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Biomedical subjects

K Kanda

Publications and source records attributed to K Kanda.

At least 163 records · Page 9Linked to original sources

RBE of a thermal neutron beam and the 10B(n, alpha)7Li reaction on cultured B-16 melanoma cells.

The RBE of a thermal neutron beam and the 10B(n, alpha)7Li reaction were determined in cultured B-16 melanoma cells. The Kyoto University Research Reactor (KUR) was used as a thermal neutron source which had a very low contamination of gamma-rays and fast neutrons. The cells were irradiated with the beam in the presence or absence of 10B-boric acid. The absorbed dose from the neutron capture reaction to the cells was calculated by a method of Kitao (1975). Survival curves in both conditions had no shoulder and Do values were 0.506 or 0.604 Gy in the presence or absence of 5 micrograms 10B/ml-medium, respectively. The Do value of the 10B(n, alpha)7Li reaction was also estimated at 0.466 Gy, assuming each component of radiation was additive. The RBEs of the KUR thermal neutron beam and the 10B(n, alpha)7Li reaction relative to 60Co gamma-rays were estimated as 4.62 and 6.01 at 0.1 surviving fraction, respectively. Using these results, we calculated the absorbed dose from the 10B-compound and estimated the specific accumulation in the cultured cells of 10B-compounds which we have previously reported but not quantified (Nakanishi et al. 1980, Ichihashi et al. 1982).

Cell Line↗

Cyclosporin A inhibits late steps of T lymphocyte activation after transmembrane signaling.

The in vitro stimulation of murine splenic T lymphocytes with concanavalin A (Con A) produced interleukin 2 (IL2). The addition of cyclosporin A (CsA) to the culture resulted in complete inhibition of IL2 production. The Con A stimulation of T lymphocytes induced the breakdown of phosphatidylinositol into inositol trisphosphate and diacylglycerol, each of which could function as the second messengers in the subsequent signal transduction pathway. CsA did not inhibit the production of inositol (poly)phosphates. Further, CsA did not affect Ca2+-calmodulin functions; a) the redistribution of various cytoskeletal proteins as well as Con A-receptor aggregation, and b) the cytosolic Ca2+-calmodulin-dependent enzyme activities. Moreover, the activity of protein kinase C, which has been accepted to be the target of diacylglycerol, was not influenced in the presence of CsA. While the above steps of signal transduction are bypassed by synergy between calcium ionophore and phorbol ester, T lymphocyte activation which was induced by such stimuli was completely inhibited by CsA. These results indicate that CsA does not influence early steps of T lymphocyte activation as bypassed by calcium ionophore and phorbol ester, but rather inhibits later step(s) subsequent to the activation of protein kinase C and Ca2+-calmodulin.

3',5'-Cyclic-AMP Phosphodiesterases↗

Calspectin (fodrin or nonerythroid spectrin)-actin interaction: a possible involvement of 4.1-related protein.

The calspectin/actin complex extracted from the bovine brain membrane crosslinks F-actin, resulting in the increasing viscosity of F-actin determined by low-shear viscometry. We demonstrated the presence of a protein factor in this complex, which regulated the calspectin-F-actin interaction in a Ca2+- and calmodulin-dependent manner. Erythrocyte protein 4.1, but not synapsin I, mimics the function of this brain factor using a reconstitution system including purified calspectin, calmodulin and F-actin. In the brain complex, the Mr 120,000 and the Mr 80,000/77,000 polypeptides were detected to crossreact with anti-protein 4.1 antibody.

Actins↗

The effects of aging on physiological properties of fast and slow twitch motor units in the rat gastrocnemius.

The mechanical properties of individual motor units were studied in the medial gastrocnemius of old (26-30 months) rats anesthetized with urethane and chloralose. The data were compared with results from motor units in the same muscle of middle-aged (11-14 months) rats. In the old rats there was a significant increase in mean tetanic tension for type S (slow twitch) units, whereas there was a slight decrease for type F (fast twitch) units. Twitch contraction time after maximum post-tetanic potentiation was shortened in type F units, but not in type S units. Conduction velocity of motor axons decreased in both unit types, but more marked change was found in type F units. These findings suggest the differential effects of aging on type F and type S motoneurons.

Aging↗

Equilibrium disorders and R-R intervals in ECG.

We have measured RR intervals on patients with equilibrium disorders to check the function of the autonomic nervous system. And we have obtained two parameters (coefficient of variation "CV%" (the static function of the autonomic nervous system] and "A-B" (the dynamic function of the autonomic nervous system). The dynamic function of the autonomic nervous system (A-B) further deteriorated in elderly patients with central nervous system disorders. We concluded that disorders in the dynamic function of the autonomic nervous system affected the autoregulation which caused the central nervous system disorders, especially in elderly patients.

Adult↗

Effects of nimodipine on cerebral blood flow in conscious rat.

The purpose of this study was to determine the effects of the calcium channel blocker nimodipine (NM) on cerebral blood flow in the rat under conscious and anesthesized conditions and to compare these to effects of the drug in other circulations. Regional blood flows were measured using the radioactive microsphere technique. Hemodynamic parameters were assessed at the time of blood flow determinations in conscious rats and in pentobarbital-anesthetized rats. NM was administered i.v. using a cumulative dosing regimen in conscious (6, 24 and 60 micrograms/kg) and anesthetized (6 and 60 micrograms/kg) rats. In conscious rats, NM dose-dependently increased coronary blood flow whereas significantly lowering pressure-rate product and slightly but significantly reduced mean arterial pressure and systemic vascular resistance. In conscious rats, NM had no significant effect on cerebral blood flow or cerebral vascular resistance. In anesthetized rats, cerebral and systemic vascular resistances were higher compared to conscious animals. In anesthetized animals, NM reduced mean arterial pressure and systemic resistance to a greater extent than in conscious rats and significantly increased cerebral blood flow. Base-line coronary resistance was similar in conscious and anesthetized rats and the coronary dilator effects of NM were also similar under the two conditions. These findings suggest that NM is not a selective cerebral vasodilator in rats and that the cerebral dilator effects of this drug are not apparent in conscious rats. Only under the high cerebral resistance condition induced by pentobarbital anesthesia was NM found to be a cerebral vasodilator in the rat.

Anesthesia↗

Histochemical study on end-plate cholinesterase in aged mice.

The cholinesterase activity at the motor end-plate of aged mice was investigated histochemically. A relative quantitative method was applied for studying the reaction at end-plates by utilizing varying concentrations of eserine. The number of end-plates reacting at a concentration of more than 5 X 10(-6) M eserine was reduced in aged diaphragm but increased in aged soleus muscle at 10(-5) M eserine in comparison with those in the young. It seems likely that there is an age-related change in cholinesterase activities in relation to muscle fiber types.

Acetylcholinesterase↗

Crosslinking of actin filaments is caused by caldesmon aggregates, but not by its dimers.

A recent report by Bretscher [(1984) J. Biol. Chem. 259, 12873-12880] showed that caldesmon prepared by his method crosslinks actin filaments to form thick bundles. This is in contrast to the results of previous work that caldesmon binds to F-actin but does not cause any gelation [(1981) Proc. Natl. Acad. Sci. USA 78, 5652-5655]. The present work clearly showed that caldesmon purified according to Bretscher does not cause any gelation of F-actin. However, caldesmon aggregates formed by concentration or by freeze-thawing gelated F-actin to form bundles.

Actins↗

Purification and characterization of caldesmon77: a calmodulin-binding protein that interacts with actin filaments from bovine adrenal medulla.

Caldesmon150, a protein composed of the Mr 150,000/147,000 doublet, alternately binds to calmodulin and actin filaments in a Ca2+-dependent "flip-flop" fashion. In all fibroblast cell lines examined, we also found a Mr 77,000 protein that crossreacts with anti-caldesmon150 antibody by using an immunoprecipitation technique [Owada, M.K., Hakura, A., Iida, K., Yahara, I., Sobue, K. & Kakiuchi, S. (1984) Proc. Natl. Acad. Sci. USA 81, 3133-3137]. In this report, we examine the tissue distribution of caldesmon by the method of immunoblotting, using caldesmon-specific antibody. Both caldesmon150 and caldesmon77 show widespread distribution in the tissues examined. Caldesmon77 is more widely distributed than caldesmon150, and we have purified caldesmon77 from bovine adrenal medulla. Its molecular weight estimated by NaDodSO4/polyacrylamide gel electrophoresis was 77,000, and a tetramer of this polypeptide may constitute the native molecule (Mr, 300,000). Caldesmon77 possesses a number of features in common with caldesmon150, including flip-flop binding to calmodulin and actin filaments depending on the concentration of Ca2+ and crossreactivity with caldesmon150-specific antibody. Analysis of caldesmon77-F actin interaction by sedimentation and electrophoresis revealed that 0.5 mg of caldesmon77 bound to 1 mg of F actin. This indicated that the molar ratio between caldesmon77 (tetramer) and actin monomer was calculated to be 1:12-14. In addition, caldesmon77 regulated the actin-myosin interaction in Ca2+-sensitive actomyosin obtained from adrenal medulla. These results suggest that caldesmon77 might be a ubiquitous actin-linked regulator of nonmuscle contractile processes, including those in adrenal medulla.

Actins↗

Phosphorylation of myosin light chain and the actin-activated ATPase activity of adrenal medullary myosin.

Myosin light chain kinase was partially purified from bovine adrenal medulla. A polypeptide of Mr 165,000 dalton was identified as kinase by using anti-gizzard myosin light chain kinase IgG on immunoreplica. Phosphorylation of medullary myosin was Ca2+- and calmodulin-dependent. The phosphorylated myosin was showed to enhance the actin-activated Mg2+-ATPase activity. In contrast, the myosin ATPase activity was dramatically decreased by dephosphorylation of myosin.

Actins↗

Rat renal cell monolayer culture: a sensitive method for investigating ADH and PTH actions on the kidney by determining adenosine 3' :5'-cyclic monophosphate.

The response of cAMP to antidiuretic hormone (ADH) was studied using rat renal medullary cells in a monolayer culture. In addition, cAMP response to parathyroid hormone (PTH) was studied in renal cortical cells. As the culture aged, an increase in basal cAMP content and a gradual decrease in the cAMP responsiveness to arginine vasopressin (AVP) were observed. After 2 days of culture, AVP and hPTH-(1-34) produced a rapid increase in intracellular cAMP with single peaks, after 10 min and 5 min, respectively. Extracellular cAMP was increased linearly by both AVP and hPTH-(1-34). The response of cAMP to AVP was markedly greater in the medulla than in the cortex, while the response to hPTH-(1-34) was remarkable only in the cortex. Outstanding sensitivity of cAMP responsiveness was observed in this system, i.e., 10(-12) M AVP (1 pg/ml) and 2.43 X 10(-10) M hPTH-(1-34) (1 ng/ml) provoked significant increases in cAMP from the basal level of 0.31 +/- 0.04 and 0.59 +/- 0.05 pmol/dish to 0.79 +/- 0.03 and 1.07 +/- 0.13 pmol/dish, respectively (P less than 0.001). In the medulla, potencies of lysine vasopressin (LVP), DDAVP and oxytocin at a concentration of 10(-9) M were 76.1%, 154.2% and 8.1% of that of AVP, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗

High mammary carcinogenicity of neutron irradiation in rats and its promotion by prolactin.

The present study was undertaken to assess the mammary carcinogenic effect of low doses of fission and thermal neutrons in female W/Fu rats. Only 2 of 62 (3.2%) rats exposed to various doses of fission radiation alone developed mammary tumors (MT) in a 12-month observation period, whereas 21 of 63 (33.3%) similarly irradiated rats developed MT if further treated with prolactin; the lowest effective dose was 4.1 rad, which contained only 1.7 rad of fission spectrum neutrons with an average energy of 2.0 MeV. The long survival of radiation-initiated potentially malignant cells was suggested by the observation that excess numbers of MT developed in irradiated rats in whom prolactin treatment was started as late as 12 months after irradiation. The negligible contribution of gamma-rays, one component of the reactor radiations, to the rat mammary carcinogenesis was proven by a simulation experiment with 60Co gamma-rays. Thermal neutrons with an average energy of 0.025 eV were less effective than fission neutrons. The rat mammary carcinogenic effects of 180 kVp X-rays, 14.1 MeV fast neutrons, 0.025 eV thermal neutrons and 2.0 MeV fission neutrons were estimated in such a way as to compare the dose of each radiation that gave an MT incidence of 40% of that in irradiated, prolactin-treated rats. The efficiency of fission spectrum neutrons is much higher than those of other radiations; the relative biological effectiveness (RBE) of fission spectrum neutrons was 17.8 against X-rays. Based on these findings, the relevance of animal data to tumor induction in atomic bomb survivors is discussed.

Animals↗

Multiple simultaneous determinations of hemodynamics and flow distribution in conscious rat.

A method for multiple simultaneous determinations of cardiocirculatory dynamics, regional blood flow, and total cardiac output distribution in the conscious rat preparation is described. The preparation allows for intravenous administration of agents and can be performed on animals either at rest or during treadmill exercise. Instrumentation procedures involve placement of fluid-filled catheters in the left ventricle, right atria, right jugular vein, and caudal artery. Left ventricular pressures are recorded via a modified 4F Millar transducer-tipped manometer containing a 10-cm extension of fluid-filled PE 50 placed into the left ventricle via the right carotid artery. Radionuclide-labeled microspheres (15 +/- 5 mu) are injected into the left ventricle through the fluid-filled PE 50 at selected times for determination of cardiac output and regional blood flows using the caudal artery catheter as the source of the reference blood sample. Details and selected validation data for procedures involving anesthesia, instrumentation, recovery from anesthesia, data gathering, and data analysis are presented. Emphasis is placed on the procedures required for use of the radioactive microsphere technique in this model with special attention given to quality control of the microsphere stock, counting procedures, and computer analysis of these data.

Animals↗