid were identified. Ethylene induced high phenylalanine ammonia-lyase (PAL) activity and RS formation in the susceptible cultivar Salinas, but not in the resistant cultivar Calmar. In the absence of ethylene neither significant PAL induction nor RS occurred. No correlation was found between the increase in polyphenol oxidase or peroxidase and the development of RS. The increase in PAL activity, however, was closely correlated with the development of RS. The increase in PAL activity preceded the development of RS, and the extent of RS was directly related to the level of PAL. Three temperatures (0.5, 5.5, and 12.5 C) were compared on the basis of their influence on both RS and PAL induction. At the lowest temperature (0.5 C) neither PAL induction nor RS occurred to a significant extent. At the highest temperature (12.5 C) an initial rapid increase in PAL activity and an earlier development of spotting were observed, but subsequently there was a decrease in both PAL activity and the rate of development of RS. At the medium temperature (5.5 C) both PAL activity and RS increased progresively with time. The decline of PAL activity at a higher temperature might be attributed to inactivation of the enzyme. Thus, a temperature favorable for induction of PAL activity by ethylene was also favorable for RS. These observations indicate a close interrelationship between the induction of PAL activity and the development of RS in response to ethylene, and suggest a causal relationship between the two events. PAL serves as a useful biochemical marker for the RS reaction.

Journal Article↗